陆地棉<em>GhSAD2</em>基因表达载体的构建、转化及抗寒性初步分析

doi:10.7668/hbnxb.2017.06.009

华北农学报 ›› 2017, Vol. 32 ›› Issue (6): 60-66. doi: 10.7668/hbnxb.2017.06.009

所属专题： 棉花；生物技术；

陆地棉GhSAD2基因表达载体的构建、转化及抗寒性初步分析

蔡曼^1,2, 李卫华², 柳延涛³, 余渝¹, 孔宪辉¹, 王娟¹, 王旭文¹, 刘丽¹

1. 新疆农垦科学院棉花研究所, 农业部西北内陆区棉花生物学与遗传育种重点实验室, 国家棉花改良中心新疆生产建设兵团分中心, 新疆石河子 832000;
2. 石河子大学, 新疆兵团绿洲生态农业重点实验室, 新疆石河子 832003;
3. 新疆农垦科学院作物研究所, 新疆石河子 832000

收稿日期:2017-09-10 出版日期:2017-12-28
通讯作者: 刘丽(1979-),女,江苏泗洪人,副研究员,博士,主要从事棉花遗传育种研究。
作者简介:蔡曼(1992-),女,陕西咸阳人,在读硕士,主要从事作物遗传育种研究。
基金资助:
国家自然科学基金项目（31360349）

Expression Vector Construction,Transformation and Cold Resistance of GhSAD2 Gene from Upland Cotton

CAI Man^1,2, LI Weihua², LIU Yantao³, YU Yu¹, KONG Xianhui¹, WANG Juan¹, WANG Xuwen¹, LIU Li¹

1. Cotton Institute, Xinjiang Academy Agricultural and Reclamation Science, Northwest Inland Region Key Laboratory of Cotton Biology and Genetic Breeding(Xinjiang), Ministry of Agriculture, Xinjiang Production & Construction Corps, Subcenter of National Cotton Improvement Center, Ministry of Agriculture, Shihezi 832000, China;
2. Shihezi University, The Key Oasis Eco-agriculture Laboratory of Xinjiang Production and Construction Group, Shihezi 832003, China;
3. Institute of Crop Research, Xinjiang Academy Agricultural and Reclamation Science, Shihezi 832000, China

Received:2017-09-10 Published:2017-12-28

摘要/Abstract

摘要： 为研究△9-硬脂酰-ACP脱氢酶基因的功能及其与植物抗寒性的相关性，以陆地棉GhSAD2基因（KX197920）为目的基因，构建植物表达载体pBI121-GhSAD2，将该表达载体通过电击法导入根癌农杆菌GV3101中，采用根癌农杆菌介导的叶盘法转化烟草。通过PCR和GUS活性染色鉴定外源GhSAD2基因是否导入并整合到烟草基因组上转录表达。将转pBI121-GhSAD2型和野生型烟草置于4℃下处理48 h后，检测其电解质渗漏率并观察其表型。研究结果证明目的基因GhSAD2已整合到烟草及棉花的基因组中，共获得转基因植株9株，转化率为75%。陆地棉GhSAD2基因能够显著增强非低温驯化植物烟草的抗寒性，从而提高植物细胞的低温防御能力，为进一步探究该基因的功能鉴定了基础。

关键词: 陆地棉, GhSAD2基因, 表达载体构建, 遗传转化

Abstract: In order to study a △9-stearoyl-ACP desaturase(GhSAD2)gene function and its correlation with the cold resistance of plants, GhSAD2 gene in Upland Cotton (KX197920) gene and construct the plant expression vector pBI121-GhSAD2,the expression vector by electroporation into Agrobacterium GV3101,tobacco leaf disc method by Agrobacterium transformation tumefaciens. Identification of exogenous GhSAD2 gene was transferred and integrated and integrated into the tobacco genome by PCR expression and GUS activity staining. The pBI121 -GhSAD2 type and wild type tobacco at 48 h treatment at 4℃,the detection rate of electrolyte leakage and to observe the phenotype. The results proved that the GhSAD2 gene had been integrated into the genome of tobacco and cotton,received a total of 9 transgenic plants,the conversion rate of 75% in Upland Cotton GhSAD2 gene could significantly enhance the cold resistance and cold acclimation non tobacco,so as to improve the low temperature the defense ability of plant cells,identification of the foundation for further research of the gene function.

Key words: Gossypium hirsutum L., GhSAD2 gene, Expression vector construction, Genetic transformation

中图分类号:

Q78
S562.03

蔡曼, 李卫华, 柳延涛, 余渝, 孔宪辉, 王娟, 王旭文, 刘丽. 陆地棉GhSAD2基因表达载体的构建、转化及抗寒性初步分析[J]. 华北农学报, 2017, 32(6): 60-66. doi: 10.7668/hbnxb.2017.06.009.

CAI Man, LI Weihua, LIU Yantao, YU Yu, KONG Xianhui, WANG Juan, WANG Xuwen, LIU Li. Expression Vector Construction,Transformation and Cold Resistance of GhSAD2 Gene from Upland Cotton[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 60-66. doi: 10.7668/hbnxb.2017.06.009.

http://www.hbnxb.net/CN/Y2017/V32/I6/60

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陆地棉GhSAD2基因表达载体的构建、转化及抗寒性初步分析

Expression Vector Construction,Transformation and Cold Resistance of GhSAD2 Gene from Upland Cotton

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