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本氏烟PP2A亚基基因的表达模式和克隆

Expression profile and cloning of genes encoding PP2A subunits of Nicotiana benthamiana
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摘要 蛋白磷酸酶2A(protein phosphatase 2A,PP2A)由结构亚基A、催化亚基C和调节亚基B组成,控制生物体内多种生命代谢过程,但目前尚不清楚其在植物抗病中的作用。本研究利用拟南芥、普通烟草、番茄和水稻PP2A的亚基序列,比对分析获得本氏烟(Nicotiana benthamiana)中对应的亚基序列。利用实时定量RT-PCR技术,分析了6个亚基基因在辣椒疫霉侵染本氏烟阶段(灌根后8、24、72 h)以及2个结构亚基基因在本氏烟不同组织部位(根、茎、叶、花和种子)中的转录水平。结果发现,在病原物侵染时,多数基因均显著上调表达,Nb PP2Ac-4在侵染阶段的表达量显著高于其他基因;2个结构亚基基因(Nb PP2Aa-1和Nb PP2Aa-2)在茎、叶、花和种子中的表达量均显著高于在根部中的表达量。克隆获得本氏烟和辣椒中2个结构亚基基因的全长c DNA序列,它们的二级结构主要为α和η螺旋,含有保守的功能域,与其他植物和人的结构亚基具有很高的同源性。 Ser/Thr protein phosphatase 2 A(PP2 A),consisting of a catalytic subunit C,a scaffold subunit A,and a highly variable regulatory subunit B,plays important roles in cellular processes in organisms.However,its role in plant disease resistance is obscure.In the present study,the genes encoding PP2 A subunits of Nicotiana benthamiana were obtained by searching its genome database using Arabidopsis thaliana,common tobacco,tomato and rice PP2 A subunit sequences as queries.Real-time RT-PCR was performed to determine the expression pattern of six selected genes during an infection period of N.benthamiana by Phytophthora capsici(0,8,24,72 h post-inoculation),and of two PP2 A-A genes in different plant tissues(root,stem,leaf,flower and seed).It turned out that most of the genes demonstrated significant up-regulation during the infection stage relative to the un-infected control.The most significant up-regulation was observed for Nb PP2 Ac-4 during the infection course.PP2 A-A genes(Nb PP2 Aa-1 and Nb PP2 Aa-2) showed significantly higher expression level in stem,leaf,flower and seed than in root.These two PP2 A-A genes were cloned with full length from N.benthamiana and its relative bell pepper.Sequence analysis showed that α-and η-helices were the main components of their secondary structure.They possess conserved domains such as HEAT_2 and showed high similarity to the homologues of PP2 A-A in other plant species and human.This study provides essential experimental data for elucidating the roles of PP2 A-A subunit in plant resistance against diseases.
作者 陈孝仁 黄沈鑫 张烨 李艳朋 车通 纪兆林 CHEN Xiao-Ren, HUANG Shen-Xin, ZHANG Ye, LI Yan-Peng, CHE Tong, JI Zhao-Lin (College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, Jiangsu 225009, China)
出处 《植物生理学报》 CSCD 北大核心 2018年第6期1073-1084,共12页 Plant Physiology Communications
基金 国家自然科学基金(31671971) 扬州市自然科学基金(YZ2016121) 公益性行业(农业)科研专项(201303018) 2016年度扬州大学优秀青年骨干教师培养对象
关键词 蛋白磷酸酶2A 植物抗病性 转录水平 功能域 辣椒疫霉 protein phosphatase 2A plant disease resistance transcriptional level domain Phytophthora capsici
作者简介 通讯作者(xrchen@yzu.edu.cn)。
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