利用VIGS技术沉默GhBES1基因对棉花幼苗生理指标的影响  

The Effects of Silenced Gh BES1 Gene on Physiological Parameters of Cotton Seedlings by VIGS Technique

作　　者：安汶铠 常丹[1] 杨艺[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《分子植物育种》2016年第3期598-603,共6页Molecular Plant Breeding

基　　金：国家自然科学基金-新疆联合基金重点项目(U1303282)资助

摘　　要：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。Silenci ng gene is helpful to study the function of gene in the adversity stress.A length of 650 bp of silencing gene fragment was determined by analyzing the silent sites of Gh BES1 gene and designed the specific primer,specifically obtained the 650 bp gene fragment from cotton‘XLZ17'(Gossypium hirsutum) by using PCR cloned technique.After Kpn Ⅰ and Xba Ⅰ digestion,the recombinant vector p TRV-Gh BES1 was obtained by connection with p TRV-RNA2 plasmid and transformed into Agrobacterium tumefaciens(GV3101).Then the Agrobacterium strain GV3101 infected the cotton seedlings with drought stress.The result showed that the proline and soluble sugar in cotton seedlings with silenced Gh BES1 gene dramatically decreased by 50.47% and 39.51%,respectively,compared with the p TRV-00.In contrast,the malondialdehyde(MDA) content increased by 1.84 times.Furthermore,the quantities of chlorophyll a,chlorophyll b,carotenoids,glycinebetaine and water content in silenced Gh BES1 seedlings also declined to some extent.To conclude,the physiology and biochemistry of cotton seedlings that was treated with drought stress expressed further descreased after Gh BES1 gene was silenced by using VIGS technique,the study indicated that Gh BES1 gene probably had a closed relationship with drought resistance of cotton seedlings.

关 键 词：棉花 干旱胁迫 VIGS 生理生化检测 基因表达 

分 类 号：S562[农业科学—作物学]