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十字花科植物CYP86MF基因同源序列的克隆与进化分析 被引量:4
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作者 王玲平 曹家树 +2 位作者 叶纨芝 向洵 周生茂 《遗传》 CAS CSCD 北大核心 2005年第3期395-402,共8页
为了从分子水平上阐明十字花科植物间的亲缘进化关系,给植物种质资源的创建提供理论依据,实验根据课题组已报道的CYP86MF基因编码的氨基酸保守区域设计特异引物,运用PCR技术分别从十字花科6个属11个物种中分离克隆到了CYP86MF基因的同... 为了从分子水平上阐明十字花科植物间的亲缘进化关系,给植物种质资源的创建提供理论依据,实验根据课题组已报道的CYP86MF基因编码的氨基酸保守区域设计特异引物,运用PCR技术分别从十字花科6个属11个物种中分离克隆到了CYP86MF基因的同源序列。经比较分析,结果表明:这些同源序列的相似性达80%以上,所推导的氨基酸序列相似性达70%以上,且两者种间差异分别为1.0%~5.7%和2.6%~7.3%,属间差异分别是5.6%~22.5%和7.3%~31.2%;由氨基酸序列构建的分子系统树可知,在亲缘进化关系上芸薹属与萝卜属较近,其他依次为蔊菜属、拟南芥属、荠菜属,而与诸葛菜属最远。因此,CYP86MF基因的核苷酸及其可能编码的氨基酸序列差异属间较种间大,它可用于属间的分类等级研究。 展开更多
关键词 十字花科植物 cyp86mf基因 克隆 进化分析
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反义CYP86MF基因植物表达载体的构建及其对白菜的转化 被引量:10
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作者 余小林 曹家树 《浙江大学学报(农业与生命科学版)》 CAS CSCD 北大核心 2004年第2期179-184,共6页
在获得一个与白菜核雄性不育相关的编码细胞色素P450(CYP450)的基因CYP86MF的基础上,采用PCR技术在该基因的内部第901个碱基至第1338个碱基之间设计引物,扩增出438bp序列作为反义基因片段.把该反义片段连接至pBI121双元载体质粒上得到Ca... 在获得一个与白菜核雄性不育相关的编码细胞色素P450(CYP450)的基因CYP86MF的基础上,采用PCR技术在该基因的内部第901个碱基至第1338个碱基之间设计引物,扩增出438bp序列作为反义基因片段.把该反义片段连接至pBI121双元载体质粒上得到CaMV35S组成型启动子的表达载体pBI35S-AMF,随后利用"三亲杂交"法将pBI35S-AMF质粒转入农杆菌LBA4404和EHA105两个菌株中.PCR扩增和酶切鉴定结果表明,所构建的反义CYP86MF基因植物表达载体pBI35S-AMF是正确的,并已成功导入了根癌农杆菌中.随后,按照已建立的遗传转化体系对'上海青'白菜进行了转化,并获得130了多株转化再生植株. 展开更多
关键词 CY86MF基因 植物表达载体 白菜 遗传转化 反义RNA技术 根癌农杆菌 核雄性不育
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Measurement of Some Physiological and Biochemical Indexes of the Antisense Fragment of CYP86MF Gene Transgenic Male Sterile Plantlets in Chinese Cabbage-Pak-Choi 被引量:8
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作者 YU Xiao-lin, CAO Jia-shu and DONG De-kun(Institute of Vegetable Science, Zhejiang University , Hangzhou 310029, P. R. China) 《Agricultural Sciences in China》 CAS CSCD 2003年第11期1239-1245,共7页
In order to study the mechanism of the transgenic male sterility induced by the antisense fragment of gene CYP86MF, some physiological and biochemical indexes were compared between the transgenic plantlets of Chinese ... In order to study the mechanism of the transgenic male sterility induced by the antisense fragment of gene CYP86MF, some physiological and biochemical indexes were compared between the transgenic plantlets of Chinese cabbage-pak-choi(Brassica campestris L. ssp. chinensis Makino)and their controls. Results showed that there was significant difference between content of the endogenous hormones in leaves and floral buds. GA3 contents in leaves of the transgenic plantlets of B. campestris L. ssp. chinensis Makino cv. Shanghai-qing and cv. Youqing were reduced by 31. 72 and 46. 88% respectively as compared with those of their controls, and GA3 contents in floral buds of them were reduced by 62. 92 and 80. 57 % respectively. In addition, ZT contents in leaves of cv. Shanghai-qing and cv. Youqing were reduced by 11. 81 and 181. 20% respectively as compared with those of their controls and those in floral buds were reduced by 105. 94 and 128.75 % respectively. But, ABA content in transgenic plantlets was significantly higher than that in non-transgenic plantlets. However, differences among respiratory rate and cyanide-resistant respiratory rate, the photosynthesic rate and the content of total flavones in floral buds were not significant. In this study, the results indicated that the gene CYP86MF might be one of the key functional genes to control fertility of Chinese cabbage. 展开更多
关键词 Brassica campestris L. ssp. chinensis Makino cyp86mf TRANSGENIC Endogenous hormones Respiratory rate Photosynthesic rate Content of total flavones
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