论文部分内容阅读
沉默基因有助于研究基因在逆境胁迫下的功能。通过对棉花Gh BES1基因进行沉默位点的分析,确定Gh BES1基因沉默片段为650 bp,设计特异性引物,利用PCR克隆从‘新陆早17号’棉花中获得650 bp的基因片段。经KpnⅠ和XbaⅠ双酶切后与p TRV-RNA2质粒连接构建成重组载体p TRV-Gh BES1,转化农杆菌(GV3101)后,侵染干旱胁迫处理的棉花幼苗。结果表明,与p TRV-00相比,携带沉默Gh BES1基因的棉花幼苗的脯氨酸和可溶性糖明显降低,分别降低了50.47%和39.51%。丙二醛(MDA)含量增加了1.84倍。叶绿素a叶绿素b和类胡萝卜素、含水量和甜菜碱也都有一定的下降。研究说明利用VIGS技术沉默Gh BES1后,干旱胁迫处理的棉花幼苗的生理生化指标均表现为进一步下降,表明Gh BES1可能与棉花抗旱性有密切的相关性。
Silencing genes help to study the function of genes under stress. Based on the analysis of the silencing sites of Gh BES1 gene in cotton, the silence fragment of Gh BES1 gene was determined to be 650 bp. The specific primers were designed and the 650 bp fragment was obtained from ’Xinluzao 17’ cotton by PCR. The recombinant plasmid pTRV-Gh BES1 was ligated with pTRV-RNA2 plasmid after being digested with KpnⅠ and XbaⅠ, and then transformed into Agrobacterium (GV3101) and then infected with cotton seedlings under drought stress. The results showed that compared with pTRV-00, proline and soluble sugar of cotton seedlings carrying silenced Gh BES1 gene were significantly reduced by 50.47% and 39.51% respectively. Malondialdehyde (MDA) content increased by 1.84 times. Chlorophyll a chlorophyll b and carotenoids, water content and betaine also have a certain decline. The results showed that the physiological and biochemical indexes of cotton seedlings treated with drought stress showed a further decline after silencing Gh BES1 by VIGS, which indicated that Gh BES1 might be closely related to the drought resistance of cotton.