紫花苜蓿MsLEA4-4基因的克

发布时间：2018-01-25 16:50

本文关键词： 紫花苜蓿胚胎晚期富集蛋白(LEA) 逆境胁迫遗传转化　出处：《草业学报》2017年12期 　论文类型：期刊论文

【摘要】：胚胎晚期富集蛋白(LEA)广泛参与植物对多种逆境胁迫的反应。本研究利用同源克隆的方法,从紫花苜蓿中克隆了一个LEA4类基因的开放阅读框(ORF),命名为Ms LEA4-4。该基因编码512个氨基酸,结构分析显示MsLEA4-4包含5个重复的由11个氨基酸TAQAAKEKTQQ组成的序列特征。利用实时荧光定量PCR检测了Ms LEA4-4在不同逆境下的表达量,结果显示,该基因受干旱、NaCl、Cu~(2+)、Zn~(2+)和外源ABA诱导表达上调,其中Na Cl胁迫2h、Cu2+和Zn2+胁迫8 h,Ms LEA4-4基因表达量最高;冷胁迫和干旱胁迫下,该基因的表达量随处理时间的延长呈逐渐上升趋势,表明该基因可能参与了紫花苜蓿的抗逆性调控。构建植物超表达载体p CAMBIA3301-Ms LEA4-4,采用农杆菌介导法侵染拟南芥花序,通过草铵膦(PPT)筛选和分子检测,7株抗性苗呈阳性,表明目的基因已成功导入拟南芥基因组中。本研究为进一步探索Ms LEA4-4基因在紫花苜蓿抗逆性调控中的作用奠定了基础。
[Abstract]:The late embryo enriched protein Lea was widely involved in the response of plants to various stresses. The method of homologous cloning was used in this study. An open reading frame of LEA4 gene was cloned from alfalfa, named MS LEA4-4. the gene encodes 512 amino acids. Structural analysis showed that MsLEA4-4 contained five repetitive sequence features consisting of 11 amino acid TAQAAKEKTQQ. Ms was detected by real-time fluorescence quantitative PCR. The expression of LEA4-4 under different stress. The results showed that the expression of the gene was up-regulated by drought induced ABA and NaCl stress for 2 hours. The expression of Ms LEA4-4 gene was the highest under the stress of Cu2 and Zn2 for 8 h. Under cold stress and drought stress, the expression of the gene increased with the prolongation of treatment time. It was suggested that the gene might be involved in the regulation of resistance to stress in alfalfa. The plant overexpression vector p CAMBIA3301-Ms LEA4-4 was constructed and infected with Arabidopsis thaliana inflorescence by Agrobacterium tumefaciens. Seven resistant seedlings were tested positive by PPTs screening and molecular detection. The results indicated that the target gene had been successfully introduced into Arabidopsis thaliana genome. This study laid a foundation for further exploring the role of Ms LEA4-4 gene in regulation of resistance to stress in alfalfa.
【作者单位】：山西省农业科学院畜牧兽医研究所;中国农业科学院北京畜牧兽医研究所;北京绿京华园林工程有限公司;山西农业大学动物科技学院;
【基金】：现代农业产业技术体系专项资金(CARS-35-25) 物种资源保护费(2130135) 山西省农业科学院畜牧兽医研究所项目(xms201506)资助
【分类号】：S541.9
【正文快照】： 由于植物的不可移动性,在生长发育过程中往往会遭受干旱、高温、低温、高盐等各种非生物胁迫,这些胁迫都会引起植物内部的水分缺失。缺水会引起植物体内的呼吸作用和光合作用等一系列生理活动失调[1]。在长期的进化过程中,植物演化出一系列从细胞到生理水平的应答反应,如基因

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