中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 利用DAD1反义片段转化创建菜薹可调控雄性不育材料 园艺学报 2009,36 (5) 685-692
Position: Home > Articles > Development of Adjustable Male Sterile Plant in Chinese Flowering Cabbage by Antisense DAD1 Fragment Transformation Acta Horticulturae Sinica 2009,36 (5) 685-692

利用DAD1反义片段转化创建菜薹可调控雄性不育材料

作  者：
陈国菊;徐飞;雷建军;曹必好
单  位：
华南农业大学园艺学院
关键词：
菜薹;雄性不育;花药不开裂基因;茉莉酸;反义RNA
摘  要：
菜薹因为没有好的雄性不育材料或自交不亲和系,至今尚无一代杂种用于生产,根据拟南芥及白菜型油菜的花药不开裂基因DAD1的保守序列设计引物,扩增菜薹的DAD1基因片段(DAD1F),构建反义DAD1F植物表达载体,用农杆菌介导法转化菜薹,对转基因植株进行分子检测,鉴定其雄性不育性并进行育性恢复试验。克隆得到的菜薹的DAD1基因片段大小为678bp,命名为BrcpDAD1F,其序列与拟南芥和白菜型油菜的DAD1高度同源,同源率分别为88%和99%;共得到了12株转基因植株,有6株在转录水平上得到表达,表现为雄性不育,花器官畸形,花粉活力低,萌发率不到10%,且开花后不能结角果或结空角果,或者得到极少种子但种子不萌发;用对照的花粉给转基因植株授粉可使其正常结实。以500μmol.L-1茉莉酸甲酯处理可使其雄性不育得到恢复,花粉可以在柱头和培养基上萌发,具有受精能力。T1代可育株与不育株的比例都呈1∶3分离,T2代不同株系的育性分离比例不同,有些株系继续呈1∶3的分离,有些株系全是可育株或全是不育株,说明反义抑制呈单基因稳定遗传。
译  名：
Development of Adjustable Male Sterile Plant in Chinese Flowering Cabbage by Antisense DAD1 Fragment Transformation
作  者：
CHEN Guo-ju1,XU Fei1,2,LEI Jian-jun1,and CAO Bi-hao1(1College of Horticulture,South China Agricultural University,Guangzhou 510642,China;2Vegetable and Floriculture Scienti-fic Research Institute of Jilin Province,Changchun 130033,China)
关键词：
Brassica campestris L.ssp.chinensis(L.)Makino var.utilis Tsen et Lee;male sterility;defective in anther dehiscence1;jasmonic acid;antisense RNA
摘  要：
There are not F1 hybrids because of no good male sterile materials or self incompatible lines in Brassica campestris L.ssp chinensis(L.)Makino var.utilis Tsen et Lee.In this experiment,in accordance with the conservative region of DAD1 gene in Arabidopsis thaliana and B.campestris,the primer was designed to amplify the DAD1 gene conservative region fragment(DAD1F)of B.campestris L.ssp chinensis var.utilis.Using this fragment to construct a antisense expression vector to develop adjustable male sterile plants.All opend flowers were removed from the inflorescence,and the remaining flowers were dipped into 500 μmol·L-1 MeJA(Wako Pure chemical,Osaka,Japan)dissolved in 0.05% aqueous Tween 20.The fragment amplified from B.campestris L.ssp chinensis var.utilis was named as BrcpDAD1F.The homologous rates were 88% and 99% with that in Arabidopsis and B.campestris,respectively.The antisense DAD1F from B.campestris L.ssp chinensis var.utilis,was connected to plant expression vector pBI121.Using cotyledons with petiole as explants,Agrobacterium tumefaciens-mediate transformation was performed with antisense BrcpDAD1F vector(pBI-antiBrcpDAD1F)into B.campestris L.ssp chinensis var.utilis.The results of Southern bloting showed that antisense DAD1F had been transferred into B.campestris L.ssp chinensis var.utilis Tsen et Lee.Twelve transformants were obtained,of which 6 the mRNA of DAD1 were inhibited and displayed male sterile,pollen germination percent of transgenic plants were very low,which were not than 10%.After flowering,transgenic plants could not set or the siliquas were empty.Occasionally we could obtain seeds,but the seeds could not germinate.Pollinated with wild type plant pollen,the siliquas of transformed plants were enlarged and the seeds could be produced.No obvious differences were observed in morphology between transformed plants and wild type plants.A few flowers in transformed plants were abnormal.The pollen treated with 500 μmol·L-1MeJA could germinate in the medium and could fertilize.The fertile and sterile plants were in the ratio of 3 to 1 in T1 population,3 to 1 in some lines,full fertile in some lines full sterile in other lines in T2 populations.It suggested that antisense RNA can inherit in Mendel model.

相似文章

导出引用
计量
文章访问数: 15
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46