CHEN Zi-qi,LI Xia,WANG Pi-wu,et al.Using RNA Interference Technology to Improve Soybean Fat Content[J].Soybean Science,2012,31(04):529-533.[doi:10.3969/j.issn.1000-9841.2012.04.004]
利用RNA干扰技术提高大豆脂肪含量
- Title:
- Using RNA Interference Technology to Improve Soybean Fat Content
- 文章编号:
- 1000-9841（2012）04-0529-05
- Keywords:
- Soybean; GY1 gene; RNAi; Genetic transformation
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 利用冻融法将大豆11S球蛋白GY1基因RNA干扰表达载体转入农杆菌EH105中，以大豆“吉农28”为受体，通过农杆菌介导的大豆子叶节转化法导入大豆，获得T1代转基因苗12株，并对得到的转基因植株进行分子生物学鉴定。PCR和Southern杂交检测表明，RNAi植物表达载体p3301-Gy1已成功插入到转基因大豆植株的基因组DNA中；RT-PCR和SDS-PAGE检测结果表明RNA干扰在转录后水平发挥了作用，11S球蛋白表达含量降低；利用BUCHI N-500型近红外谷物分析仪对转化的大豆蛋白质和脂肪含量进行检测，结果转化植株的籽粒蛋白质含量（36.07%）平均降低1.43个百分点，脂肪含量（21.28%）平均提高0.76个百分点。因此，利用RNA干扰技术提高大豆脂肪含量是可行的。
- Abstract:
- Protein was negatively correlated with fat content in soybean.We aimed to improve fat content by reducing protein content through RNA interference in this research.The RNA interference expression vector-p3301-GY1 of 11S globulin GY1 gene was transformed into cotyledon nodes of soybean Jinong 28 by Agrobacterium-mediated method.We got 12 positive transgenic plants in T1generation by PCR detection.Southern blot confirmed the RNAi expression vector of GY1 gene had been expressed in soybean genome.RT-PCR and SDS-PAGE further indicated the expression level of GY1 gene were decreased.Seed detection showed the protein content of transformed plants decreased from 37.50% to 36.07%,and fat content increased from 20.52% to 21.28%.Results suggest it is possible to improve fat content of soybean through RNA interference technology.
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备注/Memo
第一作者简介：陈子奇（1986-），男，在读硕士，研究方向为生物技术在作物遗传育种中的应用研究。E-mail：imchenziqi@yahoo.cn。