| 小麦TaPP2-A13基因的表达响应逆境胁迫并与SCF复合体接头蛋白TaSKP1相互作用 |
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| 引用本文： | 孟钰玉,魏春茹,范润侨,于秀梅,王逍冬,赵伟全,魏新燕,康振生,刘大群.小麦TaPP2-A13基因的表达响应逆境胁迫并与SCF复合体接头蛋白TaSKP1相互作用[J].作物学报,2021(2):224-236. |
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| 作者姓名： | 孟钰玉 魏春茹 范润侨 于秀梅 王逍冬 赵伟全 魏新燕 康振生 刘大群 |
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| 作者单位： | 河北农业大学生命科学学院/河北省植物生理与分子病理学重点实验室;河北省农作物病虫害生物防治技术创新中心;西北农林科技大学植物保护学院/旱区作物逆境生物学国家重点实验室 |
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| 基金项目： | 河北省高等学校科学技术研究项目(ZD2019086);河北省自然科学基金项目(C2020204050);国家自然科学基金项目(31301649);旱区作物逆境生物学国家重点实验室开放课题基金(CSBAAKF2018008)资助。 |
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| 摘 要： | 为探究韧皮部蛋白2(Phloem protein 2,PP2)基因在小麦(Triticum aestivum)响应逆境胁迫中的功能及作用机制,本文以小麦抗叶锈病近等基因系TcLr15为材料获得一个小麦韧皮部蛋白2基因TaPP2-A13。该基因编码一个由298个氨基酸组成的亲水性蛋白质,相对分子量为33.18 kD,等电点为6.36。其N端为F-box结构域,C端具有典型的PP2结构域,属于F-box/PP2(FBP)亚家族成员。TaPP2-A13与禾本科植物中PP2-A13蛋白的亲缘关系较近。表达模式分析表明,TaPP2-A13的表达受叶锈菌(Puccinia triticina)侵染的诱导,且感病组合中表达更强。用脱落酸(ABA)、水杨酸(SA)和甲基茉莉酸(MeJA)处理后,TaPP2-A13总体呈现上调表达趋势。利用聚乙二醇(PEG)和H2O2处理后,小麦TaPP2-A13显著下调表达,而NaCl处理后TaPP2-A13呈现先上调后下调的表达趋势。亚细胞定位结果表明TaPP2-A13在细胞核和细胞质均有分布。以构建的重组载体BD-TaPP2-A13为诱饵筛选酵母文库,获得11种可能与TaPP2-A13互作的蛋白,酵母双杂交(Yeast 2 Hybrid,Y2H)确定其中的5种蛋白TaPP2C5、TaSLY1、TaCHI、TaRbcS和TaSKP1分别与TaPP2-A13存在相互作用。进一步利用BiFC和Co-IP确认TaSKP1与TaPP2-A13两种蛋白质之间的相互作用关系,推测TaPP2-A13可能为SCF复合体成员。本研究结果为深入解析TaPP2-A13蛋白的功能,并探究其调控网络奠定了基础。
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| 关 键 词： | 韧皮部蛋白2 小麦叶锈病 非生物逆境 激素 表达模式 蛋白互作 |
TaPP2-A13 gene shows induced expression pattern in wheat responses to stresses and interacts with adaptor protein SKP1 from SCF complex |
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| MENG Yu-Yu,WEI Chun-Ru,FAN Run-Qiao,YU Xiu-Mei,WANG Xiao-Dong,ZHAO Wei-Quan,WEI Xin-Yan,KANG Zhen-Sheng,LIU Da-Qun.TaPP2-A13 gene shows induced expression pattern in wheat responses to stresses and interacts with adaptor protein SKP1 from SCF complex[J].Acta Agronomica Sinica,2021(2):224-236. |
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| Authors: | MENG Yu-Yu WEI Chun-Ru FAN Run-Qiao YU Xiu-Mei WANG Xiao-Dong ZHAO Wei-Quan WEI Xin-Yan KANG Zhen-Sheng LIU Da-Qun |
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| Institution: | (College of Life Sciences,Hebei Agricultural University/Hebei Key Laboratory of Plant Physiology and Molecular Pathology,Baoding 071001,Hebei,China;Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province,Baoding 071001,Hebei,China;College of Plant Protection,Northwest A&F University/State Key Laboratory of Crop Stress Biology for Arid Areas,Yangling 712100,Shaanxi,China) |
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| Abstract: | To explore the function and molecular mechanism of Phloem protein 2(PP2)gene in wheat(Triticum aestivum L.)response to stresses,a TaPP2-A13 putatively encoding a PP2 protein was obtained from TcLr15,a wheat near isogenic line against leaf rust pathogen,in the present study.The complete coding region of TaPP2-A13 encodes a hydrophilic polypeptide with molecular weight of 33.18 kD,and theoretical isoelectric point is 6.36.There is an F-box domain at N-terminal and a PP2 domain at C-terminal of the TaPP2-A13 protein sequence,which indicates that wheat TaPP2-A13 belongs to F-box/PP2(FBP)subfamily.Wheat TaPP2-A13 shared relatively higher sequence similarity with PP2-A13 from Gramineae.Quantitative real-time PCR(qRT-PCR)results indicated that TaPP2-A13 was induced by infection of leaf rust pathogen(Puccinia triticina),and showed stronger expression in susceptible combination than that in resistant one.An obvious up-regulation of TaPP2-A13 was observed after treatment with abscisic acid(ABA),salicylic acid(SA)and methyl jasmonate(MeJA)in wheat.TaPP2-A13 was significantly down-regulated after treatment with PEG and H2O2,while TaPP2-A13 striking increased first,then fell down after NaCl treatment in wheat.Subcellular localization result indicated that TaPP2-A13 distributed in both of the nucleus and cytoplasm.The recombinant vector BD-TaPP2-A13 was used as the bait to screen Yeast 2 Hybrid(Y2H)library,11 kinds of proteins were finally obtained.Further Y2H assays identified that TaPP2-A13 physically interacted with five kinds of proteins including TaPP2C5,TaSLY1,TaCHI,TaRbcS,and TaSKP1.BiFC and Co-IP results further confirmed that TaPP2-A13 interacted with TaSKP1,an adaptor protein from SKP1-Cullin-F-box(SCF)complex,which made us to speculate that TaPP2-A13 functions as a member of SCF complex by binding with TaSKP1.These findings laid some foundation for further analyzing the function of TaPP2-A13 and exploring its regulatory network. |
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| Keywords: | phloem protein 2 wheat leaf rust abiotic stresses hormone expression pattern protein interaction |
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