中国农业科学 2003,(5)557 - 560 : 36 Scientia Agricultura Sinica
利用 cDNA-AFLP 技术分析白菜核雄性 不育两用系的表达差异
王永勤,曹家树,符庆功,余小林,叶纨芝,向
(浙江大学蔬菜研究所, 杭州 310029)
王旬
摘要: 利用 cDNA-AFLP 技术, 对白菜核雄性不育两用系可育株群和不育株群分析显示: 花蕾之间基因表达存 在差异; 莲座期叶片之间, 开花期叶片之间和花茎之间基因表达无差异; 叶片, 花茎和花蕾之间基因表达存在差异. 对开花期的小蕾, 中蕾和大蕾分析显示: 对引物中, 对在可育株中蕾和大蕾中扩增出 1 条特异带, 1 对扩增 另 10 1 出 1 条在可育株中蕾中表达丰度高于不育株中蕾的条带, 其余 8 对未发现差异条带.对上述表达丰度有差异的基 因进行 Northern 验证, 结果与之一致, 表明 cDNA-AFLP 技术可以用于植物雄性不育基因表达差异的研究. 关键词:白菜; ;核雄性不育;cDNA-AFLP Brassica cam pestris L .(syn . B . rap a L . )
DifferentiaI Expression AnaIysis of Genic MaIe SteriIit y A / B Lines by cDNA-AFLP in Chinese Cabbage- pak-choi ( Brassica campes*ris ssp . c+i,e,sis Makino)
WANG Yon g-qin,CAO Jia-shu,FU Qin g- g on g,Y U Xiao-lin,YE Wan-zhi,XIANG Xun
( Institute o f Vegetable Science,Zhej iang University ,Hang zhou 310029 )
Abstract:To determine differential ex pression of genic male sterilit y A / B lines in Chinese cabbage- pak-choi ( Brassica cam p estris ssp . chinensis Makino var . com munis Tsen et Lee) cDNA-AFLP was analy zed,in dif, ferent develop ment stages and different tissues by usin g RNA fin gerprintin g technique. No obvious differential ex pressions were observed in rosette leaves,florescence leaves,and scapes. Some differential ex pressions were found in buds of A / B lines and amon g leaves,scapes and buds. Buds collected in different develop ment stages with 10 primer combinations were analy zed . With one pair primer,a unique band between middle buds and lar ge buds in B line was obtained;and with another one pair primer,one band reflected a hig her gene-ex pression level in A line than that in B line was obtained . No unique bands were found with the other primer combinations. The bands reflected different gene-ex pression level were confirmed by Northern h y bridization . All the results indicate that cDNA-AFLP is a suitable tool for study of differential ex pression of genic male sterilit y in plants. ; Key words:Chinese cabbage- pak-choi;Brassica cam p estris L .( sy n . B . ra p a L . ) Genic male sterilit y; cDNA-AFLP RNA 指纹技术可以揭示基因调控在 RNA 水平 的表现, 并进一步阐明其调控方式. Lian g 等根据 高等生物成熟 mRNA 带有 poly A 尾巴的特性, 用特 异的 锚 定 引 物 反 转 录 后, 行 PCR 扩 增, 立 了 进 建 ( mRNA 差 异 显 示 技 术 mRNA differential display, [1] 重 mRNA DD ) .但 该 技 术 存 在 假 阳 性 高, 复 性 差, 端引物倒数第二个碱基的简并性使得某些差 3' 异条带不能被扩增等缺点.1996 年 Bachem 等发展

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