竹叶花椒MYB基因家族的鉴定及其表达特性的分析

Identification and expression analysis of MYB gene family in Zanthoxylum armatum

【目的】利用测序获得的竹叶花椒Zanthoxylum armatum转录组数据,研究竹叶花椒中MYB转录因子家族的序列信息、进化模式、表达模式,探寻此家族中参与竹叶花椒皮刺生长发育的相关基因,为进一步揭示MYB转录因子家族生物学功能提供理论依据,为后续的无刺竹叶花椒品种的基因工程培育奠定理论基础。【方法】利用生物信息学方法,从测序获得的竹叶花椒转录组数据库中筛选MYB转录因子,对其蛋白基序、理化性质、系统进化关系、基因表达特性等进行分析,并利用qRT-PCR技术和基因克隆,对所选基因进行验证。【结果】从竹叶花椒中共鉴定出39条MYB转录因子序列,其中包括9个R1-MYB型基因、28个R2R3-MYB型基因、1个R3型基因和1个R4型基因。对其蛋白理化性质的分析结果表明:39个MYB转录因子所含氨基酸数量为101~621个,其均为可溶性蛋白,而其二级结构以无规则卷曲为主。亚细胞定位预测结果显示:39个MYB转录因子定位于细胞核的可能性最大,这符合转录因子调控下游基因的表达特性。基因表达情况的分析结果显示:部分MYB转录因子在竹叶花椒同一组织的不同发育时期的表达差异显著,其中ZaMYB10、ZaMYB23和ZaMYB25这3个基因的表达量随着皮刺的不断发育均呈逐渐增长的趋势,表明这3个基因可能与皮刺的生长发育相关。qRT-PCR分析结果表明:ZaMYB10基因在皮刺中有特异表达。基因克隆结果表明:ZaMYB10基因具有长为1032 bp的完整的cDNA开放阅读框,其编码343个氨基酸。【结论】鉴定出39个竹叶花椒MYB转录因子,其均包含SANT保守结构域,根据表达模式判断,其中的ZaMYB10转录因子可能参与了竹叶花椒皮刺生长发育的调控。

【Objective】Based on Zanthoxylum armatum transcriptome data to reveal MYB transcription factor family sequence information,evolutionary pattern and expression pattern.Identify the related genes that involved in growth and development process of Z.armatum aculeus in their family to provide theoretical basis for further revealing the biological functions of MYB transcription family,and to provide candidate genes for subsequent genetic engineering cultivation of prickly ash cultivars.【Method】The MYB transcription factors were identified from the whole Z.armatum transcriptome database by bioinformatics method.The protein motif,physicochemical properties,phylogenetic relationship and gene expression characteristics were analyzed.The candidate genes were verified by q-PCR technology and gene cloning.【Result】A total of 39 MYB transcription factor sequences were identified,including 9 R1-MYB genes,28 R2R3-MYB genes,1 R3 genes and 1 R4 genes.The analysis for physicochemical properties of protein revealed that this family contained 101-621 amino acids,all of which were soluble proteins,and the secondary structure was dominated by random coils.The results of subcellular localization prediction showed that 39 MYB transcription factors were most likely to locate in the nucleus,which was in line with the characteristics of transcription factors regulating the expression of downstream genes.The results of gene expression analysis showed that the expression levels of some MYB transcription factors in the same tissue was significantly different at different periods.Among them,the expression levels of ZaMYB10,ZaMYB23 and ZaMYB25 were gradually increased with the development of aculeus.This result indicated that these genes may participated in regulating the growth and development of aculeus.The results of qRT-PCR showed that ZaMYB10 gene was specifically expressed in aculeus.The results of gene cloning showed that ZaMYB10 contained a 1032bp complete cDNA open reading frame and encoded 343 amino acids.【Conclusion】This research identified 39 MYB transcription factors in Z.armatum for the first time,all of which contained SANT structural domain.Among which,ZaMYB10 transcription factors was participated in growth and development of aculeus.