热带作物学报2024，Vol.45Issue(8)：1538-1551,14.DOI:10.3969/j.issn.1000-2561.2024.08.003
香蕉A、B基因组胆碱单加氧酶基因克隆及渗透胁迫下表达特性分析
Cloning and Expression Characteristics of Choline Monooxygenase Gene in Banana A and B Genomes under Osmotic Stress
摘要
Abstract
Choline monooxygenase(CMO)is a key enzyme in the synthesis of betaine in higher plants and plays an im-portant role in the physiological process of plant resistance to stress.A CMO gene was identified in banana A and B reference genomes in banana gene database,and the biological information analysis of MaCMO and MbCMO genome sequences showed that MbCMO might be formed by a gene encoding a banana O-fucosyltransferase family protein and MbCMO gene in tandem.The CMO gene coding sequences of Zhanjiang AA(ZJ;AA genotype),Baxijiao(BX;AAA genotype),Guangdong Dajiao(GD;AAB genotype)and Jinfen(JF;ABB genotype)were cloned and compared,it was found that ZJ and BX only contained CMO-A,GD and JF both contained CMO-A,CMO-B1 and CMO-B2,and JF also contained CMO-H.Codon usage characteristics showed that there were 23 common high frequency codons among the four banana CMO gene,and the codons CUU and CCG were the most biased and the weakest codons,respectively.The physicochemical properties of CMO-A protein and CMO-H protein showed that the minimum number of amino acids was 425,the maximum number of amino acids was 470,and the secondary structure of CMO-B1 protein was the most complex.The molecular weight of CMO-B2 protein was 52.02 kDa,and the molecular weight of CMO-A protein was 47.48 kDa.All the four CMO proteins were acidic proteins,which did not have a transmembrane structure and were hydrophilic proteins.Subcellular localization prediction showed that all four CMO were localized in chloroplasts.In terms of evolution,plant CMO had obvious branches in monodicotyledonous plants during evolution,and the CMO-A,CMO-H,CMO-B1 and CMO-B2 proteins of banana were more closely related to other monocotyledonous plant CMO proteins.The results of RT-qPCR showed that CMO expression was up-regulated in the four kinds of banana roots in the early stage of osmotic stress,and the expression levels of ZJ and BX CMO in homozygous A genome were higher than those of GD and JF CMO in heterozygous A and B genomes.CMO expression in the four kinds of banana leaves was down-regulated at the early stage of osmotic stress.The expression levels of homozygous ZJ and BX CMO in A genome were up-regulated at the late stage of osmotic stress,while the expression levels of heterozygous GD and JF CMO in A and B genomes peaked at 10 days and then down-regulated again at 15 days later.It was significantly lower than that of ZJ and BX CMO.This study revealed the differences of CMO genes between the A and B genomes of bananas and the expression patterns of CMO genes in different genotypes of bananas under osmotic stress,which would lay a foundation for further research on the biological functions of CMO genes in the A and B genomes of bananas,especially the rela-tionship between CMO genes derived from different genomes and the ability of banana to resist osmotic stress.It would provide a reference for improving the stress resistance of banana by genetic engineering.关键词
香蕉/A、B基因组/胆碱单加氧酶合成基因/表达特性/抗逆性Key words
banana/A,B genome/CMO gene/characteristic of expression/stress resistance分类
农业科技引用本文复制引用
朱博为,于佳玄,李新国,刘菊华..香蕉A、B基因组胆碱单加氧酶基因克隆及渗透胁迫下表达特性分析[J].热带作物学报,2024,45(8):1538-1551,14.基金项目
海南省自然科学基金项目(No.320RC485) （No.320RC485）
国家自然科学基金项目(No.32160679) （No.32160679）
海南省研究生创新科研课题(No.Qhys2022-99). （No.Qhys2022-99）
