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  

Modification of pFGC5941 Construct and Transfer of Double Antisense Genes of NAC1 and SIP1into Arabidopsis thaliana

作　　者：梁鹏[1] 孔祥翔[2] 王春涛[2] 查向东[1] 胡向阳[2] 

机构地区：[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

出　　处：《植物分类与资源学报》2012年第4期403-408,共6页Plant Diversity

基　　金：The Major Science and Technology Program(110201101003-TS-03;2011YN02和2011YN03)

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pF-GC5941S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-1NaCl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。SIP1 encodes the protein which show interact with SOS2, the protein involving in plant responding to sa- line 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 NACl-sense and NACl-anti were inserted into pFGC5941S constructs, making the expression vectors pFGC5941S-SIP1-NACl-sense and pFGC5941S-SIP1-NACl-anti, respec- tively. Fifteen transgenic A. thaliana harboring these two constructs （pFGC5941S-SIPI-NACl-sense and pF- GC5941 S-SIP1-NAC 1-anti ） were then generated via Agrobacterium tumefaciens-mediated transformation. The pheno- types of homozygous transformants which were grown on MS medium with 75 mmol. L-~ NaC1 showed that compared with wild-type A. thaliana, pFGC5941S-SIP1-NACI-sense transgenic plants exhibited longer main root and increas- ing amounts of lateral roots, while no obvious differences were observed in pFGC5941S-SIP1-NACl-anti transgenic plants. These resuhs indicated that the development of A. thaliana lateral roots under salt stress was specifically pro- rooted by both overexpression of SIP1 gene and NAC1 gene.

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 

分 类 号：Q943.2[生物学—植物学]