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Development of plant breeding technology and new traits of Brassica crops using microspore culture

小孢子培养育种技术及芸苔属作物新性状开发

基本信息

批准号：
07556002
负责人：
TAKAHATA Yoshihito
金额：
$ 8.19万
依托单位：
Iwate University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556002/
关键词：
Brassica Raphanus Microspore culture Haploid method of breeding Mutation Transformation 形質転換 Brissica

项目摘要

1.Practical breeding : The DH lines, which had both high microspore embryogenesis ability and characteristics similar to Japanese cultivars, have been developed using microspore culture in Chinese cabbage and cabbage. The ABA-treated embryos sprouted in pots under unsterile conditions after 1 year storage at 4ﾟC.This storage system was expected to be useful for haploid breeding.2.Mechanism : The androgentic embryos were clarified to be heterotrophic at early developmental stage, but autotrophic after heart-shaped stage. Inheritance of microspore embryogenesis was studied by 4x4 diallel analysis. High embryogenesis ability was dominant to low ability, and additive gene effect was predominant. The heritability for embryogenesis was quite high. RAPD analysis showed that 3 RAPD makers were identified to be associated with embryogenesis.3.Enlargement of variation : The effect of UV irradiation on microspore culture was examined. The microspores were highly sensitive to UV.Using in vitro selection of UV irradiated-microspoes by soft rot-and black rot-filtered solution, disease tolerant-mutants of Chinese cabbage were selected. Transgenic B.napus was produced by direct gene transfer and Agrobacterium-mediated method using microspores as targets.4.Estimation of haploid breeding : The degree of genetic fixation in DH lines was examined by RAPD analysis. Although the DH lines showed much higher uniformity than parental lines, the variation among the individuals still remained.

1.实际育种：利用大白菜和结球甘蓝的小孢子培养，获得了既具有较高的小孢子胚胎发生能力，又具有与日本品种相似特性的DH系。经ABA处理的胚在4 ℃下贮藏1年后，在非无菌条件下可萌发，该贮藏体系有望用于单倍体育种。2.机理：雄核胚在发育早期为异养，心形期后为自养。采用4 × 4双列分析法研究了小孢子胚胎发生的遗传规律。高胚性对低胚性为显性，基因加性效应占主导地位。胚胎发生的遗传力较高。RAPD分析表明，有3个RAPD标记与小孢子发生有关。3.变异的抑制：研究了紫外线对小孢子培养的影响。小孢子对紫外线高度敏感，利用软腐病菌和黑腐菌滤液对紫外线辐照小孢子进行离体筛选，筛选出大白菜抗病突变体。以小孢子为目标，采用直接转基因和农杆菌介导法获得转基因B.napus。4.单倍体育种的评价：利用RAPD分析检测DH系的遗传固定程度。虽然DH系表现出比亲本系高得多的整齐度，但个体间的变异仍然存在。