摘要
【目的】鉴定金鱼草细胞壁相关受体激酶(Wall-associated kinase,WAK)及类WAK蛋白(WAK-like kinases,WAKL)基因家族成员,并挖掘抗核盘菌的候选基因,为深入解析金鱼草抗核盘菌的WAK/WAKL分子调控机制提供理论参考。【方法】以拟南芥WAK/WAKL家族蛋白为参考序列,利用生物信息学方法从金鱼草基因组数据库中鉴定出WAK/WAKL基因家族成员,并对其理化性质、系统发育、基因结构、保守基序、顺式作用元件及共线性关系等进行预测分析,并通过转录组测序和实时荧光定量PCR对该家族基因在金鱼草抗感核盘菌材料中的表达模式进行分析。【结果】从金鱼草基因组中共鉴定出27个WAK/WAKL基因家族成员,其中WAK家族基因16个(AmWAK1~AmWAK16),WAKL家族基因11个(AmWAKL1~AmWAKL11),编码的氨基酸数量为615~1085个;理论等电点(pI)为4.94~8.69,绝大多数蛋白呈酸性;所有蛋白的总平均亲水性指数(GRAVY)值小于0,均为亲水性蛋白;大多数蛋白亚细胞定位于细胞质膜区域,少部分定位于细胞外、液泡、高尔基体和细胞核。27个WAK/WAKL基因家族成员不均匀地分布在金鱼草的8条染色体上,且与拟南芥WAK/WAKL家族基因存在7对共线性同源基因;基因启动子区域含有与防御和胁迫响应、茉莉酸甲酯响应及水杨酸响应等顺式作用元件。通过叶片离体接种核盘菌试验,筛选出抗性差异明显的易感病品种Am1和抗病品种Am6。有12个WAK/WAKL家族基因在金鱼草抗感材料中显著差异表达(P<0.05),有9个基因表达模式与转录组测序结果基本一致。这9个基因中,有5个基因(AmWAK7、AmWAKL2、AmWAKL8、AmWAK6和AmWAK13)在金鱼草的根、茎和叶组织中高表达,而剩余4个基因(AmWAK2、AmWAK8、AmWAK9和AmWAK12)在根、茎和叶中基本不表达。【结论】筛选出的27个金鱼草WAK/WAKL基因家族成员,具有相似的基因结构和蛋白功能结构域,其中AmWAK6、AmWAK7、AmWAK13、AmWAKL2和AmWAKL8为�
【Objective】To identify the gene family members of the cell wall-associated kinase(WAK)and WAK-like kinases(WAKL)of Antirrhinum majus L.,and explore the candidate genes against Sclerotinia sclerotiorum.This study provided a theoretical reference for further analysis of WAK/WAKL molecular regulatory mechanism of A.majus against S.sclerotiorum.【Method】Using Arabidopsis thaliana L.WAK/WAKL family proteins as reference sequences,the WAK/WAKL gene family members were identified from the A.majus genome database by bioinformatics method,and their physicochemical properties,phylogeny,gene structure,conserved mods,cis-acting elements and collinear relationships were predicted.Transcriptome sequencing and real-time fluorescence quantitative PCR were used to analyze the expres-sion patterns of the family genes involved in S.sclerotiorum resistance.【Result】A total of 27 WAK/WAKL gene family members were identified from the A.majus genome,including 16 WAK genes(AmWAK1-AmWAK16)and 11 WAKL genes(AmWAKL1-AmWAKL11),encoding 615-1085 amino acids.The theoretical isoelectric point(pI)ranged from 4.94 to 8.69,and the most proteins were acidic.The total mean hydrophilic index(GRAVY)of all proteins was less than 0,indi-cating that all proteins are hydrophilic proteins.Most of the subcellular proteins were located in the plasma membrane re-gion of the cell,and a few were located in the extracellular,vacuole,Golgi apparatus and nucleus.There were 27 members of WAK/WAKL gene family distributed unevenly on 8 chromosomes of A.majus,and 7 pairs of collinear homologous genes with A.thaliana WAK/WAKL gene family.The gene promoter region contained cis-acting elements involved in de-fense and stress response,methyl jasmonate response and salicylic acid response.The susceptible variety Am1 and resis-tant variety Am6 were screened by inoculating the leaves of sclallodiscus in vitro.There were 12 WAK/WAKL family genes significantly differentially expressed(P<0.05)among the resistant materials,and the expression patterns of 9 genes were b
作者
杨冬梅
夏文念
宋佳怡
杨洁
严苍海
苏芸丽
汪仲毅
胡慧贞
YANG Dong-mei;XIA Wen-nian;SONG Jia-yi;YANG Jie;YAN Cang-hai;SU Yun-li;WANG Zhong-yi;HU Hui-zhen(College of Landscape and Horticulture Sciences,Southwest Forestry University/Engineering Research Center of Func-tional Flower Resources and Industrialization Technology of Yunnan Province,Kunming,Yunnan 650224,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2024年第1期24-36,共13页
Journal of Southern Agriculture
基金
国家自然科学基金青年科学项目(31901571)
云南省大学生创新创业训练计划项目(202110677078)
西南林业大学科研启动基金项目(01102-112109)。
关键词
金鱼草
WAK/WAKL基因家族
鉴定
核盘菌抗性
转录组测序
候选基因挖掘
Antirrhinum majus L.
AmWAK/WAKL gene family
identification
Sclerotinia sclerotiorum resistance
transcriptome sequencing
candidate gene mining
