VHA-c2&c4双基因沉默株系拟南芥对NaCl与ABA的响应

doi:10.13560/j.cnki.biotech.bull.1985.2020-0098

生物技术通报 ›› 2020, Vol. 36 ›› Issue (7): 48-54.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0098

VHA-c2&c4双基因沉默株系拟南芥对NaCl与ABA的响应

苏杰^1,2, 郭荣起^2,3, 高阳², 于秀敏⁴, 李国婧², 王瑞刚²

1.内蒙古农业大学职业技术学院,包头 014109;
2.内蒙古农业大学内蒙古自治区植物逆境生理与分子生物学重点实验室,呼和浩特 010018;
3.呼伦贝尔市农业科学研究所,呼伦贝尔 162650;
4.中国农业科学院草原研究所,呼和浩特 010010

收稿日期:2020-02-02 出版日期:2020-07-26 发布日期:2020-07-28
作者简介:苏杰,女,硕士研究生,研究方向：植物生物化学与分子生物学;E-mail：sujie_1546@163.com
基金资助:
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（201503004）

Response to NaCl and ABA in Arabidopsis thaliana of the Double Silent Gene VHA-c2&c4

SU Jie^1,2, GUO Rong-qi^2,3, GAO Yang², YU Xiu-min⁴, LI Guo-jing², WANG Rui-gang²

1. Vocational and Technical College of Inner Mongolia Agricultural University,Baotou 014109;
2. Inner Mongolia Agricultural University/Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology,Hohhot 010018;
3. Agricultural Sciences of Hulunbeier,Hulunbeier 162650;
4. Institute of Grassland Research of CAAS,Hohhot 010010

Received:2020-02-02 Published:2020-07-26 Online:2020-07-28

摘要/Abstract

摘要： 研究液泡H⁺-ATPase c亚基基因（VHA-c2和VHA-c4）在植物生长发育及响应非生物胁迫过程中的功能,构建拟南芥VHA-c2&c4的 RNAi表达载体,通过浸花法转化野生型拟南芥。利用卡那霉素筛选纯合体株系,进行半定量PCR鉴定,并对其进行NaCl和ABA胁迫处理。获得7个T₂代双沉默基因纯合体株系,其VHA-c2和VHA-c4的mRNA表达水平均低于野生型。挑选沉默效率较高的3个株系进行NaCl和ABA处理,在NaCl处理下,它们的主根相对伸长量和种子萌发率均小于对照,表明VHA-c2&c4沉默株系对NaCl胁迫作用敏感。ABA处理下,子叶的展开程度和主根相对伸长量均比对照高,表明VHA-c2&c4沉默株系对ABA抑制作用不敏感。NMT试验表明ABA促进了双沉默基因纯合体株系的H⁺内流能力。推测VHA-c2和VHA-c4的转录水平影响了植物对盐胁迫和ABA介导的信号转导途径的响应。

关键词: 拟南芥, 液泡H⁺-ATPase, 基因干扰技术, NaCl, ABA

Abstract: For studying the functions of vacuole H⁺- ATPase subunit c gene（VHA-c2 and VHA-c4）in plant growth and response to abiotic stress,we constructed an RNAi expression vector of VHA-c2&c4 and then transfected it into wild-type Arabidopsis thaliana plants by the floral dip method. Transgenic homozygotes were screened and identified by kanamycin,the expression levels of VHA-c2&c4 were analyzed by semi-quantitative RT-PCR,and the treatments of NaCl and ABA on them were conducted. The results showed that 7 T₂lines of double silent gene homozygous were obtained,and their mRNA expression levels of the two genes were lower than the wild type. Three lines with high silencing efficiency were selected for NaCl and ABA treatment. The relative elongation of primary roots and seed germination rate were both lower than that of the control group under NaCl treatment,indicating that the lines of VHA-c2&c4 gene silencing were sensitive to NaCl stress. The cotyledon expansion and the relative elongation of the primary root were both higher than the control under ABA treatment,indicating that the lines of the VHA-c2&c4 gene silencing were not sensitive to ABA inhibition. The results of NMT experiment showed that ABA promoted the H⁺ inflow ability of double-silenced gene homozygote lines. We hypothesized that the transcriptional levels of the genes have an impact on plant responses to salt stress and ABA-mediated signaling pathways.

Key words: Arabidopsis thaliana, vacuolar-type H⁺-ATPase, RNA interference, NaCl, ABA

苏杰, 郭荣起, 高阳, 于秀敏, 李国婧, 王瑞刚. VHA-c2&c4双基因沉默株系拟南芥对NaCl与ABA的响应[J]. 生物技术通报, 2020, 36(7): 48-54.

SU Jie, GUO Rong-qi, GAO Yang, YU Xiu-min, LI Guo-jing, WANG Rui-gang. Response to NaCl and ABA in Arabidopsis thaliana of the Double Silent Gene VHA-c2&c4[J]. Biotechnology Bulletin, 2020, 36(7): 48-54.

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VHA-c2&c4双基因沉默株系拟南芥对NaCl与ABA的响应

Response to NaCl and ABA in Arabidopsis thaliana of the Double Silent Gene VHA-c2&c4

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