苹果LRR-RLK基因家族鉴定和表达分析  

Genome-Wide Identification and Expression Pattern Analysis of LRR-RLK Gene Family in Apple

作　　者：黄金凤[1] 吕天星[1] 王寻 王颖达[1] 王冬梅[1] 闫忠业[1] 刘志[1] HUANG JinFeng;Lü TianXing;WANG Xu;WANG YingDa;WANG DongMei;YAN ZhongYe;LIU Zhi(Liaoning Institute of Pomology,Yingkou 115009,Liaoning;College of Horticultural Science and Engineering,Shangdong Agricultural University,Tai’an 271018,Shangdong)

机构地区：[1]辽宁省果树科学研究所,辽宁营口115009 [2]山东农业大学园艺科学与工程学院,山东泰安271018

出　　处：《中国农业科学》2021年第14期3097-3112,共16页Scientia Agricultura Sinica

基　　金：农业农村部园艺作物种质资源利用重点实验室开放基金(NYZS201905);辽宁省科学事业公益研究基金(20180022);辽宁省农业科学院学科建设项目(2020DD165004)。

摘　　要：【目的】从苹果全基因组中鉴定LRR-RLK家族蛋白成员,并进行生物信息学和表达模式分析,为研究苹果LRR-RLK的潜在功能提供理论基础。【方法】利用BLASTp基于GDR数据库鉴定苹果LRR-RLK家族成员,通过ExPASy Proteomics Server、Cell-PLoc、CD-Search Tool、MEGAX、MG2C等软件分析LRR-RLK蛋白序列基本信息、亚细胞定位情况、结构域组成、系统进化关系以及染色体定位情况。利用实时荧光定量PCR技术检测苹果12个LRR-RLK的组织表达和诱导表达特性。【结果】苹果LRR-RLK基因家族包含378个成员,这些LRR-RLK蛋白包括318—1 827个不等的氨基酸残基,等电点分布在5.16—9.75。亚细胞定位结果显示LRR-RLK蛋白均定位在细胞膜。系统进化分析可将其分为15类,各亚家族基因数量分布在1—111。染色体定位结果显示,LRR-RLK在苹果17条染色体上均有分布,其中第7条染色体数量最多,为40个。LRR-RLK家族基因具有2个特定的保守结构域,分别是LRR结构和蛋白激酶结构。蛋白二级结构以无规则卷曲为主,其次是α-螺旋,β-转角所占比例最小。通过定量检测发现筛选的12个家族成员在各组织中均有表达(除MD00G1105400外),且多数基因在茎中表达水平相对较高。低温条件下,7个基因上调表达,其中MD09G1153800上调最明显,最高为对照的6.8倍,而MD06G1170200和MD05G1061600均下调表达;在干旱条件下,8个基因上调表达,其中MD00G1105400上调最明显,最高为对照的9.6倍;在盐胁迫条件下,MD04G1150400、MD13G1108000和MD02G1071800始终处于上调表达状态,其中MD02G1071800上调最明显,最高为对照的14.9倍。苹果新梢接种轮纹病菌后,12个LRR-RLK家族基因表达基本上呈先上升后下降的趋势。并且在‘望山红’中,1 d时表达水平较高,而在‘鸡冠’中,3 d时表达水平较高。MD09G1153800和MD05G1065800在‘鸡冠’响应轮纹病菌侵染过程中显著上调表达,而在‘望山红’中无响应,【Objective】The study was carried out to explore the whole genome characteristics and expression patterns of LRR-RLKs in apple,to reveal the expression specificity of family members in different tissues and their responses to biological and abiotic stress,and further understand its biological function in apple.【Method】The members of LRR-RLK gene family in the whole genome of apple were identified based on the local BLAST database and Pfam database.The LRR-RLK amino acid sequence prediction,subcellular localization prediction,domain analysis,phylogenetic tree and chromosome localization were completed by software of ExPASy Proteomics Server,Cell-PLoc,CD-Search Tool,MEGAX and MG2C.In addition,the expression pattern of 12 LRR-RLK genes in different tissues and stress were analyzed by real-time fluorescent quantitative PCR(qRT-PCR).【Result】A total of 378 LRR-RLK genes were identified from apple genome.These LRR-RLK genes encoded proteins containing 318-1827 amino acid,and the theoretical isoelectric point ranged from 6.14 to 9.01.The prediction subcellular localization of apple LRR-RLK proteins was all distributed in the cell membrane.The gene family could be divided into 15 subgroups,containing 1-111 genes.The 378 genes in this family were distributed on all 17 chromosomes of the apple,and the chromosome 7 contained 40 genes.The LRR-RLK gene family had two conserved domains,namely the leucine-rich repeat structure and the protein kinase domain.Irregular curl andα-helix was the main secondary structure in the LRR-RLK gene family,and the rotation ofβ-turn was very small.It was found that the 12 selected family members were expressed in all tissues(except MD00G1105400),and most genes were expressed at relatively high levels in stem.Seven genes were up-regulated under low temperature conditions,and the expression of MD09G1153800 was the most obvious.The expression of MD09G1153800 was raised to 6.8 times of that under the control.While MD06G1170200 and MD05G1061600 were both down-regulated.Eight genes were u

关 键 词：苹果 LRR-RLK 基因家族 鉴定 表达分析 

分 类 号：S661.1[农业科学—果树学]