Management of the Diamondback Moth and Other Crucifer Insect ...

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- undertake breeding and trial activities- undertake protein based safety studies- obtain the regulatory trial permitsCIMBAA reported on the rationale for the developmentand deployment of stacked gene Cry1B/Cry1C, and theinitial import of transformed cabbage and cauliflowerlines into India for testing and selection, in theproceedings of the 5 th International Workshop in thisseries in 2006 (Russell et al. 2008).Phase I of the partnership (2002-04) dealt with countryneeds assessments, technical feasibility studies, geneselection and stakeholder agreement.Phase II (2005-2006) formally established thepartnership, demonstrated the need for the product,developed the transgenic plant material, undertook proofof concept lab and field testing and undertookpreliminary studies on biosafety.Phase III (2007-10) focused on the efficacy of the Btplants against a range of key pests, selection of plants forcommercialization and developing an IPM context forthe material.Phase IV (2011- ) would be the undertaking of formalregulatory studies in India, followed by submission of theregulatory dossier for release approval in India and forbiosafety/import approval in other major globalregulatory regions.This paper describes progress in phases II and III anddiscusses the situation vis a vis Phase IV.The CIMBAA public sector collaboration partners werethe University of Melbourne (Australia), CornellUniversity (Entomology Department and Office ofInternational Programmes) (USA), the Natural ResourcesInstitute of the University of Greenwich (UK) andAVRDC- The World Vegetable Center (Taiwan), withNunhems Pvt. (Netherlands and India), the vegetableseeds division of Bayer Crop Science, as the privatesector seed company developing the transformed plants.The 2008 paper summarized the progress of the work to2006 with the Bt proteins Cry1B and Cry1C selected in2003 on the basis of earlier research by Zhao et al.(2001) and Mohan and Gujar (2002), which suggestedgood efficacy and critically no cross resistance betweenthese two proteins. Zhao et al (2003) showed that stackedBt proteins were more effective in delaying resistancecompared to single proteins in a rotation or mosaic.Building on and expanding earlier work reported inRussell et al. (2008), Grzywacz et al. (2010) reviewedcurrent control methods for brassica pests in Asia andAfrica and concluded that effective Bt transgenics couldplay a very useful role and should be complimentary toother IPM practices including the conservation ofimportant parasitoids such as Diadegma semiclausumand Cotesia plutellae.CIMBAA STRUCTUREThe CIMBAA Steering Committee comprises thedirectors of the collaboration’s partner institutions andhas the oversight and decision making responsibility,meeting twice a year or as required. The projectmanagement team with day to day responsibility for thework comprises leading scientists from each of thepartners, with coordinators from the public and privatesectors who also act as secretaries to the SteeringCommittee. The project management team discussesmonthly by conference call. CIMBAA also set upadvisory panels to both help guide the on-going work ofthe project and to ensure that the work was to the highestinternational standards. The International Advisory Panelcomprises past and present Directors General ofConsultative Group on International AgriculturalResearch (CGIAR) or other international agriculturalresearch centres, institutes (ICIPE, IRRI, CIMMYT), andof the Indian Council for Agricultural Research (ICAR),the immediate past Director of Research of Nunhems,plus a specialist in gene flow science. The InternationalAdvisory Panel has provided advice and comment to theSteering Committee on a number of technical andprocess issues.The Indian Advisory panel has served two functions. Ithas provided detailed technical guidance on the studies tobe undertaken by CIMBAA and has provided amechanism for the project to ensure that the work isunderstood and approved of by senior Indian governmentscientists. The panel has been chaired by the ExecutiveDirector of The Energy and Resources Institute (TERI),who has had a leading role in the communication aspectsof CIMBAA. The committee’s makeup has varied overthe years but has included an Assistant Director General(Crop Science) from ICAR, Directors of the Departmentof Biotechnology (DBT), National Research Centre forPlant Biotechnology (NRCPB), Indian AgriculturalResearch Institute (Divisions of Entomology, VegetableScience and Horticulture), National Bureau for PlantGenetic Resources (NBPGR) and the National Bureau ofAgriculturally Important Insects amongst others.ACCEPTANCE OF NEED FOR AN “INTHE SEED”SOLUTIONFarmer pest control studiesA number of studies for CIMBAA, summarized inGrzywacz et al. (2010), demonstrated the potential valueto farmers in Africa and Asia of Bt brassicas (see alsoBadnes-Perez and Shelton 2006). Specifically for India, astudy undertaken in connection with CIMBAA(Weinberger and Srinivasan 2009), confirmed theunsatisfactory state of current insect control on cabbageand cauliflower. 97% of the 300 farmers interviewed inGujarat, Karnataka and West Bengal regarded insects astheir number one production problem and all of thesefarmers used chemical pesticides (72% used nothing butpesticides). Thirty seven different pesticides were in useand 66% of farmers were routinely spraying insecticide20 AVRDC - The World Vegetable Center
mixtures. One third of farmers were spraying even ifthere was no insect damage on the crop and only 8%considered pest numbers before spraying. Given thatfarmers were routinely spraying every 3-4 days and oftenmore frequently, especially on cauliflower, it is notsurprising that 75% had health issues related to pesticideuse in the preceding year and indeed 2% of farmers hadbeen hospitalized. The average cost of pest control incabbage was US$ 141/ha and US$ 316/ha for cauliflower(10-30% of total production costs).Stakeholder SupportA key workshop held in New Delhi in 2005 entitledPublic-private partnership in the use of Agribiotech forsustainable solutions to brassica pest problemsestablished the scientific commitment to geneticsolutions. Dr. Trivedi of the National Centre for IPMsaid that seed would be the most important component ofIPM, and Dr G. Kalloo, Deputy Director General ofICAR confirmed that gene pyramiding is a major drive ofICAR (Srinivasan et al. 2005). The potential role ofpublic/private partnerships in Agricultural biotechnologywas strongly endorsed by a panel including the IndianPrime Minister, Dr. Manmohan Singh, the leader of theIndian Green Revolution Prof. M.S. Swaminathan andthe Minister of Science, Sri Kapil Sibal, at theInternational Conference on Agriculture for Food,Nutritional security and Rural Growth in New Delhi inMay 2006.involved the insertion of cassette containing tightlylinked cry1B and cry1C genes driven by subterraneanclover stunt virus promoters with the bar herbicidetolerant gene and its PAT protein as a selection marker.As required under Indian regulations, Nunhems set up anInstitutional Biosafety Committee in 2005 and applied alicense for the import of Bt transgenic lines which wasreceived in April 2006 with 85 lines released fromquarantine in August that year (other importations weremade later).An initial polyhouse (plastic greenhouse) trial wasundertaken on the Nunhems field site at Bilaspur nearDelhi during autumn 2006 (Fig. 1). Laboratory reareddiamondback moth were released onto the Bt plants andthe plant performance, monitored by the India regulators,was regarded as a sufficient proof of concept to justifylarger scale trials in the following seasons.Consumer studyGiven the global concerns over GM food crops, a studyof 645 households in two large urban centres (New Delhiand Bengaluru) and two regional centres (Kolar andBarddhaman) was undertaken in 2006-07 (Krishna andQaim 2008). It found that 80% of consumers recognisedpesticide residues on vegetables as a significant problem.The concept of Bt vegetables was new to mostinterviewees, but, following a standard explanation, 68%supported the introduction of Bt vegetables if they cutresidues (and provided they were approved as safe by theIndian regulatory authorities). Most of the 17% ofhouseholds who strongly opposed to GM vegetableswould buy them if they were significantly cheaper thannon-GM alternatives. However, the more educated therespondent, the more likely they were to both be aware ofresidues and to be anti-GM foods. It seems likely that inrecent years this attitude has spread further with the antiGM political agitation by some NGOs in India.TRANSFORMATION, IMPORTATIONAND PRELIMINARY TESTING IN INDIASome 600 primary transformants for each of cabbage andcauliflower were produced at Nunhems facilities in theNetherlands. The transformation was Agrobacteriummediated into cabbage and cauliflower hypocotyls andFigure 1. Government inspection of the first CIMBAAtrial, Bilaspur, near Delhi 2006EFFICACY, SAFETY, SUSTAINABILITYAND REGISTERABILITY TESTINGExpression of Cry proteins in plantsCry protein expression levels in different tissues of theCIMBAA plants, over the growing season and indifferent parts of India were studied by N. Kaushik(paper in prep.) of The Energy and Resources Institute,new Delhi. In summary: Protein was expressed in all plant parts includingcauliflower curds. Protein expression was fairly constant in differentparts of leaves and different leaf positions on theplant. Expression was constant across plants from a givenevent but varied somewhat with the date of sampling. Expression in hybrids (single copy of each Bt gene)was 20-40% lower than in the Bt parents.The 6th International Workshop on Management of the Diamondback Moth and Other Crucifer Insect Pests 21
Page 1 and 2: PROCEEDINGSThe Sixth International
Page 3 and 4: PROCEEDINGSThe Sixth International
Page 5 and 6: Table of ContentsForewordixAcknowle
Page 7 and 8: Myco‐Jaal® - A novel formulation
Page 9 and 10: ForewordVegetable brassicas such as
Page 11: AcknowledgementsThe Sixth Internati
Page 15 and 16: Behavioral and geneticcomponents of
Page 17 and 18: more wandering behavior in the pres
Page 19 and 20: influenced whether larvae remained
Page 21 and 22: CLIMEX calculates a growth index (G
Page 23 and 24: Table 1 (continued)LimitingprocessW
Page 25 and 26: Figure 5. Observed DBM light trap c
Page 27 and 28: Rising to the challenge: anational
Page 29 and 30: Table 1. List of insecticides for w
Page 31: Progress and challengesin the Bt br
Page 35 and 36: Table 1. Mortality of lepidopteran
Page 37 and 38: o The Genetics Engineering Approval
Page 39 and 40: AVRDC; A. Rauf of Bogor University
Page 41: SESSION 2Biology, ecology and behav
Page 44 and 45: fields of Thondamuthur village, Coi
Page 46 and 47: Table 2. Morphometrics of C. binota
Page 48 and 49: On the basis of LC 50 values, the s
Page 50 and 51: Natural mortality ofPlutella xylost
Page 52 and 53: (x10 magnification) and excess eggs
Page 54 and 55: Figure 2. Lifetables for insect coh
Page 56 and 57: Diadegma semiclausum is well establ
Page 58 and 59: Diurnal behavior ofnaturally micros
Page 60 and 61: changed after the eggs were counted
Page 62 and 63: Geden, C. J., Long, S. J., Rutz, D.
Page 64 and 65: Scaptomyza flava (GP Walker, unpubl
Page 66 and 67: longer distance migrations into the
Page 68 and 69: FIGURESFigures 1A & B. Mean number
Page 70 and 71: Monitoring of cabbagelooper, Tricho
Page 72 and 73: populations of British Columbia col
Page 75 and 76: Importance ofglucosinolates indeter
Page 77 and 78: the importance of glucosinolates in
Page 79 and 80: Olfactory responses ofPlutella xylo
Page 81 and 82: Figure. 1: Percentage of adult P. x
Page 83 and 84:
Moorthy, 1992) have reported that p
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P
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egg masses and more eggs in total o
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SESSION 4Biological and non‐chemi
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on cabbage cultivation in one seaso
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four stages at day 7 post treatment
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Table 5. Biological effects of 17 e
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Kelly A Cook. K.a., S.T. Ratcliffe.
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applied only once every three weeks
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South Africa was assigned to supply
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Waladde SM, Leutle MF, Villet MH. 2
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MATERIALS AND METHODSThe study site
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Several studies have demonstrated t
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approach would continue until it ev
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ecorded in Hod 3 (19.1%), followed
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plutellae. Many adults of C. plutel
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litura (SL) which normally live in
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Table 2. Sizes of m-PX and m-SL fro
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helping us with the in vivo study.
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Since temperature is one of the mos
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Figure 4. Effect of different conce
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Predators in early seasonbrassica c
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Table 2. Predators encountered duri
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Table 5. Sticky trap captures for c
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dwelling predators, although it is
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Predators per trap109876543Lycosida
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Mortality was calculated using Abbo
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DBM fed on the leaf discs treated w
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Quality aspects of Bacillusthuringi
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five applications one week apart fo
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Antifeedant effect ofAcorus calamus
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Figure 1. Concentration-antifeedant
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CONCLUSIONA. calamus extracted in f
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of D. grandiflora on the growth kin
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etween antifeedant activity and con
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Table 2. Antifeedant index (AFI) an
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Field evaluation of insectexclusion
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Three main Lepidoptera pests were o
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price (SBD)30.0025.0020.0015.0010.0
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STATISTICAL ANALYSISThe data collec
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Myco-Jaal: a novelformulation of Be
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asexual conidium. The conidia adher
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Table 1. Efficacy of Myco-Jaal on y
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Susceptibility ofdiamondback moth a
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Table 1. Toxicity of Cry1Ac to fiel
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ReferencesAbbott WS. 1925. A method
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76°58′17″E), Tamil Nadu, India
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International Workshop, Talekar NS,
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Table 3. Effect of Neem limonoids o
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Figure 1. Structure of neem limonoi
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catch up with the farming community
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All treatments were imposed by usin
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The above results indicate that the
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A strong negative relationship obse
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ReferencesAnonymous 2000. Package o
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egularly causes epizootics in popul
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treatments. It is evident that DBM
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Table 1. Effects of three aqueous s
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egion (Adams, 1959); it is a native
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Effects of Cry toxins and Bt formul
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Hilbeck A, Baumgartner M, Fried MP,
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Table 3. Effect of Cry 1Ba2 on Cote
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SESSION 5Insecticides and insectici
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time. Within four years of the firs
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Update on DBM diamideresistance fro
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In year 2011, Sai Noi and Tha Muang
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Lahm GP, Cordova D, Barry JD. 2009.
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Fipronil acts by blocking chloride
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Table 2. Slope, LC 50 (ppm) and let
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Figure 2. Lethal dose ratios for fi
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y the inherent lower susceptibility
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Diamondback mothresistance to commo
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of the relative potency of each ins
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Table 3: Susceptibility of six fiel
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Spinetoram, a newspinosyn insectici
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Spinetoram is also highly effective
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Norte del Estado areas of Guanajuat
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Insecticide resistancemanagement: s
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still exist. The second source of i
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they organized the so called “Dia
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Recent developments inmanagement of
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is the reciprocal of the relative p
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developing thresholds for new brass
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Table 3: Spinosad (Success®) and i
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Insecticide sprays are generally ap
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may still benefit from doing so, ir
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generations” to Group 28 insectic
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Management of insecticideresistance
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Stop cultivation of crucifer crops
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252 AVRDC - The World Vegetable Cen
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Impact of reduced-riskinsecticides
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mines) on the 10 plants in each plo
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Table 4. Follow up with growers aft
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Guided by such objective, the Thai
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establishment rapidly achieved by D
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Malaysian Agricultural Research and
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the farmers. We report the developm
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Table 3. Number of DBM larvae in th
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Pest status and damage potentialThe
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Life tableLife table of DBM reveals
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Bacillus thuringiensis (Bt)Currentl
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CHEMICAL INSECTICIDESPeter et al. (
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Manjunath TM. 1972. Biological stud
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Armyworm, Spodoptera litura Armywo
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Adoption and perceptionTable 5 show
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Control of brassica pests -an examp
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Table 2. A Comparison of IPM and Pe
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Occurrence and control ofPlutella x
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Apopul at i on dynami c ( l ar va/
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Diadegma semiclausumD. semiclausum,
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Diamondback moth(Plutella xylostell
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SESSION 7Genomic and other novel ap
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management (IPM) systems (Shelton e
Page 314 and 315:
Kalra VK, Sharma SS, Chauhan R, Bha
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Table 2. Segregation analysis based
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current paper provides detail on th
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Proteins do not act antagonisticall
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CONCLUSIONThe transformation events
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Enhancement of the sterileinsect te
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optimisation of transformation prot
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Workshop PresidentsDr. Jacqueline d
Page 330 and 331:
Participant e‐mail Organization C
Page 332 and 333:
Author IndexAbuzid, I..............
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