AtJAR1基因在拟南芥耐盐性中的功能分析  

Functional analysis of AtJAR1 gene in salt tolerance of Arabidopsis thaliana

作　　者：李丹丹 林蓉 李新国[3] 郑月萍 LI Dandan;LIN Rong;LI Xinguo;ZHENG Yueping(College of Advanced Agricultural Sciences,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;Institute of Crop Germplasm Resources,Shandong Academy of Agricultural Sciences,Ji’nan 250100,Shandong,China)

机构地区：[1]浙江农林大学现代农学院,浙江杭州311300 [2]浙江农林大学林业与生物技术学院,浙江杭州311300 [3]山东省农业科学院农作物种质资源研究所,山东济南250100

出　　处：《浙江农林大学学报》2022年第5期998-1009,共12页Journal of Zhejiang A&F University

基　　金：国家重点研发计划项目(2018YFD1000906-3);浙江省自然科学基金青年基金项目(Q21C020003)。

摘　　要：【目的】茉莉酰氨基酸结合物合成酶(jasmonoyl amino acid conjugate synthase,JAR1)可以催化茉莉酸(jasmonic acid,JA)形成茉莉酸的活性形式茉莉酸异亮氨基酸复合体(jasmonic acid-isoleucine,JA-Ile),从而激活JA信号途径。JA信号途径在介导植物盐胁迫的响应中发挥重要作用,因此,探究AtJAR1在植物耐盐性中的功能对于研究JA信号途径影响植物耐盐性的机制具有重要作用。【方法】运用CRISPR/Cas9基因编辑技术,创建了2个不同的拟南芥Arabidopsis thaliana AtJAR1基因突变体,并对这2个突变体进行地上部生物量的统计分析和JA信号标记基因的表达分析,以确定AtJAR1基因功能缺失。之后,观察分析不同浓度氯化钠和脱落酸(ABA)处理对jar1突变体的种子萌发和幼苗建成的影响,明确AtJAR1基因对拟南芥耐盐性的影响。最后,通过比较分析盐处理前后野生型和突变体的钾离子(K^(+))和钠离子(Na^(+))质量摩尔浓度,以及高亲和力K~+转运蛋白基因AtHAK5的表达变化情况,初步探究AtJAR1基因在拟南芥耐盐性中的功能。【结果】JA信号标记基因AtVSP1和AtVSP2的表达量大幅下调,表明AtJAR1基因功能丧失。与点突变产生的jar1-1突变体不同的是,这2个突变体表现为前3周生长加快,之后逐渐减缓并出现叶片萎蔫的表型。同时,AtJAR1突变可以缓解盐胁迫和ABA对种子萌发和根系生长产生的抑制作用。此外,盐胁迫下AtJAR1突变可以促进AtHAK5的表达和根系对K~+的吸收转运。【结论】JA信号途径可能通过与ABA交互作用影响AtHAK5的表达量,以调节植物根系对K+的吸收转运,进而改变细胞内K^(+)/Na^(+)平衡,最终影响植物耐盐性。[Objective] Jasmonal amino acid conjugate synthase(JAR1) can catalyze jasmonic acid(JA) to form jasmonic acid-soleucine(JA-Ile), an active form of jasmonic acid, and activate the JA signal pathway. JA signaling pathway plays an important role in mediating the response of plants to salt stress. This study aims to explore the function of AtJAR1 in plant salt tolerance, which plays an important role in studying the mechanism of JA signal pathway affecting plant salt tolerance. [Method] CRISPR/cas9 gene editing technology was used to create two different Arabidopsis thaliana AtJAR1 gene mutants. The aboveground biomass of the two mutants and the expression of JA signal marker gene were analyzed to determine the loss of AtJAR1 gene function. Then, the effects of different concentrations of NaCl and ABA treatment on seed germination and seedling establishment of jar1 mutants were observed and analyzed to clarify the effect of AtJAR1 gene on salt tolerance of A. thaliana. Finally, the role of AtJAR1 gene in salt tolerance of A. thaliana was investigated by comparing the content of potassium and sodium ions in wild type and mutants before and after salt treatment, as well as the expression of AtHAK5, a high affinity potassium ion transporter gene. [Result] The expression of JA signal marker genes AtVSP1 and AtVSP2 decreased significantly, indicating the loss of AtJAR1 gene function. Different from the jar1-1 mutant produced by point mutation, the growth of the two mutants accelerated in the first three weeks, then gradually slowed down and the leaves wilted. At the same time,AtJAR1 mutation can alleviate the inhibition of salt stress and ABA on seed germination and root growth. In addition, AtJAR1 mutation can promote the expression of AtHAK5 and the absorption and transport of potassium ions in roots under salt stress. [Conclusion] JA signaling pathway may affect the expression of AtHAK5 through interaction with ABA, so as to regulate the absorption and transport of Kby plant roots,change the intracellular K/Nabalance,

关 键 词：拟南芥 CRISPR/Cas9 茉莉酰氨基酸结合物合成酶 耐盐性 K^(+)/Na^(+)平衡 

分 类 号：S722.3[农业科学—林木遗传育种] Q781[农业科学—林学]