ZHU Hong-lin,SHA Ai-hua,FU Xiu-mei,et al.Cloning and Transformation Study of Transcription Factor GmLEC1 in Soybean[J].Soybean Science,2010,29(01):7-12.[doi:10.11861/j.issn.1000-9841.2010.01.0007]
转录调控基因GmLEC1转化大豆及转化方法的比较
- Title:
- Cloning and Transformation Study of Transcription Factor GmLEC1 in Soybean
- 文章编号:
- 1000-9841（2010）01-0007-06
- Keywords:
- Soybean; LEC1gene; RNA interference vector; Transformation; Cotyledon node; Embryonic tips
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 以高油大豆“中豆30”开花后30 d的发育种子为材料，根据已报道的拟南芥脂肪酸合成相关转录因子LEC1序列设计简并引物，采用同源序列法从大豆种子中分离了大小为850 bp的cDNA片段；采用Gateway技术构建RNAi表达载体pHGlec；分别利用大豆子叶节和胚尖为外植体进行农杆菌介导的遗传转化；以抗性芽获得率，PCR检测阳性率为指标，对转化系统进行优化。确定了6.0 mg·L-1的6-BA为胚尖转化法最佳激素配比；经PCR鉴定，优化后的胚尖转化法阳性苗获得率为7.58%，子叶节转化法转化率为1.86%。
- Abstract:
- The developing seeds of high-oil soybean“Zhongdou 30” were employed in this study.Degenerate primers were designed according to the reported sequence of transcription factor LEC1 related to fatty acid synthesis in Arabidopsis.A 850 bp cDNA fragment was isolated from soybean seed by homology-based candidate gene method.And RNAi vector pHGlec was constructed by Gateway technology.And then the vector was delivered into soybean via Agrobacterium-mediated transformation，using cotyledonary nodes and embryonic tips of soybean as explants.The rate of resistant shoots and the positive rate of PCR was used as criteria for selection the proper conditions.The results indicated that the optimal system of shoots elongation was 6.0 mg?L-1 of 6-BA for embryonic tips.The positive rate of PCR by optimized embryonic tips transformation system was 7.58%，and it was 1.86% by cotyledon nodes transformation system.
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备注/Memo
基金项目：中国农业科学院油料作物研究所资助项目；国家高技术研究发展计划资助项目(2006AA100104)。