辣椒CcBBX20基因克隆和表达分析  

Cloning and Expression Analysis of Gene CcBBX20 in pepper

作　　者：邓巧灵 华玮 王华素 张明先 张芮豪 吕俊恒 邓明华[1] DENG Qiaoling;HUA Wei;WANG Huasu;ZHANG Mingxian;ZHANG Ruihao;LV Junheng;DENG Minghua(College of Landscape and Horticulture,Yunnan Agricultural University,Key Laboratory of Vegetable Biology of Yunnan Province,Vegetable Seed Industry Engineering Research Center of the Universities of Yunnan Province,Kunming 650201,China)

机构地区：[1]云南农业大学园林园艺学院,云南省蔬菜生物学重点实验室,云南省高校蔬菜种业工程研究中心,昆明650201

出　　处：《辣椒杂志》2024年第1期1-8,共8页Journal of China Capsicum

基　　金：国家自然科学基金(32160708);云南省科技计划项目(202102AE090005,202205AR070001,202402AE090012);云南省专家基层科研工作站(云人社通[2022]17号)。

摘　　要：锌指蛋白在植物生长发育和对非生物胁迫因子的响应中发挥着重要的调节作用。BBX蛋白属于锌指蛋白,CcBBX20基因是锌指蛋白家族的重要成员。为探讨CcBBX20在辣椒生长发育和响应非生物胁迫中的生物学功能,本研究利用基因克隆获得基因的编码序列;应用生物信息学分析方法预测该基因编码蛋白质的结构和功能;根据PepperHub数据库相关数据分析该基因在10种非生物胁迫处理下辣椒幼苗根和叶的表达量模式。结果表明:辣椒CcBBX20所编码的蛋白有209个氨基酸残基,相对分子质量为23.12 kD,分子式为C984H1551N285O329S15,理论等电点为5.78,是不稳定的亲水性蛋白;有2个典型的Bbox-SF结构域,没有信号肽和跨膜结构域,主要定位在细胞核中;其二级结构主要由无规则卷曲组成(占58.85%),α螺旋占比22.01%,β-转角占比3.83%。辣椒CcBBX20和马铃薯、番茄、茄子BBX20的亲缘关系最近。基因表达分析表明CcBBX20在辣椒幼苗根和叶受到不同胁迫后表达差异显著,在脱落酸、赤霉素、生长素、水杨酸、茉莉酸甲酯以及高温处理下表现为促进作用,其中高温42℃,处理1.5 h后表达量最高;在过氧化氢、甘露醇和盐处理下表现为抑制作用。因此辣椒CcBBX20基因作为BBX基因家族成员,在响应高温胁迫中发挥作用,可能会提高幼苗对高温的耐受力,可为辣椒耐高温育种目标提供候选基因。Zine finger protein plays an important regulatory role in plant growth and development,as well as abiotic stress response.BBX protein belongs to a subgroup of zinc finger proteins,and CcBBX20 gene in pepper is an important member of zinc finger protein family.To investigate the biological function of CcBBX20 in the growth and development of pepper and response to abiotic stress,the structure and function of the CcBBX20 were analyzed by using gene cloning and bioinformatic methods.Expressions of the gene in the roots and leaves of pepper seedlings under 10 types of abiotic stress treatments were analyzed by using the PepperHub database.The protein coded by CcBBX20 is an unstable hydrophobic protein and has 201 amino acids with a relative molecular mass of 23.12 kD,molecule formula of C984H1551N285O329S15 and theoretical electrical point of 5.78.Moreover,the protein has two typical Bbox-SF structural domains with no signal peptide and transmembrane domain and is mainly located in the cell nucleus.Its secondary structure is mainly composed of irregular coil(58.85%)with 22.01%ofα-helix and 3.83%ofβ-turn.The sequence of CcBBX20 in pepper is highly similar with that of potato,tomato and eggplant.Gene expression data showed that expressions of CcBBX20 were different in the roots and leaves of pepper seedlings suffering different stresses.Treatments of abscisic acid,gibberellin,auxin,salicylic acid,methyl jasmonate and high temperature increased the expressions of CcBBX20,which reached up to the highest at temperature of 42℃for 1.5 h.However,its expressions were decreased under the treatments of hydrogen peroxide,mannitol and salinity.As a result,as a member of the BBX gene family,pepper CcBBX20 gene plays a role in response to high temperature stress,potentially improving the tolerance of seedlings to high temperature and providing a candidate gene for breeding new heat-tolerant pepper cultivars.

关 键 词：辣椒 BBX基因家族 基因克隆 生物信息分析 表达分析 高温胁迫 

分 类 号：S641.3[农业科学—蔬菜学]