中国农业科学2009，Vol.42Issue(11)：3804-3811,8.DOI:10.3864/j.issn.0578-1752.2009.11.006
应用RNA干扰技术创造低脂肪氧化酶活性大豆新种质
Development of Novel Soybean Germplasms with Low Activity of Lipoxygenases by RNAi Method
摘要
Abstract
[Objective] The objective of this study was try to find a new avenue of breeding technique in soybean to improve the quality of soybeans and create new superordinary germplasm resource of soybeans by RNAi, which prohibits the gene expression of iipoxygenase in soybean seed. [Method] The consensus sequence of lipoxygenase gene (Lx1, Lx2, Lx3) in soybean seeds was cloned by RT-PCR, the length of the sequence was 357 bp. An RNAi expression vector of soybean lipoxygenase gene was reconstructed by recombinant PCR, and transformed the vector in soybean (Jinong 18) by pollen tube pathway. [Result] Twelve transgenic strains were obtained and 10 of which demonstrated significant differences. SDS-PAGE and activity of iipoxygenase demonstrated that the iipoxygenase activity of transgenic soybeans decreased significantly, the degree of decreasing was 64.2% on average. The near-infrared analyses demonstrated that the fat content of the transgenic soybeans increased by 0.8%-1.5% and 1.2%-3.0% the total protein decreased by 1.2%-3.0% compared with the control group. The results of RT-PCR demonstrated that the mRNA of lipoxygenase in transgenic soybeans was significantly inhibited. In addition, the activity of lipoxygenase decreased significantly in the progenies of transgenic soybean seeds and the seeds had more fat. [Conclusion] The RNAi technology can effectively inhibit the expression of lipoxygenase gene in soybean seed, decrease the activity of lipoxygenase, increase the amount of oil content in its products, improve the quality of soybeans, and create new superordinary germplasm resources of soybean.关键词
大豆/种质资源/脂肪氧化酶/RNA干扰/重组PCRKey words
soybean/germplasm resource/iipoxygenase/RNAi/recombinant PCR分类
农业科技引用本文复制引用
马建,张君,曲静,王云鹏,魏益凡,王丕武..应用RNA干扰技术创造低脂肪氧化酶活性大豆新种质[J].中国农业科学,2009,42(11):3804-3811,8.基金项目
教育部博士点基金(20070193005) （20070193005）
