| 根癌农杆菌介导的fad2 RNA干扰体基因转化甘蓝型油菜的研究 |
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| 引用本文： | 梁会娟,曹刚强,苗利娟,田保明,翁海波,黄冰艳,陈占宽.根癌农杆菌介导的fad2 RNA干扰体基因转化甘蓝型油菜的研究[J].河南农业科学,2007(1):34-38. |
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| 作者姓名： | 梁会娟 曹刚强 苗利娟 田保明 翁海波 黄冰艳 陈占宽 |
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| 作者单位： | 1. 郑州大学,生物工程系,河南,郑州,450001
2. 河南省农业科学院,生物技术研究所,河南,郑州,450002 |
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| 摘 要： | Δ12-油酸去饱和酶(Delta-12 oleate desaturase FAD2)是植物产生多聚不饱和脂肪酸的关键酶。对fad2基因表达进行抑制,可提高油菜种子油酸的相对含量。依据hpRNA介导的RNAi技术的原理,构建了以甘蓝型油菜fad2基因为靶标的RNAi植物表达载体pCAMBIA3301-fad2i。以甘蓝型油菜Y14-1的带柄子叶作为转化受体,利用根癌农杆菌介导法,将fad2RNA干扰体基因导入甘蓝型油菜中,获得了PPT抗性植株。同时还探讨了影响Y14-1品种转化频率的几个要素,为甘蓝型油菜的高效转化提供参考依据。对获得的再生植株进行GUS组织化学染色检测和PCR检测,证明外源基因已整合到油菜基因组中。
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| 关 键 词： | 甘蓝型油菜 根癌农杆菌 fad2 RNAi 转化频率 |
| 文章编号： | 1004-3268(2007)01-0034-05 |
| 修稿时间： | 2006年6月2日 |
The fad2 Gene Transformation of Brassica napus Mediated by Agrobacterium |
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| LIANG Hui-juan,CAO Gang-qiang,MIAO Li-juan,TIAN Bao-ming,WENG Hai-bo,HUANG Bing-yan,CHEN Zhan-kuan.The fad2 Gene Transformation of Brassica napus Mediated by Agrobacterium[J].Journal of Henan Agricultural Sciences,2007(1):34-38. |
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| Authors: | LIANG Hui-juan CAO Gang-qiang MIAO Li-juan TIAN Bao-ming WENG Hai-bo HUANG Bing-yan CHEN Zhan-kuan |
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| Abstract: | Delta-12 oleate desaturase gene(fad2)is a rate-limiting gene for multi-unsaturated fatty acid synthesis in rapeseed(Brassica napus).Blocking the expression of fad2 gene could increase the comparative content of the linolic acid in rapeseed.The authors constructed the RNAi expression vector-pCAMBIA3301-fad2i targeting the gene fad2 by RNAi strategy.Using the cotyledons with petiole of rapeseed cultivar Y141 as material,the gene fad2 RNAi constructs was introduced into rapeseed by.A.tumefaciens-mediated method.Some PPT-resistant green shoots were obtained.At the same time,some factors which affected the transformation of cotyledons with petiole were studied.The results of GUS assay and PCR amplification indicated that the foreign gene was stably integrated and expressed in the genome of rapeseed. |
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| Keywords: | Brassica napus L A tumefaciens fad2 RNAi Trans-frequency |
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