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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。
The soybean cryptochrome genes GmCRY1 and GmCRY2 were constructed on pEGAD vector containing green fluorescent protein (GFP) at the N-terminus and transformed into Agrobacterium tumefaciens strain EHA105. The EHA105 bacteria containing the above genes were injected into the leaves of single leaf and compound leaf respectively. The plants were observed under long-day conditions (16 h light / 8 h dark) and short-day conditions (8 h light / 16 h dark) Blossom time. Results The leaves of plants containing GmCRY1 gene were prematurely flowered under long-term conditions. The expression of GmCRY1 in the injected leaves was confirmed by RT-PCR. However, the plants infected with GmCRY2 were also detected by RT-PCR Expression, but there was no change in the flowering time under long-day and short-day conditions. The results confirmed that the injection of leaf method can quickly identify GmCRY1 and GmCRY2 gene on soybean flowering time.