粉蕉MbACS7基因的克隆及表达分析  

Cloning and Expression Analysis of MbA CS7 Gene in Musa ABB Group,cv Pisang Awak,FJ

作　　者：符思颖 李美英[2] 颜彦[2] 田丽波[3] 商桑[1] 胡伟[2] Fu Siying;Li Meiying;Yan Yan;TTian Libo;Shang Sang;Hu Wei(School of Life Sciences,Hainan University,Haikou,570228;Institute for Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou,571101;Key Laboratory for Quality Regulation of Tropical Horticultural Crops of Hainan Province,Horticulture CollegeofHainanUniversity,Haikou,570228)

机构地区：[1]海南大学生命科学学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海口571101 [3]海南大学园艺学院,海南省热带园艺作物品质调控重点实验室,海口570228

出　　处：《分子植物育种》2023年第16期5247-5254,共8页Molecular Plant Breeding

基　　金：海南省自然科学基金项目(2019RC295)资助。

摘　　要：乙烯合成及其效应是决定香蕉果实成熟的核心要素,其中1-氨基环丙烷-1-羧酸(ACC)合成酶(ACS)是乙烯合成途径的限速酶。粉蕉果实成熟速度较快,保鲜难度较大。研究粉蕉ACS家族基因的特征和表达模式可为控制香蕉果实成熟进程和延长货架期提供理论依据。本研究从粉蕉中克隆获得一个全长序列为1458 bp的目的片段,编码485个氨基酸,分子量为54.751 kD,具有PLN02450保守结构域(37~1377氨基酸),属于ACS基因家族蛋白,因此被命名为MbACS7。理化性质分析表明MbACS7蛋白亲水且不稳定。进化分析表明MbACS7蛋白预测的氨基酸序列与苦瓜(Momordica charantia)ACS蛋白的氨基酸序列(CAE53271.1)遗传关系较近。瞬时表达分析表明MbACS7蛋白在烟草表皮细胞的细胞核中强烈表达,表明该蛋白可能在细胞核中发挥着重要的生理功能。实时荧光定量表达分析表明MbACS7基因在粉蕉果实成熟期表达量最高,为果实发育初期的900多倍。本研究为进一步解析MbACS7基因在粉蕉果实乙烯生物合成中的功能奠定基础,也为改良香蕉果实贮藏性提供基因资源。Ethylene synthesis and its effects are the core elements that determine the ripening of banana fruit,of which 1-aminocyclopropane-1-carboxylic acid(ACC)synthase(ACS)is the rate-limiting enzyme in the ethylene synthesis pathway.The fruit of Musa ABB Group,cv Pisang Awak,FJ ripens faster,and it is more difficult to keep fresh.Studying the characteristics and expression patterns of FJ ACS family genes can provide theoretical basis for controlling banana fruit ripening process and prolonging shelf life.In this study,a target fragment with a full-length sequence of 1458 bp was cloned from FJ,encoding 485 amino acids,with a molecular weight of 54.751 kD and a conserved domain of PLN02450(37~1377 amino acids),belonging to the ACS gene family protein,so it was named as MbACS7.Physicochemical property analysis showed that MbACS7 protein was hydrophilic and unstable.Evolutionary analysis showed that the predicted amino acid sequence of MbACS7 protein was closely related to the amino acid sequence of ACS protein of bitter gourd(Momordica charantia)(CAE53271.1).Transient expression analysis showed that MbACS7 protein was strongly expressed in the nucleus of tobacco epidermal cells,suggesting that this protein may play an important physiological function in the nucleus.Real-time fluorescence quantitative expression analysis showed that the expression level of MbA CS7 gene was the highest at the ripening stage of FJ fruit,which was more than 900 times higher than that at the early stage of fruit development.This study lays a foundation for further analysis of the function of MbA CS7 gene in ethylene biosynthesis of banana fruit,and also provides genetic resources for improving banana fruit storage.

关 键 词：粉蕉 1-氨基环丙烷-1-羧酸合成酶(ACS) 乙烯生物合成 表达分析 

分 类 号：S668.1[农业科学—果树学]