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小麦茉莉酸诱导蛋白基因TaJIP的克隆和功能鉴定 |
王飞1, 田雪军1, 赵月1, 臧新山1, 耿晓丽1, 倪中福1, 姚颖垠1, 胡兆荣1, 辛明明1, 张金波2, 丛花2, 孙其信1, 彭惠茹1
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(1.中国农业大学 农学与生物技术学院/农业生物技术国家重点实验室/杂种优势研究与利用教育部重点实验室, 北京 100193;2.新疆农业科学院 农作物品种资源研究所, 乌鲁木齐 830091) |
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摘要: |
为探讨高温影响小麦品质和产量的主要非生物胁迫因素,从小麦(Triticum aestivum)中克隆了1个受热胁迫诱导的茉莉酸诱导蛋白基因,命名为TaJIP,该蛋白具有Dirigent和Jacalin-like 2个保守结构域。通过生物信息学分析发现TaJIP基因的启动子区域包含多种激素响应元件。采用荧光实时定量PCR的方法,分析TaJIP在转录水平的表达模式,发现TaJIP不仅受茉莉酸甲酯(Methyl jasmonate,MeJA)的诱导,还受高温、干旱和高盐胁迫的诱导。通过农杆菌介导的方法将TaJIP超表达转化拟南芥并对其功能进行鉴定,发现TaJIP转基因株系高温胁迫后的成活率和下胚轴生长活力明显高于野生对照(Wild type,WT)。但是在含有甘露醇或NaCl的MS培养基中生长时,转基因株系的表型与WT基本没有差异。以上结果说明TaJIP基因可以提高植物对高温的适应性,但不能提高植物的耐盐和抗旱能力。 |
关键词: 小麦 高温 茉莉酸诱导蛋白 拟南芥 耐热性 |
DOI:10.11841/j.issn.1007-4333.2015.02.007 |
投稿时间:2014-04-22 |
基金项目:国家自然科学基金(31171549,30930058); 转基因重大专项(2011ZX08002-002); 国家"863"计划(2012AA10A309); 自治区高新技术项目(201311103) |
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Isolation and characterization of Jasmonate induced protein TaJIP in wheat (Triticum aestivum) |
WANG Fei1, TIAN Xue-jun1, ZHAO Yue1, ZANG Xin-shan1, GENG Xiao-li1, NI Zhong-fu1, YAO Ying-yin1, HU Zhao-rong1, XIN Ming-ming1, ZHANG Jin-bo2, CONG Hua2, SUN Qi-xin1, PENG Hui-ru1
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(1.College of Agronomy and Biotechnology/State Key Laboratory for Agrobiotechnology/Key Laboratory ofCrop Heterosis and Utilization (MOE), China Agricultural University, Beijing 100193, China;2.Institute of Crop Germplasm Resource, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China) |
Abstract: |
Climate change and higher temperature become more detrimental pressures on wheat production and quality.In this study, a Jasmonate induced gene (TaJIP) was cloned from wheat genotype Chinese Spring(CS).TaJIP was strongly induced by heat stress.TaJIP protein contained two conserved domains of Dirigent and Jacalin-like domain.In addition, the promoter region of TaJIP was cloned by insilico cloning method, several cis-elements involved in phytohormone response were found in this region.The expression pattern of TaJIP was analyzed by real-time PCR, and it was strongly induced by heat, drought, NaCl and Methyl jasmonate(MeJA).In order to investigate the biological function of TaJIP, the expression vector of TaJIP gene was constructed and transformed into Arabidopsis by Agrobacterium-mediation.The overexpressed Arabidopsis showed higher survival rate and longer hypocotyls than WT when heat stressed at 45 ℃.When cultivated on MS medium containing mannitol or NaCl, no difference was found between overexpressed plants and the wild type(WT).These results indicated that TaJIP can confer thermotolerance to plants, but can’t improve the resistance to drought or salt. |
Key words: wheat high temperature Jasmonate induced protein Arabidopsis thermotolerance |