新疆野苹果NAC基因分析及抗腐烂病基因筛选  

Identification, classification and expression analysis of NAC transcriptional factor genes in Malus sieversii during canker disease response

作　　者：周童 李小双[1,3] 刘晓洁[1,4] 丁雨 文雪静 张道远 ZHOU Tong;LI Xiaoshuang;LIU Xiaojie;DING Yu;WEN Xuejing;ZHANG Daoyuan(Xinjiang Institute of Ecology and Geography,Chinese Academy of Sciences/State Key Laboratory of Desert and Oasis Ecology,Urumqi 830011,Xinjiang,China;College of Life Sciences,Shihezi University,Shihezi 832003,Xinjiang,China;Turpan Desert Botanical Garden,Chinese Academy of Sciences,Turpan 838008,Xinjiang,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院新疆生态与地理研究所·荒漠与绿洲生态国家重点实验室,乌鲁木齐830011 [2]石河子大学生命科学学院,新疆石河子832003 [3]中国科学院吐鲁番沙漠植物园,新疆吐鲁番838008 [4]中国科学院大学,北京100049

出　　处：《果树学报》2020年第8期1111-1121,共11页Journal of Fruit Science

基　　金：中国科学院西部之光项目(2016-QNXZ-B-17);国家重点研发计划(2016YFC0501505)。

摘　　要：【目的】探明新疆野苹果(Malus sieversii)中NAC基因资源,筛选潜在的抗腐烂病NAC基因。【方法】以模式植物拟南芥(Arabidopsis thaliana)的NAC蛋白为种子构建隐马可夫模型,从新疆野苹果转录组数据库中鉴定NAC转录因子,利用Protparam及qRT-PCR等手段对其理化性质、亚细胞定位、保守基序、系统进化及腐烂病菌侵染后的表达谱进行分析。【结果】从新疆野苹果中鉴定到165个NAC基因,根据蛋白系统进化关系分为12个亚组,且MsNACs在各亚组间分布不均。蛋白理化性质分析结果表明,MsNACs大多属酸性,且多为亲水性蛋白。保守基序分析结果表明,大多数MsNAC转录因子拥有由7个保守基序组成的完整NAC结构域,少数存在亚结构域的缺失。膜结合域分析结果表明,有18个MsNACs蛋白具有膜结合域,且大多定位于细胞核,少数定位于内质网、线粒体等其他细胞器。表达分析结果表明,34个MsNACs在腐烂病菌侵染后差异表达,可根据表达模式分为A、B、C三个亚簇,其中MsNAC073和MsNAC102表达变化剧烈,分别在5 dpi下调近6倍和上调近140倍。【结论】基于新疆野苹果转录组数据,首次对MsNAC家族基因进行鉴定、分类和表达量分析,为新疆野苹果抗病响应关键NAC基因的功能研究奠定了基础,也为其他物种中抗病基因的筛选提供了参考。【Objective】The ork aimed at identification and analysis of NAC gene family in order to explore the potential canker-resistance gene resources in Malus sieversii,the ancestor of domestic apple.【Methods】The sequences of AtNACs were downloaded from the PlantTFDB v4.0 database to establish Hidden Markov Model(HMM)for identifying the MsNACs from the full-length transcriptome database of M.sieversii.MEME suite.11.2 was used to analyze the conserved motif with the maximum search value(7).Phylogenetic tree of MsNACs and AtNACs was generated by MEGA 6.0 with neighbor-joining(NJ)method.The physicochemical properties,membrane-bound domain and subcellular localization of MsNACs were analyzed by Protparam,TMHMM and WoLF PSORT software,seperately.The qRT-PCR was used to validate the MsNACs gene expression patterns under the infection of canker pathogen(Cytospora mali).【Results】165 MsNACs of M.sieversii were obtained,which differed from each other in molecular weight(MW),theoretical isoelectric point(IP)and hydrophobicity.The maximum MW was 95.86 ku(859 aa)and the minimum was 6.77 ku(58 aa).These MsNACs were divided into 12 subgroups(Ⅰ-Ⅻ)according to the phylogenetic analysis,and AtNACs and MsNACs were both included in each subgroup.However,the proportion of MsNACs and AtNACs in subgroups were dramatically different.The 113 of 165 MsNACs had integral NAC domain and the others had one or more subdomains missed.Conserved motifs analysis showed that NAC domains of MsNACs were composed of 7 conserved motifs.We identified 5 subdomains A(composed of motif 1),B(composed of motif 5),C(composed of motif 4 and 3),D(composed of motif 2 and 7),E(composed of motif 6).In addition,membrane-bound domain analysis showed that 18 of 165 MsNACs had the membrane bound domain,which belonged to subgroupⅢ(1),Ⅶ(1),Ⅷ(6),Ⅸ(9)andⅩ(1).Subcellular location analysis indicated that most of the MsNACs were located in nucleus while the other MsNACs were located in different organelles such as chloroplast(MsNAC034),endoplasmic reticulum

关 键 词：新疆野苹果 NAC转录因子 苹果腐烂病 抗病基因 

分 类 号：S661.1[农业科学—果树学]