椰子CnRADIALIS-like转录因子的克隆与表达分析  

Cloning and Expression Analysis of Coconut CnRADIALIS-like Transcription Factor

作　　者：郭树宽 曾春茹 吴翼[2,3] 李静 李新国[1] 杨耀东 GUO Shukuan;ZENG Chunru;WU Yi;LI Jing;LI Xinguo;YANG Yaodong(College of Horticulture,Hainan University,Haikou,Hainan 570228,China;Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wenchang,Hainan 571339,China;Hainan Provincial Key Laboratory of Tropical Oil Crops Biology,Wenchang,Hainan 571339,China)

机构地区：[1]海南大学园艺学院,海南海口570228 [2]中国热带农业科学院椰子研究所,海南文昌571339 [3]海南省热带油料作物生物学重点实验室,海南文昌571339

出　　处：《热带作物学报》2021年第9期2478-2486,共9页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.32071805);中国热带农业科学院基本科研业务费项目(No.17CXTD-28,1630152017019)。

摘　　要：本研究依据前期已经完成的4个品种椰子外果皮转录组测序数据结果,筛选获得在4个品种椰子外果皮中表达水平显著差异的MYB类基因CnRADIALIS-like。从椰子(Cocos nucifera L.)外果皮中克隆获得CnRADIALIS-like,序列分析显示,CnRADIALIS-like基因的开放阅读框(ORF)长288 bp,编码95个氨基酸。结构域分析显示,CnRADIALIS-like含有1个典型的SANT/MYB结构域,属于MYB转录因子家族中的I-box-binding-MYB型。多序列比对发现,CnRADIALIS-like氨基酸序列与油棕MYB-RADIALIS(RAD)氨基酸序列相似度最高。理化性质、亲/疏水性和无序化分析显示,CnRADIALIS-like转录因子是亲水性蛋白且偏碱性,存在无序化区域。信号肽和跨膜结构域分析表明,其不存在信号肽和跨膜结构域。空间结构分析显示,CnRADIALIS-like有典型的α-螺旋结构。实时荧光定量PCR(qRT-PCR)显示,CnRADIALIS-like基因在4个品种椰子外果皮中的表达量存在显著差异,在红矮椰子中表达量最高,红矮椰子不同组织中在外果皮表达量最高。该基因的表达在红矮椰果从幼果至成熟果的3个发育阶段依次降低,套袋条件下表达量低于正常光照条件下,表明该基因可能与光形态下细胞发育及代谢产物合成存在相关作用。通过对该基因的生物信息功能进行分析,为深入研究其功能及作用机制提供参考,同时以期为深入剖析椰子的外果皮颜色形成机制提供重要的理论依据。Based on the results of the transcriptome sequencing data of the four coconut exocarps that have been completed previously,the expression levels of a MYB gene,CnRADIALIS-like,show significant difference in four coconut exocarps with different color.CnRADIALIS-like was cloned from coconut(Cocos nucifera L.)exocarp.Sequence analysis showed that the open reading frame(ORF)of CnRADIALIS-like was 288 bp long,encoding 95 amino acids.The domain analysis showed that CnRADIALIS-like contains a typical SANT/MYB domain,which belongs to the I-box-binding-MYB type in the MYB transcription factor family.Multi-sequence comparison found that the amino acid sequence of CnRADIALIS-like had the highest similarity with the amino acid sequence of oil palm MYB-RADIALIS(RAD).Physicochemical properties,affinity/hydrophobicity and disorder analysis showed that the CnRADIALIS-like transcription factor is a hydrophilic protein and alkalescent,with disordered regions.Analysis of signal peptide and transmembrane domain showed that there was no signal peptide and transmembrane domain.Spatial structure analysis shows that CnRADIALIS-like had a typicalα-helical structure.Real-time fluorescent quantitative PCR(qRT-PCR)showed that the expression levels of CnRADIALIS-like were significantly different in the epicarp with different color,and the highest expression was observed in the red dwarf coconut.This gene showed the highest expression in the epicarp in comparison with other tissues of the red dwarf coconut.The expression of the gene decreased in the three developmental stages of red dwarf coconut from young fruit to mature fruit.The expression level under bagging conditions was lower than under normal light conditions,indicating that this gene may be related to cell development and metabolite synthesis under light morphology effect.The bioinformatic analysis of this gene would provide a reference for in-depth study of its functiona,and at the same time,it would also provide an important theoretical basis for the role of this gene in the forma

关 键 词：椰子 外果皮颜色 MYB家族 RAD蛋白 

分 类 号：S667.4[农业科学—果树学]