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Plant Diversity ›› 2012, Vol. 34 ›› Issue (4): 403-.DOI: 10.3724/SP.J.1143.2012.11179

• 研究论文 • 上一篇    下一篇

pFGC5941的改造及拟南芥NAC1和SIP1双基因反义表达载体的构建和遗传转化

梁鹏1，孔祥翔2，王春涛2，查向东1**，胡向阳2   

  1. 1 安徽大学生命科学学院，安徽 合肥230031；2 中国科学院昆明植物研究所，云南 昆明650201
  • 收稿日期:2011-12-28 出版日期:2012-08-25 发布日期:2012-04-19
  • 基金资助:

    The Major Science and Technology Program (110201101003TS03, 2011YN02和2011YN03)

Modification of pFGC5941 Construct and Transfer of Double Antisense Genes of NAC1 and SIP1 into Arabidopsis thaliana

LIANG  Peng-1, KONG  Xiang-Xiang-2, WANG  Chun-Tao-2, ZHA  Xiang-Dong-1, HU  Xiang-Yang-2   

  1. 1 Institute of Life Sciences, Anhui University, Hefei 230031, China; 2 Kunming Institute of Botany,
    Chinese Academy of Sciences, Kunming 650201, China
  • Received:2011-12-28 Online:2012-08-25 Published:2012-04-19
  • Supported by:

    The Major Science and Technology Program (110201101003TS03, 2011YN02和2011YN03)

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摘要：

拟南芥中的SIP1基因编码的蛋白与拟南芥盐胁迫应答中的关键蛋白SOS2存在互作关系，而NAC1为拟南芥中介导生长素信号促进其侧根发生的蛋白。本研究中我们将SIP1基因和NAC1正义基因以及SIP1基因和NAC1反义基因分别整合到一个经改造的具有2个35S启动子的可用于双基因表达的载体pFGC5941S中，构建了两个双基因表达载体pFGC5941SSIP1NAC1sense和pFGC5941SSIP1NAC1anti。并将这两个载体通过农杆菌介导的方法转化到野生型拟南芥中，共获得15株转基因植株。对这些转基因植株进行盐胁迫实验发现，在含75mmol·L-1 NaCl的MS培养基上，相比于野生型，pFGC5941SSIP1NAC1sense转基因植株主根增长，侧根数量明显增多，而pFGC5941SSIP1NAC1anti转基因植株长势与野生型苗相似。由此我们推测可能只有当SIP1和NAC1同时过表达时，才会促进盐胁迫下拟南芥侧根的发育。

关键词: NAC1基因, SIP1基因, 反义表达载体, 双基因表达载体, 拟南芥转化, 表型

Abstract:

SIP1 encodes the protein which show interact with SOS2, the protein involving in plant responding to saline stress while NAC1 encodes the protein which involves in auxin signal and promoting the development of lateral root of Arabidopsis thaliana. In present study SIP1 and NAC1-sense and NAC1-anti were inserted into pFGC5941S constructs, making the expression vectors pFGC5941S-SIP1-NAC1-sense and pFGC5941S-SIP1-NAC1-anti, respectively. Fifteen transgenic A.thaliana harboring these two constructs (pFGC5941S-SIP1-NAC1-sense and pFGC5941S-SIP1-NAC1-anti) were then generated via Agrobacterium tumefaciens-mediated transformation. The phenotypes of homozygous transformants which were grown on MS medium with 75mmol·L-1 NaCl showed that compared with wild-type A.thaliana, pFGC5941S-SIP1-NAC1-sense transgenic plants exhibited longer main root and increasing amounts of lateral roots, while no obvious differences were observed in pFGC5941S-SIP1-NAC1-anti transgenic plants. These results indicated that the development of A.thaliana lateral roots under salt stress was specifically promoted by both overexpression of SIP1 gene and NAC1 gene.

Key words: NAC1 gene, SIP1 gene, Antisense-expession vector, Double genes-expession vector, Arabidopsis thaliana transformation, Phenotypes

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