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三种芹菜品种中Agvip1基因的分离与鉴定及其对非生物胁迫和金属离子胁迫的响应

Isolation and characterization of the Agvip1 gene and response to abiotic and metal ions stresses in three celery cultivars.

作者信息

Li Yan, Chen Yi-Yun, Wang Feng, Xu Zhi-Sheng, Jiang Qian, Xiong Ai-Sheng

机构信息

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China of Ministry of Agriculture, College of Horticulture, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, China.

出版信息

Mol Biol Rep. 2014 Sep;41(9):6003-11. doi: 10.1007/s11033-014-3478-x. Epub 2014 Jun 27.

Abstract

VIP1, a VirE2-interacting protein 1, specifically interacts with VirE2 and acts as a molecular adaptor in Agrobacterium-mediated genetic transformation. This protein is widely used in plant genetic engineering. In this study, we cloned the Agvip1 gene that encodes the AgVIP1 protein from three celery (Apium graveolens) cultivars, namely, "Liuhe Huangxinqin", "Jinnan Shiqin", and "Ventura". The sequence analysis indicated that the Agvip1 gene from the three celery cultivars contained 768 bp Open Reading Frame and encoded with 255 amino acid residues. The N-terminal of AgVIP1 contained RNA recognition motif superfamily, a conserved domain. The Agvip1 gene in three cultivars had very high homology. The phylogenetic tree of VIP1-like proteins was constructed among celery and other plant species, showing that VIP1-like proteins from Solanum lycopersicum and Solanum tuberosum in Solanaceae had the shortest evolutionary relationship with AgVIP1 from A. graveolens in Apiaceae. Quantitative real-time PCR demonstrated that the Agvip1 gene had tissue-specific expression, mainly in the celery root. The expression analysis showed that the Agvip1 gene was induced by abiotic stresses differently in three celery cultivars. In "Liuhe Huangxinqin", the Agvip1 gene was up-regulated under hot, cold stresses. In "Jinnan Shiqin", the Agvip1 gene was up-regulated obviously under cold, drought treatments. However, in "Ventura", the Agvip1 gene was up-regulated under salt stress. The Agvip1 was also induced after metal ions treatments in three celery cultivars. These findings will provide more information on the Agvip1 gene and AgVIP1 protein, and enhance the understanding of the Agvip1 gene regulatory mechanisms under abiotic and metal ions stresses in celery.

摘要

VIP1是一种与VirE2相互作用的蛋白1,它特异性地与VirE2相互作用,并在农杆菌介导的遗传转化中作为分子衔接子发挥作用。该蛋白广泛应用于植物基因工程。在本研究中,我们从三个芹菜(Apium graveolens)品种,即“六合黄心芹”、“津南实芹”和“文图拉”中克隆了编码AgVIP1蛋白的Agvip1基因。序列分析表明,来自这三个芹菜品种的Agvip1基因包含768 bp的开放阅读框,编码255个氨基酸残基。AgVIP1的N端包含RNA识别基序超家族,这是一个保守结构域。三个品种中的Agvip1基因具有非常高的同源性。构建了芹菜和其他植物物种中类VIP1蛋白的系统发育树,结果表明茄科的番茄(Solanum lycopersicum)和马铃薯(Solanum tuberosum)中的类VIP1蛋白与伞形科的芹菜中的AgVIP1进化关系最近。实时定量PCR表明,Agvip1基因具有组织特异性表达,主要在芹菜根中。表达分析表明,在三种芹菜品种中,Agvip1基因受非生物胁迫的诱导情况不同。在“六合黄心芹”中,Agvip1基因在高温、低温胁迫下上调。在“津南实芹”中,Agvip1基因在低温、干旱处理下明显上调。然而,在“文图拉”中,Agvip1基因在盐胁迫下上调。在三种芹菜品种中,金属离子处理后Agvip1也被诱导。这些发现将为Agvip1基因和AgVIP1蛋白提供更多信息,并增进对芹菜在非生物和金属离子胁迫下Agvip1基因调控机制的理解。

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