摘要
为研究液泡H+-ATPase c亚基VHA-c4和VHA-c5基因在植物生长发育过程中的作用,本研究构建了拟南芥VHA-c4和VHA-c5过表达载体并转化野生型拟南芥,分别获得9个和7个T2代转基因纯合体株系。采用半定量RT-PCR方法对过表达VHA-c4和VHA-c5的转基因纯合体进行阳性鉴定,发现其mRNA表达量均高于对照。对转基因纯合体进行暗培养和正常光照培养,结果显示,黑暗条件下,所有VHA-c4转基因株系的主根变短,而在正常光照下,所有VHA-c5转基因株系的主根变短,推测VHA-c4和VHA-c5分别在黑暗和光照条件下影响植物根的生长。用ABA和糖(葡萄糖和蔗糖)处理转基因纯合体,结果显示它们与野生型的表型无明显差异,表明VHA-c4和VHA-c5基因的过表达没有影响拟南芥对ABA和糖的响应。
To study the role of vacuole H~＋-ATPase subunit c genes on plant growth and development,over-expression vectors of VHA-c4 and VHA-c5 from Arabidopsis thaliana were constructed and introduced into wild-type A. thaliana,and 9 and 7 homozygous T2 generations of VHA-c4 and VHA-c5 overexpressed transgenic lines were obtained,respectively. Semi-quantitative RT-PCR methods were used for identification of the positive VHA-c4 and VHA-c5 transgenic homozygotes,and their corresponding transcript levels were increased in every transgenic line. When cultivated under different light conditions,the VHA-c4 transgenic plants exhibited a reduction in primary root length compared with that of the wild-type when cultivated in the dark,whereas the VHA-c5 transgenic plants showed reduced primary root length under normal light conditions. These results indicated that VHA-c4 and VHA-c5 affected the growth of roots during light / dark cycles. Following ABA and sugar（glucose and sucrose） treatment,no significant differences were found between the transgenic homozygote phenotype and wildtype,indicating that VHA-c4 and VHA-c5 overexpression did not affect the response of Arabidopsis to ABA or sugar.
作者
苏杰
郭荣起
李国婧
王瑞刚
SU Jie GUO Rong-Qi LI Guo-Jing WANG Rui-Gang(Vocational and Technical College of Inner Mongolia Agricultural University, Tuyouqi, Inner Mongolia 014109, China College of Life Sciences Inner Mongolia Agricultural University, Hohhot 010018, China Agricultural Sciences of Hulunbeier, Yakeshi, Inner Mongolia 162650, China)
出处
《植物科学学报》
CAS
CSCD
北大核心
2016年第5期765-774,共10页
Plant Science Journal
基金
国家自然科学基金项目(30460018)
内蒙古自治区科技创新团队项目(201503004)~~
