摘要
为了研究植物液泡H+-ATPase c亚基VHA-c4基因在拟南芥响应非生物胁迫中的作用,本研究构建了拟南芥VHA-c4基因沉默载体,利用农杆菌介导,将线性化的沉默载体dsDNA片段整合在拟南芥基因组上,筛选获得了能够不同程度特异沉默VHA-c4基因的7个转基因株系c4-1^c4-7,并对VHA-c4基因沉默效果最好的株系c4-2进行NaCl、ABA、6%葡萄糖胁迫处理。结果显示:在特定浓度梯度的NaCl处理后,沉默株系c4-2的主根相对伸长量和种子萌发率都被明显抑制,并且被抑制效果远远高于野生型株系。然而,在特定浓度梯度的ABA和葡糖糖处理后,沉默株系c4-2的主根相对伸长量和种子的萌发率虽然也都有一定程度的抑制,但是其被抑制效果远远低于野生型拟南芥。结果显示沉默了VHA-c4基因后,显著降低了拟南芥对NaCl的耐受性,影响了其的生长发育,然而对ABA和葡糖糖的抑制作用却表现为很不敏感,这可能是因为H^+-ATPase在盐胁迫信号通路和ABA信号通路中起着不同的(正负)调控功能。
In order to elucidate the roles of vacuole H+-ATPase subunit VHA-c4 genes against abiotic stress in Arabidopsis thaliana,in this study,the plasmid of VHA-c4 gene silence was constructed,and the designed double strand DNA fragments was integrated into the genome of Arabidopsis thaliana by Agrobacterium mediation,and then seven transgenic lines c4-1-c4-7 was screened to specifically silence the coding gene of VHA-c4 subunit.It was found that under the stress of NaCl,the relative root length and the germination rate of seeds of c4-2 line were inhibited to some extent and decreased more obviously than wild strain.Under the stress of ABA or glucose,the relative root length and the germination rate of seeds of c4-2 line were inhibited to some extent;however,they did not decreased obviously as wild strain.The results showed that silenced VHA-c4 gene significantly reduced the tolerance of Arabidopsis to NaCl and affected its growth and development.However,VHA-c4 gene silenced lines showed insensitivity to ABA and glucose.This may be due to the different(positive or negative)regulatory functions of H+-ATPase in salt stress signaling pathway and ABA signaling pathway.
作者
武春燕
苏杰
郭荣起
高阳
李国婧
王瑞刚
Wu Chunyan;Su Jie;Guo Rongqi;Gao Yang;Li Guojing;Wang Ruigang(Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology,Inner Mongolia Agricultural University,Hohhot,010018;Vocational and Technical College of Inner Mongolia Agricultural University,Tuyouqi,014109;Agricultural Sciences of Hulunbeier,Yakeshi,162650)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第4期1785-1790,共6页
Genomics and Applied Biology
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建设计划项目(201503004)共同资助。
关键词
液泡ATP酶
基因干扰技术
拟南芥
脱落酸
Vacuolar type H+-ATPase
RNA interference
Arabidopsis thaliana
Abscisic acid
