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小麦倒春寒响应基因TaJAZ6的克隆、表达模式及亚细胞定位分析OA北大核心CSTPCD

Cloning,Expression Pattern and Subcellular Localization Analysis of Late Spring Cold Response Gene TaJAZ6 in Wheat

中文摘要英文摘要

JAZ蛋白在植物生长发育和胁迫信号通路中具有关键作用.为探究JAZ蛋白在小麦倒春寒中的调控机制,从小麦幼穗中克隆了TaJAZ6基因,并对其分子特征、表达特性与亚细胞定位进行分析.结果表明,该基因CDS序列全长为549 bp,编码178个氨基酸,预测编码蛋白质的分子质量为18.376 ku,理论等电点为9.37,不稳定系数为62.44,属于不稳定蛋白.该基因编码蛋白质具有1个TIFY结构域和1个CCT_2结构域.系统进化树分析发现,该蛋白质与野生二粒小麦和乌拉尔图小麦TIFY 11b蛋白亲缘关系最近.TaJAZ6基因启动子区除含有CAAT-box、TA-TA-box等基础作用元件外,还含有激素类响应元件、光响应元件、低温响应元件、防御和应激反应元件等.实时荧光定量PCR分析发现,TaJAZ6基因在根、茎、叶和幼穗中均有表达,根系中表达量最高.TaJAZ6基因还受低温和茉莉酸甲酯(MeJA)的诱导表达,低温胁迫下,TaJAZ6在矮抗58(耐倒春寒)和郑麦366(倒春寒敏感)根、茎和叶中表达趋势相同,均显著升高;喷施300,350 μmol/L的MeJA之后再低温处理TaJAZ6在2个小麦品种中表达量均显著下降.TaJAZ6在低温胁迫后的幼穗中表达量出现相反的趋势,在矮抗58幼穗中表达量显著下降,在郑麦366幼穗中则显著上升,推测该基因可能负调控了小麦倒春寒胁迫防御反应.通过喷施MeJA显著降低了低温胁迫下2个小麦品种幼穗中TaJA26基因的相对表达量,同时提高了小麦的结实粒数.亚细胞定位试验显示,TaJAZ6蛋白定位于细胞核中.以上研究结果表明,TaJAZ6可能在小麦响应倒春寒胁迫过程中发挥重要作用.

JAZ protein plays a key role in plant growth and development and stress signaling pathways.In or-der to explore the regulatory mechanism of JAZ protein in wheat late spring cold,TaJAZ6 gene was cloned from young spike of wheat,and its molecular characteristics,expression characteristics and subcellular localization were analyzed.The results showed that the full-length CDS sequence of the gene was 549 bp,encoding 178 amino acids.The predicted molecular weight of the encoded protein was 18.376 ku,the theoretical isoelectric point was 9.37,and the instability coefficient was 62.44,so it was an unstable protein.The protein encoded by this gene had a TIFY domain and a CCT_2 domain.Phylogenetic tree analysis showed that the protein had the closest relationship with TIFY 11b proteins of Triticum dicoccoides and Triticum urartu.In addition to the basic response elements such as CAAT-box and TATA-box,the promoter region of TaJAZ6 gene also contained hormone response elements,light re-sponse elements,low temperature response elements,defense and stress response elements.Real-time Quantitative PCR analysis showed that TaJAZ6 gene was expressed in roots,stems,leaves and young panicles,with the highest expression in roots.The expression of TaJAZ6 gene was also induced by low temperature and methyl jasmonate(MeJA).Under low temperature stress,the expression trends of TaJAZ6 in roots,stems and leaves of Aikang 58(tolerant to late spring cold)and Zhengmai 366(sensitive to late spring cold)were the same trend,which signif-icantly increased.After spraying 300,350 μmol/L MeJA,the expression of TaJAZ6 in plants treated with low tem-perature decreased significantly in both wheat varieties.The expression level of TaJAZ6 in the young panicles after low temperature stress showed an opposite trend.The expression level of TaJAZ6 in the young panicles of Aikang 58 decreased significantly,and increased significantly in the young panicles of Zhengmai 366.It was speculated that the gene might negatively regulate the defense response of wheat to late spring cold stress.By spraying MeJA,the rela-tive expression of TaJAZ6 gene in young spikes of two wheat varieties under low temperature stress was significantly reduced,and the grain number of wheat was increased.Subcellular localization assay showed that TaJAZ6 protein was localized in the nucleus.The results above indicate that TaJAZ6 may play an important role in the response of wheat to late spring cold stress.

张自阳;周谦;王依;王志伟;朱启迪;茹振钢;刘明久

河南科技学院农学院,河南新乡 453003

农业科学

小麦TaJAZ6基因表达分析低温胁迫亚细胞定位

WheatTaJAZ6Expression analysisCold stressSubcellular localization

《华北农学报》 2024 (001)

27-36 / 10

河南省科技攻关项目(222102110114;222102110236;232102111093)

10.7668/hbnxb.20194434

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