四倍体大白菜小孢子植株的获得与倍性鉴定

Plant Regeneration from Isolated Microspore Culture of Autotetraploid Chinese Cabbage Brassica campestris L.ssp pekinensis (Lour.)Olsson

以同源四倍体大白菜为试材，进行了游离小孢子培养的胚胎诱导条件及植株再生研究，并对再生株的染色体数进行了鉴定。结果表明：单核靠边期小孢子是诱导胚胎发生的最佳时期；不同基因型的四倍体大白菜小孢子胚胎发生能力不同，其中９４０５品系产胚率达８１胚／蕾；适于小孢子胚胎发生的培养基为ＮＬＮ附加蔗糖１３％～１７％、ＢＡ０．０５～０．２ｍｇ／Ｌ和ＮＡＡ０～０．５ｍｇ／Ｌ；ＮＬＮ中添加活性炭０．０５～０．１ｇ／Ｌ可有效提高小孢子胚胎产率及发育同步性；３３℃２４ｈ热激处理是诱导小孢子胚胎发生的必要条件；子叶型胚适宜植株再生，在Ｂ５附加琼脂１．２％或０．８％并铺有滤纸的培养基上子叶型胚植株再生可达４４％～６２％。共获得７３２个小孢子株系，其中三体株系１０９个。四倍体小孢子培养获得植株为大白菜三体系的创建找到了一条有效的途径。

Microspores culture and plant regeneration of autotetraploid Chinese cabbage were stu died .The results showed that it was necessary to isolate microspores in the late uninucleate stage.The optimal response was obtained at 33℃ for 24h at initial culture period,the frequency of embryogenesis was the highest.Embryogenesis were genotype dependent,embryos were induced from 8 of 10 genotypes,of which line 9405 produced 81 embryos per bud which was the highest yield.Medium NLN supplemented with 13%-17% sucrose,BA 0.05-0.2mg/L and NAA 0-0.5mg/L induced high frequency of embryogenesis.Addition of 0.05-0.1g/L activated charcoal to microspore suspension had significant effect upon the yield of embryos and percentage of cotyledary embryos.Compared with other type embryos,cotyledary embryo had a higher plant regeneration rate.The sui table medium for embryo germination was B 5 with 1.2% agar.The number of chromosome of 732 microspore plant lines were identified,the result showed that 109 lines were trisomic lines.