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2.14 1988). This is accomplished by providing an in Mm culture environment for the haploid em bryos similar to that provided by the ernbryo sac for zygotic embryos in vivo. for example, by inducing desiccation toleranœ and dehydration of the embryos. In v h embryos do not naturally develop desiccation tolerance or undergo dehydration or domancy. To induce these conditions, abscisic acid (ABA). cold and heat treatments along with desiccation have been implemented to increase the frequency of conversion from embryos to plants (Kott and Beversdorf. 1990; Senaratna et al.. 1991 ; Brown et al., 1993). Self-pollination or inbreeding to produce homozygosity in obligate outcrossing crops leads to inbreeding depre&on. seen as a loss in population performance, due to the expression of deleteflous homozygous recessive alleles. lnbreeding depression has been reported in DH lines of B. rapa expressed as poor germination rates, spindly branching, reduced height, late floweting. lack of pollen, low seed set and poor seed quality (Dewan et al., 1995). OH production does not avoid inbreeding by minimizing the repeated cycles of inbreeding. In order to exploit the benefits of DH development in cross pollinating crops, there must be a recovery of performance to at least the level of the original heterozygous population. Genotype Analysis Traditional plant breeding techniques are based on the selection of a genotype using phenotypic analysis. Phenotype is not an accurate indicator of genotype when
2.1 5 traits have low heritability. are quantively inherïted or are influenced by the environment. Direct analysis of genotype is a more accurate method of seledion. This could be accomplished through the identification of genetic markers linked to desirable traits (Tanhuanpaa et al., 1996). development of genetic maps (Zhang et al.. 1995) and establishment of genetic relationships between and among species (Mailer et al.. 1994; Bagheri et al.. 1995; Demeke et al., 1996). The use of genetic markers may help reduce the amount of field screening and therefore avoid the necessity of carrying plants to maturity which is often neœssary with the use of morphological traits such as seed colour or seed quality. Genotype analysis used along with phenotypic or morphological information could reduce time required to develop a desirable variety. Procedures such as restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) based assays have been used to characterize a genotype through DNA analysis. Restriction Fragment Length Polymorphisms (RFLPs) RFLP detects variation between genotypes using restriction endonudeases to fragment DNA along with blot hybridization to visualize polymorphisrn. RFLP analysis has been used successfully in the mapping of B. napus and B. rapa (Landry et al., 1991 ; Chyi, et al., 1992), detemining genetic relationships within and among Brassica species (Song and Osbom, 1992; Dien and Osbom, 1994) and predicting heterosis in tetraploid alfalfa (Kidwell at al.. 1994). RFLP markers segregate codominantly so that
Page 1 and 2: The Application of Ooubled Haploid
Page 3 and 4: COPYRIGET P1ERbIISSION PAGE A Thesu
Page 5 and 6: 111 application in 8. rapa cultivar
Page 7 and 8: v TABLE OF CONTENTS ABSTWT ACKNOWLE
Page 9 and 10: VI1 LIST OF TABLES TABLE Page EFI',
Page 11 and 12: lx LlST OF FIGURES FIGURE Page Prod
Page 13 and 14: 1.2 performance. Production of DH p
Page 15 and 16: History of Canola Produdion of Bras
Page 17 and 18: Economic Importance and Distributio
Page 19 and 20: 2.5 genes involved in recognizhg an
Page 21 and 22: 2.7 There is potential for loss of
Page 23 and 24: 2.9 their stability under different
Page 25 and 26: 2.1 1 selected and selfed to create
Page 27: 2.1 3 production (Mathias and Wbbel
Page 31 and 32: 2.17 opposite strands of template D
Page 33 and 34: Figure 2.3. The AFLP procedure usin
Page 35 and 36: 2.21 non-specifïc arüfacts which
Page 37 and 38: 2.23 maintenance of DNA extradion p
Page 39 and 40: Cornparison of Bud Pollination and
Page 41 and 42: INTRODUCTION Production of doubled
Page 43 and 44: Effkiency lndex = total number of s
Page 46 and 47: Application of doubfeâ haploid dev
Page 48 and 49: Brassita rapa represents approximat
Page 50 and 51: 4.5 and a daylnight temperature of
Page 52 and 53: 4-7 plants within each tent using a
Page 54 and 55: 4.9 Days to Flowanng Reward OH Iine
Page 56 and 57: 4-11 In Canan, Reward Cja and Cl,,
Page 58 and 59: 4.13 counterparts. The extra genera
Page 60 and 61: 4-15 composite population can eithe
Page 62: Table 4.2. Mean parameter values fo
Page 68 and 69: Table 4.8. Correlation of days to f
Page 72 and 73: Detedion of Genetic Variation in Bm
Page 74: INTRODUCTION The production of doub
Page 77 and 78: 5-6 represented one sample from C,
Page 79 and 80:
5-8 the percent of polymorphism pre
Page 81 and 82:
5-10 and C, and a similar level of
Page 83 and 84:
Table 5.2. Polymorphic levels in Cl
Page 86 and 87:
5-15 Table 5.5. Proportion of polyr
Page 88 and 89:
6. GENERAL DlSCUSSlON AND CONCLUSIO
Page 90 and 91:
6-3 B. rapa OH Iines. Seleded DH Ii
Page 92 and 93:
Abramson, RD. 1995. Thermostable DN
Page 94 and 95:
Diers, B.W. and Osbom, T.C. 1994. G
Page 96 and 97:
Jasieniuk, M., Brill&Babel, AL. and
Page 98 and 99:
Narasimhulu, SBw and Chopra, U.L. 1
Page 100 and 101:
Schuler, T.J., Hutcheson, O.S. and
Page 102 and 103:
Williams, J.G.K., HanMey, M.K., Raf
Page 104 and 105:
Appendix 12. Data matrix of RAPO pr
Page 106 and 107:
Apperidi 1. (-nueû) Month D~Y Temp
Page 108 and 109:
Mean Max [mm)
Page 110 and 111:
2 3 4 5 6 7 8 9 10 11 12 13 14 15 1
Page 112:
Append'bc 4. FM measummmts fw param
Page 116 and 117:
Locaüon Plant Id. RedTcate €MG D
Page 118 and 119:
C24 C24 C2-4 C2-8 C2-8 C2-8 C2-8 c2
Page 120 and 121:
Location Plant Id. Replicate €MG
Page 122 and 123:
Locaüori Plant ld. Replicate EMG O
Page 124 and 125:
Appendix 10. The mean performance o
Page 127:
f i . I 1 t a I 1 I I t t c i r' !m
Page 130 and 131:
Appendii 12. Data matrix of RAPO pm
Page 133:
Appendix 13. @mtïnued) Primer Comp
Page 138:
Appendk 14. (continuedl Primer Comp
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