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Genetic Analyses of Intertribal Brassica Species×Capsella Bursa-pastoris Hybrids and B. Napus Nullisomics

Author: ChenHaiFeng
Tutor: LiZaiYun
School: Huazhong Agricultural University
Course: Crop Genetics and Breeding
Keywords: Distant hybridization Rapeseed quality Sclerotinia Penetrate Genomic in situ hybridization (GISH) Amplified fragment length polymorphism (AFLP) Nullisomic Cytology
CLC: S565.4
Type: PhD thesis
Year: 2008
Downloads: 171
Quote: 3
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Abstract


Distant hybridization is an important way for crop genetic improvement, good traits of the wild species can be transferred to the cultivated species. The Cruciferous in many excellent wild germplasm resources, provides many useful nucleus and cytoplasm gene for rapeseed breeding. China is now a large area of ??cultivated Brassica napus (Brassic napus, 2n = 38, AACC) in the 20th century, the 30 and 40's from abroad, the paucity of genetic variation, it is through a variety of ways to create new germplasm resources of their genetic improvement has important significance. Dishes family Cruciferae lone shepherd's purse (Capsellabursa-pastoris (L.) Medic, 2n = 4x = 32) widely distributed in the world, in China and some other countries as vegetables and medicinal. Shepherd's purse seed erucic acid (0.68%) and glucosinolates (15.68μmol / g) in line with the double-low (erucic acid <1% the glucosinolate <30μmol / g) standard for the natural genetic improvement rape double low germplasm resources, and high resistance to Sclerotinia. This research synthesis of Brassica campestris (B.rapa, 2n = 20), the family of Brassica napus and shepherd's purse between hybrids and hybrids and their offspring morphology, cytology, molecular biology, fatty acids and anti-Sclerotinia analysis. In addition, previously obtained from the genus Brassica napus Orychophrogmus interspecific hybridization nullisomic plants (2n = 36) and offspring cytogenetic studies. The main results are as follows: 1. Rape × shepherd's purse. Rapa and Brassica napus as female parent, shepherd's purse the most F parent in the family of sexual hybridization between a large number of artificial pollination 1 plant morphology and maternal similar, only The few performance for intermediate. F 1 plant has some of the characteristics of the male parent, shepherd's purse, such as leaves, dark green leaves crack depth, small plant, base branching, white petals. Somatic cells (ovary cells) chromosome number, F 1 plants are divided into five categories: The first category 2n = 27-29, the second type of 2n = 20, these two types of plants from the Brassica napus The hybridization with shepherd's purse; Brassica napus haploid, 2n = 19, fourth class 2n = 29, the fifth category 2n = 38, three types of plants from Brassica napus and shepherd's purse hybridization. Genomic in situ hybridization (GISH) analysis shows that in the first class of hybrid plants pollen mother cells (pollenmother cells, PMCs) 1-2 shepherd's purse chromosome, while the other detected in some plant ovary cells and PMCs shepherd's purse chromosome fragment. AFLP (amplified fragment length polymorphism) analysis showed that, in addition to three F 1 plants (Nos.13, 24,32) shepherd's purse specific bands, all plants, shepherd's purse specific band, rape mother the missing band and parental no new band, but not the same as the number and composition of the polymorphic bands between the various monoclonal showed that the infiltration of the exogenous genetic material at different levels, while causing the receptor genome structure changes. By fatty acid analysis and Sclerotinia identification, erucic acid and glucosinolate content found some female parent progenies reduced Sclerotinia resistance increased significantly. 2 male sterile material. A female parent plants from Brassica napus Oro and shepherd's purse hybridization (No.30) small stamens, male infertility. Oro backcross generation plants full fertile, since the good seed. Proportion the 359 BC 1 F 1 plants, 276 fertile and 83 infertile, fertile plants and sterile plants of 3:1 (χ c 2 = 0.58 <χ 0.05 2 ); in F 1 × BC 1 109 plants, 58 fertile and 51 infertile, fertile strains infertility strain ratio of 1:1 (the χ C / sub> . 2 < / sup> = 0.33 the <χ 0.05 2 ), the male sterility of a nuclear recessive gene infertile. Anther slice observations suggest that the sterile plants abnormal anthers sporogenous cell stage, the tapetum multilayer, mast, but its the subtrahend separation is normal. The tetrad stage tapetum further deterioration, multi-layer, hypertrophy, vacuolization, started to degrade, and squeeze the tetrad tetrad can not form mononucleate pollen, the small spores begin degradation. The most progeny plants significantly reduced glucosinolate content, some plants reach double low standard, and Sclerotinia resistance increased significantly. The Brassica napus nullisomic. Brassica napus Oro and Orychophrogmus of a hybrid mixed ploidy microspore culture, obtained a nullisomic (2n = 36) plants, plant height of only 70-80cm, flowering about two months earlier than in Brassica napus . PMCs at diakinesis the nullisomic plants with 18 bivalents late Ⅰ normal separation performance 18:18, 14 selfing generation plants, six plants were 31.2% -43.8% PMCs at a later stage I. 17:19 separation, which led to the formation of the offspring nullisomic - four-body (2n = 38). Brassica campestris L. the × nullisomic the PMCs at diakinesis has 10 Ⅱ 8 Ⅰ, the nullisomic missing from Brassica C genome chromosome. The Oro × nullisomic offspring plant height and flowering time of Brassica napus close, but did not find the missing body plants in the progeny. Plant height Biao Luo of nullisomic - four-body dwarf 5cm, flowering as early as a month or so longer than Brassica napus Oro, 59% of PMCs at diakinesis 17 Ⅱ 1 Ⅳ, the remaining 19 Ⅱ, better self-seed. Finally, we propose a hybrid formation mechanism of rapeseed and shepherd's purse. We think that chromosome elimination in the early development of the zygote formation or hybrid embryos, doubling and introgressed. Female parent type plants by hybridization, shepherd's purse can be quickly transferred to rapeseed good traits (double low resistance to Sclerotinia sclerotiorum). At the same time, we discussed in rapeseed breeding, gene mapping and functional analysis of the missing body and lack of body - four-body.

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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Rapeseed ( Brassica )
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