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发明公开
CN107988251A 植物不饱和脂肪酸含量相关蛋白GhSAD在抗寒性鉴定过程中的应用 无效 - 驳回

植物不饱和脂肪酸含量相关蛋白GhSAD在抗寒性鉴定过程中的应用
摘要：
本发明公开了植物不饱和脂肪酸含量相关蛋白GhSAD在抗寒性鉴定过程中的应用，以陆地棉GhSAD2基因为目的基因，构建植物表达载体PBI121-GhSAD2，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48h后，检测其电解质渗漏率并观察其表型。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。
摘要（英）：

The invention discloses application of a protein GhSAD relevant with the content of plant unsaturated fatty acid in an authentication process of the cold resistance. The application comprises the following steps: constructing a plant expression vector PBI121-GhSAD2 by utilizing a GhSAD2 gene of upland cotton as a target gene, introducing the plant expression vector PBI121-GhSAD2 into agrobacteriumtumefaciens GV3101 by virtue of an electroporation method, and converting tobaccos by virtue of an agrobacterium tumefaciens medicated leaf disc method; authenticating whether an exogenous GhSAD2 gene is introduced or integrated into genomes of the tobaccos for transcription and expression by virtue of PCR and GUS vital staining; and processing transgenic PBI121-GhSAD2 type tobaccos and wild tobaccos at 4 DEG C for 48 hours, detecting the electrolyte leakage rates, and observing the phenotypes. The target gene GhSAD2 is integrated into the genomes of the tobaccos and the cotton, 9 transgenicplants are obtained, and the conversion rate is 75%. The GhSAD2 gene of the upland cotton is capable of enhancing the low-temperature defense capability of plant cells, so that the foundation is laidfor the further exploration of the functions of the gene.

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