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作物学报 ›› 2011, Vol. 37 ›› Issue (04): 571-578.doi: 10.3724/SP.J.1006.2011.00571

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

GmCHS8GmIFS2基因共同决定大豆中异黄酮的积累

易金鑫1,徐照龙1,2,王峻峰4,张大勇1,何晓兰1,ZULFIQAR Ali1,3,朱虹润1,2,马鸿翔1,SANGEETA Dhaubhadel5   

  1. 1 江苏省农业科学院农业生物技术研究所,江苏南京 210014; 2 南京农业大学农学院,江苏南京 210095; 3 Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad 38040, Pakistan; 4 江苏省连云港职业技术学院,江苏连云港 222006; 5 Agriculture and Agri-Food Canada, London, ON N5V-4T3, Canada
  • 收稿日期:2010-08-30 修回日期:2011-01-06 出版日期:2011-04-12 网络出版日期:2011-02-24
  • 基金资助:

    本研究由国家自然科学基金项目(30971798),江苏省自然科学基金项目(BK2010474)和江苏省农业科技自主创新基金项目[CX(10)433]资助。

GmCHS8 and GmIFS2 Gene Co-Determine Accumulation of Isoflavonoid in Soybean

YI Jin-Xin1,XU Zhao-Long1,2,WANG Jun-Feng4,ZHANG Da-Yong1,HE Xiao-Lan1,ZULFIQAR Ali1,3,ZHU Hong-Run1,2,MA Hong-Xiang1,SANGEETA Dhaubhadel 5   

  1. 1 Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210094, China; 2 Nanjing Agricultural University, Nanjing 210095, China; 3 Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad 38040, Pakistan; 4 Liangyungang Technical College, Lianyungang 222006, China; 5 Agriculture and Agri-Food Canada, London, ON N5V-4T3, Canada
  • Received:2010-08-30 Revised:2011-01-06 Published:2011-04-12 Published online:2011-02-24

摘要: 大豆异黄酮受多基因控制,采用传统育种方法提高大豆异黄酮含量比较困难。我们之前的研究中表明,CHS8基因在异黄酮生物合成过程中发挥着重要作用,但CHS8基因过量表达并不能显著提高异黄酮含量。本研究利用Microarray技术,检测了高异黄酮品种RCAT Angra (RCAT)和低异黄酮品种Harovinton (HVNT)的18 362基因在种子发育过程中表达水平的变化以及大豆种子中异黄酮累积的趋势,利用RT-PCR证实CHS8IFS2分别是CHSsIFSs基因家族中的主要基因;证实CHS8是类苯基丙醇主路径中的主基因,并发现异黄酮支路中的IFS2基因在RCAT和HVNT品种中表达差异亦达到显著水平。进一步利用发根农杆菌转化系统,在大豆上分别过量表达CHS8IFS2CHS8+IFS2,前两者异黄酮含量较对照分别提高65.9%和34.4%,但增幅未达显著水平;而后者则提高了82.3%,增幅达显著水平(P<0.0001),因而证实大豆中异黄酮的积累由CHS8IFS2基因共同决定。

关键词: CHS8, IFS2, 过量表达, 共同决定, 大豆异黄酮

Abstract: Soybean isoflavonoids (isoFLVs) are natural secondary compondsproduced in Phenylpropanol metabolism Pathway. In past ten years, the isoFLVs were proved with anti-cancer activity to human health and therefore intensively included in nowadays food industry. However, the low content in natural soybean germplasm is a major limit to be weed in desired population. Herein we report that expression patterns of 18 362 genes were monitored by using Microarray between 30 to 70 days after pollination (DAP) during the soybean seed development. The expression profile of CHS8 and IFS2 revealed by both Microarray and RT-PCR indicated these two genes may play critical roles in determination of the accumulation of isoFLV in soybean seeds. According to Agrobacterium rhizogenes transformation system, over-expression of individuals of CHS8 and IFS2 resulted in 65.9% and 34.4% isoFLV increase respectively, but not significant in Duncan’s multiple test at 0.05 probability level. However, co-expression of CHS8+ IFS2 significantly increased (P<0.0001) the isoFLV content up to 82.3% when comparing to non-transgenic control. Conclusively, both IFS2 and CHS8 gene co-determine the accumulation of isoFLV in soybean.

Key words: CHS8, IFS2, Over-expression, Co-determination, Soybean isoflavonoid content

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