Bibliography Golden Rice and Biofortification

Bibliography Golden Rice and
Biofortification
Klaus Ammann, Version 20091008
Abbo, S., C. Molina, et al. (2005). "Quantitative trait loci governing carotenoid concentration and weight in seeds of chickpea (Cicer arietinum
L.)." Theoretical and Applied Genetics 111(2): 185-195.
://000230689800001 AND http://www.botanischergarten.ch/Golden-Rice/Abbo-Quantitative-Trait-Loci-2005.pdf
Chickpea is a staple protein source in many Asian and Middle Eastern countries. The seeds contain carotenoids such as betacarotene,
cryptoxanthin, lutein and zeaxanthin in amounts above the engineered beta-carotene-containing "golden rice" level. Thus,
breeding for high carotenoid concentration in seeds is of nutritional, socio-economic, and economic importance. To study the
genetics governing seed carotenoids in chickpea, we studied the relationship between seed weight and concentrations of betacarotene
and lutein by means of high-performance liquid chromatography in segregating progeny from a cross between an Israeli
cultivar and wild Cicer reticulatum Ladiz. Seeds of the cross progeny varied with respect to their carotenoid concentration
(heritability estimates ranged from 0.5 to 0.9), and a negative genetic correlation was found between mean seed weight and
carotenoid concentration in the F-3. To determine the loci responsible for the genetic variation observed, the population was
genotyped using 91 sequence tagged microsatellite site markers and two CytP450 markers to generate a genetic map consisting of
nine linkage groups and a total length of 344.6 cM. Using quantitative data collected for beta-carotene and lutein concentration and
seed weight of the seeds of the F-2 population, we were able to identify quantitative trait loci (QTLs) by interval mapping. At a LOD
score of 2, four QTLs for beta-carotene concentration, a single QTL for lutein concentration and three QTLs for seed weight were
detected. The results of this investigation may assist in improving the nutritional quality of chickpea.
Adeoti, J. O. and A. A. Adekunle (2007). "Awareness of and attitudes towards biotechnology and GMOs in Southwest Nigeria: a survey of people
with access to information." International Journal of Biotechnology 9(2): 209-230.
://BIOSIS:PREV200700402349 AND http://www.botanischergarten.ch/Golden-Rice/Adeoti-Awareness-Attitudes-Nigeria-2007.pdf
In this paper, we examine awareness of biotechnology and the attitudes towards products and services derived from Genetically
Modified Organisms (GMOs) in Southwest Nigeria. For this purpose, we surveyed 180 respondents across selected segments of the
regional population. The paper demonstrates that while respondents have some awareness of biotechnology techniques, there is
little awareness about specific GM crops that include Bt maize, Bt cotton and golden rice. Most of the respondents tend to be in
favour of the introduction of GM crops and would eat GM foods if they are more nutritious than non-GM foods. The attitudes of the
respondents towards GMOs suggest that though more people are in favour of the introduction of GM crops, they do not consider
the current state of Nigeria's institutional preparedness satisfactory for the approval and release of GMOs.
Al-Babili, S. and P. Beyer (2005). "Golden Rice - five years on the road - five years to go?" Trends in Plant Science 10(12): 565-573.
://000234155300005 AND http://www.botanischergarten.ch/Rice/Babili-Golden-Rice-5years-2005.pdf
Provitamin A accumulates in the grain of Golden Rice as a result of genetic transformation. In developing countries, where vitamin A
deficiency prevails, grain from Golden Rice is expected to provide this important micronutrient sustainably through agriculture. Since
its original production, the prototype Golden Rice has undergone intense research to increase the provitamin A content, to establish
the scientific basis for its carotenoid complement, and to better comply with regulatory requirements. Today, the current focus is on
how to get Golden Rice effectively into the hands of farmers, which is a novel avenue for public sector research, carried out with the
aid of international research consortia. Additional new research is underway to further increase the nutritional value of Golden Rice.
Al-Babili, S., T. T. C. Hoa, et al. (2006). "Exploring the potential of the bacterial carotene desaturase CrtI to increase the beta-carotene content in
Golden Rice." Journal of Experimental Botany 57(4): 1007-1014.
://000235771700028 AND http://www.botanischergarten.ch/Golden-Rice/Al-Babili-Exploring-the-Potential-2006.pdf
To increase the beta-carotene (provitamin A) content and thus the nutritional value of Golden Rice, the optimization of the enzymes
employed, phytoene synthase (PSY) and the Erwinia uredovora carotene desaturase (CrtI), must be considered. CrtI was chosen for
this study because this bacterial enzyme, unlike phytoene synthase, was expressed at barely detectable levels in the endosperm of
the Golden Rice events investigated. The low protein amounts observed may be caused by either weak cauliflower mosaic virus 35S
promoter activity in the endosperm or by inappropriate codon usage. The protein level of CrtI was increased to explore its potential
for enhancing the flux of metabolites through the pathway. For this purpose, a synthetic CrtI gene with a codon usage matching that
of rice storage proteins was generated. Rice plants were transformed to express the synthetic gene under the control of the
endosperm-specific glutelin B1 promoter. In addition, transgenic plants expressing the original bacterial gene were generated, but
the endosperm-specific glutelin B1 promoter was employed instead of the cauliflower mosaic virus 35S promoter. Independent of
codon optimization, the use of the endosperm-specific promoter resulted in a large increase in bacterial desaturase production in
the T-1 rice grains. However, this did not lead to a significant increase in the carotenoid content, suggesting that the bacterial
enzyme is sufficiently active in rice endosperm even at very low levels and is not rate-limiting. The endosperm-specific expression of
CrtI did not affect the carotenoid pattern in the leaves, which was observed upon its constitutive expression. Therefore, tissuespecific
expression of CrtI represents the better option.

Aluru, M., Y. Xu, et al. (2008). "Generation of transgenic maize with enhanced provitamin A content." Journal of Experimental Botany 59(13):
3551-3562.
://WOS:000259973800006 AND http://www.botanischergarten.ch/Golden-Rice/Aluru-Generation-Maize-ProvitaminA-2008.pdf
Vitamin A deficiency (VAD) affects over 250 million people worldwide and is one of the most prevalent nutritional deficiencies in
developing countries, resulting in significant socio-economic losses. Provitamin A carotenoids such as beta-carotene, are derived
from plant foods and are a major source of vitamin A for the majority of the world's population. Several years of intense research
has resulted in the production of 'Golden Rice 2' which contains sufficiently high levels of provitamin A carotenoids to combat VAD.
In this report, the focus is on the generation of transgenic maize with enhanced provitamin A content in their kernels.
Overexpression of the bacterial genes crtB (for phytoene synthase) and crtI (for the four desaturation steps of the carotenoid
pathway catalysed by phytoene desaturase and zeta-carotene desaturase in plants), under the control of a 'super gamma-zein
promoter' for endosperm-specific expression, resulted in an increase of total carotenoids of up to 34-fold with a preferential
accumulation of beta-carotene in the maize endosperm. The levels attained approach those estimated to have a significant impact
on the nutritional status of target populations in developing countries. The high beta-carotene trait was found to be reproducible
over at least four generations. Gene expression analyses suggest that increased accumulation of beta-carotene is due to an upregulation
of the endogenous lycopene beta-cylase. These experiments set the stage for the design of transgenic approaches to
generate provitamin A-rich maize that will help alleviate VAD.
Anderson, K., L. Jackson, et al. (2004). GM Rice Adoption: Implications for Welfare and Poverty Alleviation. Washington, World Bank Policy
Research Working Paper, 2004: 24.
http://www.botanischergarten.ch/Golden-Rice/Anderson-Implications-Welfare-WHO-2004.pdf
The first generation of genetically modified (GM) crop varieties sought to increase farmer profitability through cost reductions or
higher yields. The next generation of GM food research is focusing also on breeding for attributes of interest to consumers,
beginning with ‘golden rice’, which has been genetically engineered to contain a higher level of vitamin A and thereby boost the
health of poor people in developing countries. This paper analyses empirically the potential economic effects of adopting both types
of innovation in Asia, including its impact on rice producers and other poor households. It does so using the global economy-wide
computable general equilibrium model known as GTAP. The results suggest the very considerable farm productivity gains (even if
extended beyond GM rice to include those from adopting other GM grains and oilseeds) could be exceeded by the welfare gains
resulting from the potential health-enhancing attributes of golden rice, which would boost the productivity of unskilled workers
among Asia’s poor.
Anderson, K. and L. A. Jackson (2005). "Some implications of GM food technology policies for Sub-Saharan africa." Journal of African Economies
14(3): 385-410.
://000231362700003 AND http://www.botanischergarten.ch/Golden-Rice/Anderson-Implications-GM-Food-Sub-Saharan-2005.pdf
The first generation of genetically modified (GM) crop varieties sought to increase farmer profitability through cost reductions or
higher yields. The next generation of GM food research is focusing also on breeding for attributes of interest to consumers,
beginning with 'golden rice', which has been genetically engineered to contain a higher level of vitamin A and thereby boost the
health of unskilled labourers in developing countries. This paper analyses empirically the potential economic effects of adopting
both types of innovation in Sub-Saharan Africa (SSA). It does so using the global economy-wide computable general equilibrium
model known as GTAP. The results suggest the welfare gains are potentially very large, especially from golden rice and that-contrary
to the claims of numerous interests-those estimated benefits are diminished only slightly by the presence of the European Union's
current barriers to imports of GM foods. In particular, if SSA countries impose bans on GM crop imports in an attempt to maintain
access to EU markets for non-GM products, the loss to domestic consumers due to that protectionism boost to SSA farmers is far
more than the small gain in terms of greater market access to the EU.
Anonymous (2001). "'Golden Rice' comes to Asia." Outlook on Agriculture 30(2): 141-142.
://000169355200018 AND http://www.botanischergarten.ch/Golden-Rice/Anonymous-GR-comes-to-Acia-2001.pdf
Anonymous (2005). "Reburnishing Golden Rice." Nature Biotechnology 23(4): 395-395.
://000228197300001 AND http://www.botanischergarten.ch/Golden-Rice/Anonymous-Reburnishing-Golden-Rice-2005.pdf
Apgar, J., D. Makdani, et al. (1996). "Serum carotenoid concentrations and their reproducibility in children in Belize." American Journal of
Clinical Nutrition 64(5): 726-730.
://A1996VP75300009 AND http://www.botanischergarten.ch/Golden-Rice/Apgar-Serum-Carotenoid-Belize-1996.pdf
Suggestions that carotenoid-containing foods are beneficial in maintaining health have led to Several studies of circulating
carotenoid concentrations of adults. Because few data are available for children, we report serum carotenoid concentrations of 493
children in Belize. Carotenoid concentrations were determined as part of a survey of vitamin A status of children, most between 65
and 89 mo of age. Reproducibility was tested by collecting a second blood sample 2 wk after the first collection from a subset of
children (n = 23) who consumed their habitual diet with no treatment during the interim. Predominant serum carotenoids were
lutein/zeaxanthin and beta-carotene; which ac counted fur 26% and 24% of median total carotenoids, respectively. The three
provitamin A carotenoids, alpha- and beta-carotene and beta-cryptoxanthin, constituted 51% of median total carotenoid
concentrations. Partial correlations of each carotenoid with lasting retinol concentration indicated that beta-carotene had the
highest correlation. Concordance correlation coefficients (r(c)) for lasting carotenoid concentrations determined 2 wk apart were
greater than or equal to 0.89 for lycopene, beta-cryptoxanthin, and alpha- and beta-carotene. The r(c) for lutein/zeaxanthin and
total carotenoids was lower, 0.59 and 0.68 respectively, because of higher lutein/zeaxanthin concentrations at the second sampling
than at the first. The reproducibility of the concentrations suggests both that individuals have characteristic profiles and that serum
carotenoid concentrations ran be measured randomly over greater than or equal to 2 wk without significant bias.

Baisakh, N., S. Rehana, et al. (2006). "Marker-free transgenic (MFT) near-isogenic introgression lines (NIILs) of 'golden' indica rice (cv. IR64) with
accumulation of provitamin A in the endosperm tissue." Plant Biotechnology Journal 4(4 %R doi:10.1111/j.1467-7652.2006.00196.x): 467-475.
http://www.blackwell-synergy.com/doi/abs/10.1111/j.1467-7652.2006.00196.x AND http://www.botanischergarten.ch/Golden-Rice/Baisakh-
Marker-Free-Transgenic-Golden-2006.pdf
Summary We have developed near-isogenic introgression lines (NIILs) of an elite indica rice cultivar (IR64) with the genes for betacarotene
biosynthesis from dihaploid (DH) derivatives of golden japonica rice (cv. T309). A careful analysis of the DH lines indicated
the integration of the genes of interest [phytoene synthase (psy) and phytoene desaturase (crtI)] and the selectable marker gene
(hygromycin phosphotransferase, hph) in two unlinked loci. During subsequent crossing, progenies could be obtained carrying only
the locus with psy and crtI, which was segregated independently from the locus containing the hph gene during meiotic segregation.
The NIILs (BC2F2) showed maximum similarity with the recurrent parent cultivar IR64. Further, progenies of two NIILs were devoid
of any fragments beyond the left or right border, including the chloramphenicol acetyltransferase (cat) antibiotic resistance gene of
the transformation vector. Spectrophotometric readings showed the accumulation of up to 1.06 ug total carotenoids, including betacarotene,
in 1 g of the endosperm. The accumulation of beta-carotene was also evident from the clearly visible yellow colour of the
polished seeds.
Becker, D. J. and R. Y. Gordon (2008). "Low-dose statin concentration in red yeast rice: A condfounding effect on outcome (vol 83, pg 1187,
2008)." Mayo Clinic Proceedings 83(11): 1301-1301.
://WOS:000260743900023 AND http://www.botanischergarten.ch/Golden-Rice/Becker-Red-Yeast-Rice-Suitable-Reply-1-correction-
2008.pdf AND http://www.botanischergarten.ch/Golden-Rice/Becker-Low-Dose-Statin-Confounding-2008.pdf
Becker, D. J., R. Y. Gordon, et al. (2008). "Is Red Yeast Rice a Suitable Alternative for Statins? Reply." Mayo Clinic Proceedings 83(11): 1294-1296.
://WOS:000260743900016 AND http://www.botanischergarten.ch/Golden-Rice/Becker-Red-Yeast-Rice-Suitable-Reply-1-2008.pdf
Becker, D. J., R. Y. Gordon, et al. (2009). "Red Yeast Rice for Dyslipidemia in Statin-Intolerant Patients: A Randomized Trial." Ann Intern Med
150(12): 830-839.
http://www.annals.org/cgi/content/abstract/150/12/830 AND http://www.botanischergarten.ch/Golden-Rice/Becker-Red-Yeast-Rice-2009.pdf
Background: Red yeast rice is an herbal supplement that decreases low-density lipoprotein (LDL) cholesterol level. Objective: To
evaluate the effectiveness and tolerability of red yeast rice and therapeutic lifestyle change to treat dyslipidemia in patients who
cannot tolerate statin therapy. Design: Randomized, controlled trial. Setting: Community-based cardiology practice. Patients: 62
patients with dyslipidemia and history of discontinuation of statin therapy due to myalgias. Intervention: Patients were assigned by
random allocation software to receive red yeast rice, 1800 mg (31 patients), or placebo (31 patients) twice daily for 24 weeks. All
patients were concomitantly enrolled in a 12-week therapeutic lifestyle change program. Measurements: Primary outcome was LDL
cholesterol level, measured at baseline, week 12, and week 24. Secondary outcomes included total cholesterol, high-density
lipoprotein (HDL) cholesterol, triglyceride, liver enzyme, and creatinine phosphokinase (CPK) levels; weight; and Brief Pain Inventory
score. Results: In the red yeast rice group, LDL cholesterol decreased by 1.11 mmol/L (43 mg/dL) from baseline at week 12 and by
0.90 mmol/L (35 mg/dL) at week 24. In the placebo group, LDL cholesterol decreased by 0.28 mmol/L (11 mg/dL) at week 12 and by
0.39 mmol/L (15 mg/dL) at week 24. Low-density lipoprotein cholesterol level was significantly lower in the red yeast rice group than
in the placebo group at both weeks 12 (P < 0.001) and 24 (P = 0.011). Significant treatment effects were also observed for total
cholesterol level at weeks 12 (P < 0.001) and 24 (P = 0.016). Levels of HDL cholesterol, triglyceride, liver enzyme, or CPK; weight loss;
and pain severity scores did not significantly differ between groups at either week 12 or week 24. Limitation: The study was small,
was single-site, was of short duration, and focused on laboratory measures. Conclusion: Red yeast rice and therapeutic lifestyle
change decrease LDL cholesterol level without increasing CPK or pain levels and may be a treatment option for dyslipidemic patients
who cannot tolerate statin therapy. Primary Funding Source: Commonwealth of Pennsylvania.
Becker, D. J., R. Y. Gordon, et al. (2009). "A Novel Approach to Lipid-Lowering in Patients With Statin-Associated Myalgias: A Randomized,
Placebo-Controlled, Double-Blind Trial." Journal of the American College of Cardiology 53(10): A209-A210.
://WOS:000263864200870 AND http://www.botanischergarten.ch/Golden-Rice/Gordon-Marked-Variability-Monacolin-2009.pdf
Becker, D. J., R. Y. Gordon, et al. (2008). "Simvastatin vs therapeutic lifestyle changes and supplements: Randomized primary prevention trial."
Mayo Clinic Proceedings 83(7): 758-764.
://WOS:000257198000004 AND http://www.botanischergarten.ch/Golden-Rice/Becker-Simvastatin-vs-Therapeutic-2008.pdf
OBJECTIVE: To compare the lipid-lowering effects of an alternative regimen (lifestyle changes, red yeast rice, and fish oil) with a
standard dose of a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin). PATIENTS AND METHODS: This randomized
trial enrolled 74 patients with hypercholesterolemia who met Adult Treatment Panel III criteria for primary prevention using statin
therapy. All participants were randomized to an alternative treatment group (AG) or to receive simvastatin (40 mg/d) in this openlabel
trial conducted between April 1, 2006, and June 30, 2006. The alternative treatment included therapeutic lifestyle changes,
Ingestion of red yeast rice, and fish oil supplements for 12 weeks. The simvastatin group received medication and traditional
counseling. The primary outcome measure was the percentage change in low-density lipoprotein cholesterol (LDL-C). Secondary
measures were changes in other lipoproteins and weight loss. RESULTS: There was a statistically significant reduction in LDL-C levels
in both the AG (-42.4%+/- 15%) (P

Bellovino, D., M. Apreda, et al. (2003). "Vitamin A transport: In vitro models for the study of RBP secretion." Molecular Aspects of Medicine
24(6): 411-420.
://BIOSIS:PREV200400031051 AND http://www.botanischergarten.ch/Golden-Rice/Bellovino-Vitamin-A-Transport-2003.pdf
Beyer, P., S. Al-Babili, et al. (2002). Golden Rice: introducing the beta-carotene biosynthesis pathway into rice endosperm by genetic
engineering to defeat vitamin A deficiency. The Journal of Nutrition: Symposium: Plant Breeding: A New Tool for Fighting Micronutrient
Malnutrition. H. G. E. Bouis. Presented as part of the symposium “Plant Breeding: A New Tool for Fighting Micronutrient Malnutrition” given at
the Experimental Biology 2001 meeting, Orlando, Florida, on April 1, 2001. 132.
http://www.gramene.org/perl/pub_search?ref_id=6871 AND http://www.botanischergarten.ch/Golden-Rice/Beyer-Golden-Rice-Introducing-B-
Carotene-2002.pdf
To obtain a functioning provitamin A (beta-carotene) biosynthetic pathway in rice endosperm, we introduced in a single, combined
transformation effort the cDNA coding for phytoene synthase (psy) and lycopene beta-cyclase (beta-lcy) both from Narcissus
pseudonarcissus and both under the control of the endosperm-specific glutelin promoter together with a bacterial phytoene
desaturase (crtI, from Erwinia uredovora under constitutive 35S promoter control). This combination covers the requirements for
beta-carotene synthesis and, as hoped, yellow beta-carotene-bearing rice endosperm was obtained in the T(0)-generation.
Additional experiments revealed that the presence of beta-lcy was not necessary, because psy and crtI alone were able to drive
beta-carotene synthesis as well as the formation of further downstream xanthophylls. Plausible explanations for this finding are that
these downstream enzymes are constitutively expressed in rice endosperm or are induced by the transformation, e.g., by
enzymatically formed products. Results using N. pseudonarcissus as a model system led to the development of a hypothesis, our
present working model, that trans-lycopene or a trans-lycopene derivative acts as an inductor in a kind of feedback mechanism
stimulating endogenous carotenogenic genes. Various institutional arrangements for disseminating Golden Rice to research
institutes in developing countries also are discussed.
Beyer, P., S. Al-Babili, et al. (2002). "Golden rice: Introducing the beta-carotene biosynthesis pathway into rice endosperm by genetic
engineering to defeat vitamin A deficiency." Journal of Nutrition 132(3): 506S-510S.
://000174189800032 and http://www.botanischergarten.ch/Rice/Beyer-Nutrition-2002.pdf
To obtain a functioning provitamin A (beta-carotene) biosynthetic pathway in rice endosperm, we introduced in a single, combined
transformation effort the cDNA coding for phytoene synthase (psy) and lycopene beta-cyclase (beta-/cy) both from Narcissus
pseudonarcissus and both under the control of the endosperm-specific glutelin promoter together with a bacterial phytoene
desaturase (crtl, from Erwinia uredovora under constitutive 35S promoter control). This combination covers the requirements for
betaP-carotene synthesis and, as hoped, yellow beta-carotene-bearing rice endosperm was obtained in the T-o-generation.
Additional experiments revealed that the presence of beta-/cy was not necessary, because psy and crtl alone were able to drive
beta-carotene synthesis as well as the formation of further downstream xanthophylls. Plausible explanations for this finding are that
these downstream enzymes are constitutively expressed in rice endosperm or are induced by the transformation, e.g., by
enzymatically formed products. Results using N. pseudonarcissus as a model system led to the development of a hypothesis, our
present working model, that trans-lycopene or a trans-lycopene derivative acts as an inductor in a kind of feedback mechanism
stimulating endogenous carotenogenic genes. Various institutional arrangements for disseminating Golden Rice to research
institutes in developing countries also are discussed.
Bhargava, A., H. E. Bouis, et al. (2001). "Dietary Intakes and Socioeconomic Factors Are Associated with the Hemoglobin Concentration of
Bangladeshi Women." J. Nutr. 131(3): 758-764.
http://www.nutrition.org/cgi/content/abstract/131/3/758 and http://www.botanischergarten.ch/Rice/Bhargava-Intakes-2001.pdf
Iron deficiency anemia affects a large number of women in developing countries, especially during child-bearing years. The
hemoglobin concentration is useful for identifying iron deficiency anemia. The main objectives of this study were, first, to extend
algorithms for calculating bioavailable iron from mixed diets, taking into account the enhancers and inhibitors of iron absorption
under alternative assumptions on body iron stores. Second, a comprehensive longitudinal model was developed for the proximate
determinants of hemoglobin concentration that included the subjects' dietary intakes, nutritional status, morbidity and
socioeconomic factors and the unobserved between-subject differences. The model for hemoglobin concentration was estimated
using three repeated observations on 514 free living women in Bangladesh. Socioeconomic factors affecting the iron intake from
meat, fish and poultry and from all animal sources were also modeled. The main results were that bioavailable iron, women's height
and mid upper arm circumference and intake of iron tablets were significant predictors of hemoglobin concentration. Increases in
household incomes were associated with higher intake of iron from meat, fish and poultry and from all animal sources. The
algorithms for estimating bioavailable iron showed the importance of assumptions regarding body iron stores and underscored the
need to develop suitable algorithms for subjects in developing countries.
Bhaskarachary, K., D. S. S. Rao, et al. (1995). "Carotene Content of Some Common and Less Familiar Foods of Plant-Origin." Food Chemistry
54(2): 189-193.
://A1995RN64500013 AND http://www.botanischergarten.ch/Golden-Rice/Bhaskarachary-Carotene-Content-Plants-1995.pdf
A number of commonly consumed plant foods including green leafy vegetables, roots and tubers, other vegetables and fruits were
analysed for their total carotenes spectrophotometrically and by separation of provitamin A carotenoids on HPLC. beta-Carotene is
the predominant carotenoid in all foods. Green leafy vegetables (GLV) were found to be the best source of provitamin A, and also
yellow fruits such as mango and papaya. Carrot contains significant amounts of alpha-carotene while, in papaya, beta-cryptoxanthin
is the predominant carotenoid. Incorporation of these vegetables in the diets of the children would be the most effective approach
to combat vitamin A deficiency. A few lesser-known GLV were also analysed. Among them, chennangiaku and botla benda appeared

to be good sources of beta-carotene. Organised cultivation of these vegetables and fruits would help in overcoming the problem of
vitamin A deficiency.
Biesalski, H. K. and D. Nohr (2003). "Importance of vitamin-A for lung function and development." Molecular Aspects of Medicine 24(6): 431-
440.
http://www.sciencedirect.com/science/article/B6T9P-49202PF-2/2/77f8555e0d619c3225e8d91aa21ba25d AND
http://www.botanischergarten.ch/Golden-Rice/Biesalski-Importance-Vitamin-A-2003.pdf
Vitamin-A is essential for growth and development of cells and tissues. In its active form, retinoic acid, it controls the regular
differentiation as a ligand for retinoic acid receptors (RAR, RXR) and is involved in the integration (gap junction formation) of cell
formations [Nature 37 (1994) 528; International Review of Cytology. San Diego Academic Press, 1-31]. Vitamin-A plays a substantial
role, especially in the respiratory epithelium and the lung. During moderate vitamin-A-deficiency, the incidence for diseases of the
respiratory tract is considerably increased and repeated respiratory infections can be influenced therapeutically by a moderate
vitamin-A-supplementation [Aust. Paediatr. J. 22 (1986) 95; Lancet 338 (1991) 67]. In addition to the importance of the vitamin for
the lung function, vitamin-A is also responsible for the development of many tissues and cells as well as for the embryonic lung
development. Recent studies proved that the control occurs by different expressions of retinoid receptors as well as by timedependent
changes of the vitamin-A-metabolism respectively via cellular vitamin-A-binding proteins (CRBP: cytoplasmatic retinol
binding protein; CRABP: cytoplasmatic retinoic acid binding protein).
Bloem, M. W., N. Huq, et al. (1996). "Production of fruits and vegetables at the homestead is an important source of vitamin A among women
in rural Bangladesh." European Journal of Clinical Nutrition 50: S62-S67.
://A1996VA87500013 AND http://www.botanischergarten.ch/Golden-Rice/Bloem-Production-Homestead--Source-Vitamin-1996.pdf
Background: Vitamin A deficiency is considered to be an important public health problem in Bangladesh. A universal biannual
distribution of high-dose vitamin A. capsules has been in place for over the past two decades. This supplementation has been
beneficial for preschool children. Bangladesh has been exploring more sustainable approaches for all segments of the population. To
support this initiative, Helen Keller International has implemented a home-gardening promotion project since 1993. This project is
executed on a large scale and currently reaches an estimated 244 000 families. Methods: This paper presents data from 7341
women of reproductive age which were collected as part of the baseline census of a community monitoring system whose objective
is to track progress and measure the impact of home-gardening activities. Results: Vitamin A intake in this population derived almost
entirely from the consumption of fruits and vegetables. Logistic regression analyses showed that maternal vitamin A intake was
determined by qualitative indicators of homestead gardens (type of home garden, the total quantity of provitamin A-rich foods
produced and the number of fruits and vegetables varieties grown in the garden) after adjusting for socio-economic status.
Conclusions: These results indicate that traditional production of provitamin A-rich fruits and vegetables in the homestead may
provide a valuable contribution to vitamin A intake in communities where alternative dietary sources of vitamin A are scarce.
Botella-Pavia, P. and M. Rodriguez-Concepcion (2006). "Carotenoid biotechnology in plants for nutritionally improved foods." Physiologia
Plantarum 126(3): 369-381.
://000235591400008 AND http://www.botanischergarten.ch/Rice/Botella-Carotenoid-2006.pdf
Carotenoids participate in light harvesting and are essential for photoprotection in photosynthetic plant tissues. They also furnish
non-photosynthetic flowers and fruits with yellow to red colors to attract animals for pollination and dispersal of seeds. Although
animals can not synthesize carotenoids de novo, carotenoid-derived products such as retinoids (including vitamin A) are required as
visual pigments and signaling molecules. Dietary carotenoids also provide health benefits based on their antioxidant properties. The
main pathway for carotenoid biosynthesis in plants and microorganisms has been virtually elucidated in recent years, and some of
the identified biosynthetic genes have been successfully used in metabolic engineering approaches to overproduce carotenoids of
interest in plants. Alternative approaches that enhance the metabolic flux to carotenoids by upregulating the production of their
isoprenoid precursors or interfere with light-mediated regulation of carotenogenesis have been recently shown to result in
increased carotenoid levels. Despite spectacular achievements in the metabolic engineering of plant carotenogenesis, much work is
still ahead to better understand the regulation of carotenoid biosynthesis and accumulation in plant cells. New genetic and genomic
approaches are now in progress to identify regulatory factors that might significantly contribute to improve the nutritional value of
plant-derived foods by increasing their carotenoid levels.
Bouis, H. E. (2003). "Micronutrient fortification of plants through plant breeding: can it improve nutrition in man at low cost?" Proceedings of
the Nutrition Society 62(2): 403-411.
://000184036700017 and http://www.botanischergarten.ch/Rice/Bouis-Nutrition-2003.pdf
Can commonly-eaten food staple crops be developed that fortify their seeds with essential minerals and vitamins? Can farmers be
induced to grow such varieties? If so, would this result in a marked improvement in human nutrition at a lower cost than existing
nutrition interventions? An interdisciplinary international effort is underway to breed for mineral- and vitamin-dense varieties of
rice, wheat, maize, beans and cassava for release to farmers in developing countries. The biofortification strategy seeks to take
advantage of the consistent daily consumption of large amounts of food staples by all family members, including women and
children as they are most at risk for micronutrient malnutrition. As a consequence of the predominance of food staples in the diets
of the poor, this strategy implicitly targets low-income households. After the one-time investment is made to develop seeds that
fortify themselves, recurrent costs are low and germplasm may be shared internationally. It is this multiplier aspect of plant
breeding across time and distance that makes it so cost-effective. Once in place, the biofortified crop system is highly sustainable.
Nutritionally-improved varieties will continue to be grown and consumed year after year, even if government attention and
international funding for micronutrient issues fades. Biofortification provides a truly feasible means of reaching malnourished
populations in relatively remote rural areas, delivering naturally-fortified foods to population groups with limited access to
commercially-marketed fortified foods that are more readily available in urban areas. Biofortification and commercial fortification

are, therefore, highly complementary. Breeding for higher trace mineral density in seeds will not incur a yield penalty. Mineralpacked
seeds sell themselves to farmers because, as recent research has shown, these trace minerals are essential in helping plants
resist disease and other environmental stresses. More seedlings survive and initial growth is more rapid. Ultimately, yields are
higher, particularly in trace mineral-'deficient' soils in arid regions.
Bouis, H. E. (2007). "The potential of genetically modified food crops to improve human nutrition in developing countries." Journal of
Development Studies 43(1): 79-96.
://WOS:000244690800004 AND http://www.botanischergarten.ch/Developing/Bouis-Potential-GM-crops-2007.pdf
Because of poor dietary quality and consequent widespread micronutrient malnutrition in low income countries, children and their
mothers, who have higher requirements for vitamins and minerals due to rapid growth and reproduction respectively, have higher
mortality, become sick more often, have their cognitive abilities compromised for a lifetime, and are less productive members of the
workforce. Their quality of life and aggregate economic growth are unnecessarily compromised. One way that biotechnology can
help to improve the nutrition and health of consumers in developing countries is by increasing the vitamin and mineral content and
their bioavailability in staple foods.
Burkhardt, P. K., P. Beyer, et al. (1997). "Transgenic rice (Oryza sativa) endosperm expressing daffodil (Narcissus pseudonarcissus) phytoene
synthase accumulates phytoene, a key intermediate of provitamin A biosynthesis." Plant Journal 11(5): 1071-1078.
://A1997XB79000015 and http://www.botanischergarten.ch/Rice/Burkhardt-et-al-Daffodill-1997.pdf
Rice (Oryza sativa L.), the major food staple for more than two billion people, contains neither beta-carotene (provitamin A) nor C-40
carotenoid precursors thereof in its endosperm. To improve the nutritional value of rice, genetic engineering was chosen as a means
to introduce the ability to make beta-carotene into rice endosperm tissue. Investigation of the biochemical properties of immature
rice endosperm using [C-14]-labelled substrates revealed the presence of geranyl geranyl diphosphate, the C-20 general isoprenoid
precursor necessary for C-40 carotenoid biosynthesis. Phytoene synthase, which condenses two molecules of geranyl geranyl
diphosphate, is the first of four specific enzymes necessary for beta-carotene biosynthesis in plants. Therefore, the Japonica rice
model variety Taipei 309 was transformed by microprojectile bombardment with a cDNA coding for phytoene synthase from daffodil
(Narcissus pseudonarcissus) under the control of either a constitutive or an endosperm-specific promoter. In transgenic rice plants,
the daffodil enzyme is active, as measured by the in vivo accumulation of phytoene in rice endosperm. Thus, it is demonstrated for
the first time that it is in principle possible to engineer a critical step in provitamin A biosynthesis in a non-photosynthetic,
carotenoid-lacking plant tissue. These results have important implications for longterm prospects of overcoming worldwide vitamin
A deficiency.
Burri, B. J. (1997). "Beta-carotene and human health: A review of current research." Nutrition Research 17(3): 547-580.
://A1997WJ75200015 AND http://www.botanischergarten.ch/Golden-Rice/Burri-Beta-Carotene-Health-Review-1997.pdf
Beta-carotene research has progressed rapidly in the past few years. New techniques have allowed accurate measurements of betacarotene
concentrations in plasma, tissues, and diet. New animal models, stable isotope techniques, and human depletion and
repletion experiments are leading an explosion in the information about beta-carotene utilization in humans. These advances are
leading to a better understanding of the possibilities and limitations of the influence of beta-carotene on human health. This review
summarizes important new developments in beta-carotene research, and provides an interpretation of how these results reflect the
impact of beta-carotene and human health. 1997 Elsevier Science Inc.
Byrne, P. F. (2006). "Safety and public acceptance of transgenic products." Crop Science 46(1): 113-117.
://000235103700016 AND http://www.botanischergarten.ch/Golden-Rice/Byrne-Safety-Public-Acceptance-2005.pdf
Public acceptance of transgenic (genetically engineered, GE) products is influenced by the perception of direct or indirect risks and
benefits and the credibility of regulatory agencies that evaluate food and environmental safety. In North America acceptance of GE
foods is holding steady, while knowledge about them remains low. Development of transgenic foods with improved nutritional
properties or other quality factors will likely be better received than products that primarily benefit the grower or developer of the
product. There is continuing unease about biopharming and the association, by some members of the public, of GE crops with
corporate agriculture. Several recent reports have reviewed the U.S. regulatory system for transgenic crops and called for a more
coordinated and transparent process that allows for greater public participation. For society to benefit from GE crops, we must
move away from the polarized positions that have defined the transgenic debate in the past, to positions of mutual respect that
allow a rational discussion of the technology's merits and risks.
Canfield, L. M., R. G. Kaminsky, et al. (2001). "Red palm oil in the maternal diet increases provitamin A carotenoids in breastmilk and serum of
the mother-infant dyad." European Journal of Nutrition 40(1): 30-38.
://000167668500005 AND http://www.botanischergarten.ch/Golden-Rice/Canfield-Red-Palm-Oil-2001.pdf
Background Despite vitamin A supplementation programs, vitamin A deficiency in children remains a public health concern in
Honduras. Aim of the Study We investigated the effectiveness of short-term dietary supplementation of mothers with red palm oil
as a strategy for improving the vitamin A status of the mother-infant dyad. Methods Lactating mothers in Colonia Los Pines, a barrio
of Tegucigalpa, Honduras, consumed a total of 90-mg beta -carotene as red palm oil (n = 32) supplements (n = 36) or placebo (n =
18) in six equal doses over 10 days. Carotenoids and retinol in maternal and infant serum, and breastmilk carotenoids and retinol
were measured before and after supplementation. Maternal diet was evaluated by 24-hour recall. Results Maternal serum alpha -
carotene and beta -carotene concentrations were increased 2 fold by palm oil compared with 1.2 fold by beta -carotene
supplements. Changes were significantly different in infant serum alpha -carotene but not beta -carotene among the three
experimental groups. Increases in breastmilk beta -carotene were greater for the palm oil group (2.5 fold) than for the beta -
carotene supplement group (1.6 fold) and increases in milk alpha -carotene concentrations (3.2 fold) were slightly greater than those
of beta -carotene. There were also small but significant changes among groups in breastmilk lutein and lycopene. Breastmilk retinol

was not significantly different among the groups over the treatment period. Conclusions Red palm oil in the maternal diet increases
provitamin A carotenoids in breastmilk and serum of the mother-infant dyad. The use of dietary red palm oil to improve the vitamin
A status of this population should be further investigated.
Carter, M. and J. H. O'Keefe (2008). "Is Red Yeast Rice a Suitable Alternative for Statins?" Mayo Clinic Proceedings 83(11): 1294-1294.
http://www.mayoclinicproceedings.com/content/83/11/1294.1.short AND http://www.botanischergarten.ch/Golden-Rice/Carter-Red-Yeast-
Rice-Suitable-2008.pdf
Chandler, V. L. and S. Wessler (2001). "Grasses. A Collective Model Genetic System." Plant Physiol. 125(3): 1155-1156.
http://www.plantphysiol.org and http://www.botanischergarten.ch/Rice/ChandPlantPys200103.pdf
Chandrika, U. G., E. R. Jansz, et al. (2005). "Analysis of carotenoids in ripe jackfruit (Artocarpus heterophyllus) kernel and study of their
bioconversion in rats." Journal of the Science of Food and Agriculture 85(2): 186-190.
://000226266300002 AND http://www.botanischergarten.ch/Golden-Rice/Chandrika-Analysis-of-Carotenoid-2005.pdf
Vitamin A deficiency is of public health importance in Sri Lanka. Carotenoids are a significant source of provitamin A. The objective of
this study was to analyse the carotenoid composition of jackfruit (Artocarpus heterophyllus sinhala: Waraka) kernel using MPLC and
visible spectrophotometry and to determine the bioavailability and bioconversion of carotenoids present in jackfruit kernel by
monitoring (i) the growth and (ii) levels of retinol and carotenoids in the liver and serum of Wistar rats provided with jackfruit
incorporated into a standard daily diet. Carotenoid pigments were extracted using petroleum ether/methanol and saponified using
10% methanolic potassium hydroxide. Six carotenoids were detected in jackfruit kernel. The carotenes beta-carotene, alphacarotene,
beta-zeacarotene, alpha-zeacarotene and beta-carotene-5,6-epoxide and a dicarboxylic carotenoid, crocetin, were
identified, corresponding theoretically to 141.6 retinol equivalents (RE) per 100g. Our study indicated that jackfruit is a good source
of provitamin A carotenoids, though not as good as papaya. Serum retinol concentrations in rats supplemented with jackfruit
carotenoids were significantly higher (p = 0.008) compared with the control group. The same was true for liver retinol (p = 0.006).
Quantification was carried out by RP-HPLC. These results show that the biological conversion of provitamin A in jackfruit kernel
appears satisfactory. Thus increased consumption of ripe jackfruit could be advocated as part of a strategy to prevent and control
vitamin A deficiency in Sri Lanka. (C) 2004 Society of Chemical Industry.
Chandrika, U. G., E. R. Jansz, et al. (2003). "Carotenoids in yellow- and red-fleshed papaya (Carica papaya L)." Journal of the Science of Food and
Agriculture 83(12): 1279-1282.
://000185120800014 AND http://www.botanischergarten.ch/Golden-Rice/Chandrika-Carotenoids-Papaya-2003.pdf
Vitamin A deficiency is a disorder of public health importance in Sri Lanka. A recent national survey revealed that 36% of preschool
children in Sri Lanka have vitamin A deficiency (serum retinol

eadily accept uncertainty about safety. The major risks related to food are food borne illness and the consequences of poor dietary
choices rather than biotechnology. In general, good availability of reliable information reduces uncertainty. Biotechnologists and
food safety experts are therefore faced with a science communication challenge.
Chen, B., J. Zhang, et al. (1995). "Discovery Of Rice Phytoliths In The Neolithic Site At Jiahu Of Henan Province And Its Significance." Chinese
Science Bulletin 40(14, July).
http://www.carleton.ca/~bgordon/Rice/papers/chen95.htm
Neolithic rice sites play an important role in the study of the origin, evolution and propagation of cultivated rice (1). Of >100 Chinese
Neolithic rice sites, 80% and all earlier ones (Hemudu & Luojiajiao, ca. 7 ka B.P.; Pengtoushan, ca. 7.5-9 ka B.P.) are in the Yangtze
River area. Is there earlier, cultivated rice near the Yellow or Huai Rivers? This problem has received great attention academically.
Jiahu is on the alluvial plain east of Funiu Mountain in central Henan. Based on the analysis of excavated stone and pottery, early
Jiahu people may have lived mainly by agriculture (2), but cultivated type and features remain a mystery.
In recent years, phytolith analysis has found wide archaeological and Quaternary application. Many world scholars concentrate on rice phytolith
study (3-5) and now rice species, even subspecies, are identified by phytolith morphological traits (5-8).
This finding is very significant. It is not only the earliest cultivated rice along the Yellow and Huai Rivers, but among the world’s earliest. Jiahu’s
first period is the same as Pengtoushan culture in Hunan Province[2], considered the earliest evidence of Chinese cultivated rice.
Jiahu people can be considered more advanced than others in the same period, as seen in primitive script and a 7-tone bone flute.
Considering palaeoclimatic edge effects, we deduce early Jiahu people planted rice earlier than Pengtoushan, and the Huai River and
Jiahu may be the cradle of Chinese cultivated rice or one of the cradles.
Much needed data were lacking because little has been preserved and analysis difficult. Many farm tools were excavated in many Chinese
Neolithic sites without crops, a regrettable obstacle to the study of early cultivation. Phytoliths of crops and other plants were much
more easily preserved in sediments and other sites than sporopollens, and easily identified. For this reason, phytolith analysis has
become an important technique in agricultural and environmental archaeology, and will play a very important role in future studies
of the origin and propagation of cultivated rice.
Chong, M. (2003). "Acceptance of golden rice in the Philippine 'rice bowl'." Nature Biotechnology 21(9): 971-972.
://000185051000012 AND http://www.botanischergarten.ch/Golden-Rice/Chong-Acceptance-2003.pdf
Christian, P., K. P. West, et al. (1998). "Night blindness of pregnancy in rural Nepal - nutritional and health risks." International Journal of
Epidemiology 27(2): 231-237.
://000073623600011 AND http://www.botanischergarten.ch/Golden-Rice/Christian-Night-Blindness-Pregnant-1998.pdf
Background Night blindness (XN) is the most common clinical symptom of vitamin A deficiency among children in developing
countries. Yet little is known about the aetiology or associated risks of maternal XN. Emerging evidence from South East Asia
suggests that it may be more frequent than previously thought in women of reproductive age, especially during pregnancy. Methods
A population-based, case-control study was conducted to reveal the epidemiology of XN among pregnant Nepali women. Night blind
cases were identified by history through a weekly community surveillance system. Controls were randomly selected from a pool of
pregnant women without XN and pair-matched for gestational age of the cases. A home-based assessment was done within a week
of selection, at which 7-day food frequency and morbidity histories were collected, anthropometry measured, and capillary blood
drawn for serum retinol, beta-carotene and haemoglobin (Hb) estimation. Results Cases and controls did not differ by age or
number of previous pregnancies. How ever, cases were more likely to be from the lower castes, be illiterate, live in poorer quality
homes, and own no land. The mean serum retinol level of cases was similar to 0.30 mu mol/l lower than controls (P < 0.001),
indicating a low vitamin A status of night blind pregnant women. Mean Hb level was significantly lower (by 0.7 g/dl, P < 0.004), and
the risk of severe anaemia (Hb

Corrigan, J. R., D. P. T. Depositario, et al. (2009). "Comparing Open-Ended Choice Experiments and Experimental Auctions: An Application to
Golden Rice." American Journal of Agricultural Economics 91(3): 837-853.
://WOS:000266796600021 AND http://www.botanischergarten.ch/Golden-Rice/Corrigan-Comparing-Open-ended-2009.pdf
We use two experimental valuation methods to estimate consumer demand for genetically modified golden rice. The first is an
open-ended choice experiment (OECE) where participants name the quantities of golden rice and conventional rice demanded at
each of several price combinations, one of which will be randomly chosen as binding. This allows us to estimate market demand by
aggregating demand across participants. This estimate of market demand also allows us to estimate own-price elasticity and
consumer surplus for golden rice. Comparing willingness-to-pay (WTP) estimates from the OECE with those from a uniform-price
auction, we find that OECE WTP estimates exhibit less affiliation across rounds, and the effects of positive and negative information
under the OECE are more consistent with prior expectations and existing studies. We also find that, while auction WTP estimates
more than double across five rounds, OECE WTP estimates are stable across rounds and are always roughly equal to those from the
final auction round.
Daniell, H. and A. Dhingra (2002). "Multigene engineering: dawn of an exciting new era in biotechnology." Current Opinion in Biotechnology
13(2): 136-141.
://000174643900010 AND http://www.botanischergarten.ch/Golden-Rice/Daniell-Multigene-Engineering-2002.pdf
Development of a rice variety enriched in provitamin A, the accumulation of polyhydroxybutyrate polyester in Arabidopsis nuclear
transgenic plants (with enzymes targeted to chloroplasts in both), and the expression of bacterial operons via the chloroplast
genome are recent landmark achievements in multigene engineering. Hyper-expression of transgenes has resulted in the formation
of insecticidal protein crystals or inclusion bodies of pharmaceutical proteins in transgenic chloroplasts, achieving the highest level of
transgene expression ever reported in transgenic plants. These achievements illustrate the potential of multigene engineering to
realize benefits of the post-genomic revolution.
Datta, K., N. Baisakh, et al. (2003). "Bioengineered 'golden' indica rice cultivars with beta-carotene metabolism in the endosperm with
hygromycin and mannose selection systems." Plant Biotechnology Journal 1(2): 81-90.
://000188439600002 and http://www.botanischergarten.ch/Rice/Dattan-et-al-2003.pdf
Vitamin-A deficiency (VAD) is a major malnutrition problem in South Asia, where indica rice is the staple food. Indica-type rice
varieties feed more than 2 billion people. Hence, we introduced a combination of transgenes using the biolistic system of
transformation enabling biosynthesis of provitamin A in the endosperm of several indica rice cultivars adapted to diverse
ecosystems of different countries. The rice seed-specific glutelin promoter (Gt-1 P) was used to drive the expression of phytoene
synthase (psy), while lycopene beta-cyclase (Icy) and phytoene desaturase (crtl), fused to the transit peptide sequence of the pea-
Rubisco small subunit, were driven by the constitutive cauliflower mosaic virus promoter (CaMV35S P). Transgenic plants were
recovered through selection with either CaMV35S P driven hph (hygromycin phosphotransferase) gene or cestrum yellow leaf
curling virus promoter (CMP) driven pmi (phophomannose isomerase) gene. Molecular and biochemical analyses demonstrated
stable integration and expression of the transgenes. The yellow colour of the polished rice grain evidenced the carotenoid
accumulation in the endosperm. The colour intensity correlated with the estimated carotenoid content by spectro photometric and
HPLC analysis. Carotenoid level in cooked polished seeds was comparable (with minor loss of xanthophylls) to that in non-cooked
seeds of the same transgenic line. The variable segregation pattern in T-1 selfing generation indicated single to multiple loci insertion
of the transgenes in the genome. This is the first report of using nonantibiotic pmi driven by a novel promoter in generating
transgenic indica rice for possible future use in human nutrition.
Datta, K., M. Rai, et al. (2006). "Improved 'golden' indica rice and post-transgeneration enhancement of metabolic target products of
carotenoids (beta-carotene) in transgenic elite cultivars (IR64 and BR29)." Current Science 91(7): 935-939.
://000241392800025 AND http://www.botanischergarten.ch/Rice/Datta-Improved-Golden-Rice-2006.pdf
Transgene stability and post-translational expression levels of genes are of tremendous interest for developing value-added
transgenic crops. Transgenic high-yielding indica rice cultivars (11164 and BR29) with enhanced level of carotenoid accumulation
have been developed by Agrobacterium-mediated transformation. Genetic transformation was done using non-antibiotic Positech
(TM) marker system. Selectable marker gene, phosphomannose isomerase (pmi), and two carotenogenic pathway genes, phytoene
synthase (psy) and phytoene desaturase (crtI) were introduced in two popular Asian rice cultivars, IR64 and BR29. The highest level
of total carotenoids obtained in progenies of trangenic BR29 was 9.34 mu g/g and beta-carotene level alone reached to 3.92 mu g/g
in polished grains. Whereas the highest accumulation of total carotenoids obtained in transgenic progenies of IR64 was 2.32 mu g/g
in polished grains. T2 seeds showed higher carotenoid content than the original parental line which might be attributed to posttransgeneration
effect.
Datta, S. K., K. Datta, et al. (2007). "Golden rice: introgression, breeding, and field evaluation." Euphytica 154(3): 271-278.
://WOS:000244781500003 AND http://www.botanischergarten.ch/Golden-Rice/Datta-Golden-Rice-introgression-2007.pdf
Considerable progress has been made on the genetic engineering of rice for improved nutritional content involving micronutrients
and carotenoid content. Golden Rice, developed by genetic engineering (Agrobacterium and biolistic transformation) was used in
rice breeding for the transfer of high-nutritional value to the local rice cultivars. Simultaneously, commercial Asian indica rice
cultivars were also developed with expression of high-carotenoid levels. The lines were developed based on POSITECH (PMI)
selection system or made marker free by segregating out the marker gene from the gene of interest. Anther culture was used to
develop the homozygous stable lines, which could be of much use in further introgress-breeding and in farmer's field. Enhanced
carotenoids levels (up to T3 generation) were observed in a number of lines compared to the T0-T1 seeds which could be due to
transgeneration effect of growing under greenhouse versus field conditions. However, a few introgressed lines showed less
carotenoid levels than the original lines used in the breeding process. Agronomic performance of introgressed lines, non-transgenic

controls, and transgenic golden rice (IR64 and BR29) developed at IRRI showed acceptable and comparable data under identical
limited field conditions (screenhouse data). Syngenta generated a new Golden Rice (US cultivar) containing high level of carotenoids
grown in the field at Louisiana, USA is expected to be available to the public domain. Incorporation of genes for carotenogenesis in
seeds by transgenesis or by introgression did not change any significant agronomic characteristics in rice plants. The ongoing and
future study of bioavailability, quality, larger field testing and freedom to operate will ensure the benefit of Golden Rice to the
people who need them most.
Davies, K. M. (2007). "Genetic modification of plant metabolism for human health benefits." Mutation Research-Fundamental and Molecular
Mechanisms of Mutagenesis 622(1-2): 122-137.
://WOS:000249319100013 AND http://www.botanischergarten.ch/Rice/Davis-Genetic-Modification-2007.pdf
There has been considerable research progress over the past decade on elucidating biosynthetic pathways for important human
health components of crops. This has enabled the use of genetic modification (GM) techniques to develop crop varieties with
increased amounts of essential vitamins and minerals, and improved profiles of 'nutraceutical' compounds. Much of the research
into vitamins and minerals has focused on generating new varieties of staple crops to improve the diet of populations in developing
nations. Of particular note is the development of new rice lines with increased amounts of provitamin A and iron. Research on
modifying production of nutraceuticals has generally been aimed at generating new crops for markets in the developed nations,
commonly to deliver distinctive cultivars with high consumer appeal. Most progress on nutraceuticals has been made with just a few
types of metabolites to date, in particular in the production of novel long-chain polyunsaturated fatty acids in oil-seed crops and to
increase amounts of flavonoids and carotenoids in tomato and potato. However, given the rapid progress on elucidating plant
metabolite biosynthetic pathways, wide-ranging success with metabolic engineering for levels of human health-related compounds
in plants would be expected in the near future. A key aspect for future success will be better medical information to guide metabolic
engineering endeavors. Although the desired levels of many vitamins are known, detailed information is lacking for most of the
nutraceuticals that have attracted much interest over the past few years. (C) 2007 Elsevier B.V. All rights reserved.
Dawe, D., R. Robertson, et al. (2002). "Golden rice: what role could it play in alleviation of vitamin A deficiency?" Food Policy 27(5-6): 541-560.
://000181303300007 http://www.botanischergarten.ch/Rice/Dawe-Alleviatioin-GR-2002.pdf
Golden rice (GR) is a new rice variety that has been genetically modified to contain beta-carotene, a source of vA. This modification
was undertaken as a strategy to address VAD, which is widespread in less developed countries of Asia. Children's food intake data
from a poor rural region of the Philippines are used to simulate the potential impact of GR on vA intake. The potential impact,
coverage of deficient subpopulations, and costs of GR are compared to two other interventions, food fortification and
supplementation. While investments in future development of GR compare favorably with other interventions in terms of costs and
coverage, GR would deliver amounts of vA that are modest, and unlikely to fulfill requirements. Thus, it should be viewed as a
complement to existing interventions. To have greatest impact at a cost comparable with wheat fortification, GR varieties should be
suited for widespread adoption in Asia and should deliver as much beta-carotene as possible. (C) 2002 Elsevier Science Ltd. All rights
reserved.
DellaPenna, D. (1999). "Carotenoid synthesis and function in plants: Insights from mutant studies in Arabidopsis." Pure and Applied Chemistry
71(12): 2205-2212.
://WOS:000088728300002 AND http://www.botanischergarten.ch/Golden-Rice/DellaPenna-Carotenoid-Synthesis-Plants-1999.pdf
From a molecular and genetic perspective, the decade of the 1990s was truly unparalleled in the study of carotenoids. A
combination of new technologies and approaches allowed the isolation of bacterial carotenoid biosynthetic genes and the
subsequent isolation of higher plant homologs based on colour complementation in Escherichia coil. These genes provided a basis
for molecular and transgenic studies of the carotenoid pathway in various eukaryotic organisms which, although not detailed in this
article, critically and synergistically impacted the work described. For in-depth information of plant carotenoid molecular biology,
readers are directed to a recent comprehensive review [F.X. Cunningham, E. Gantt. Annu. Rev. Plant Physiol. Plant Mel. Biol. 49, 557-
583 (1998)]. The primary focus on this article is to review the identification and characterization of novel mutations in Arabidopsis,
defining genes essential for xanthophyll synthesis in photosynthetic plant tissues. These mutations, both singly and in combination,
allow dramatic alterations in the carotenoid composition of Arabidopsis photosystems in vivo, in the most extreme cases eliminating
the synthesis of all wild-type xanthophylls (lutein, violaxanthin and neoxanthin). This work demonstrates a surprising plasticity in the
carotenoid compositions that plant photosystems will accept in vivo and provides an insight into the functional role(s) of individual
carotenoids in the photosystems.
DellaPenna, D. and B. J. Pogson (2006). "Vitamin synthesis in plants: Tocopherols and carotenoids." Annual Review of Plant Biology 57: 711-738.
://WOS:000239807700027 AND http://www.botanischergarten.ch/Golden-Rice/DellaPenna-Vitamin-Synthesis-2006.pdf
Carotenoids and tocopherols are the two most abundant groups of lipid-soluble antioxidants in chloroplasts. In addition to their
many functional roles in photosynthetic organisms, these compounds are also essential components of animal diets, including
humans. During the past decade, a near complete set of genes required for the synthesis of both classes of compounds in
photosynthetic tissues has been identified, primarily as a result of molecular genetic and biochemical genomics-based approaches in
the model organisms Arabidopsis thaliana and Synecboiystis sp. PCC6803. Mutant analysis and transgenic studies in these and other
systems have provided important insight into the regulation, activities, integration, and evolution of individual enzymes and are
already providing a knowledge base for breeding and transgenic approaches to modify the types and levels of these important
compounds in agricultural crops.
Deodhar, S. Y., S. Ganesh, et al. (2008). "Emerging markets for GM foods: an Indian perspective on consumer understanding and the willingness
to pay." International Journal of Biotechnology 10(6): ISSN 0963-6048(print)|1741-5020(electronic).
://BIOSIS:PREV200900213319 AND http://www.botanischergarten.ch/Golden-Rice/Deodhar-Emerging-Markets-2007.pdf

A random utility approach is used to estimate the logit equations which indicate what factors affect the likelihood of consumption of
non-Genetically Modified (GM) and GM foods and whether or not consumers are willing to pay a premium for non-GM/GM foods.
Ceteris paribus, as the price difference between non-GM and GM foods rises, people arc more likely to consume GM foods. The
likelihood of GM food consumption is higher in the middle income brackets. Consumers are willing to pay an expected premium of
19.5% and 16.12% for golden rice and GM edible oil, respectively. In the case of chicken, consumers seem to pay a very negligible
premium for non-GM-fed chicken. Overall, it appears that GM foods may be acceptable in the Indian market. However, basic
awareness about GM foods may have to be created among the consumers through government ministries, consumer interest
groups and biotechnology food crop companies.
Depee, S., C. E. West, et al. (1995). "Lack of Improvement in Vitamin-a Status with Increased Consumption of Dark-Green Leafy Vegetables."
Lancet 346(8967): 75-81.
://A1995RH22000009 http://www.botanischergarten.ch/Rice/DePee-Lack-1995.pdf
There is little evidence to support the general assumption that dietary carotenoids can improve vitamin A status. We investigated in
Bogor District, West Java, Indonesia, the effect of an additional daily portion of dark-green leafy vegetables on vitamin A and iron
status in women with low haemoglobin concentrations (

that beta-carotene added to soybean oil used in the preparation of rice is absorbed, heated or not, and could be a practical source of
provitamin A. Developing countries looking for strategies to increase vitamin A intake could use fortification of vegetable oils with
synthetic beta-carotene as a simple method.
Englberger, L., W. Aalbersberg, et al. (2003). "Provitamin A carotenoid content of different cultivars of edible pandanus fruit." Journal of Food
Composition and Analysis 16(2): 237-247.
http://www.sciencedirect.com/science/article/B6WJH-489YXJF-1/2/2910142666bb12b184d806207f108af7 AND
http://www.botanischergarten.ch/Golden-Rice/Englberger-Provitamin-A-Pandanus-237-2003.pdf
As part of an overall study to identify vitamin A-rich foods, a study was carried out in the Federated States of Micronesia (FSM) to
provide information on production, acquisition, consumption and cultural acceptability of edible pandanus cultivars, Pandanus
tectorius, and to identify their carotenoid content. Samples of five pandanus cultivars were collected and analyzed for [alpha]- and
[beta]-carotene by HPLC. The results showed that the two cultivars with yellow fruit coloration contained low levels of carotenoids,
while the orange fruits, which were also well liked as a food in the community, contained higher levels at maxima of
190 [mu]g/100 g and 393 [mu]g/100 g for [alpha]- and [beta]-carotene, respectively. Common patterns of intake when the fruit is
available show that pandanus can provide a large proportion of estimated requirements of retinol equivalents. Local people were
generally unaware that pandanus had health benefits, although the food was very popular. Nevertheless, key informants report that
production had greatly decreased in recent years. To reverse this trend, those acceptable cultivars high in carotenoid content should
be promoted both for their general enjoyment and their health benefits.
Englberger, L., W. Aalbersberg, et al. (2003). "Further analyses on Micronesian banana, taro, breadfruit and other foods for provitamin A
carotenoids and minerals." Journal of Food Composition and Analysis 16(2): 219-236.
://000182673500012 AND http://www.botanischergarten.ch/Golden-Rice/Engtberger-Further-Analysis-Provitamin-219-2003.pdf
Few Micronesian foods have been analyzed for nutrient content. Information is needed on locally grown, culturally acceptable foods
that could be promoted to alleviate, vitamin A deficiency in the Federated States of Micronesia. Using an ethnographic approach
that included key informant interviews and observation, Micronesian cultivars with potential for high-carotenoid content according
to their coloration were identified. These cultivars of banana, giant swamp taro, breadfruit and other foods were analyzed for alpha-
and beta-carotene using high-performance liquid chromatography (HPLC) and for nine minerals using inductively coupled plasma
(ICP). A wide range of provitamin A carotenoid levels was found in banana, taro, and breadfruit cultivars, some containing very high
levels (beta-carotene content from 515 to 6360 mug/100 g in banana, 260 to 1651 mug/100 g in taro, and 295 to 868 mug/100 g in
breadfruit, edible portion). Other cultivars contained moderate levels, but as they can be eaten in large quantities, they may
contribute significantly to vitamin A status. The taro samples contained very high levels of zinc (mean 5.9 mg/100 g) and significant
levels of other minerals (mean content of calcium was 120 mg/100 g). These staples with cultural acceptability and high availability
potentially could play a role in vitamin A, micronutrient, and chronic disease programs in the Pacific. (C) 2003 Elsevier Science Ltd. All
rights reserved.
Englberger, L., G. C. Marks, et al. (2004). "Factors to consider in Micronesian food-based interventions: a case study of preventing vitamin A
deficiency." Public Health Nutrition 7(3): 423-431.
://000222034100008 AND http://www.botanischergarten.ch/Golden-Rice/Englberger-Micronesian-Food-Based-2004.pdf
Background: Many factors need to be considered in a food-based intervention. Vitamin A deficiency and chronic diseases, such as
diabetes, heart disease and cancer, have become serious problems in the Federated States of Micronesia (FSM) following the
decreased production and consumption of locally grown foods. However, agricultural and social conditions are still favourable for
local food production. Aim: To identify key factors to consider in a Micronesian food-based intervention focusing on increased
production and consumption of four major Micronesian staple foods: banana, breadfruit, giant swamp taro and pandanus. Methods:
Ethnographic methods including key informant interviews and a literature review. Results: Pacific and Micronesian values, concepts
of food and disease, and food classifications differ sharply from Western concepts. There are few FSM professionals with nutrition
expertise. Traditional foods and food cultivars vary in nutrient content, consumption level, cost, availability, status, convenience in
growing, storing and cooking, and organoleptic factors. Conclusions: A systematic consideration of the factors that relate to a foodbased
intervention is critical to its success. The evaluation of which food and cultivar of that food that might be most effectively
promoted is also critical. Regional differences, for example FSM inter-island differences between the staple foods and cultivars, must
be considered carefully. The evaluation framework presented here may be relevant to Pacific island and other countries with similar
foods where food-based interventions are being planned. An ethnographic approach was found to be essential in understanding the
cultural context and in data collection and analysis.
Englberger, L., J. Schierle, et al. (2003). "Micronesian banana, taro, and other foods: newly recognized sources of provitamin A and other
carotenoids." Journal of Food Composition and Analysis 16(1): 3-19.
://000181886400002 AND http://www.botanischergarten.ch/Golden-Rice/Englberger-Micronesian-Banana-2003.pdf
Vitamin A deficiency (VAD) and chronic diseases are serious problems in the Federated States of Micronesia and other Pacific island
countries. Nutrition education programs to address these in Micronesia have had limited success, partly due to lack of information
on nutrient content in local foods. The study objective was to identify local plant foods rich in provitamin A and other carotenoids
that have high levels of cultural acceptability. Food cultivars likely to be carotenoid-rich (suggested by coloration) were identified
using an ethnographic approach including key informant interviews. Raw and cooked samples (mostly cultivars previously not
analyzed) of 12 banana, 13 giant swamp taro, 10 breadfruit cultivars and four other local foods were analyzed by high-performance
liquid chromatography. Many banana and taro cultivars were found with significant levels of beta- and alpha-carotene; the betacarotene
levels ranged from 30 to 2780 mug/100 g (banana) and. 50 to 2040 mug/100 g (taro). The results highlight the potential
significance of cultivar differences in human nutrition, important for evaluation of the diet, establishment of locally relevant dietary

guidelines, and research on the relationship between diet, health, and disease. These highly acceptable food cultivars could play an
important role in VAD and chronic disease prevention programs in the Pacific. (C) 2003 Elsevier Science Ltd. All rights reserved.
Enserink, M. (2008). "Tough lessons from golden rice." Science 320(5875): 468-471.
://WOS:000255249300029 AND http://www.botanischergarten.ch/Golden-Rice/Enserink-Tough-Lessons-2008.pdf
Golden rice is a rice variety, which produces a group of molecules called pro-vitamin A in its seeds and researchers believe this rice
can be used to fight vitamin A deficiency. By using genetic engineering techniques, it is possible to develop this type of rice and by
normal breeding techniques this is not possible. Golden rice is yet in the field trials and the vitamin A efficiency is yet to be tested.
Favaro, R. M. D., N. V. Desouza, et al. (1986). "VITAMIN-A STATUS OF YOUNG-CHILDREN IN SOUTHERN BRAZIL." American Journal of Clinical
Nutrition 43(5): 852-858.
://A1986C409700018 AND http://www.botanischergarten.ch/Golden-Rice/Favaro-Vitamin-A-Status-Brazil-1986.pdf
Ferry, M. (2002). "Golden rice: what for?" Biofutur(220): 62-62.
://000176658600013 AND http://www.botanischergarten.ch/Golden-Rice/Ferry-A-Quoi-Sert-GR-2002.pdf
Gamble, M. V., N. A. Palafox, et al. (2004). "Carotenoid status among preschool children with vitamin A deficiency in the Republic of the
Marshall Islands." Asia Pacific Journal of Clinical Nutrition 13(4): 336-340.
://000225836900004 AND http://www.botanischergarten.ch/Golden-Rice/Gamble-Carotenoid-Preschool-2004.pdf
Although carotenoids are known to be important dietary sources of vitamin A, there have been few epidemiological studies that
have characterized the serum concentrations of major dietary carotenoids among preschool children with vitamin A deficiency. We
conducted a population-based, cross-sectional study of serum provitamin A carotenoids (alpha-carotene, beta-carotene, betacryptoxanthin),
non-provitamin A carotenoids (lutein/ zeaxanthin, and lycopene), and retinol among 278 children, aged 1-5 y, in the
Republic of the Marshall Islands. Vitamin A deficiency was defined as serum retinol

78 +/- 18% of iron was soluble in the presence of 3.3 mu mol of vitamin A, whereas with no vitamin addition, only 26 +/- 13% of iron
was soluble (

the effects of undemutrition. As newly developed crops with nutritionally improved traits come closer to being available to
producers and consumers, scientifically sound and efficient processes are needed to assess the safety and nutritional quality of
these crops. In 2004, a Task Force of international scientific experts, convened by the International Food Biotechnology Committee
(IFBiC) of ILSI, published recommendations for the safety and nutritional assessment of foods and feeds nutritionally improved
through modem biotechnology (J. Food Science, 2004, 69:CRH62-CRH68). The comparative safety assessment process is a basic
principle in this publication and is the starting point, not the conclusion, of the analysis. Significant differences in composition are
expected to be observed in the case of nutritionally enhanced crops and must be assessed on a case-by-case basis. The Golden Rice
2 case study will be presented as an example of a food crop nutritionally enhanced through the application of modem biotechnology
(i.e., recombinant DNA techniques) to illustrate how the 2004 recommendations provide a robust paradigm for the safety
assessment of "real world" examples of improved nutrition crops.
Goff, S. A., D. Ricke, et al. (2002). "A Draft Sequence of the Rice Genome (Oryza sativa L. ssp. japonica)." Science 296(5565): 92-100.
http://www.sciencemag.org/cgi/content/abstract/296/5565/92 AND http://www.botanischergarten.ch/Rice/Goff-Rice-Genome-2003.pdf AND
http://www.botanischergarten.ch/Rice/Science-Riceposter.pdf
Gomez-Porras, J. L., D. M. Riano-Pachon, et al. (2007). "Genome-wide analysis of ABA-responsive elements ABRE and CE3 reveals divergent
patterns in Arabidopsis and rice." BMC Genomics 8.
://WOS:000250054400001 AND 10.1186/1471-2164-8-260 AND http://www.botanischergarten.ch/Golden-Rice/Gomez-Porras-
Genome-wide-Analysis-ABA-2007.pdf
Background: In plants, complex regulatory mechanisms are at the core of physiological and developmental processes. The
phytohormone abscisic acid ( ABA) is involved in the regulation of various such processes, including stomatal closure, seed and bud
dormancy, and physiological responses to cold, drought and salinity stress. The underlying tissue or plant-wide control circuits often
include combinatorial gene regulatory mechanisms and networks that we are only beginning to unravel with the help of new
molecular tools. The increasing availability of genomic sequences and gene expression data enables us to dissect ABA regulatory
mechanisms at the individual gene expression level. In this paper we used an insilico-based approach directed towards genome-wide
prediction and identification of specific features of ABA-responsive elements. In particular we analysed the genome-wide occurrence
and positional arrangements of two well-described ABA-responsive cis-regulatory elements ( CREs), ABRE and CE3, in thale cress (
Arabidopsis thaliana) and rice ( Oryza sativa). Results: Our results show that Arabidopsis and rice use the ABA-responsive elements
ABRE and CE3 distinctively. Earlier reports for various monocots have identified CE3 as a coupling element ( CE) associated with
ABRE. Surprisingly, we found that while ABRE is equally abundant in both species, CE3 is practically absent in Arabidopsis. ABRE-
ABRE pairs are common in both genomes, suggesting that these can form functional ABA-responsive complexes ( ABRCs) in
Arabidopsis and rice. Furthermore, we detected distinct combinations, orientation patterns and DNA strand preferences of ABRE
and CE3 motifs in rice gene promoters. Conclusion: Our computational analyses revealed distinct recruitment patterns of ABAresponsive
CREs in upstream sequences of Arabidopsis and rice. The apparent absence of CE3s in Arabidopsis suggests that another
CE pairs with ABRE to establish a functional ABRC capable of interacting with transcription factors. Further studies will be needed to
test whether the observed differences are extrapolatable to monocots and dicots in general, and to understand how they contribute
to the fine-tuning of the hormonal response. The outcome of our investigation can now be used to direct future experimentation
designed to further dissect the ABA-dependent regulatory networks.
Goodner, B., G. Hinkle, et al. (2001). "Genome Sequence of the Plant Pathogen and Biotechnology Agent Agrobacterium tumefaciens C58."
Science 294(5550): 2323-2328.
http://www.sciencemag.org/cgi/content/abstract/294/5550/2323 AND http://www.botanischergarten.ch/Genomics/Goodner-Genome-
Agrobacterium-tumefaciens-2001.pdf
Gordon, R. Y. and D. J. Becker (2009). "Significant Difference in LDL-Cholesterol Lowering Using Two Readily Available Red Yeast Rice
Formulations." Journal of the American College of Cardiology 53(10): A206-A206.
://WOS:000263864200855 AND http://www.botanischergarten.ch/Golden-Rice/Gordon-Marked-Variability-Monacolin-2009.pdf
Gordon, R. Y., W. Obermeyer, et al. (2009). "Marked Variability of Monacolin and Citrinin Content of 12 Readily Available Red Yeast Rice
Formulations." Journal of the American College of Cardiology 53(10): A198-A198.
://WOS:000263864200822 AND http://www.botanischergarten.ch/Golden-Rice/Gordon-Marked-Variability-Monacolin-2009.pdf
Goyal, K. C., Y. K. Sharma, et al. (1984). "Restoration of Vitamin-a-Deficiency Induced Lesions by Provitamin-a Carotenoids in Chicks - Studies on
Tissue DNA, Rna and Protein." Nutrition Reports International 30(2): 501-504.
://A1984TG87000029 AND http://www.botanischergarten.ch/Golden-Rice/Goyal-Restoration-Vitamin-Deficiency-.pdf
Graebner, I. T., E. M. A. Siqueira, et al. (2004). "Carotenoids from native Brazilian dark-green vegetables are bioavailable: a study in rats."
Nutrition Research 24(8): 671-679.
://000223728900010 AND http://www.botanischergarten.ch/Golden-Rice/Graebner-Carotenoids-Native-Brazilian-2004.pdf
One of the major dietary sources of vitamin A in the human diet is provitamin A carotenoids. However, the activity of these
carotenoids as a vitamin A source is uncertain due to concerns about the bioavailability of the carotenoids from vegetables. This
study evaluated the bioavailability of provitamin A carotenoids from native Brazilian dark-green vegetables. Vitamin A-depleted rats
were fed a basal diet (AIN-93G) in which the synthetic vitamin A content was replaced by non conventional leaves. At the end of the
30 day-repletion period, 1 mug of retinol accumulated in the liver after the intake of 43.1, 95.3 or 178.9 mug beta-carotene from
Sonchus oleraceus (So), Amaranthus viridis (Av) and Xanthosoma sagittifolium (Xs) leaves, respectively. The relative bioavailability of

eta-carotene from the leaves was 36%, 16%, and 9% for So, Av, and Xs leaf, respectively. The results showed that the carotenoids
from the three dark-green leaves were absorbed, converted to retinol, and stored in the liver of rats. Because they are pest-resistant
and widely distributed vegetables, they may be an inexpensive alternative source of vitamin A to reduce vitamin A deficiency. (C)
2004 Elsevier Inc. All rights reserved.
Graham, J. M., M. J. Haskell, et al. (2007). "Supplementation with iron and riboflavin enhances dark adaptation response to vitamin A-fortified
rice in iron-deficient, pregnant, nightblind Nepali women." American Journal of Clinical Nutrition 85(5): 1375-1384.
://WOS:000246511400028 AND http://www.botanischergarten.ch/Golden-Rice/Graham-Supplementation-Iron-2007.pdf
Background: Nightblindness affects 16-52% of pregnant women in areas of Nepal and in some cases persists after vitamin A
treatment. Iron and riboflavin affect vitamin A utilization and photoreceptor function, respectively, and pilot data in the study
population showed a high prevalence of iron and riboflavin deficiencies. Objective: The objective was to assess the effect of
supplemental iron and riboflavin on pupillary threshold (PT) and plasma retinol in nightblind, pregnant Nepali women given vitamin
A-fortified rice. Design: Nightblind pregnant women were randomly assigned to receive, 6 d/wk under supervision for 6 wk, a
vitamin A-fortified rice curry dish providing 850 jig retinal activity equivalents/d with either a 30-mg Fe and 6-mg riboflavin (FeR +
VA) capsule or a placebo control (VA only) capsule. Hemoglobin, erythrocyte riboflavin, and plasma ferritin and retinol were
measured before and after the intervention. Dark adaptation was assessed by PT score. Results: Women who were iron deficient at
baseline (n = 38) had significantly greater improvement in PT score with iron and riboflavin supplementation than without (P = 0.05).
Iron and riboflavin supplements significantly reduced the prevalences of riboflavin deficiency (from 60% to 6%; P < 0.0001), iron
deficiency anemia (from 35% to 15%; P < 0.007), and abnormal PT (from 87% to 30%; P < 0.05) from baseline. Mean increases in
erythrocyte riboflavin (P < 0.0001) and plasma ferritin (P = 0.01) were greater in the FeR + VA group than in the VA only group.
Conclusions: Iron deficiency may limit the efficacy of vitamin A to normalize dark adaptation in pregnant Nepali women. Further
studies are needed to assess the effect of simultaneous delivery of iron and vitamin A for the treatment of nightblindness.
Graham, R. D., R. M. Welch, et al. (2007). "Nutritious subsistence food systems." Advances in Agronomy, Vol 92 92: 1-74.
://WOS:000243626600001 AND http://www.botanischergarten.ch/Biofortification/Graham-Subsistence-Food-Systems-2008.pdf
The major subsistence food systems of the world that feed resource-poor populations are identified and their capacity to supply
essential nutrients in reasonable balance to the people dependent on them has been considered for some of these with a view to
overcoming their nutrient limitations in sound agronomic and sustainable ways. The approach discusses possible cropping system
improvements and alternatives in terms of crop combinations, external mineral supply, additional crops, and the potential for
breeding staples in order to enhance their nutritional balance while maintaining or improving the sustainability and dietary,
agronomic, and societal acceptability of the system. The conceptual framework calls for attention first to balancing crop nutrition
that in nearly every case will also increase crop productivity, allowing sufficient staple to be produced on less land so that the
remaining land can be devoted to more nutrient-dense and nutrient-balancing crops. Once this is achieved, the additional
requirements of humans and animals (vitamins, selenium, and iodine) can be addressed. Case studies illustrate principles and
strategies. This chapter is a proposal to widen the range of tools and strategies that could be adopted in the HarvestPlus Challenge
Program to achieve its goals of eliminating micronutrient deficiencies in the food systems of resource-poor countries. (c) 2007,
Elsevier Inc.
Greco, R., P. B. F. Ouwerkerk, et al. (2001). "Transposon Insertional Mutagenesis in Rice." Plant Physiol. 125(3): 1175-1177.
http://www.plantphysiol.org and http://www.botanischergarten.ch/Rice/GrecoPlantPhys200103.pdf
Greger, J. (2001). "Genetically engineered "golden" rice unlikely to overcome vitamin A deficiency - Response." Journal of the American Dietetic
Association 101(3): 290-290.
://000170312000006 AND http://www.botanischergarten.ch/Golden-Rice/Greger-Response-Nestle-2001.pdf
Grotto, I., M. Mimouni, et al. (2003). "Vitamin A supplementation and childhood morbidity from diarrhea and respiratory infections: A metaanalysis."
142(3): 297-304.
http://www.sciencedirect.com/science/article/B6WKR-4864GF5-P/2/5f9404331d37b48b9a2663b466bbabff
Objectives To perform an updated meta-analysis of the effect of vitamin A supplementation on childhood morbidity from respiratory
tract infections and diarrhea. Study design A comprehensive search of the 1966 to 2000 MEDLINE database and review of the
reference lists of relevant articles identified 9 randomized controlled trials dealing with morbidity from respiratory infections and
diarrhea in children 6 months to 7 years of age and provided “intention-to-treat” data. Results The combined results indicated that
vitamin A supplementation has no consistent overall protective effect on the incidence of diarrhea (relative risk, 1.00; 95% CI, 0.94-
1.07) and that it slightly increases the incidence of respiratory tract infections (relative risk, 1.08; 95% CI, 1.05-1.11). Conclusions
High-dose vitamin A supplements are not recommended on a routine basis for all preschool children and should be offered only to
individuals or populations with vitamin A deficiency.
Grusak, M. A. (2005). "Golden Rice gets a boost from maize." Nature Biotechnology 23(4): 429-430.
://000228197300017 AND http://www.botanischergarten.ch/Golden-Rice/Grusak-Golden-Rice-Boost-2005.pdf
A more nutritious version of Golden Rice may offer a practical solution to vitamin A deficiency.
Gura, T. (1999). "New genes boost rice nutrient." Science 285: 994 - 995.
http://www.botanischergarten.ch/Golden-Rice/Gura-new-Genes-1999.pdf
Hargrove, T. and W. R. Coffman (2006). "Breeding History." Rice Today October - December: 34-39.
http://www.botanischergarten.ch/Golden-Rice/Hargrove-Breeding-History-Golden-Rice-2006.pdf

Forty years ago, a remarkable rice-breeding project culminated in the release of a rice variety under an unremarkable name—
IR8.This is the story of the research that would ultimately change the face of agriculture across Asia
Haskell, M. J., P. Pandey, et al. (2005). "Recovery from impaired dark adaptation in nightblind pregnant Nepali women who receive small daily
doses of vitamin A as amaranth leaves, carrots, goat liver, vitamin A-fortified rice, or retinyl palmitate." American Journal of Clinical Nutrition
81(2): 461-471.
://000226943100019 and http://www.botanischergarten.ch/Rice/Haskell-Clin-Nutr-Small-Dos-2005.pdf
Background: It is not known whether daily consumption of vitamin A-containing foods is efficacious for treating nightblindness.
Objective: We assessed the effect of supplementation with vitamin A from food or synthetic sources on dark adaptation and plasma
retinol concentrations in nightblind pregnant Nepali women. Design: Nightblind pregnant women were randomly assigned to 1 of 6
treatment groups to receive 6 d/wk for 6 wk either 850 mug retinol equivalents/d as retinyl palmitate, vitamin A-fortified rice, goat
liver, amaranth leaves, or carrots or 2000 mug retinol equivalents/d as retinyl palmitate. Dark adaptation was assessed weekly by
using the pupillary threshold (PT) test; plasma retinol concentrations were measured before and after the intervention. These
outcomes were also assessed in a comparison group of nonnightblind pregnant women. Results: In the nightblind women, the mean
PT improved significantly (P < 0.0001) from -0.71 +/- 0.04 to -1.42 +/- 0.02 log cd/m(2), and the final mean PT did not differ
significantly from that in the nonnightblind women (-1.43 +/- 0.04; P = 0.55). Improvement in dark adaptation was greater in the
liver group than in the vitamin A-fortified rice group (P < 0.02). Plasma retinol concentrations increased significantly (P < 0.0001)
from 0.95 +/- 0.05 to 1.07 +/- 0.05 mumol/L. The plasma retinol response was greater in the higher-dose capsule and liver groups
than in the vegetable groups and significantly greater in the liver group than in the vitamin A-fortified rice group (both: P < 0.05).
Conclusion: Improvement in dark adaptation did riot differ significantly between women who received vitamin A as liver, amaranth
leaves, carrots, or retinyl palmitate.
Hirschberg, J. (2001). "Carotenoid biosynthesis in flowering plants." Genome studies and Molecular Genetics, edited by Juliette de Meaux and
Maarten Koornneef / Plant Biotechnology, edited by Andy Greenland and Jan Leach 4(3): 210-218.
http://www.sciencedirect.com/science/article/B6VS4-42T4DVS-9/2/9d4b62c49e42668ee8b27b5dace34c81 AND
http://www.botanischergarten.ch/Golden-Rice/Hirschberg-Carotenoid-Biosynthesis-2001.pdf
The general scheme of carotenoid biosynthesis has been known for more than three decades. However, molecular description of the
pathway in plants began only in the 1990s after the genes for the carotenogenic enzymes were cloned. Recent data on the
biochemistry of carotenogenesis and its regulation in vivo present the possibility of genetically manipulating this pathway in crop
plants.
Hirschi, K. (2008). "Nutritional improvements in plants: time to bite on biofortified foods." Trends in Plant Science 13(9): 459-463.
://WOS:000259747500001 AND http://www.botanischergarten.ch/Golden-Rice/Hirschi-Nutritional-Improvement-2008.pdf
Modern breeding, molecular genetic and biotechnology studies frequently describe changes in plant metabolism to improve
nutritional content; however, this is often where the process of assessing biofortification ends. Ideally, these modified plants need
to be used in controlled animal and human feeding studies to assess nutritional impact. Such bioavailability studies are crucial if any
claims are to be made regarding health benefits and might be an important component in public acceptance of biofortified foods.
Hoa, T. T. C., S. Al-Babili, et al. (2003). "Golden indica and Japonica rice lines amenable to deregulation." Plant Physiology 133(1): 161-169.
://000185429700018 AND http://www.botanischergarten.ch/Golden-Rice/Hoa-Golden-Indica-Deregulation-2003.pdf
As an important step toward free access and, thus, impact of GoldenRice, a freedom-to-operate situation has been achieved for
developing countries for the technology involved. Specifically, to carry the invention beyond its initial "proof-of-concept" status in a
Japonica rice (Oryza sativa) cultivar, we report here on two transformed elite Indica varieties (IR64 and MTL250) plus one Japonica
variety Taipei 309. Indica varieties are predominantly consumed in the areas with vitamin A deficiency. To conform with regulatory
constraints, we changed the vector backbone, investigated the absence of beyond-border transfer, and relied on Agrobacterium
tumefaciens-mediated transformation to obtain defined integration patterns. To avoid an antibiotic selection system, we now rely
exclusively on phosphomannose isomerase as the selectable marker. Single integrations were given a preference to minimize
potential epigenetic effects in subsequent generations. These novel lines, now in the T, generation, are highly valuable because they
are expected to more readily receive approval for follow-up studies such as nutritional and risk assessments and for breeding
approaches leading to locally adapted variety development.
Horton, S. (2006). "The economics of food fortification." Journal of Nutrition 136(4): 1068-1071.
://WOS:000236456900037 AND http://www.botanischergarten.ch/Golden-Rice/Horton-Economics-Food-Fortification-2006.pdf
This paper summarizes some of the literature on the cost effectiveness and cost benefit of food fortification with selected
micronutrients most relevant for developing countries. Micronutrients covered include iron, iodine, vitamin A, and zinc. The main
focus is on commercial fortification, although home fortification and biofortification are mentioned. Fortification with iron, vitamin
A, and zinc averts significant numbers of infant and child deaths and is a very attractive preventive health-care intervention.
Fortification with iron, iodine, and potentially zinc provides significant economic benefits and the low unit cost of food fortification
ensures large benefit:cost ratios, with effects via cognition being very important for iron and iodine. Fortification will not reach all
individuals and is most attractive as an investment where there is a convenient food vehicle, where processing is more centralized,
and where either the deficiency is widespread or the adverse effects are very costly even though only a small group is affected.
Houdebine, L. M. (2006). "Genetically modified plants (GMPs) and developing countries." Cahiers Agricultures 15(2): 227-231.
://000237307200008 AND http://www.botanischergarten.ch/Golden-Rice/Houdebine-Plantes-genetiquement-development-
2006.pdf

A significant part of agrofood now comes from genetically modified plants (GMPs). Essentially four plants-maize, cotton, soybean,
and rapeseed-share the GMP market. These Cultures tend to plateau in the Countries where they were initially used, namely the
USA and Canada. These new varieties provide farmers with limited but significant advantages on a case by case basis: simplification
of cultures, enhancement of income, reduction of environmental Pollution. Essentially plants resistant to diseases or herbicides are
currently being used. Moreover, a vast majority of available GMPs are used for animal feeding. Papaye resistant to a virus is one of
the only PGM generated for, and used by, human consumption. Logically, the new varieties obtained by transgenesis should be able
to bring more substantial and mainly more rapid advantages to human communities suffering from food deficiency. GMPs currently
under Study aim at improving food composition, by increasing their content in vitamins, in antioxidants, in polyunsaturated fatty
acids, etc. Some projects currently under way indicate that the GMP approach is beginning to pay off. A more extensive use of GMPs
in some developing Countries remains limited because of an insufficient transfer of techniques, because of industrial property
issues, but more directly because local biosafety committees are unable to define how GMPs can be used with no particular risk.
This article aims at showing in which conditions developing Countries can take advantage of GMPs.
Humphrey, J. H., T. Agoestina, et al. (1998). "Neonatal vitamin A supplementation: effect on development and growth at 3 y of age." American
Journal of Clinical Nutrition 68(1): 109-117.
://000074534300017 AND http://www.botanischergarten.ch/Rice/Humphrey-Neonatal-Vitamin-A-1998.pdf
We reported recently that neonatal supplementation with 52 mu mol vitamin A reduced infant mortality by 64%; acute side effects
were limited to a 3% excess rate of a bulging fontanelle. The current study was conducted to identify developmental changes at 3 y
of age associated with neonatal vitamin A supplementation or a bulging fontanelle. Children who had a bulging fontanelle (n = 91)
and 432 children who had normal fontanelles after receiving vitamin A or placebo were evaluated with the Bayley Scales of Infant
Development. Mean scores for the mental, psychomotor, and behavioral rating scale (BRS) plus 3 subscales of the BRS were not
significantly different for treatment-fontanelle-specific groups. In regression models predicting each score, a bulging fontanelle had
a small negative effect in all models; when 1 child who was injured from birth was removed from the analysis the effect of a bulging
fontanelle was not significant in any model (P greater than or equal to 0.35). Vitamin A supplementation had a small beneficial effect
on all developmental scores, which was significant for one of the BRS subscales (orientation-engagement) and also for a second
(motor quality) when the outlier child was removed. Compared with children with normal fontanelles in the placebo group, children
with a bulging fontanelle in the vitamin A group tended to grow less (-0.5 cm, P = 0.33), whereas those with normal fontanelles in
the vitamin A group grew significantly more (0.68 cm, P < 0.05), over the first 3 y of life. This study provides no evidence that
neonatal vitamin A supplementation is associated with biologically significant adverse growth or developmental sequelae.
Humphrey, J. H., T. Agoestina, et al. (1996). "Impact of neonatal vitamin A supplementation on infant morbidity and mortality." Journal of
Pediatrics 128(4): 489-496.
://A1996UF24400007 AND http://www.botanischergarten.ch/Golden-Rice/Humphrey-Impact-Neonatal-Vitamin-A-1996.pdf
Objective: To determine whether vitamin A supplementation at birth could reduce infant morbidity and mortality. Study design: We
conducted a placebo-controlled trial among 2067 Indonesian neonates who received either 52 mu mol (50,000 IU) orally
administered vitamin A or placebo on the first day of life. Infants were followed up at 1 year to determine the impact of this
intervention on infant mortality, A subgroup (n = 470) was also examined at 4 and 6 months of age to examine the impact on
morbidity. Results: Vital status was confirmed in 89% of infants in both groups at 1 year, There were 19 deaths in the control group
and 7 in the vitamin A group (relative risk = 0.36; 95% confidence interval = 0.16, 0.87), The impact wets stronger among boys,
infants of normal compared with low birth weight, and those of greater ponderal index, Among infants examined at 4 months of
age, the 1-week period prevalence of common morbidities was similar for vitamin A and control infants. However, during this same
4-month period, 73% and 51% more control infants were brought for medical treatment for cough (p = 0.008) and fever (p = 0.063),
respectively. Conclusions: Neonatal vitamin A supplementation may reduce the infant mortality rate and the prevalence of severe
respiratory infection among young infants.
Humphrey, J. H., P. J. Iliff, et al. (2008). "Erratum to: Effects of a single large dose of vitamin A, given during the postpartum period to HIVpositive
women and their infants, on child HIV infection, HIV-free survival, and mortality (vol 193, pg 860, 2006)." Journal of Infectious Diseases
197(10): 1485-1485.
://WOS:000255437800019 AND http://www.botanischergarten.ch/Golden-Rice/Humphrey-Erratum-Effects-2008.pdf
Humphrey, J. H., P. J. Iliff, et al. (2006). "Effects of a single large dose of vitamin A, given during the postpartum period to HIV-positive women
and their infants, on child HIV infection, HIV-free survival, and mortality." Journal of Infectious Diseases 193(6): 960-971.
://WOS:000235536200017 AND http://www.botanischergarten.ch/Golden-Rice/Humphrey-Effect-Single-Large-Dose-2006.pdf AND
http://www.botanischergarten.ch/Golden-Rice/Humphrey-Erratum-Effects-2008.pdf
Background. Low maternal serum retinol level is a risk factor for mother-to-child transmission (MTCT) of human immunodeficiency
virus (HIV). Multiple-large-dose vitamin A supplementation of HIV-positive children reduces mortality. The World Health
Organization recommends single-large-dose vitamin A supplementation for postpartum women in areas of prevalent vitamin A
deficiency; neonatal dosing is under consideration. We investigated the effect that single-large-dose maternal/neonatal vitamin A
supplementation has on MTCT, HIV-free survival, and mortality in HIV-exposed infants. Methods. A total of 14,110 mother-infant
pairs were enrolled

infants who were PCR negative at 6 weeks, all 3 vitamin A regimens were associated with similar to 2-fold higher mortality (P

allow comparative analysis with the genome sequence of other organisms. As of October 2004, the database contains a total of 215
Mb sequence with relevant annotation results including 30 000 manually curated genes. The database can provide the latest
information on manual annotation as well as a comprehensive structural analysis of various features of the rice genome. The
database can be accessed at http://rad.dna.affrc.go.jp/.
Jain, M., A. K. Tyagi, et al. (2008). "Genome-wide identification, classification, evolutionary expansion and expression analyses of homeobox
genes in rice." FEBS Journal 275(11 %R doi:10.1111/j.1742-4658.2008.06424.x): 2845-2861.
http://www.blackwell-synergy.com/doi/abs/10.1111/j.1742-4658.2008.06424.x AND http://www.botanischergarten.ch/Rice/Jain-Genome-
.wide-2008.pdf
Homeobox genes play a critical role in regulating various aspects of plant growth and development. In the present study, we
identified a total of 107 homeobox genes in the rice genome and grouped them into ten distinct subfamilies based upon their
domain composition and phylogenetic analysis. A significantly large number of homeobox genes are located in the duplicated
segments of the rice genome, which suggests that the expansion of homeobox gene family, in large part, might have occurred due to
segmental duplications in rice. Furthermore, microarray analysis was performed to elucidate the expression profiles of these genes
in different tissues and during various stages of vegetative and reproductive development. Several genes with predominant
expression during various stages of panicle and seed development were identified. At least 37 homeobox genes were found to be
differentially expressed significantly (more than two-fold; P < 0.05) under various abiotic stress conditions. The results of the study
suggest a critical role of homeobox genes in reproductive development and abiotic stress signaling in rice, and will facilitate the
selection of candidate genes of agronomic importance for functional validation.
Jauhar, P. P. (2006). "Modern biotechnology as an integral supplement to conventional plant breeding: The prospects and challenges." Crop
Science 46(5): 1841-1859.
://WOS:000240821800001 AND http://www.botanischergarten.ch/Golden-Rice/Jauhar-Modern-Biotechnology-Supplement-
2006.pdf
The art of plant breeding was developed long before the laws of genetics became known. The advent of the principles of genetics at
the turn of the last century catalyzed the growth of breeding, making it a science-based technology that has been instrumental in
substantial improvements in crop plants. Largely through exploitation of hybrid vigor, grain yields of several cereal crops were
substantially increased. Intervarietal and interspecific hybridizations, coupled with appropriate cytogenetic manipulations, proved
useful in moving genes for resistance to diseases and insect pests from suitable alien donors into crop cultivars. Plant improvement
has been further accelerated by biotechnological tools of gene transfer, to engineer new traits into plants that are very difficult to
introduce by traditional breeding. The successful deployment of transgenic approaches to combat insect pests and diseases of
important crops like rice (Oryza sativa L.), wheat (Triticum aestivum L.), maize (Zea mays L.), barley (Hordeum vulgare L.), and cotton
(Gossypium hirsutum L.) is a remarkable accomplishment. Biofortification of crops constitutes another exciting development in
tackling global hunger and malnutrition. Golden Rice, genetically enriched with vitamin A and iron, has, for example, the real
potential of saving millions of lives. Yet another exciting application of transgenic technology is in the production of edible vaccines
against deadly diseases. How these novel approaches to gene transfer can effectively supplement the conventional breeding
programs is described. The current resistance to acceptance of this novel technology should be assessed and overcome so that its
full potential in crop improvement can be realized.
Ji, J., G. Wang, et al. (2009). "Functional analysis of multiple carotenogenic genes from Lycium barbarum and Gentiana lutea L. for their effects
on beta-carotene production in transgenic tobacco." Biotechnology Letters 31(2): 305-312.
://WOS:000262123500020 AND http://www.botanischergarten.ch/Golden-Rice/Ji-Functional-analysis-Multiple-carotenogenic-
2009.pdf
Carotenoids are red, yellow and orange pigments, which are widely distributed in nature and are especially abundant in yelloworange
fruits and vegetables and dark green leafy vegetables. Carotenoids are essential for photosynthesis and photoprotection in
plant life and also have different beneficial effects in humans and animals (van den Berg et al. 2000). For example, beta-carotene
plays an essential role as the main dietary source of vitamin A. To obtain further insight into beta-carotene biosynthesis in two
important economic plant species, Lycium barbarum and Gentiana lutea L., and to investigate and prioritize potential genetic
engineering targets in the pathway, the effects of five carotenogenic genes from these two species, encoding proteins including
geranylgeranyl diphosphate synthase, phytoene synthase and delta-carotene desaturase gene, lycopene beta-cyclase, lycopene
epsilon-cyclase were functionally analyzed in transgenic tobacco (Nicotiana tabacum) plants. All transgenic tobacco plants
constitutively expressing these genes showed enhanced beta-carotene contents in their leaves and flowers to different extents. The
addictive effects of co-ordinate expression of double transgenes have also been investigated.
Kawahigashi, H., S. Hirose, et al. (2006). "Phytoremediation of the herbicides atrazine and metolachlor by transgenic rice plants expressing
human CYP1A1, CYP2B6, and CYP2C19." Journal of Agricultural and Food Chemistry 54(8): 2985-2991.
://000237067800028 AND http://www.botanischergarten.ch/Rice/Kawahigashi-Atrazine-2006.pdf
This study evaluated the expression of human cytochrome P450 genes CYP1A1, CYP2B6, and CYP2C19 in rice plants (Oryza sativa cv.
Nipponbare) introduced using the plasmid pIKBACH. The transgenic rice plants (pIKBACH rice plants) became more tolerant toward
various herbicides than nontransgenic Nipponbare rice plants. Rice plants expressing pIKBACH grown in soil showed tolerance to the
herbicides atrazine, metolachlor, and norflurazon and to a mixture of the three herbicides. The degradation of atrazine and
metolachlor by pIKBACH rice plants was evaluated to confirm the metabolic activity of the introduced P450s. Although both
pIKBACH and nontransgenic Nipponbare rice plants could decrease the amounts of the herbicides in plant tissue and culture
medium, pIKBACH rice plants removed greater amounts in greenhouse experiments. The ability of pIKBACH rice plants to remove
atrazine and metolachlor from soil was confirmed in large-scale experiments. The metabolism of herbicides by pIKBACH rice plants
was enhanced by the introduced P450 species. Assuming that public and commercial acceptance is forthcoming, pIKBACH rice plants

may become useful tools for the breeding of herbicide-tolerant crops and for phytoremediation of environmental pollution by
organic chemicals.
Kennedy, D. (2002). "The Importance of Rice." Science 296(5565): 13-.
http://www.sciencemag.org and
http://www.botanischergarten.ch/Rice/Kennedy-Importance-Rice-2003.pdf
Key, S., J. K. C. Ma, et al. (2008). "Genetically modified plants and human health." Journal of the Royal Society of Medicine 101(6): 290-298.
://WOS:000257095000009 AND http://www.botanischergarten.ch/Golden-Rice/Key-Genetically-modified-plants-2008.pdf
Genetically modified (or GM) plants have attracted a large amount of media attention in recent years and continue to do so. Despite
this, the general public remains largely unaware of what a GM plant actually is or what advantages and disadvantages the
technology has to offer, particularly with regard to the range of applications for which they can be used. From the first generation of
GM crops, two main areas of concern have emerged, namely risk to the environment and risk to human health. As GM plants are
gradually being introduced into the European Union there is likely to be increasing public concern regarding potential health issues.
Although it is now commonplace for the press to adopt 'health campaigns', the information they publish is often unreliable and
unrepresentative of the available scientific evidence. We consider it important that the medical profession should be aware of the
state of the art, and, as they are often the first port of call for a concerned patient, be in a position to provide an informed opinion.
This review will examine how GM plants may impact on human health both directly - through applications targeted at nutrition and
enhancement of recombinant medicine production - but also indirectly, through potential effects on the environment. Finally, it will
examine the most important opposition currently facing the worldwide adoption of this technology: public opinion.
King, J. C. (2002). "Biotechnology: A solution for improving nutrient bioavailability." International Journal for Vitamin and Nutrition Research
72(1): 7-12.
://000174524200002 AND http://www.botanischergarten.ch/Golden-Rice/King-Nutrient-Bioavailability-2002.pdf
Biotechnology strategies are now available to improve the amount and availability of nutrients in plant crops: Those strategies
include simple plant selection for varieties with high nutrient density in the seeds, crossbreeding for incorporating a desired trait
within a plant, and genetic engineering to manipulate the nutrient content of the plant. In plant cross-breeding, all genes of the
parent plants are combined and the progeny have both desirable and undesirable traits. To eliminate undesirable traits, plant
breeders back- cross the new plant varieties with other plants over several generations. This technique, called hybridization; has
been used to create varieties of low-phytate corn, barley, and rice. Using the techniques of genetic engineering, the gene(s)
encoding; for a desired trait(s) in a plant are introduced in a precise and controlled manner within a relatively short period of time.
Golden rice, containing carotenoids, and rice with higher amounts of iron, are two examples of genetically engineered plants for
improved nutrition. Genetic engineering has tremendous potential for revolutionizing nutrition. However, public concerns regarding
safety, appearance, and ethics must be overcome before these products can be effectively introduced into the food, supply.
Knight, J. G., D. K. Holdsworth, et al. (2008). "GM food and neophobia: connecting with the gatekeepers of consumer choice." Journal of the
Science of Food and Agriculture 88(5): 739-744.
://WOS:000254212500001 AND http://www.botanischergarten.ch/Golden-Rice/Knight-Perspective-Neophobia-2008.pdf
Resistance to importation of genetically modified (GM) foods in rich countries has deterred governments in many food-exporting
countries from approving the planting of GM food crops for fear of damaging export markets for conventional food. Apart from
governments that have actively imposed barriers to entry, another level of resistance can arise from food distribution channel
members deciding not to import foods which they believe consumers in their markets will not want. GM foods fall in this category in
some markets, particularly in Europe. In China and India, the two most populous consumer markets, pragmatic considerations
appear likely to overcome neophobia regarding this technology, provided that benefits are adequately communicated to consumers.
Choice-modelling experiments show that the same may well be true in Europe. (C) 2008 Society of Chemical Industry.
Kochetkova, T. (2006). "The transatlantic conflict over GM food: Cultural background." Ethics and the Politics of Food: 325-329.
://WOS:000240055000053 AND http://www.botanischergarten.ch/Regulation/Kochetkova-Transatlantic-Conflict-2006.pdf
During the last two decades, an unexpected conflict over GM food arose, with particularly strong transatlantic consequences. Both
the defenders and the opponents of GM food are found on both sides of the Atlantic, however, the opponents, affiliated with NGO's
and ecological parties, have more success in Europe, while the defenders, bound to GM multinationals, are stronger on the
American side. This paper explores the cultural grounds of this conflict. It examines the academic, legal and political argumentation
on GM food to uncover the underlying metaphors. It will claim the current debate to be a rationalization of the top of an iceberg,
the underwater part of which includes visions on food, nature and human relation to Earth. The talk includes the analysis of the
metaphors on GM food, like 'Frankenstein foods' or 'golden rice', and food as a 'program' or 'mechanism'. The conflict is seen as
existing between, on the one hand, the Enlightenment ideology, equating living entities to mechanical ones, i.e., objects of
engineering, and, on the other hand, the organic approach, rooted in traditional world views, which attributes importance not only
to the final result but to the actual processing of organisms into food as well. The debate involves the metaphors of 'pure' versus
'impure' food, 'natural' versus 'artificial', with overtones traceable to alternative visions of morality. A contrast is evident between
Europeans, inclined to accept the safety of traditional foodstuffs like raw milk cheese, and Americans, more attracted to
experiments with new food processing technologies.
Krawinkel, M. B. (2007). "What we know and don't know about Golden Rice." Nature Biotechnology 25(6): 623-623.
://WOS:000247077500011 AND http://www.botanischergarten.ch/Golden-Rice/Krawinkel-What-Know-not-Know-2007.pdf

Kusaba, S., Y. Kano-Murakami, et al. (1998). "Alteration of hormone levels in transgenic tobacco plants overexpressing the rice homeobox gene
OSH1." Plant Physiology 116(2): 471-476.
http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?db=m&form=6&dopt=r&uid=9489007
The rice (Oryza sativa L.) homeobox gene OSH1 causes morphological alterations when ectopically expressed in transgenic rice,
Arabidopsis thaliana, and tobacco (Nicotiana tabacum L.) and is therefore believed to function as a morphological regulator gene. To
determine the relationship between OSH1 expression and morphological alterations, we analyzed the changes in hormone levels in
transgenic tobacco plants exhibiting abnormal morphology. Levels of the plant hormones indole-3- acetic acid, abscisic acid,
gibberellin (GA), and cytokinin (zeatin and trans-zeatin [Z]) were measured in leaves of OSH1-transformed and wild- type tobacco.
Altered plant morphology was found to correlate with changes in hormone levels. The more severe the alteration in phenotype of
transgenic tobacco, the greater were the changes in endogenous hormone levels. Overall, GA1 and GA4 levels decreased and
abscisic acid levels increased compared with wild-type plants. Moreover, in the transformants, Z (active form of cytokinin) levels
were higher and the ratio of Z to Z riboside (inactive form) also increased. When GA3 was supplied to the shoot apex of
transformants, internode extension was restored and normal leaf morphology was also partially restored. However, such GA3treated
plants still exhibited some morphological abnormalities compared with wild-type plants. Based on these data, we propose
the hypothesis that OSH1 affects plant hormone metabolism either directly or indirectly and thereby causes changes in plant
development.
Laget, P. and M. Cantley (2001). "European responses to biotechnology: Research, regulation, and dialogue." Issues in Science and Technology
17(4): 37-42.
://000172172100022 AND http://www.botanischergarten.ch/Regulation/Laget-Cantley-European-Responses-2001.pdf
Lampert, J. M., J. Holzschuh, et al. (2003). "Provitamin A conversion to retinal via the beta,beta-carotene-15,15 '-oxygenase (bcox) is essential
for pattern formation and differentiation during zebrafish embryogenesis." Development 130(10): 2173-2186.
://000183143800014 AND http://www.botanischergarten.ch/Golden-Rice/Lampert-Provitamin-A-Conversion-2003.pdf
The egg yolk of vertebrates contains carotenoids, which account for its characteristic yellow color in some species. Such plantderived
compounds, e.g. beta-carotene, serve as the natural precursors (provitamins) of vitamin A, which is indispensable for
chordate development. As egg yolk also contains stored vitamin A, carotenoids have so far been solely discussed as pigments for the
coloration of the offspring. Based on our recent molecular identification of the enzyme catalyzing provitamin A conversion to
vitamin A, we address a possible role of provitamin A during zebrafish (Danio rerio) development. We cloned the zebrafish gene
encoding the vitamin A-forming enzyme, a beta,beta-carotene-15,15'-oxygenase. Analysis of its mRNA expression revealed that it is
under complex spatial and temporal control during development. Targeted gene knockdown using the morpholino antisense
oligonucleotide technique indicated a vital role of the provitamin A-converting enzyme. Morpholino-injected embryos developed a
morphological phenotype that included severe malformation of the eyes, the craniofacial skeleton and pectoral fins, as well as
reduced pigmentation. Analyses of gene expression changes in the morphants revealed that distinct retinoic acid-dependent
developmental processes are impaired, such as patterning of the hindbrain and differentiation of hindbrain neurons, differentiation
of neural crest derivatives (including the craniofacial skeleton), and the establishment of the ventral retina. Our data provide strong
evidence that, for several developmental processes, retinoic acid generation depends on local de novo formation of retinal from
provitamin A via the carotene oxygenase, revealing an unexpected, essential role for carotenoids in embryonic development.
Lewinsohn, E. and M. Gijzen (2009). "Phytochemical diversity: The sounds of silent metabolism." Plant Science 176(2): 161-169.
://WOS:000262566600001 AND http://www.botanischergarten.ch/Golden-Rice/Lewinsohn-Silent-Metabolism-2009.pdf
Plants produce tens of thousands of different natural products also referred to as secondary metabolites. These metabolites were
once thought to be the result of aberrant metabolism, or a form of transient storage of byproducts and intermediates thereof.
Although the true role of such metabolites in plants remains mostly unknown, it is evident that plants invest a great deal of
resources in synthesizing, accumulating and sorting such metabolites. often produced through complex and highly regulated
biosynthetic pathways operating in multiple cellular and sub-cellular compartments. There is also growing evidence indicating that
many biosynthetic pathways leading to the accumulation of plant natural products are not fully active. Thus, occult enzymes exist,
sometimes without any apparent endogenous substrate or function, suggesting that plants have a reservoir of metabolic capabilities
that normally remains hidden or unused. It is often difficult to accurately guess what are the actual biological roles of such enzymes
solely based on bioinformatics, due to promiscuity towards substrates and the relatively ease to change substrate or product
specificity by introducing minor changes in sequence of the enzymes. It could be that such orphan enzyme activities are relics of a
recent past that have not been fully eliminated through selection and evolution. Additionally, it could be that such occult activities
possess unknown biochemical roles and coincidentally are able to accept novel substrates. We have coined the term "silent
metabolism" to describe occult metabolic capacities present or induced in plants. A few examples illustrating silent metabolism in
the terpenoid and phenylpropanoid pathways, as well as their repercussion in the metabolic engineering of plant secondary
metabolism are discussed in this review. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
Li, H. W., C. H. Hu, et al. (1962). "Progress report on the studies of utilization of X-ray radiation for rice improvement, 1961." The International
Journal of Applied Radiation and Isotopes 13(7-8): 455-465.
http://www.sciencedirect.com/science/article/B6X3R-46WMW8B-SN/2/6407711920a4620d8cc775c79d549252
Lidén, M., K. Tryggvason, et al. (2003). "Structure and function of retinol dehydrogenases of the short chain dehydrogenase/reductase family."
Molecular Aspects of Medicine 24(6): 403-409.
http://www.sciencedirect.com/science/article/B6T9P-494C34R-1/2/2f4c8faf04dd047dddb64c193fa3ad0c AND
http://www.botanischergarten.ch/Golden-Rice/Liden-Structure-Function-Retinol-2003.pdf

All-trans-retinol is the common precursor of the active retinoids 11-cis-retinal, all-trans-retinoic acid (atRA) and 9-cis-retinoic acid
(9cRA). Genetic and biochemical data supports an important role of the microsomal members of the short chain
dehydrogenases/reductases (SDRs) in the first oxidative conversion of retinol into retinal. Several retinol dehydrogenases of this
family have been reported in recent years. However, the structural and functional data on these enzymes is limited. The prototypic
enzyme RDH5 and the related enzyme CRAD1 have been shown to face the lumen of the endoplasmic reticulum (ER), suggesting a
compartmentalized synthesis of retinal. This is a matter of debate as a related enzyme has been proposed to have the opposite
membrane topology. Recent data indicates that RDH5, and presumably other members of the SDRs, occur as functional
homodimers, and need to interact with other proteins for proper intracellular localization and catalytic activity. Further analyses on
the compartmentalization, membrane topology, and functional properties of microsomal retinol dehydrogenases, will give
important clues about how retinoids are processed.
Lorch, A. (2001). "Is this the way to solve malnutrition?" Biotechnology and Development Monitor(44-45): 18-22.
://000168291200007 AND http://www.botanischergarten.ch/Golden-Rice/Lorch-Golden-Rice-is-this-the-way-2001.pdf
Vitamin A deficiency (VAD) is a serious form of malnutrition that weakens the immune system and may cause blindness. Several
measures address VAD and have shown positive results. Genetically modified rice containing beta-carotene is a new approach in an
early experimental stage. Golden Rice demonstrates the problems of public research in an area where both plant and technology are
heavily protected by patents.
Lowe, G. M., K. Vlismas, et al. (2003). "Carotenoids as prooxidants?" Molecular Aspects of Medicine 24(6): 363-369.
http://www.sciencedirect.com/science/article/B6T9P-49202PF-1/2/bb7c010673c533ee202fd077c7da800f AND
http://www.botanischergarten.ch/Golden-Rice/Stahl-Antioxidant-Activity-Carotenoids-2003.pdf
Lubzens, E., L. Lissauer, et al. (2003). "Carotenoid and retinoid transport to fish oocytes and eggs: what is the role of retinol binding protein?"
Molecular Aspects of Medicine 24(6): 441-457.
http://www.sciencedirect.com/science/article/B6T9P-496FMKX-1/2/380b39de60acb58684d9fad6cc66ee70 AND
http://www.botanischergarten.ch/Golden-Rice/Lubzens-Carotenoid-Retinoid-2003.pdf
Fish eggs contain carotenoids, retinals (retinal and dehydroretinal) and retinols (retinol, dehydroretinol and retinyl-esters) that are
utilized during embryonic development, after fertilization. The carotenoids (mainly astaxanthins) are transported in the plasma by
the low density lipoproteins, high density lipoproteins, and very high density lipoproteins (VHDL) and were found to be associated
also with serum albumin. Retinals were found to be associated vitellogenin (VTG), a component of the plasma VHDL fraction that is
internalized by oocytes during vitellogenesis. However, the transport of retinols and retinyl-esters that were located in the oil
droplet fraction of homogenized eggs, has yet to be elucidated. Retinols are more abundant in freshwater fish eggs than in eggs of
marine fish species. Since retinol is transported in the plasma of vertebrates in association with retinol binding protein (RBP), recent
studies on the molecular characterization and expression sites of RBP, could contribute to determining the involvement of RBP in
transporting retinol to developing oocytes in vertebrates. Recently, results from our laboratory show that RBP mRNA levels in the
liver and RBP plasma levels did not significantly change with the onset and during vitellogenesis in the Rainbow trout. These results
were in contrast with a dramatic elevation in the mRNA levels of VTG in the liver and an increase in VTG plasma levels that was
observed in the same females. Moreover, 17[beta]-estradiol treatment of immature fish, resulted in relatively lower mRNA levels of
RBP in the liver, concomitantly with an increase in the level of VTG transcripts and the appearance of VTG in the plasma of treated
fish. In addition, RBP was localized in the cytosol of ovulated oocytes. These results for Rainbow trout are similar to those reported
for the chicken but differ from those of Xenopus, where an increase in RBP mRNA was reported in the liver and higher levels of
retinal and retinol were found in the plasma of 17[beta]-estradiol treated animals. The results, reported here for the first time in
Rainbow trout, showing RBP transcripts in the ovary, oviduct (the ovarian tissue adjacent to the gonopore) and oocytes, suggest a
modulating role for RBP in follicular development, as has been suggested for the bovine ovary.
Lucca, P., S. Poletti, et al. (2006). "Genetic engineering approaches to enrich rice with iron and vitamin A." Physiologia Plantarum 126(3): 291-
303.
http://www.blackwell-synergy.com/doi/abs/10.1111/j.1399-3054.2006.00609.x AND http://www.botanischergarten.ch/Golden-Rice/Lucca-
Genetic-engineering-rice-2006.pdf
Major staple crops are often deficient in some of the nutrients required in human diet. Thus, malnutrition is a major problem,
especially in developing countries, where a diversified diet is not affordable for the majority. Several strategies have been adopted
to improve nutrition. However, micronutrient deficiency is still widely spread. Rice is one of the most important staple foods for a
large part of the world's population. Therefore, even a small improvement in nutritional content of rice seeds can have a dramatic
impact on human health. Different approaches are being exploited to produce rice enhanced in nutrients with iron and provitamin
A.
Lusk, J. L. (2003). "Effects of Cheap Talk on Consumer Willingness-to-Pay for Golden Rice." American Journal of Agricultural Economics 85(4 %R
doi:10.1111/1467-8276.00492): 840-856.
http://www.blackwell-synergy.com/doi/abs/10.1111/1467-8276.00492 AND http://www.botanischergarten.ch/Rice/Lusk-Cheaptalk-2003.pdf
A large body of literature suggests willingness-to-pay is overstated in hypothetical valuation questions as compared to when actual
payment is required. Recently, "cheap talk" has been proposed to eliminate the potential bias in hypothetical valuation questions.
Cheap talk refers to process of explaining hypothetical bias to individuals prior to asking a valuation question. This study explores the
effect of cheap talk in a mass mail survey using a conventional value elicitation technique. Results indicate that cheap talk was
effective at reducing willingness-to-pay for most survey participants; however, consistent with previous research, cheap talk did not
reduce willingness-to-pay for knowledgeable consumers.

Manley, T. (2005). "Golden rice." Chemistry & Industry(10): 14-14.
://000229294800020 AND http://www.botanischergarten.ch/Golden-Rice/Manley-GoldenRice-Letter-2005.pdf
Matsuo, T. (1962). "Review of Research on Use of Radiation-Induced Mutations in Crop Breeding with Special Reference to Rice in Japan."
International Journal of Applied Radiation and Isotopes 13(JUL-A): 477-&.
://A19622688B00013 AND http://www.botanischergarten.ch/Mutations/Matsuo-Rice-Radiation.pdf
Mayer, J. E. (2005). "The Golden Rice controversy: Useless science or unfounded criticism?" Bioscience 55(9): 726-727.
://000231524700003 AND http://www.botanischergarten.ch/Rice/Mayer-Controversy-2005.pdf
Mayer, J. E. (2007). "Delivering Golden Rice to developing countries." Journal of Aoac International 90: 1445-1449.
://WOS:000249752800031 AND http://www.botanischergarten.ch/Golden-Rice/Mayer-Delivering-Golden-Rice-2007.pdf
Micronutrient deficiencies create a vicious circle of malnutrition, poverty, and economic dependency that we must strive to break.
Golden Rice offers a sustainable solution to reduce the prevalence of vitamin A deficiency-related diseases and mortality, a problem
that affects the health of millions of children in all developing countries. The technology is based on the reconstitution of the
carotenoid biosynthetic pathway by addition of 2 transgenes. The outcome of this high-tech approach will be provided to end users
as nutrient-dense rice varieties that are agronomically identical to their own, locally adapted varieties. This intervention has the
potential to reach remote rural populations without access to fortification and supplementation programs. As part of our delivery,
strategy, we are partnering with government and nongovernment, national and international agricultural institutions to navigate
through cumbersome and expensive regulatory regimes that affect the release of genetically modified crops, and to create local
demand for the biofortified rice varieties.
Mayer, J. E., W. H. Pfeiffer, et al. (2008). "Biofortified crops to alleviate micronutrient malnutrition." Genome studies and Molecular Genetics,
edited by Juliette de Meaux and Maarten Koornneef / Plant Biotechnology, edited by Andy Greenland and Jan Leach 11(2): 166-170.
http://www.sciencedirect.com/science/article/B6VS4-4S0R701-1/1/e12139b40ae67abc932e4bdb46069503 AND
http://www.botanischergarten.ch/Rice/Mayer-Biofortified-COPB-2008..pdf
Micronutrient malnutrition affects more than half of the world population, particularly in developing countries. Concerted
international and national fortification and supplementation efforts to curb the scourge of micronutrient malnutrition are showing a
positive impact, alas without reaching the goals set by international organizations. Biofortification, the delivery of micronutrients via
micronutrient-dense crops, offers a costeffective and sustainable approach, complementing these efforts by reaching rural
populations. Bioavailable micronutrients in the edible parts of staple crops at concentrations high enough to impact on human
health can be obtained through breeding, provided that sufficient genetic variation for a given trait exists, or through transgenic
approaches. Research and breeding programs are underway to enrich the major food staples in developing countries with the most
important micronutrients: iron, provitamin A, zinc and folate
McCarthy, E., J. Liu, et al. (2002). "Long terminal repeat retrotransposons of Oryza sativa." Genome Biology 3(10): research0053.1 -
research0053.11.
http://genomebiology.com/2002/3/10/research/0053
BACKGROUND:Long terminal repeat (LTR) retrotransposons constitute a major fraction of the genomes of higher plants. For
example, retrotransposons comprise more than 50% of the maize genome and more than 90% of the wheat genome. LTR
retrotransposons are believed to have contributed significantly to the evolution of genome structure and function. The genome
sequencing of selected experimental and agriculturally important species is providing an unprecedented opportunity to view the
patterns of variation existing among the entire complement of retrotransposons in complete genomes.RESULTS:Using a new datamining
program, LTR_STRUC, (LTR retrotransposon structure program), we have mined the GenBank rice (Oryza sativa) database as
well as the more extensive (259 Mb) Monsanto rice dataset for LTR retrotransposons. Almost two-thirds (37) of the 59 families
identified consist of copia-like elements, but gypsy-like elements outnumber copia-like elements by a ratio of approximately 2:1. At
least 17% of the rice genome consists of LTR retrotransposons. In addition to the ubiquitous gypsy- and copia-like classes of LTR
retrotransposons, the rice genome contains at least two novel families of unusually small, non-coding (non-autonomous) LTR
retrotransposons.CONCLUSIONS:Each of the major clades of rice LTR retrotransposons is more closely related to elements present in
other species than to the other clades of rice elements, suggesting that horizontal transfer may have occurred over the evolutionary
history of rice LTR retrotransposons. Like LTR retrotransposons in other species with relatively small genomes, many rice LTR
retrotransposons are relatively young, indicating a high rate of turnover.
Miller, H. I. (2009). "A golden opportunity, squandered." Trends in Biotechnology 27(3): 129-130.
http://www.sciencedirect.com/science/article/B6TCW-4VGTGRX-1/2/89948850e8a0e9a03b66f54ad6257a6b AND
http://www.botanischergarten.ch/Golden-Rice/Miller-Golden-Opportunity-2009.pdf
Minorsky, P. V. (2001). "Introduction Cereal Grains." Plant Physiol. 125(3): 1153-1154.
http://www.plantphysiol.org and http://www.botanischergarten.ch/Rice/Minorsky1PlantPhys200103.pdf
Minorsky, P. V. (2001). " Molecular biology of starch-branching and -debranching enzymes." Plant Physiol. 125(3): 1162-1163.
http://www.plantphysiol.org and http://www.botanischergarten.ch/Rice/Minorsky2PlantPhys200103.pdf
Monastra, G. and L. Rossi (2003). "Transgenic foods as a tool for malnutrition elimination and their impact on agricultural systems." Rivista Di
Biologia-Biology Forum 96(3): 363-383.
://000189259600002 AND http://www.botanischergarten.ch/Golden-Rice/Monastra-Transgenic-Foods-Tools-2003.pdf

GMO crops were introduced for commercial production in 1996 Since then, their use has increased rapidly. GMOs have primarily
benefited large farms and multinational companies in Industrialised Countries and now is more and more debating their utilisation in
Developing World. The objective of the present review is an analysis of this subject from a comprehensive point of view; in addition
to that, the changes related to the nutritional content of transgenic foods will be treated. Despite the progress that has been made,
the world food situation is still marked by mass hunger and chronic malnutrition. In particular micronutrient malnutrition, that
means vitamin and mineral deficiencies, represents an important public health problem in several areas of the world. The "golden
rice", bioengineered to contain beta-carotene, as a source of vitamin A is the most famous example of GM food used for reduction
(or even to solve) of a public health problem. The expected results of this approach have presently not been achieved. Further
studies are necessary to increase the general knowledge about GMOs and their long-term effects on human health. Collaborative
attitude of different research sectors (private and public) and involvement of different sectors of society will be an added value for
comprehension of the real impact of the application of modern biotechnology to food and agriculture systems.
Nandakumar, N., A. K. Singh, et al. (2004). "Molecular fingerprinting of hybrids and assessment of genetic purity of hybrid seeds in rice using
microsatellite markers." Euphytica 136(3): 257-264.
://000222236000004 AND http://www.botanischergarten.ch/Golden-Rice/Nandakumar-Molecular-Fingerprinting-2004.pdf
Microsatellite markers were used for fingerprinting of hybrids, assessing variation within parental lines and testing the genetic purity
of hybrid seed lot in rice. Ten sequence tagged microsatellite sites (STMS) markers were employed for fingerprinting 11 rice hybrids
and their parental lines. Nine STMS markers were found polymorphic across the hybrids and produced unique fingerprint for the 11
hybrids. A set of four markers (RM 206, RM 216, RM 258 and RM 263) differentiated all the hybrids from each other, which can be
used as referral markers for unambiguous identification and protection of these hybrids. Cluster analysis based on Jaccard's
similarity coefficient using UP-GMA grouped the hybrids into three clusters. Within the cluster all the hybrids shared a common
cytoplasmic male sterile line as female parent. The genetic similarity between the hybrids ranged from 0.33 to 0.92 with an average
similarity index of 0.63. The analysis of plant-to-plant variation within the parental lines of the hybrid Pusa RH 10, using informative
markers indicated residual heterozygosity at two marker loci. This highlights the importance of STMS markers in maintaining the
genetic purity of the parental lines. The unique value of the restorer gene linked marker for testing the genetic purity of hybrid seeds
is demonstrated for the first time.
Nandakumar, N., N. K. Tyagi, et al. (2008). "Molecular analysis of transgene (psy) inheritance in a golden rice line developed in the genetic
background of IR64." Journal of Plant Biochemistry and Biotechnology 17(2): 127-132.
://WOS:000258336900002 AND http://www.botanischergarten.ch/Golden-Rice/Nandakumar-Molecular-Analysis-Golden-2008.pdf
Stability in transgene inheritance and expression is an important consideration in utilizing transgenic germplasm for the
development of commercially viable transgenic crops. In the present study, inheritance of the carotenogenic transgene encoding
enzyme phytoene synthase (psy), was studied in rice through PCR analysis in F-2, BC1F1 (Swarna X F-1), its reciprocal cross (F-1 X
Swarna) and BC2F2 populations., Segregation distortion was observed for the transgene in F-2 and BC1F1 (Swarna X F-1)
populations. However, in the case of reciprocal BC1F1 (F-1 X Swarna) cross, the transgene showed normal Mendelian segregation
ratio 1:1. This trend was also observed in the BC2F2 where the transgene followed an expected seregation ratio of 3:1. Segregation
distortion of the transgene has been explained on the basis of the position effect of gametophytic selection locus ga2. The results
obtained here are significant for the molecular marker assisted transfer of the golden rice transgene to widely grown Indian rice
varieties.
Nasiruddin, K. M. and A. Nasim (2007). "Development of agribiotechnology and biosafety regulations used to assess safety of genetically
modified crops in Bangladesh." Journal of Aoac International 90: 1508-1512.
://WOS:000249752800039 AND http://www.botanischergarten.ch/Golden-Rice/Nasiruddin-Development-Agribiotechnology-
2007.pdf
Bangladesh is on the verge of adopting genetically modified (GM) crops for commercial cultivation and consumption as feed and
food. Most of the laboratories are engaged in tissue culture and molecular characterization on plants, whereas some have started
living modified organism research with shortages of trained manpower, infrastructure, and funding. Nutritionally improved Golden
Rice, biotech brinjal, and late blight-resistant potato are in contained trials in a greenhouse, and potato ring spot virus-resistant
papaya is in the process of approval for a field trial. The government has taken some initiative in support of GM organism research,
which include the formation of a Biotechnology Department in all institutes and the formation of the apex body, the National Task
Force Committee on Biotechnology of Bangladesh under the chairpersonship of the Prime Minister. Biosafety policy guidelines and
related aspects of biotechnology issues have been approved, and the laws are in the process of being promulgated. Being a party to
the Cartagena Protocol, proper biosafety measures are regulated by the appropriate authority as stated. Although there are no laws
made yet directly for biosafety of GM crops/foods, the relevant laws on agriculture, medicine, food, import, trade, environment, etc.
may suffice and explain the situation.
Neidecker-Gonzales, O., P. Nestel, et al. (2007). "Estimating the global costs of vitamin A capsule supplementation: A review of the literature."
Food and Nutrition Bulletin 28: 307-316.
://WOS:000249801500007 AND http://www.botanischergarten.ch/Golden-Rice/Neidecker-Gonzales-Estimating-costs-Vitamin-Acapsule-2007.pdf
Background. Vitamin A supplementation reduces child mortality. It is estimated that 500 million vitamin A capsules are distributed
annually. Policy recommendations have assumed that the supplementation programs offer a proven technology at a relatively low
cost of around US$0.10 per capsule. Objectives. To review data on costs of vitamin A supplementation to analyze the key factors
that determine program costs, and to attempt to model these costs as a function of per capita income figures. Methods. Using data
from detailed cost studies in seven countries, this study generated comparable cost categories for analysis, and then used the
correlation between national incomes and wage rates to postulate a simple model where costs of vitamin A supplementation are

egressed on per capita incomes. Results. Costs vary substantially by country and depend principally on the cost of labor, which is
highly correlated with per capita income. Two other factors driving costs are whether the program is implemented in conjunction
with other health programs, such as National Immunization Days (which lowers costs), and coverage in rural areas (which increases
costs). Labor accounts for 70% of total costs, both for paid staff and for volunteers, while the capsules account for less than 5%.
Marketing, training, and administration account for the remaining 25%. Conclusions. Total costs are lowest (roughly US$0.50 per
capsule) in Africa, where wages and incomes are lowest, US$l in developing countries in Asia, and US$1.50 in Latin America. Overall,
this study derives a much higher global estimate of costs of around US$l per capsule.
Nestel, P., H. E. Bouis, et al. (2006). "Biofortification of Staple Food Crops." Journal of Nutrition 136(4): 1064-1067.
http://jn.nutrition.org/cgi/content/abstract/136/4/1064 AND http://www.botanischergarten.ch/Golden-Rice/Nestel-Biofortification-Staple-
Crop-2006.pdf
Deficiencies of vitamin A, iron, and zinc affect over one-half of the world's population. Progress has been made to control
micronutrient deficiencies through supplementation and food fortification, but new approaches are needed, especially to reach the
rural poor. Biofortification (enriching the nutrition contribution of staple crops through plant breeding) is one option. Scientific
evidence shows this is technically feasible without compromising agronomic productivity. Predictive cost-benefit analyses also
support biofortification as being important in the armamentarium for controlling micronutrient deficiencies. The challenge is to get
producers and consumers to accept biofortified crops and increase their intake of the target nutrients. With the advent of good seed
systems, the development of markets and products, and demand creation, this can be achieved.
Nestle, M. (2001). "Genetically engineered "golden" rice unlikely to overcome vitamin A deficiency." Journal of the American Dietetic
Association 101(3): 289-290.
://000170312000005 AND http://www.botanischergarten.ch/Golden-Rice/Greger-Response-Nestle-2001.pdf
Normile, D. (2000). "Agriculture biotechnology - Monsanto donates its share of golden rice." Science 289(5481): 843-845.
://000088701200002 AND http://www.botanischergarten.ch/Rice/Normile-Monsanto.pdf
Normile, D. and E. Pennisi (2002). "THE RICE GENOME: Rice: Boiled Down to Bare Essentials." Science 296(5565): 32-36.
http://www.sciencemag.org
Nwilene, F., A. Togola, et al. (2008). "Parasitoid biodiversity conservation for sustainable management of the African rice gall midge, Orseolia
oryzivora (Diptera: Cecidomyiidae) in lowland rice." Biocontrol Science and Technology 18(10): 1075-1081.
://WOS:000261802200009 AND http://www.botanischergarten.ch/Rice/Nwilene-Parasitoid-Biodiversity-2008.pdf
Platygaster diplosisae and Aprostocetus procerae attack both the African rice gall midge (AfRGM), Orseolia oryzivora, and Orseolia
bonzii, a closely related gall midge that feeds on Paspalum scrobiculatum. Recent research has shown that managing this weed at
the edge of rice fields offers African farmers, low-cost non-chemical control of AfRGM.
Oyarzun, M. T., R. Uauy, et al. (2001). "Food based approaches to improve vitamin and mineral nutritional adequacy." Archivos
Latinoamericanos De Nutricion 51(1): 7-18.
://000173167800001 AND http://www.botanischergarten.ch/Golden-Rice/Oyarzun-Enfoque-Alimentario-2001.pdf
This paper, using the nutrient density concept and the development of Food-Based Dietary Guidelines (FBDG), explores the
nutritional adequacy of food patterns based on the main staples around the world -maize, wheat, rice and potato. The nutritional
adequacy of the vitamins A, C, folates and the minerals iron, zinc and calcium in diets composed by these staples is analyzed.
Projected diets modified by the addition of foods high in vitamins and minerals, based on food table composition information, are
analyzed for changes in nutritional adequacy, in order to cover the nutritional requirements of all family members. This theoretical
exercise shows that diets diversified by the addition of relatively small quantities (27 - 70 g) of foods like fruits, vegetables, legumes
and meat improve substantially the nutritional adequacy of the vitamins A, C, folates, iron and zinc. In the case of calcium, the
addition of a dairy source becomes essential. From the practical point of view, the implementation of the food based approach
requires a national effort in order to support the production of nutrient rich foods and to promote consciousness of nutrient
bioavailability, so that people may obtain all nutrient potential from foods. Simultaneously, the need of fortification programs at the
national level is recognized. This is the case for critical nutrients, where requirements might be above the potential in available
foods. A similar situation occurs in the case of vulnerable groups like pregnant women and infants, who need supplementation to
ensure nutritional adequacy of their diets. In conclusion, the use of FBDG is a good alternative for improving the nutritional
adequacy of diets based predominantly on staple foods. However, food fortification and supplementation programs directed to
vulnerable groups are necessary and complementary practices to the FBDG implementation.
Paine, J. (2005). "Improving the nutritional value of Golden Rice through increased pro-vitamin A content." Nature Biotechnology Published
online 27 March 2005; doi:10.1038/nbt1082: 1-6.
http://www.botanischergarten.ch/Rice/Paine-et-al-2005-NB-Golden-Rice.pdf
‘Golden Rice’ is a variety of rice engineered to produce bcarotene
(pro-vitamin A) to help combat vitamin A deficiency1,
and it has been predicted that its contribution to alleviating
vitamin A deficiency would be substantially improved through
even higher b-carotene content2. We hypothesized that the
daffodil gene encoding phytoene synthase (psy), one of the
two genes used to develop Golden Rice, was the limiting step
in b-carotene accumulation. Through systematic testing

of other plant psys, we identified a psy from maize that
substantially increased carotenoid accumulation in a model
plant system. We went on to develop ‘Golden Rice 2’
introducing this psy in combination with the Erwinia uredovora
carotene desaturase (crtI) used to generate the original Golden
Rice1. We observed an increase in total carotenoids of up to
23-fold (maximum 37 lg/g) compared to the original Golden
Rice and a preferential accumulation of b-carotene.
Paine, J. A., C. A. Shipton, et al. (2005). "Improving the nutritional value of Golden Rice through increased pro-vitamin A content." Nature
Biotechnology 23(4): 482-487.
://000228197300033 AND http://www.botanischergarten.ch/Golden-Rice/Paine-Improving-Nutritional-Value-2005.pdf
'Golden Rice' is a variety of rice engineered to produce beta-carotene(pro-vitamin A) to help combat vitamin A deficiency 1, and it
has been predicted that its contribution to alleviating vitamin A deficiency would be substantially improved through even higher
beta-carotene content(2). We hypothesized that the daffodil gene encoding phytoene synthase (psy), one of the two genes used to
develop Golden Rice, was the limiting step in beta-carotene accumulation. Through systematic testing of other plant psys, we
identified a psy from maize that substantially increased carotenoid accumulation in a model plant system. We went on to develop
'Golden Rice 2' introducing this psy in combination with the Erwinia uredovora carotene desaturase (crtI) used to generate the
original Golden Rice(1). We observed an increase in total carotenoids of up to 23-fold (maximum 37 mu g/g) compared to the
original Golden Rice and a preferential accumulation of beta-carotene.
Paiva, S. A. R. and R. M. Russell (1999). "{beta}-Carotene and Other Carotenoids as Antioxidants." J Am Coll Nutr 18(5): 426-433.
http://www.jacn.org/cgi/content/abstract/18/5/426 AND http://www.botanischergarten.ch/Golden-Rice/Paiva-B-Carotene-Antioxidants-
1999.pdf
Carotenoids are natural pigments which are synthesized by plants and are responsible for the bright colors of various fruits and
vegetables. There are several dozen carotenoids in the foods that we eat, and most of these carotenoids have antioxidant activity.
{beta}-carotene has been best studied since, in most countries it is the most common carotenoid in fruits and vegetables. However,
in the U.S., lycopene from tomatoes now is consumed in approximately the same amount as {beta}-carotene. Antioxidants (including
carotenoids) have been studied for their ability to prevent chronic disease. {beta}-carotene and others carotenoids have antioxidant
properties in vitro and in animal models. Mixtures of carotenoids or associations with others antioxidants (e.g. vitamin E) can
increase their activity against free radicals. The use of animals models for studying carotenoids is limited since most of the animals
do not absorb or metabolize carotenoids similarly to humans. Epidemiologic studies have shown an inverse relationship between
presence of various cancers and dietary carotenoids or blood carotenoid levels. However, three out of four intervention trials using
high dose {beta}-carotene supplements did not show protective effects against cancer or cardiovascular disease. Rather, the high
risk population (smokers and asbestos workers) in these intervention trials showed an increase in cancer and angina cases. It
appears that carotenoids (including {beta}-carotene) can promote health when taken at dietary levels, but may have adverse effects
when taken in high dose by subjects who smoke or who have been exposed to asbestos. It will be the task of ongoing and future
studies to define the populations that can benefit from carotenoids and to define the proper doses, lengths of treatment, and
whether mixtures, rather than single carotenoids (e.g. {beta}-carotene) are more advantageous.
Palozza, P., S. Serini, et al. (2003). "Prooxidant effects of [beta]-carotene in cultured cells." Molecular Aspects of Medicine 24(6): 353-362.
http://www.sciencedirect.com/science/article/B6T9P-4932392-2/2/7eb45eddd7c135e37c9b239c167d35b1 AND
http://www.botanischergarten.ch/Golden-Rice/Palozza-Prooxydant-Effects-B-Carotene-2003.pdf
There is a growing body of interest on the role of [beta]-carotene and other carotenoids in human chronic diseases, including cancer.
While epidemiological evidence shows that people who ingest more dietary carotenoids exhibit a reduced risk for cancer, results
from intervention trials indicate that supplemental [beta]-carotene enhances lung cancer incidence and mortality among smokers. A
possible mechanism which can explain the dual role of [beta]-carotene as both a beneficial and a harmful agent in cancer as well as
in other chronic diseases is its ability in modulating intracellular redox status. [beta]-Carotene may serve as an antioxidant or as a
prooxidant, depending on its intrinsic properties as well as on the redox potential of the biological environment in which it acts. This
review summarizes the available evidence for a prooxidant activity of [beta]-carotene in cultured cells, focusing on biochemical and
molecular markers of oxidative stress, which have been reported to be enhanced by the carotenoid.
Park, H., S. S. Kreunen, et al. (2002). "Identification of the carotenoid isomerase provides insight into carotenoid biosynthesis, prolamellar body
formation, and photomorphogenesis." Plant Cell 14(2): 321-332.
://WOS:000174516200004 AND http://www.botanischergarten.ch/Golden-Rice/Park-Identification-Carotenoid-Isomerase-2002.pdf
Carotenoids are essential photoprotective and antioxidant pigments synthesized by all photosynthetic organisms. Most carotenoid
biosynthetic enzymes were thought to have evolved independently in bacteria and plants. For example, in bacteria, a single enzyme
(Crtl) catalyzes the four desaturations leading from the colorless compound phytoene to the red compound lycopene, whereas
plants require two desaturases (phytoene and zeta-carotene desaturases) that are unrelated to the bacterial enzyme. We have
demonstrated that carotenoid desaturation in plants requires a third distinct enzyme activity, the carotenoid isomerase (CRTISO),
which, unlike phytoene and zeta-carotene desaturases, apparently arose from a progenitor bacterial desaturase. The Arabidopsis
CRTISO locus was identified by the partial inhibition of lutein synthesis in light-grown tissue and the accumulation of poly-ciscarotene
precursors in dark-grown tissue of crtlSO mutants. After positional cloning, enzymatic analysis of CRTISO expressed in
Escherichia coli confirmed that the enzyme catalyzes the isomerization of poly-cis-carotenoids to all-trans-carotenoids. Etioplasts of
dark-grown crtlSO mutants accumulate acyclic poly-cis-carotenoids in place of cyclic all-trans-xanthophylls and also lack prolamellar
bodies (PLBs), the lattice of tubular membranes that defines an etioplast. This demonstrates a requirement for carotenoid

iosynthesis to form the PLB. The absence of PLBs in crtISO mutants demonstrates a function for this unique structure and
carotenoids in facilitating chloroplast development during the first critical days of seedling germination and photomorphogenesis.
Park, H., S. S. Kreunen, et al. (2001). "Identification of the elusive carotenoid isomerase enzyme redefines carotenoid biosynthesis and reveals a
role for carotenoids in the membrane dynamics of plastid development." Photosynthesis Research 69(1-3): 23.
://BIOSIS:PREV200200226191 AND http://www.botanischergarten.ch/Golden-Rice/Park-Identification-Carotenoid-Isomerase-
2002.pdf
Pearce, F. (2001). "Protests take the shine off golden rice." New Scientist 169(2284): 15-15.
://000167905400014 ANDhttp://www.botanischergarten.ch/Golden-Rice/Pearce-Golden-Rice-Worse-2001.pdf
Pfeiffer, W. H. and B. McClafferty (2007). "HarvestPlus: Breeding crops for better nutrition." Crop Science 47: S88-S105.
://WOS:000253124800008 AND http://www.botanischergarten.ch/Biofortification/Pfeiffer-HarvestPlus-Nutrition-2007.pdf
Micronutrient malnutrition, the so-called hidden hunger, affects more than one-half of the world’s population, especially women
and preschool children in developing countries. Despite past progress in controlling micronutrient decencies through
supplementation and food fortification, new approaches are needed to expand the reach of food-based interventions.
Biofortification, a new approach that relies on conventional plant breeding and modern biotechnology to increase the micronutrient
density of staple crops, holds great promise for improving the nutritional status and health of poor populations in both rural and
urban areas of the developing world. HarvestPlus, a research program implemented with the international research institutes of the
CGIAR, targets a multitude of crops that are a regular part of the staple-based diets of the poor and breeds them to be rich in iron,
zinc, and provitamin A. This paper emphasizes the need for interdisciplinary research and addresses the key research issues and
methodological considerations for success. The major activities to be undertaken are broadly grouped into research related to
nutrition research and impact analysis, and research considerations for delivering biofortifi ed crops to end-users effectively. The
paper places particular emphasis on the activities of the plant breeding and genetics component of this multidisciplinary program.
The authors argue that for biofortifi cation to succeed, product profi les developed by plant breeders must be driven by nutrition
research and impact objectives and that nutrition research must understand that the probability of success for biofortifi ed crops
increases substantially when product concepts consider farmer adoption and, hence, agronomic superiority.
Potrykus, I. (2001). "Golden rice and beyond." Plant Physiology 125(3): 1157-1161.
://000168179800002 and http://www.plantphysiol.org/cgi/reprint/125/3/1157.pdf and
http://www.botanischergarten.ch/Rice/PotrykusPlantPhys200103.pdf
Potrykus, I. (2001). "The 'Golden Rice' Tale." In Vitro Cellular & Developmental Biology-Plant 37(2): 93-100.
://000171551900002 AND http://www.botanischergarten.ch/Golden-Rice/Potrykus-Golden-Rice-Tale-2001.pdf
Potrykus, I. (2003). "Nutritionally enhanced rice to combat malnutrition disorders of the poor." Nutrition Reviews 61(6): S101-S104.
://000189308300002 and http://www.botanischergarten.ch/Rice/Potrykus-Nutrition-2003.pdf
Major deficiency disorders, including vitamin A deficiency, are especially common in countries in which rice is the staple food. In
response to the devastating effects of vitamin A deficiency, which may include blindness and, even death, "Golden Rice" has been
developed to deliver this nutrient to those populations who need it most. The case of Golden Rice is used to demonstrate the
challenges of radical GMO opposition, consumer acceptance, and regulation of biotechnology-derived foods.
Potrykus, I., P. K. Burkhardt, et al. (1995). "Genetic-Engineering of Indica Rice in Support of Sustained Production of Affordable and High-Quality
Food in Developing-Countries." Euphytica 85(1-3): 441-449.
://A1995TF37600055 AND http://www.botanischergarten.ch/Golden-Rice/Potrykus-Genetic-Engineering-India-Rice-1995.pdf
Indica-type rice provides the staple food for two billion people in Third World countries. Several problems involved in the stable and
sustained production of high quality food cannot be solved by traditional breeding. Methods have been established for gene transfer
to Indica rice breeding lines to study possible contributions from genetic engineering. Experiments are in progress on the
development of transgenic resistance towards Yellow Stem Borer, resistance towards Rice Tungro Virus, accumulation of provitamin
A in the endosperm, increase of essential amino acids in the endosperm such as lysine, cysteine and methionine and resistance
towards fungal pests such as Rice Blast and Sheath Blight. Transgenic clones from Indica rice breeding lines have been recovered
from several of the approaches mentioned, some of which have been regenerated to plants.
Qaim, M., C. E. Pray, et al. (2008). "Economic and Social Considerations in the Adoption of Bt Crops." Integration of Insect-Resistant Genetically
Modified Crops within IPM Programs: 329-356.
://BIOSIS:PREV200900077886 AND http://www.botanischergarten.ch/Bt/Qaim-Economic-Considerations-2008.pdf
This chapter provides an overview of the economics of Bt crop adoption, with a special emphasis on Bt cotton. On average, the
technology reduces insecticide applications and pest-related crop losses. In spite of higher seed prices, farmers realize substantial
gains in cotton incomes. In India and China alone, Bt cotton produces annual welfare gains of several hundred million US dollars,
with farmers being the main beneficiaries. In the USA, stronger IPR protection leads to larger benefit shares for biotechnology
companies, but agricultural producers and consumers profit as well. More preliminary studies for maize, rice, and eggplant suggest
that similar results can also be expected for other Bt crops. Furthermore, health benefits and aspects of consumer acceptance are
analyzed. Finally, several institutional and regulatory issues are discussed, as these might have important ramifications for
technology access, benefit distribution, seed market structures, and biodiversity.

Qaim, M. and A. J. Stein (2008). Economic consequences of Golden Rice. Invited presentation at the Fourth Conference of the European Plant
Science Organisation. Toulon (Cote d'Azur), France.
http://www.epsoweb.org/catalog/conf2008.htm AND http://www.botanischergarten.ch/Golden-Rice/Qaim-Economic-Consequences-Epso-
Abstract-2008.pdf
Qaim, M. and A. J. Stein (2009). "Biologische Anreicherung von Grundnahrungspflanzen: Wirksamkeit und Wirtschaftlichkeit." Ernahrungs-
Umschau 56(5): 274.
http://www.ernaehrungs-umschau.de/themen/wissenschaft_aktuell/?id=3846 AND http://www.botanischergarten.ch/Golden-Rice/Qaim-
Biologische-Anreicherung-2009.pdf
Biologische Anreicherung von Grundnahrungspflanzen ist ein neuer Ansatz zur Bekämpfung von Mikronährstoffmangel. Hierbei
werden Pflanzen so gezüchtet, dass sie höhere Mengen an Mikronährstoffen enthalten. Dieser Ansatz zielt vor allem auf
Entwicklungsländer ab, wo viele Menschen in abgelegenen ländlichen Gebieten kaum von anderen Mikronährstoffprogrammen
erreicht werden. Bisherige Studien zeigen, dass dieser Ansatz sowohl wirksam als auch wirtschaftlich sein kann.
Biologische Anreicherung von Grundnahrungspflanzen ist ein neuer Ansatz zur Bekämpfung von Mikronährstoffmangel. Hierbei werden
Pflanzen so gezüchtet, dass sie höhere Mengen an Mikronährstoffen enthalten. Dieser Ansatz zielt vor allem auf Entwicklungsländer
ab, wo viele Menschen in abgelegenen ländlichen Gebieten kaum von anderen Mikronährstoffprogrammen erreicht werden. Ersten
Studien zufolge scheint eine Akzeptanz dieser neuen Pflanzen in den Zielgruppen erreichbar zu sein. Analysen deuten ferner darauf
hin, dass diese Pflanzen bei entsprechenden Rahmenbedingungen wirtschaftlich effizient sind und eine deutliche Reduzierung der
Krankheitslast von Mikronährstoffmangel bewirken können. Auch in Industrieländern könnten biologisch angereicherte Pflanzen in
einigen Fällen eine interessante Alternative darstellen.
Qaim, M., A. J. Stein, et al. (2007). "Economics of biofortification." Contributions of Agricultural Economics to Critical Policy Issues: 119-133.
://WOS:000252133200010 AND http://www.botanischergarten.ch/Biofortification/Qaim-Economics-Biofortification.pdf
Micronutrient malnutrition is a serious public health problem in many developing countries. Different interventions are currently
used, but their overall coverage is relatively limited. Biofortification-that is, breeding staple food crops for higher micronutrient
contents-is a new agriculture-based approach, but relatively little is known about its ramifications. Here, the main factors influencing
success are discussed and a methodology for economic impact assessment is presented. Ex ante studies from India and other
countries suggest that biofortified crops can reduce the problem of micronutrient malnutrition in a cost-effective way, when
targeted to specific situations. Further research is needed to corroborate these findings and address certain issues still unresolved.
Qaim, M., A. J. Stein, et al. (2007). "Economics of biofortification." Agricultural Economics 37: 119-133.
://WOS:000251940700010 AND http://www.botanischergarten.ch/Biofortification/Qaim-Economics-Biofortification.pdf
Micronutrient malnutrition is a serious public health problem in many developing countries. Different interventions are currently
used, but their overall coverage is relatively limited. Biofortification-that is, breeding staple food crops for higher micronutrient
contents-is a new agriculture-based approach, but relatively little is known about its ramifications. Here, the main factors influencing
success are discussed and a methodology for economic impact assessment is presented. Ex ante studies from India and other
countries suggest that biofortified crops can reduce the problem of micronutrient malnutrition in a cost-effective way, when
targeted to specific situations. Further research is needed to corroborate these findings and address certain issues still unresolved.
Qaim, M., A. J. Stein, et al. (2007). "Economics of biofortification." Agricultural Economics 37: 119-133.
://WOS:000251940700010 AND http://www.botanischergarten.ch/Biofortification/Qaim-Economics-Biofortification.pdf
Micronutrient malnutrition is a serious public health problem in many developing countries. Different interventions are currently
used, but their overall coverage is relatively limited. Biofortification-that is, breeding staple food crops for higher micronutrient
contents-is a new agriculture-based approach, but relatively little is known about its ramifications. Here, the main factors influencing
success are discussed and a methodology for economic impact assessment is presented. Ex ante studies from India and other
countries suggest that biofortified crops can reduce the problem of micronutrient malnutrition in a cost-effective way, when
targeted to specific situations. Further research is needed to corroborate these findings and address certain issues still unresolved.
Quadro, L., L. Hamberger, et al. (2003). "Understanding the physiological role of retinol-binding protein in vitamin A metabolism using
transgenic and knockout mouse models." Molecular Aspects of Medicine 24(6): 421-430.
http://www.sciencedirect.com/science/article/B6T9P-49202PF-7/2/d5c49c78d46c27e6a536f2b543759368 AND
http://www.botanischergarten.ch/Golden-Rice/Quadro-Understanding-Physiological-Role-2003.pdf
Retinoids (vitamin A and its derivatives) play an essential role in many biological functions. However mammals are incapable of de
novo synthesis of vitamin A and must acquire it from the diet. In the intestine, dietary retinoids are incorporated in chylomicrons as
retinyl esters, along with other dietary lipids. The majority of dietary retinoid is cleared by and stored within the liver. To meet
vitamin A requirements of tissues, the liver secretes retinol (vitamin A alcohol) into the circulation bound to its sole specific carrier
protein, retinol-binding protein (RBP). The single known function of this protein is to transport retinol from the hepatic stores to
target tissues. Over the last few years, the generation of knockout and transgenic mouse models has significantly contributed to our
understanding of RBP function in the metabolism of vitamin A. We discuss below the role of RBP in maintaining normal vision and a
steady flux of retinol throughout the body in times of need.
Rodriquez-Amaya, D. and M. Kimura (2004). Harvestplus Handbook for Carotenoid Analysis. C. IFPRI. Washington and Cali, International Food
Policy Research Institute (IFPRI) and International Center for Tropical Agriculture (CIAT). 2: 63.
http://www.botanischergarten.ch/Golden-Rice/Rodriguez-Amaya-Handbook-Carotenoid-Analysis-2004.pdf

Carotenoid analysis is inherently difficult and error prone. Despite substantial improvements and refinements in the methods for
analyzing these fascinating but complicated compounds, discrepancies in analytical data can still be perceived in the literature.
Analysts must be well informed about the nature and properties of carotenoids and the problems associated with their identification
and quantification.
Carotenoid analysis consists of several steps that can be carried out efficiently in various ways. Thus, in this handbook several procedures are
given for some steps to enable the analyst to choose the procedure that best suits his/her laboratory conditions and experience.
These procedures have been put together and evaluated for HarvestPlus crops in our laboratory, but we recognize that other
procedures may also produce reliable results. It is important that analysts test the methods in their own laboratory prior to
performing the actual analyses. Knowledge of the purpose and possible sources of error in each step will assist the analyst in
appraising the performances of the methods. Indeed, the key element in the accurate determination of carotenoids is undoubtedly
the analyst. Even a well-validated method may perform badly in the hands of an analyst who does not seek the appropriate
information and does not pay attention to the many important details that are necessary to ensure the accuracy of the analytical
results.
The authors wish to thank HarvestPlus for the opportunity to publish this handbook. A special thanks is extended to Penelope Nestel, Nutrition
Coordinator of HarvestPlus, without whom this handbook would not exist. We also gratefully acknowledge the financial support of
the Brazilian Ministry of Science and Technology (MCT) through the National Council of Scientific and Technological Development
(CNPq), under the Program for Research Groups of Excellence (PRONEX CNPq 662307/1996-8), which has enabled us to gain the
knowledge and experience we are sharing through this handbook. Thanks are also due to Barbara Underwood, Jing Tan, and Julia
Humpries for reviewing the manuscript.
Delia B. Rodriguez-Amaya
Mieko Kimura
Rogers, R. (2000). "Golden rice: For sale, or free." Chemical & Engineering News 78(21): 9-9.
://000087225900008 AND http://www.botanischergarten.ch/Golden-Rice/Rogers-Golden-Rice-Sale-2000.pdf
Römer, S. and P. D. Fraser (2005). "Recent advances in carotenoid biosynthesis, regulation and manipulation." Planta 221(3): 305-308.
http://dx.doi.org/10.1007/s00425-005-1533-5 AND http://www.botanischergarten.ch/Golden-Rice/Roemer-Recent-Advances-Carotenoid-
2005.pdf
Rong, J., B.-R. Lu, et al. (2006). "Dramatic reduction of crop-to-crop gene flow within a short distance from transgenic rice fields." New
Phytologist 0(0 %R doi:10.1111/j.1469-8137.2006.01906.x): ???-???
http://www.blackwell-synergy.com/doi/abs/10.1111/j.1469-8137.2006.01906.x AND http://www.botanischergarten.ch/Rice/Rong-Distance-
Dramaticfall-2006.pdf
Summary . Genetically modified (GM) rice with enhanced agronomic traits and pharmaceutical uses are ready for widespread
adoption. Little is known about isolation requirements for achieving stringent transgene confinement in rice. To investigate the
extent of pollen-mediated crop-to-crop transgene flow, we conducted a field experiment with four plot-size treatments of adjacent
GM and nonGM rice (Oryza sativa) in China. . Three insect-resistant GM rice (Bt/CpTI) and nonGM isogenic lines were used in the
study. The hygromycin-resistance transgene (hpt) marker was used to screen seeds from the nonGM rice rows at different distance
intervals from GM rice plots. . Based on the examination of > 2.1 million germinated seeds, we found a dramatic reduction in
transgene frequencies with increasing distance from the GM crop, ranging from c. 0.28% at 0.2 m to < 0.01% at 6.2 m. In addition,
different plot size did not significantly affect the frequencies of gene flow. . In conclusion, pollen-mediated crop-to-crop transgene
flow in rice can be maintained at negligible levels with short spatial isolation. The model can also be applied to other crops with self-
and wind-pollination. New Phytologist (2006) doi: 10.1111/j.1469-8137.2006.01906.x c The Authors (2006). Trustees of New
Phytologist 2006
Rozan, A., J. L. Lusk, et al. (2007). "Consumer acceptance of a genetically modified organism of the second generation: The Golden Rice." Revue
D Economie Politique 117: 843-852.
://WOS:000252201100014 AND http://www.botanischergarten.ch/Golden-Rice/Rozan-Consumers-Acceptance-2007.pdf
In this paper we present the results of a study which focuses on a second generation genetically modified organism - golden rice
genetically modified to contain b-carotene. We report the results of a contingent valuation survey conducted with a
demographically representative sample of French households. If the majority of the sample are not aware of Golden Rice and judge
this food product unacceptable, however we observe a quarter of the sample would be willing to buy this product if the price is less
than or equal to that of non-genetically modified rice.
Sakata, K., Y. Nagamura, et al. (2002). "RiceGAAS: an automated annotation system and database for rice genome sequence." Nucleic Acids
Research 30(1): 98-102.
://000173077100025
An extensive effort of the International Rice Genome Sequencing Project (IRGSP) has resulted in rapid accumulation of genome
sequence, and >137 Mb has already been made available to the public domain as of August 2001. This requires a high-throughput
annotation scheme to extract biologically useful and timely information from the sequence data on a regular basis. A new
automated annotation system and database called Rice Genome Automated Annotation System (RiceGAAS) has been developed to
execute a reliable and up-to-date analysis of the genome sequence as well as to store and retrieve the results of annotation. The
system has the following functional features: (i) collection of rice genome sequences from GenBank; (ii) execution of gene prediction
and homology search programs; (iii) integration of results from various analyses and automatic interpretation of coding regions; (iv)
re-execution of analysis, integration and automatic interpretation with the latest entries in reference databases; (v) integrated

visualization of the stored data using web-based graphical view. RiceGAAS also has a data submission mechanism that allows public
users to perform fully automated annotation of their own sequences. The system can be accessed at
http://RiceGAAS.dna.affrc.go.jp/.
Saleh, N. T. and Z. S. Ahmed (1998). "Impact of natural sources rich in provitamin A on cooking characteristics, color, texture and sensory
attributes of beef patties." Meat Science 50(3): 285-293.
://000077201600002 AND http://www.botanischergarten.ch/Golden-Rice/Saleh-Impact-Natural-Sources-1998.pdf
Eight ground beef patty formulations, based on replacing meat with a natural source rich in provitamin A (i.e. carrot and sweet
potato) were prepared. All treatments displayed better moisture retention than the controls and hence yield was improved. L, a, and
b Hunter values increased and instron texture profile analysis indicated lower peak forces, increased springiness, and, in some cases
cohesiveness (i.e. juicier patties) for formulations with boiled carrot or sweet potato. However, adding these ingredients in a dry
form increased hardness, springiness and cohesiveness. Improvement in the content of provitamin A was achieved by incorporating
carrot or sweet potato into the patties. Mixing meat with a natural rich source of provitamin A could be a long-term solution for
combating vitamin A deficiency. (C) 1998 Published by Elsevier Science Ltd. All rights reserved.
Sasaki, T. (2002). "Rice genomics to understand rice plant as an assembly of genetic codes." Current Science 83(7): 834-839.
://000178662800021and http://www.botanischergarten.ch/Rice/Sasaki-Rice-Genomics-2002.pdf and
http://www.botanischergarten.ch/Rice/Sasaki-Rice-Genomics-ABIC2004_sasaki.pdf
Rice genome research is a topic and an attractive theme in plant genomics. Because of its practical importance to feed about half of
the world's population, and because of predicted unbalance of food supply and population increase in the near future, breeders,
geneticists and molecular biologists in plant fields have strong concern in the results of rice genome research. In this review, recent
research results of the rice genome are presented to offer information to develop ideas for understanding and improvement of rice
plants.
Sautter, C., S. Poletti, et al. (2006). "Biofortification of essential nutritional compounds and trace elements in rice and cassava." Proceedings of
the Nutrition Society 65(2): 153-159.
://000237233200002 AND http://www.botanischergarten.ch/Rice/Sautter-Biofortification-Essential-2006.pdf
Plant biotechnology can make important contributions to food security and nutritional improvement. For example, the development
of 'Golden Rice' by Professor Ingo Potrykus was a milestone in the application of gene technology to deliver both increased
nutritional qualities and health improvement to wide sections of the human population. Mineral nutrient and protein deficiency as
well as food security remain the most important challenges for developing countries. Current projects are addressing these issues in
two major staple crops, cassava (Manihot esculenta Crantz) and rice. The tropical root crop cassava is a major source of food for
approximately 600 million of the population worldwide. In sub-Saharan Africa > 200 million of the population rely on cassava as their
major source of dietary energy. The nutritional quality of the cassava root is not sufficient to meet all dietary needs. Rice is the
staple food for half the world population, providing approximately 20% of the per capita energy and 13% of the protein for human
consumption worldwide. In many developing countries the dietary contributions of rice are substantially greater (29 center dot 3%
dietary energy and 29 center dot 1% dietary protein). The current six most popular 'mega' rice varieties (in terms of popularity and
acreage), including Chinese hybrid rice, have an incomplete amino acid profile and contain limited amounts of essential
micronutrients. Rice lines with improved Fe contents have been developed using genes that have functions in Fe absorption,
translocation and accumulation in the plant, as well as improved Fe bioavailability in the human intestine. Current developments in
biotechnology-assisted plant improvement are reviewed and the potential of the technology in addressing human nutrition and
health are discussed.
Schaub, P., S. Al-Babili, et al. (2005). "Why is Golden Rice golden (yellow) instead of red?" Plant Physiology 138(1): 441-450.
://000229023100043 AND http://www.botanischergarten.ch/Golden-Rice/Schaub-Why-Golden-Rice-Yellow-2005.pdf
The endosperm of Golden Rice (Oryza sativa) is yellow due to the accumulation of &beta;-carotene (provitamin A) and xanthophylls.
The product of the two carotenoid biosynthesis transgenes used in Golden Rice, phytoene synthase (PSY) and the bacterial carotene
desaturase (CRTI), is lycopene, which has a red color. The absence of lycopene in Golden Rice shows that the pathway proceeds
beyond the transgenic end point and thus that the endogenous pathway must also be acting. By using TaqMan real-time PCR, we
show in wild-type rice endosperm the mRNA expression of the relevant carotenoid biosynthetic enzymes encoding phytoene
desaturase, &zeta;-carotene desaturase, carotene cis-trans-isomerase, &beta;-lycopene cyclase, and &beta;-carotene hydroxylase;
only PSY mRNA was virtually absent. We show that the transgenic phenotype is not due to up-regulation of expression of the
endogenous rice pathway in response to the transgenes, as was suggested to be the case in tomato (Lycopersicon esculentum) fruit,
where CRTI expression resulted in a similar carotenoid phenomenon. This means that &beta;-carotene and xanthophyll formation in
Golden Rice relies on the activity of constitutively expressed intrinsic rice genes (carotene cis-trans-isomerase, &alpha;/&beta;lycopene
cyclase, &beta;-carotene hydroxylase). PSY needs to be supplemented and the need for the CrtI transgene in Golden Rice is
presumably due to insufficient activity of the phytoene desaturase and/or &zeta;-carotene desaturase enzyme in endosperm. The
effect of CRTI expression was also investigated in leaves of transgenic rice and Arabidopsis (Arabidopsis thaliana). Here, again, the
mRNA levels of intrinsic carotenogenic enzymes remained unaffected; nevertheless, the carotenoid pattern changed, showing a
decrease in lutein, while the &beta;-carotene-derived xanthophylls increased. This shift correlated with CRTI-expression and is most
likely governed at the enzyme level by lycopene-cis-trans-isomerism. Possible implications are discussed.
Schubert, D. R. (2008). "The Problem with Nutritionally Enhanced Plants." Journal of Medicinal Food 11(4): 601-605.
://WOS:000261384700001 AND http://www.botanischergarten.ch/Golden-Rice/Schubert-Problem-Nutritionally-Enhanced-Food-
2008.pdf

Among the next generation of genetically modified (GM) plants are those that are engineered to produce elevated levels of
nutritional molecules such as vitamins, omega-3 fatty acids, and amino acids. Based upon the U. S. current regulatory scheme, the
plants and their products may enter our food supply without any required safety testing. The potential risks of this type of GM plant
are discussed in the context of human health, and it is argued that there should be very careful safety testing of plants designed to
produce biologically active molecules before they are commercially grown and consumed. This will require a mandatory,
scientifically rigorous review process.
Scott, K. J. and D. Rodriquez-Amaya (2000). "Pro-vitamin A carotenoid conversion factors: retinol equivalents - fact or fiction?" Food Chemistry
69(2): 125-127.
://WOS:000085901300002 AND http://www.botanischergarten.ch/Golden-Rice/Scott-Pro-Vitamin-A-Carotinoid-1999.pdf
The vitamin A potential of a food is conventionally expressed as retinol equivalents, i.e. retinol plus the corresponding retinol
equivalent of the provitamin A carotenoids. 1 mg of beta-carotene, for example, is taken to have the same biological activity as
0.167 mg of retinol. However, this figure is based on certain assumptions regarding the amount of beta-carotene absorbed and
subsequently converted to retinol. This commentary considers the numerous food and host related factors which may ultimately
determine carotenoid bioavailability and conversion to retinol. The authors endorse suggestions that until more definitive data on
the potential absorption of carotenoids from foods is available, any calculated value of the retinol equivalent of the provitamins
must be treated with caution. Furthermore, we would suggest that the use of current data on retinol equivalents in tables of food
composition should be abandoned. (C) 2000 Elsevier Science Ltd. All rights reserved.
Shankar, A. V., K. P. West, et al. (1996). "Chronic low intakes of vitamin A-rich foods in households with xerophthalmic children. A case-control
study in Nepal." American Journal of Clinical Nutrition 64(2): 242-248.
://A1996UZ12900018 AND http://www.botanischergarten.ch/Golden-Rice/Shankar-Chronic-Low-Intakes-1996.pdf
Dietary patterns in 81 rural Nepali households with a 1-6-y-old child with a history of xerophthalmia were compared with dietary
patterns of 81 households with an age matched nonxerophthalmic control subject. Weekly food-frequency questionnaires were
collected from case and control ''focus'' children, a younger sibling (if present), and the household 1-2 y after recruitment and
treatment of cases. Control households and children were more likely than case households and children to consume vitamin A-rich
foods during the monsoon (July-September) and major rice harvesting (October-December) seasons. Cases were less likely to
consume preformed vitamin A-rich foods throughout the year [odds ratio (OR) = 1.2-4.5] with the strongest differences observed
from October to December (OR = 2.0-4.2). Dietary risks were generally shared by younger siblings of cases, suggesting that
infrequent intake of beta-carotene and preformed vitamin-A rich foods begins early in life and clusters among siblings within
households, a pattern that is consistent with their higher risk of xerophthalmia and mortality. In developing countries where vitamin
A deficiency is endemic, dietary counseling for children with xerophthalmia should be extended to their younger siblings. Moreover,
dietary intake of preformed vitamin A may be as, or more, important as carotenoid-containing food consumption in protecting
children and other members of households from vitamin A deficiency.
Sharoni, Y., R. Agbaria, et al. (2003). "Modulation of transcriptional activity by antioxidant carotenoids." Molecular Aspects of Medicine 24(6):
371-384.
http://www.sciencedirect.com/science/article/B6T9P-494HKHN-1/2/7ee07724879981349531e956a2b07e65 AND
http://www.botanischergarten.ch/Golden-Rice/Sharoni-Modulation-Activity-Carotenoids-2003.pdf
It is widely accepted that diet changes are a powerful means to prevent cancer. The possible involvement of transcriptional activity
in the anticancer activity of carotenoids will be the focus of this review. Carotenoids function as potent antioxidants, and this is
clearly a major mechanism of their action. In addition carotenoids action involves interference in several pathways related to cancer
cell proliferation and includes changes in the expression of many proteins participating in these processes such as connexins, phase
II enzymes, cyclins, cyclin-dependent kinases and their inhibitors. These changes in protein expression suggest that the initial effect
involves modulation of transcription by ligand-activated nuclear receptors or by other transcription factors. It is feasible to suggest
that carotenoids and their oxidized derivatives interact with a network of transcription systems that are activated by different
ligands at low affinity and specificity and that this activation leads to the synergistic inhibition of cell growth.
Song, Z., B.-R. Lu, et al. (2004). "Pollen flow of cultivated rice measured under experimental conditions." Biodiversity and Conservation 13(3):
579-590.
http://springerlink.metapress.com/index/10.1023/B:BIOC.0000009491.24573.1d and http://www.botanischergarten.ch/Rice/Song-Rice-
Geneflow-2004.pdf
Stahl, W. and H. Sies (2003). "Antioxidant activity of carotenoids." Molecular Aspects of Medicine 24(6): 345-351.
http://www.sciencedirect.com/science/article/B6T9P-49202PF-3/2/d8b73bcb6a3e3ea1528b98e9e5b29b9d AND
http://www.botanischergarten.ch/Golden-Rice/Stahl-Antioxidant-Activity-Carotenoids-2003.pdf
Carotenoids are pigments which play a major role in the protection of plants against photooxidative processes. They are efficient
antioxidants scavenging singlet molecular oxygen and peroxyl radicals. In the human organism, carotenoids are part of the
antioxidant defense system. They interact synergistically with other antioxidants; mixtures of carotenoids are more effective than
single compounds. According to their structure most carotenoids exhibit absorption maxima at around 450 nm. Filtering of blue light
has been proposed as a mechanism protecting the macula lutea against photooxidative damage. There is increasing evidence from
human studies that carotenoids protect the skin against photooxidative damage.
Stein, A. J., J. V. Meenakshi, et al. (2008). "Potential impacts of iron biofortification in India." Social Science & Medicine 66(8): 1797-1808.
://WOS:000255600900014 AND http://www.botanischergarten.ch/Golden-Rice/Stein-Potential-Impacts-Iron-2008.pdf

Iron deficiency is a widespread nutrition and health problem in developing countries, causing impairments in physical activity and
cognitive development, as well as maternal mortality. Although food fortification and supplementation programmes have been
effective in some countries, their overall success remains limited. Biofortification, that is, breeding food crops for higher
micronutrient content, is a relatively new approach, which has been gaining international attention recently. We propose a
methodology for ex ante impact assessment of iron biofortification, building on a disability-adjusted life years (DALYs) framework.
This methodology is applied in an Indian context. Using a large and representative data set of household food consumption, the
likely effects of iron-rich rice and wheat varieties are simulated for different target groups and regions. These varieties, which are
being developed by an international public research consortium, based on conventional breeding techniques, might be ready for
local distribution within the next couple of years. The results indicate sizeable potential health benefits. Depending on the
underlying assumptions, the disease burden associated with iron deficiency could be reduced by 19-58%. Due to the relatively low
institutional cost to reach the target population, the expected cost-effectiveness of iron biofortification compares favourably with
other micronutrient interventions. Nonetheless, biofortification should not be seen as a substitute for other interventions. Each
approach has its particular strengths, so they complement one another. (C) 2008 Elsevier Ltd. All rights reserved.
Stein, A. J., P. Nestel, et al. (2007). "Plant breeding to control zinc deficiency in India: how cost-effective is biofortification?" Public Health
Nutrition 10(5): 492-501.
://WOS:000246108600012 AND http://www.botanischergarten.ch/Golden-Rice/Stein-Plant-Breeding-Zinc-2007.pdf
Objective: To estimate the potential impact of zinc biofortification of rice and wheat on public health in India and to evaluate its
cost-effectiveness compared with alternative interventions and international standards. Design: The burden of zinc deficiency (ZnD)
in India was expressed in disability-adjusted life years (DALYs) lost. Current zinc intakes were derived from a nationally
representative household food consumption survey (30-day recall) and attributed to household members based on adult equivalent
weights. Using a dose-response function, projected increased zinc intakes from biofortified rice and wheat were translated into
potential health improvements for pessimistic and optimistic scenarios. After estimating the costs of developing and disseminating
the new varieties, the cost-effectiveness of zinc biofortification was calculated for both scenarios and compared with alternative
micronutrient interventions and international reference standards. Setting: India. Subjects: Representative household survey (n =
119 554). Results: The calculated annual burden of ZnD in India is 2.8 million DALYs lost. Zinc biofortification of rice and wheat may
reduce this burden by 20-51% and save 0.6-1.4 million DALYs each year, depending on the scenario. The cost for saving one DALY
amounts to $US 0.73-7.31, which is very cost-effective by standards of the World Bank and the World Health Organization, and is
lower than that of most other micronutrient interventions. Conclusions: Not only may zinc biofortification save lives and prevent
morbidity among millions of people, it may also help accommodate the need to economise and to allocate resources more
efficiently. Further research is needed to corroborate these findings.
Stein, A. J. and M. Qaim (2007). "The human and economic cost of hidden hunger." Food and Nutrition Bulletin 28(2): 125-134.
://WOS:000246969900001 AND http://www.botanischergarten.ch/Golden-Rice/Stein-Human-Economic-Cost-2007.pdf
Background. Micronutrient malnutrition is a public health problem in many developing countries. Its negative impact on income
growth is recognized in principle, but there are widely varying estimates of the related economic cost. Objective. To discuss available
studies that quantify the cost of micronutrient malnutrition, and to develop an alternative framework and apply it to India. Methods.
Detailed burden of disease calculations are used to estimate the economic cost of micronutrient malnutrition based on disability-
adjusted life years (DALYs) lost. Results. The short-term economic cost of micronutrient malnutrition in India amounts to 0.8% to
2.5% of the gross domestic product. Conclusions. Although the results confirm that micronutrient malnutrition is a huge economic
problem, the estimates are lower titan those of most previous studies. The differences may be due to differences in underlying
assumptions, quality of data, and precision of calculation but also to dynamic interactions between nutrition, health, and economic
productivity, which are difficult to capture. Clear explanation of all calculation details would be desirable for future studies in order
to increase credibility and transparency.
Stein, A. J., H. P. S. Sachdev, et al. (2006). "Potential impact and cost-effectiveness of Golden Rice." Nature Biotechnology 24(10): 1200-1201.
://WOS:000241191700014 AND http://www.botanischergarten.ch/Golden-Rice/Stein-Potential-Impact-2006.pdf
Stein, A. J., H. P. S. Sachdev, et al. (2007). "What we know and don't know about Golden Rice." Nature Biotechnology 25(6): 624-624.
http://dx.doi.org/10.1038/nbt0607-624a AND http://www.botanischergarten.ch/Rice/Stein_NatureBiotech_response-2007.pdf
Stein, A. J., H. P. S. Sachdev, et al. (2008). "Genetic engineering for the poor: Golden Rice and public health in India." World Development 36:
144-158.
://WOS:000253005000008 AND http://www.botanischergarten.ch/Golden-Rice/Stein-Genetic-Engineering-Poor-2008.pdf
Vitamin A deficiency (VAD) affects millions of people, causing serious health problems. Golden Rice (GR), which has been genetically
engineered to produce beta-carotene, is being proposed as a remedy. While this new technology has aroused controversial debates,
its actual impact remains unclear. We develop a methodology for ex ante evaluation, taking into account health and nutrition
details, as well as socioeconomic and policy factors. The framework is used for empirical analyses in India. Given broad public
support, GR could more than halve the disease burden of VAD. Juxtaposing health benefits and overall costs suggests that GR could
be very cost-effective. (C) 2007 Elsevier Ltd. All rights reserved.
Tan, J., N. Baisakh, et al. (2005). "The screening of rice germplasm, including those transgenic rice lines which accumulate beta-carotene in their
polished seeds, for their carotenoid profile." International Journal of Food Science and Technology 40(5): 563-569.
://000228640200011 AND http://www.botanischergarten.ch/Golden-Rice/Tan-Screening-Germplasm-2005.pdf

Tanaka, Y. and A. Ohmiya (2008). "Seeing is believing: engineering anthocyanin and carotenoid biosynthetic pathways." Current Opinion in
Biotechnology 19(2): 190-197.
://WOS:000255982100017 AND http://www.botanischergarten.ch/Golden-Rice/Tanaka-Seeing-is-Believing-2008.pdf
The biosynthetic pathways of flavonoids and carotenoids have been well established, and the biosynthetic genes have been mostly
isolated. Metabolic engineering of their biosynthetic pathways has provided not only novel colored or health-beneficial plants but
also excellent models to study the efficacy of such engineering. In order to achieve a specific color by accumulating a corresponding
compound, it is necessary to upregulate the pathway leading to the compound and downregulate, the competing pathway. The
regulation of gene expression has to be optimized in a target crop as well.
Tang, G. W., J. Qin, et al. (2009). "Golden Rice is an effective source of vitamin A." American Journal of Clinical Nutrition 89(6): 1776-1783.
://WOS:000266245500011 AND http://www.botanischergarten.ch/Golden-Rice/Tang-Golden-Rice-Effective-Source-2009.pdf
Background: Genetically engineered "Golden Rice'' contains up to 35 mu g beta-carotene per gram of rice. It is important to
determine the vitamin A equivalency of Golden Rice beta-carotene to project the potential effect of this biofortified grain in riceconsuming
populations that commonly exhibit low vitamin A status. Objective: The objective was to determine the vitamin A value
of intrinsically labeled dietary Golden Rice in humans. Design: Golden Rice plants were grown hydroponically with heavy water
(deuterium oxide) to generate deuterium-labeled [H-2]beta-carotene in the rice grains. Golden Rice servings of 65-98 g (130 200 g
cooked rice) containing 0.99-1.53 mg beta-carotene were fed to 5 healthy adult volunteers (3 women and 2 men) with 10 g butter. A
reference dose of [C-13(10)] retinyl acetate (0.4-1.0 mg) in oil was given to each volunteer 1 wk before ingestion of the Golden Rice
dose. Blood samples were collected over 36 d. Results: Our results showed that the mean (+/- SD) area under the curve for the total
serum response to [H-2] retinol was 39.9 +/- 20.7 mu g . d after the Golden Rice dose. Compared with that of the [C-13(10)] retinyl
acetate reference dose (84.7 +/- 34.6 mu g . d), Golden Rice beta-carotene provided 0.24-0.94 mg retinol. Thus, the conversion
factor of Golden Rice beta-carotene to retinol is 3.8 +/- 1.7 to 1 with a range of 1.9-6.4 to 1 by weight, or 2.0 +/- 0.9 to 1 with a range
of 1.0-3.4 to 1 by moles. Conclusion: beta-Carotene derived from Golden Rice is effectively converted to vitamin A in humans. This
trial was registered at clinicaltrials. gov as NCT00680355. Am J Clin Nutr 2009; 89: 1776-83.
Tanumihardjo, S. A., H. Bouis, et al. (2008). "Biofortification of staple crops: An emerging strategy to combat hidden hunger." Comprehensive
Reviews in Food Science and Food Safety 7(4): 329-334.
://WOS:000259308200008 http://www.botanischergarten.ch/Golden-Rice/Tanumihardjo-Biofortification-Staple-Crops-2008.pdf
Diverse diets rich in micronutrients offer the ultimate sustainable solution to undernutrition. Unfortunately, poverty drives food
consumption habits. For the poor, a simple meal consisting mostly of staple foods make up the daily diet. A diet based
predominantly on staple food lacks adequate essential nutrients and thus can lead to hidden hunger. The biofortification strategy
targets the poor by naturally adding nutrients to these staple foods through plant breeding. Biofortified crops offer a rural-based
intervention that, by design, initially reach these more remote populations, which comprise a majority of the undernourished in
many countries, and then extend to urban populations as production surpluses are marketed. In this way, biofortification
complements fortification and supplementation programs, which currently work best in centralized urban and peri-urban areas and
then reach into rural areas only with good infrastructure. Initial investments in agricultural research at a central location can
generate high recurrent benefits at low cost as adapted biofortified varieties become available in country after country across time
at low recurrent costs. HarvestPlus is working to develop and distribute varieties of food staples that are high in iron, zinc, and
provitamin A through an interdisciplinary, global alliance of scientific institutions and implementing agencies in developing and
developed countries. HarvestPlus, comanaged by the International Center for Tropical Agriculture and the International Food Policy
Research Institute, collaborates with the India Biofortification Project to develop biofortified varieties of rice, wheat, and maize for
India.
Taylor, M. and G. Ramsay (2005). "Carotenoid biosynthesis in plant storage organs: recent advances and prospects for improving plant food
quality." Physiologia Plantarum 124(2): 143-151.
http://dx.doi.org/10.1111/j.1399-3054.2005.00509.x AND http://www.botanischergarten.ch/Golden-Rice/Taylor-Carotenoid-Biosynthesis-
2005.pdf
Carotenoids are components of the photosynthetic machinery, intermediates in the biosynthesis of abscisic acid and other
apocarotenoids and act as coloured pigments particularly in floral and fruit tissue. Humans benefit in a number of ways from dietary
carotenoids present in green leaf tissue and many fruits, seeds, roots and tubers. Carotenoids with a 03B2-ring end group are
required for the synthesis of vitamin A, and deficiency for this vitamin remains a major health problem in some parts of the world.
Epidemiological studies suggest that carotenoids also have important roles in a range of diseases including age-related macular
degradation and some cancers. The isoprenoid pathway is described, which leads to the carotenoids via the condensation of fivecarbon
isoprenoid units to form a 40-carbon chain in phytoene. The early part of the biosynthetic pathway involves crosstalk
between plastidic and cytosolic components. Desaturation, isomerization, cyclization, hydroxylation and epoxidation of the 40carbon
phytoene gives the range of carotenoids found in plants. The nuclear-encoded enzymes for these stages are targeted to
plastids, and in some cases, different members of a gene family are active in plastids in different tissue types. Studies on the
transgenic manipulation of the pathway, natural and induced mutants, gene silencing and gene and quantitative trait locus mapping
are increasingly unravelling the pathway and its control, giving opportunities for directed manipulation of the types and quantities of
these nutritionally important compounds in crop tissues used in human and animal diets.
Thomson, J. A. (2002). "Research needs to improve agricultural productivity and food quality, with emphasis on biotechnology." Journal of
Nutrition 132(11): 3441S-3442S.
://000179109300034 AND http://www.botanischergarten.ch/Golden-Rice/Thomson-Research-Needs-Improve-2002.pdf
Research into agricultural productivity, especially for crops in the developing world, should include resistance to plant viruses, fungi
and the parasitic weed Striga. It must also include research into the development of resistance to Bacillus thuringiensis (Bt) toxin-

expressing crops. Drought- and heat- tolerant crops, and those that can combat the problems of soil deficiencies, are required, and
vaccine production in plants should be a high priority. Research into food quality should include the equivalent of "golden rice" in
maize, the enhancement of the production of phytosterols and improved qualities of vegetable oils.
Tyssandier, V., C. Feillet-Coudray, et al. (2004). "Effect of Tomato Product Consumption on the Plasma Status of Antioxidant Microconstituents
and on the Plasma Total Antioxidant Capacity in Healthy Subjects." J Am Coll Nutr 23(2): 148-156.
http://www.jacn.org/cgi/content/abstract/23/2/148 AND http://www.botanischergarten.ch/Golden-Rice/Tyssandier-Tomato-Antioxidant-
Capacity-2004.pdf
Objectives: to identify the plasma antioxidant microconstituents mainly affected by tomato product consumption, to check whether
tomato product consumption can affect antioxidant status, and to identify tomato-product antioxidant-microconstituents mainly
involved in the effect of these products on oxidative stress. Design: Medium-term dietary supplementation study. Setting: Human
Nutrition Laboratory, Clermont-Ferrand, France. Subjects: Twenty healthy young (20 < years < 40), non obese (18 < BMI (kg/m2) <
25), females were recruited by advertisement. All of them completed the study. Intervention: The usual diet of the subjects was
supplemented for three weeks with 96 g/day tomato puree. The volunteers then avoided tomato-product-rich foods for a
subsequent three-week period. Measures of Outcome: Fasting blood samples were collected the day before supplementation, the
day after the supplementation period, and the day after the depletion period. The status of several antioxidant microconstituents
(plasma microconstituent concentrations), and the antioxidant status (plasma total antioxidant capacity) were assessed. Results:
Supplementation with tomato puree significantly increased plasma lycopene, {beta}-carotene and lutein. Conversely it did not
significantly affect plasma vitamin C and E, plasma antioxidant trace metals (Cu, Zn and Se), and plasma total antioxidant capacity.
Avoidance of tomato-product-rich foods for three weeks significantly (p < 0.05) decreased plasma lycopene, {beta}-carotene, lutein
and vitamin C, as well as plasma total antioxidant capacity. Plasma total antioxidant capacity, as measured by chemiluminescence,
was positively related (p < 0.05) to the status of lycopene, vitamin C and {beta}-carotene. Conclusions: Tomato product consumption
can affect not only the lycopene status, but also that of other antioxidant microconstituents ({beta}-carotene and lutein). Lycopene,
but also {beta}-carotene, are apparently the main tomato microconstituents responsible for the effect of tomato products on
antioxidant status.
van Lieshout, M., C. E. West, et al. (2003). "Isotopic tracer techniques for studying the bioavailability and bioefficacy of dietary carotenoids,
particularly beta-carotene, in humans: a review." American Journal of Clinical Nutrition 77(1): 12-28.
://000179965700003 and http://www.botanischergarten.ch/Rice/Lieshout-et-al-tracer-corotinoids.pdf
Vitamin A deficiency is a serious health problem in many developing countries. Provitamin A carotenoids in fruit and vegetables are
the major source of vitamin A for a large proportion of the world's population. However, the contribution of plant foods is
substantial only when both the consumption and provitamin A content of such food is high and, at the same time, the bioefficacy of
the provitamin A is high. With respect to provitamin A carotenoids, the term bioefficacy is defined as the product of the fraction of
the ingested amount that is absorbed (bioavailability) and the fraction of that which is converted to retinol in the body
(bioconversion). Isotopic tracer techniques can meet the need for accurate and precise estimates of the bioavailability,
bioconversion, and bioefficacy of dietary carotenoids in humans. Use of such techniques will enable proper evaluation of food-based
approaches to eliminating vitamin A deficiency. In addition, the putative antioxidant capacities of carotenoids can be better
understood if their bioavailability is known. Here, we discuss how tracer techniques can be applied to obtain reliable and
representative data. A step-by-step discussion of aspects related to these techniques is provided, including study design, choice of
isotopic tracers, dosing regimen, collection of samples, chemical analysis of samples, and data analysis.
Weil, J. H. (2005). "Are genetically modified plants useful and safe?" Iubmb Life 57(4-5): 311-314.
://000230120400033 AND http://www.botanischergarten.ch/Golden-Rice/Weil-GM-Plants-Useful-2005.pdf
So far, plants have been genetically modified essentially to achieve resistance to herbicides, or to pathogens (mainly insects, or
viruses), but resistance to abiotic stresses (such as cold, heat, drought, or salt) is also being studied. Genetically modified (GM) plants
with improved nutritional qualities have more recently been developed, such as plants containing higher proportions of unsaturated
fatty acids (omega-3 and omega-6) in their oil (to prevent cardio-vascular diseases), or containing beta-carotene as in the golden rice
(to prevent vitamin A deficiency). Possible risks for human health (such as the production of allergenic proteins), or for the
environment (such as the appearance of superweeds as a result from gene flow), should be carefully studied, and a science-based
assessment of benefits vs. risks should be made on a csae by case basis, both for GM plants and for plants obtained by conventional
breeding methods.
Welch, R. M. and R. D. Graham (1999). "A new paradigm for world agriculture: meeting human needs - Productive, sustainable, nutritious."
Field Crops Research 60(1-2): 1-10.
://WOS:000078617900002 AND http://www.botanischergarten.ch/Golden-Rice/Welch-New-Paradigm-World-Agriculture-1999.pdf
Micronutrient malnutrition ('Hidden Hunger') now afflicts over two billion people worldwide, resulting in poor health, low worker
productivity, high rates of mortality and morbidity, increased rates of chronic diseases (coronary heart disease, cancer, stroke, and
diabetes), and permanent impairment of cognitive abilities of infants born to micronutrient-deficient mothers. The consequences of
food system failures include lethargic national development efforts, continued high population growth rates, and a vicious cycle of
poverty for massive numbers of underprivileged people in all nations, Our food systems are failing us globally by not providing
enough balanced nutrient output to meet all the nutritional needs of every person, especially resource-poor women, infants and
children in developing countries, Agriculture is partly responsible because it has never held nutrient output as an explicit goal of its
production systems. Indeed, many agricultural policies have fostered a decline in nutrition and diet diversity for the poor in many
countries, Nutrition and health communities are also partly responsible because they have never considered using agriculture as a
primary tool in their programs directed at alleviating poor nutrition and ill health globally. Now is the time for a new paradigm for
agriculture and nutrition, We must consider ways in which agriculture can contribute to finding sustainable solutions to food system

failures through holistic food-based system approaches, thereby closely linking agricultural production to improving human health,
livelihood and well being. Such action will stimulate support for agricultural research in many developed countries because it
addresses consumer issues as well as agricultural production issues and is, therefore, politically supportable. (C) 1999 Elsevier
Science B.V. All rights reserved.
Welch, R. M. and R. D. Graham (2004). "Breeding for micronutrients in staple food crops from a human nutrition perspective." Journal of
Experimental Botany 55(396): 353-364.
://000188634500007 AND http://www.botanischergarten.ch/Golden-Rice/Welch-Breeding-Micronutrients-2004.pdf
Over three billion people are currently micronutrient (i.e. micronutrient elements and vitamins) malnourished, resulting in egregious
societal costs including learning disabilities among children, increased morbidity and mortality rates, lower worker productivity, and
high healthcare costs, all factors diminishing human potential, felicity, and national economic development. Nutritional deficiencies
(e.g. iron, zinc, vitamin A) account for almost two-thirds of the childhood death worldwide. Most of those afflicted are dependent on
staple crops for their sustenance. Importantly, these crops can be enriched (i.e. 'biofortified') with micronutrients using plant
breeding and/or transgenic strategies, because micronutrient enrichment traits exist within their genomes that can to used for
substantially increasing micronutrient levels in these foods without negatively impacting crop productivity. Furthermore, 'proof of
concept' studies have been published using transgenic approaches to biofortify staple crops (e.g. high beta-carotene 'golden rice'
grain, high ferritin-Fe rice grain, etc). In addition, micronutrient element enrichment of seeds can increase crop yields when sowed
to micronutrient-poor soils, assuring their adoption by farmers. Bioavailability issues must be addressed when employing plant
breeding and/or transgenic approaches to reduce micronutrient malnutrition. Enhancing substances (e.g. ascorbic acid, S-containing
amino acids, etc) that promote micronutrient bioavailability or decreasing antinutrient substances (e.g. phytate, polyphenolics, etc)
that inhibit micronutrient bioavailability, are both options that could be pursued, but the latter approach should be used with
caution. The world's agricultural community should adopt plant breeding and other genetic technologies to improve human health,
and the world's nutrition and health communities should support these efforts. Sustainable solutions to this enormous global
problem of 'hidden hunger' will not come without employing agricultural approaches.
Welch, R. M. and R. D. Graham (2005). "Agriculture: the real nexus for enhancing bioavailable micronutrients in food crops." Journal of Trace
Elements in Medicine and Biology 18(4): 299-307.
://WOS:000230515000002 AND http://www.botanischergarten.ch/Golden-Rice/Welch-Agriculture-Real-Nexus-2005.pdf
Human existence requires that agriculture provide at least 50 nutrients (e.g., vitamins, minerals, trace elements, amino acids,
essential fatty acids) in amounts needed to meet metabolic demands during all seasons. If national food systems do not meet these
demands, mortality and morbidity rates increase, worker productivity declines, livelihoods are diminished and societies suffer.
Today, many food systems within the developing world cannot meet the nutritional needs of the societies they support mostly due
to farming systems that cannot produce enough micronutrients to meet human needs throughout the year. Nutrition transitions are
also occurring in many rapidly developing countries that are causing chronic disease (e.g., cancer, heart disease, stroke, diabetes,
and osteoporosis) rates to increase substantially. These global developments point to the need to explicitly link agricultural
technologies to human health. This paper reviews some ways in which agriculture can contribute significantly to reducing
micronutrient malnutrition globally. It concludes that it is imperative that close linkages be forged between the agriculture, nutrition
and health arenas in order to find sustainable solutions to micronutrient malnutrition with agriculture becoming the primary
intervention toot to use in this fight. Published by Elsevier GmbH.
Welsch, R., F. Wust, et al. (2008). "A third phytoene synthase is devoted to abiotic stress-induced abscisic acid formation in rice and defines
functional diversification of phytoene synthase genes." Plant Physiology 147(1): 367-380.
://WOS:000256419400033 AND http://www.botanischergarten.ch/Golden-Rice/Welsch-Third-Phytoene-Synthase-2008.pdf
We here report on the characterization of a novel third phytoene synthase gene (PSY) in rice (Oryza sativa), OsPSY3, and on the
differences among all three PSY genes with respect to the tissue-specific expression and regulation upon various environmental
stimuli. The two already known PSYs are under phytochrome control and involved in carotenoid biosynthesis in photosynthetically
active tissues and exhibit different expression patterns during chloroplast development. In contrast, OsPSY3 transcript levels are not
affected by light and show almost no tissue-specific differences. Rather, OsPSY3 transcripts are up-regulated during increased
abscisic acid (ABA) formation upon salt treatment and drought, especially in roots. The simultaneous induction of genes encoding 9cis-epoxycarotenoid
dioxygenases (NCEDs), involved in the initial steps of ABA biosynthesis, indicate that decreased xanthophyll
levels are compensated by the induction of the third PSY gene. Furthermore, OsPSY3 and the OsNCEDs investigated were also
induced by the application of ABA, indicating positive feedback regulation. The regulatory differences are mirrored by cis-acting
elements in the corresponding promoter regions, with light-responsive elements for OsPSY1 and OsPSY2 and an ABA-response
element as well as a coupling element for OsPSY3. The investigation of the gene structures and 5' untranslated regions revealed that
OsPSY1 represents a descendant of an ancient PSY gene present in the common ancestor of monocots and dicots. Since the genomic
structures of OsPSY2 and OsPSY3 are comparable, we conclude that they originated from the most recent common ancestor,
OsPSY1.
West, C. E., A. Eilander, et al. (2002). "Consequences of revised estimates of carotenoid bioefficacy for dietary control of vitamin A deficiency in
developing countries." Journal of Nutrition 132(9): 2920S-2926S.
://000178033300011 AND http://www.botanischergarten.ch/Golden-Rice/West-Consequences-Revised-2002.pdf
According to existing recommendations of the Food and Agriculture Organization (FAO)/World Health Organization (WHO), the
amount of provitamin A in a mixed diet having the same vitamin A activity as 1 mug of retinol is 6 mug of beta-carotene or 12 mug of
other provitamin A carotenoids. The efficiency of this conversion is referred to as bioefficacy. Recently, using data from healthy
people in developed countries and based on a two-step process, the U.S. Institute of Medicine (IOM) derived new conversion
factors. The first step established the bioefficacy of beta-carotene in oil at 2 mug having the same vitamin A activity as 1 mug of

etinol; the second step established the bioavailability of beta-carotene in foods relative to that of beta-carotene in oil at 1:6. Thus, 2
mug of p-carotene in oil or 12 mug of beta-carotene in mixed foods has the same vitamin A activity as 1 mug of retinol. Based on
existing FAO food balance sheets and the FAO/WHO conversion rates, all populations should be able to meet their vitamin A
requirements from existing dietary sources. However, using the new IOM conversion rates, populations in developing countries
could not achieve adequacy. Additionally, field studies suggest that, instead of 12 mug, 21 mug of beta-carotene has the same
vitamin A activity as 1 mug of retinol, which implies that effective vitamin A intake is even lower. Therefore, controlling vitamin A
deficiency in developing countries requires not only vitamin A supplementation but also food-based approaches, including food
fortification, and possibly the introduction of new strains of plants with enhanced vitamin A activity.
Winklhofer-Roob, B. M., E. Rock, et al. (2003). "Effects of vitamin E and carotenoid status on oxidative stress in health and disease. Evidence
obtained from human intervention studies." Molecular Aspects of Medicine 24(6): 391-402.
http://www.sciencedirect.com/science/article/B6T9P-49JPMJB-1/2/9e27129463b09fc03b1fa7b03d666b75 AND
http://www.botanischergarten.ch/Golden-Rice/Winklhofer-Roob-Effects-Vitamin-E-2003.pdf
Vitamin E and carotenoids are known to act as antioxidants both in vitro and in vivo. In this review we present a series of studies in
healthy subjects and in patients who exhibit either acute or chronic oxidative stress. In the EU-Commission funded VITAGE project
we investigated the status and effects of vitamin E and carotenoids on oxidative stress in 300 healthy volunteers. Depletion studies
limiting dietary vitamin E or carotenoid intake to ~25% of the dietary reference intakes and subsequent repletion by
supplementation with either large doses of vitamin E or intermediate doses of carotenoids showed significant changes in ex vivo LDL
oxidizability, total plasma peroxide concentrations and urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine excretion. Patients on chronic
hemodialysis present with oxidative stress in the presence of normal vitamin E but impaired vitamin C status and, due to anemia,
need to be treated with parenteral iron. We studied the effects of a single oral dose of vitamin E taken 6 h prior to intravenous
infusion of 100 mg iron, which exceeded the iron-binding capacity of transferrin. Vitamin E significantly reduced and in combination
with a single dose of vitamin C completely abrogated acute oxidative stress induced by the iron load. Patients with cystic fibrosis are
exposed to chronic oxidative stress due to an overproduction of reactive oxygen species as a result of neutrophil-dominated lung
inflammation and impaired antioxidant status. Biochemical vitamin E and carotenoid deficiencies could be fully corrected even in the
presence of fat malabsorption using intermediate doses of either RRR [alpha]-tocopherol or all-rac [alpha]-tocopheryl acetate and
water-miscible all-trans [beta]-carotene. Long-term supplementation reduced ex vivo LDL oxidizability, in vivo lipid peroxidation and
lung inflammation.
Wurbs, D., S. Ruf, et al. (2007). "Contained metabolic engineering in tomatoes by expression of carotenoid biosynthesis genes from the plastid
genome." Plant Journal 49(2): 276-288.
://WOS:000243864500010 AND http://www.botanischergarten.ch/Golden-Rice/Wurbs-Contained-Metablolic-Carotenoid-2007.pdf
Applications of chloroplast engineering in agriculture and biotechnology will depend critically on success in extending the crop range
of chloroplast transformation, and on the feasibility of expressing transgenes in edible organs (such as tubers and fruits), which
often are not green and thus are much less active in chloroplast gene expression. We have improved a recently developed
chloroplast-transformation system for tomato plants and applied it to engineering one of the central metabolic pathways in fruits:
carotenoid biosynthesis. We report that plastid expression of a bacterial lycopene beta-cyclase gene results in herbicide resistance
and triggers conversion of lycopene, the main storage carotenoid of tomatoes, to beta-carotene, resulting in fourfold enhanced provitamin
A content of the fruits. Our results demonstrate the feasibility of engineering nutritionally important biochemical pathways
in non-green plastids by transformation of the chloroplast genome.
Ye, X. D., S. Al-Babili, et al. (2000). "Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm."
Science 287(5451): 303-305.
://000084769600039 AND http://www.botanischergarten.ch/Golden-Rice/Ye-Engineering-Provitamin-A-2000.pdf
Rice (Oryza sativa), a major staple food, is usually milled to remove the oil-rich aleurone layer that turns rancid upon storage,
especially in tropical areas. The remaining edible part of rice grains, the endosperm, Lacks several essential nutrients, such as
provitamin A. Thus, predominant rice consumption promotes vitamin A deficiency, a serious public health problem in at least 26
countries, including highly populated areas of Asia, Africa, and Latin America. Recombinant DNA technology was used to improve its
nutritional value in this respect. A combination of transgenes enabled biosynthesis of provitamin A in the endosperm.
Yonekura-Sakakibara, K. and K. Saito (2006). "Review: genetically modified plants for the promotion of human health." Biotechnology Letters
28(24): 1983-1991.
://WOS:000243861500002 AND http://www.botanischergarten.ch/Golden-Rice/Yonekura-Sakakibara-Review-GM-Plants-2006.pdf
Plants are attractive biological resources because of their ability to produce a huge variety of chemical compounds, and the
familiarity of production in even the most rural settings. Genetic engineering gives plants additional characteristics and value for
cultivation and post-harvest. Genetically modified (GM) plants of the "first generation" were conferred with traits beneficial to
producers, whereas GM plants in subsequent "generations" are intended to provide beneficial traits for consumers. Golden Rice is a
promising example of a GM plant in the second generation, and has overcome a number of obstacles for practical use. Furthermore,
consumer-acceptable plants with health-promoting properties that are genetically modified using native genes are being developed.
The emerging technology of metabolomics will also support the commercial realization of GM plants by providing comprehensive
analyzes of plant biochemical components.
Yu, J., S. Hu, et al. (2002). "A Draft Sequence of the Rice Genome (Oryza sativa L. ssp. indica)." Science 296(5565): 79-92.
http://www.sciencemag.org/cgi/content/abstract/296/5565/79 and http://www.botanischergarten.ch/Rice/Yu-et-al-Draft-Rice-Genome-
2002.pdf

Yuan, Q., J. Quackenbush, et al. (2001). "Rice Bioinformatics. Analysis of Rice Sequence Data and Leveraging the Data to Other Plant Species."
Plant Physiol. 125(3): 1166-1174.
http://www.plantphysiol.org/cgi/content/abstract/125/3/1166 and http://www.botanischergarten.ch/Rice/YuanPlantPhys200103.pdf
Rice (Oryza sativa) is a model species for monocotyledonous plants, especially for members in the grass family. Several attributes
such as small genome size, diploid nature, transformability, and establishment of genetic and molecular resources make it a
tractable organism for plant biologists. With an estimated genome size of 430 Mb (Arumuganathan and Earle, 1991), it is feasible to
obtain the complete genome sequence of rice using current technologies. An international effort has been established and is in the
process of sequencing O. sativa spp. japonica var "Nipponbare" using a bacterial artificial chromosome/P1 artificial chromosome
shotgun sequencing strategy. Annotation of the rice genome is performed using prediction-based and homology-based searches to
identify genes. Annotation tools such as optimized gene prediction programs are being developed for rice to improve the quality of
annotation. Resources are also being developed to leverage the rice genome sequence to partial genome projects such as expressed
sequence tag projects, thereby maximizing the output from the rice genome project. To provide a low level of annotation for rice
genomic sequences, we have aligned all rice bacterial artificial chromosome/P1 artificial chromosome sequences with The Institute
of Genomic Research Gene Indices that are a set of nonredundant transcripts that are generated from nine public plant expressed
sequence tag projects (rice, wheat, sorghum, maize, barley, Arabidopsis, tomato, potato, and barrel medic). In addition, we have
used data from The Institute of Genomic Research Gene Indices and the Arabidopsis and Rice Genome Projects to identify putative
orthologues and paralogues among these nine genomes.
Zeigler, R. S. (2001). "Agricultural biotechnology - Reducing poverty in developing countries." Plant Disease 85(6): 568-579.
://000168777500002 AND http://www.botanischergarten.ch/Golden-Rice/Zeigler-Reducing-Poverty2001.pdf
Zhao, F.-J. and S. P. McGrath (2009). "Biofortification and phytoremediation." Genome studies and Molecular Genetics, edited by Juliette de
Meaux and Maarten Koornneef / Plant Biotechnology, edited by Andy Greenland and Jan Leach 12(3): 373-380.
http://www.sciencedirect.com/science/article/B6VS4-4WCFV3B-1/2/e0a412ba259b8e9c5bd35cdf2ce05d84 AND
http://www.botanischergarten.ch/Golden-Rice/Zhao-Biofortification-Bioremediation-2009.pdf
Producing nutritious and safe foods sufficiently and sustainably is the ultimate goal of modern agriculture. Past efforts have focused
on increasing crop yields, but enhancing the concentrations of mineral micronutrients has become an urgent task because about half
of the world population suffers from the malnutrition of iron, zinc, and selenium. Biofortification of these trace elements can be
achieved through fertilization, crop breeding or biotechnology. On the other hand, soils contaminated with metals or metalloids may
be cleaned up by phytoextraction that combines hyperaccumulation with high biomass production. Progress has been made in
identifying inter-species and intra-species variation in trace element accumulation, and mechanistic understanding of some aspects
of trace element transport and homeostasis in plants, but much remains to be elucidated.
Zhu, C. F., S. Naqvi, et al. (2008). "Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid
pathway in maize." Proceedings of the National Academy of Sciences of the United States of America 105(47): 18232-18237.
://WOS:000261489300037 AND http://www.botanischergarten.ch/Golden-Rice/Zhu-Combinatorial-Genetic-2008.pdf
Combinatorial nuclear transformation is a novel method for the rapid production of multiplex-transgenic plants, which we have used
to dissect and modify a complex metabolic pathway. To demonstrate the principle, we transferred 5 carotenogenic genes controlled
by different endosperm-specific promoters into a white maize variety deficient for endosperm carotenoid synthesis. We recovered a
diverse population of transgenic plants expressing different enzyme combinations and showing distinct metabolic phenotypes that
allowed us to identify and complement rate-limiting steps in the pathway and to demonstrate competition between beta-carotene
hydroxylase and bacterial beta-carotene ketolase for substrates in 4 sequential steps of the extended pathway. Importantly, this
process allowed us to generate plants with extraordinary levels of beta-carotene and other carotenoids, including complex mixtures
of hydroxycarotenoids and ketocarotenoids. Combinatorial transformation is a versatile approach that could be used to modify any
metabolic pathway and pathways controlling other biochemical, physiological, or developmental processes.
Zimmermann, M. B. and R. F. Hurrell (2002). "Improving iron, zinc and vitamin A nutrition through plant biotechnology." Current Opinion in
Biotechnology 13(2): 142-145.
http://www.sciencedirect.com/science/article/B6VRV-45SHW2J-F/2/a6f301d2b4a27095d6616d1c076a505b AND
http://www.botanischergarten.ch/Golden-Rice/Zimmermann-Improving-Iron-Zinc-Vitamin-A-2002.pdf
Recent understanding of plant metabolism has made it possible to increase the iron, zinc and [beta]-carotene (provitamin A) content
in staple foods by both conventional plant breeding and genetic engineering. Improving the micronutrient composition of plant
foods may become a sustainable strategy to combat deficiencies in human populations, replacing or complementing other strategies
such as food fortification or nutrient supplementation.
Zimmermann, R. and M. Qaim (2004). "Potential health benefits of Golden Rice: a Philippine case study." Food Policy 29(2): 147-168.
://000221924100003 and http://www.botanischergarten.ch/Rice/Zimmermann-Benefit-Goldenrice.pdf
Golden Rice has been genetically modified to produce beta- carotene in the endosperm of grain. It could improve the vitamin A
status of deficient food consumers, especially women and children in developing countries. This paper analyses potential impacts in
a Philippine context. Since the technology is still at the stage of R&D, benefits are simulated with a scenario approach. Health effects
are quantified using the methodology of disability-adjusted life years (DALYs). Golden Rice will not completely eliminate the
problems of vitamin A deficiency, such as blindness or increased mortality. Therefore, it should be seen as a complement rather than
a substitute for alternative micronutrient interventions. Yet the technology could bring about significant benefits. Depending on the
underlying assumptions, annual health improvements are worth between US$ 16 and 88 million, and rates of return on R&D
investments range between 66% and 133%. Due to the uncertainty related to key parameters, these results should be treated as
preliminary. (C) 2004 Elsevier Ltd. All rights reserved.