利用注射叶片法快速鉴定大豆GmCRY1和GmCRY2基因功能  

Rapid Identification of GmCRY1 and GmCRY2 Function by Leaf Injection in Soybean

作　　者：秦正睿[1,2] 孙磊[1,2] 李宏宇[2] 

机构地区：[1]吉林大学植物科学学院,吉林长春130062 [2]中国农业科学院作物科学研究所,北京100081

出　　处：《大豆科学》2012年第1期13-16,共4页Soybean Science

基　　金：转基因专项资助项目(2009ZX08004-010B;2008ZX08010-004);国家高技术研究发展计划资助项目(2006AA10Z107)

摘　　要：将大豆隐花色素基因GmCRY1、GmCRY2构建在N端含有绿色荧光蛋白(GFP)的pEGAD载体上,并转化农杆菌EHA105菌株。将含有上述基因的EHA105菌液分别注射到大豆单叶及复叶叶片中,观察长日条件下(16 h光照/8 h黑暗)和短日条件下(8 h光照/16 h黑暗)注射植株的开花时间。结果叶片注射含GmCRY1基因菌液的植株在长日条件下早开花,通过RT-PCR检测表明注射叶片中GmCRY1成功表达,而叶片注射含GmCRY2基因菌液的植株虽然也通过RT-PCR检测到基因表达,但无论长日和短日条件下开花时间均无改变。结果证实注射叶片法可快速鉴定GmCRY1和GmCRY2基因对大豆开花时间的影响。Soybean（Glycine max）Cryptochrome 1（GmCRY1）and Cryptochrome 2（GmCRY2）genes were constructed into expression vector pEGAD,and injected into soybean leaves by Agrobacterium tumefaciens EHA105 mediated transformation.The Agrobacterium tumefaciens EHA105 with vector pEGAD was used as control.The RT-PCR method was carried on to detect if the GmCRY1 and GmCRY2 were transient expressed in the leaves.The transformed plants were grown in different photoperiod light conditions（SD：8 h light/16 h darkness;LD：16 h light/ 8 h darkness）and the flowering data was calculated to conclude the effect of transient expression of GmCRY1 and GmCRY2 on the flowering time in soybean.The results indicated that the transient expressed GmCRY1 in soybean flowered earlier in LD,and the GmCRY1 expression was detected in the injected leaves through RT-PCR analysis.Although the expression of GmCRY2 was detected,it had no effect on flowering time in both LD and SD conditions.This result confirmed that the leaves injection method can quickly identify the function of GmCRY1 and GmCRY2 genes on soybean flowering time.

关 键 词：大豆 隐花色素 根癌农杆菌 开花时间 

分 类 号：S565.1[农业科学—作物学]