甘蓝型油菜小孢子培养影响因素研究及再生苗早期倍性鉴定

doi:10.7668/hbnxb.2011.05.020

摘要/Abstract

摘要： 对不同年份、不同地点种植的油菜育种材料进行小孢子产胚率鉴定,并以11种不同基因型的甘蓝型油菜品系为供体材料,在武汉田间,分别于初花期和盛花期分离培养小孢子,研究基因型、生态条件、开花时间对小孢子产胚率的影响及秋水仙碱处理的加倍效果,通过采用流式细胞仪测定小孢子再生苗的DNA含量鉴定其倍性,并种植验证。结果表明:相同基因型在不同生态条件下种植及不同基因型间产胚率差异很大,西宁春播种植产胚率显著高于武汉秋播种植;大田条件下,初花期分离的小孢子产胚率显著高于盛花期。不同基因型油菜小孢子再生苗的自然加倍率也有差异,变异幅度为8.62%～20.88%。用50 mg/L秋水仙碱处理小孢子可以显著增加二倍体再生苗数量,加倍率最高可提高3倍以上,达到60%～77.59%。利用流式细胞仪测定小孢子再生苗的DNA含量鉴定再生植株的倍性与田间鉴定结果一致,可以作为小孢子再生植株早期倍性鉴定的有效手段。

关键词: 油菜, 小孢子培养, DNA含量, 倍性鉴定

Abstract: The breeding materials of Brassica napus grown in different ecological regions and years were used to determine their microspore embryogenesis capacity.The microspores of 11 genotypes grown in field condition of Wuhan,Hubei Province,were isolated at different flowering stages and cultured.The aim of this study was to investigate the effects of genotype,ecological condition and sampling time on microspore embryogenesis,as well as the colchicine treatment on chromosome doubling of regenerated seedlings.Ploidy level of regenerated plants was determined according to DNA content measured by flow cytometric analysis and confirmed by field observation.The results showed that the microspore embryogenesis was significantly different among the genotypes and in different ecological conditions.The plants sown in the summer in Xining,Qinghai Province,had significantly higher microspore embryogenesis than those sown in the spring in Wuhan,Hubei Province.The microspores isolated at early flowering stage(1-3 days)gave rise to significantly higher embryogenesis than those isolated at full flowering stage(12-15 days after flowering).The treatment of 50 mg/L colchicine directly on isolated microspores for 24 h could significantly increase the number of diploids in regenerated plants.The spontaneous chromosome doubling rate of regenerated plants ranged from 8.62% to 20.88% for different genotypes,whereas the colchicine treatment could raise the chromosome doubling rate to 60%-77.59%.The ploidy level of regenerated plants determined by flow cytometry was consistent with the result observed in the field,so the flow cytometry provided a useful means for early dihaploid discrimination of microspore-derived plants.

Key words: Oilseed rape, Microspore culture, DNA content, Ploidy identification

中图分类号:

S565.4

米哲, 李云昌, 梅德圣, 李英德, 徐育松, 陈玉峰, 胡琼. 甘蓝型油菜小孢子培养影响因素研究及再生苗早期倍性鉴定[J]. 华北农学报, 2011, 26(5): 97-102. doi: 10.7668/hbnxb.2011.05.020.

MI Zhe, LI Yun-chang, MEI De-sheng, LI Ying-de, XU Yu-song, CHEN Yu-feng, HU Qiong. Factors Affecting Microspore Embryogenesis and Early Ploidy Level Determination of Regenerated Plants in Brassica napus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(5): 97-102. doi: 10.7668/hbnxb.2011.05.020.

http://www.hbnxb.net/CN/Y2011/V26/I5/97

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