亚洲棉NCED3基因克隆及其抗旱功能分析  

Cloning and Functional Analysis of NCED3 on Drought Resistance in Gossypium arboreum

作　　者：蔡肖[1] 王海涛 李兴河 甄军波[1] 刘琳琳[1] 刘迪 迟吉娜[1] 张建宏[1] CAI Xiao;WANG Haitao;LI Xinghe;ZHEN Junbo;LIU Linlin;LIU Di;CHI Jina;ZHANG Jianhong(Institute of Cotton,Hebei Academy of Agriculture and Forestry Sciences/Key Laboratory of Cotton Biology and Genetic Breeding in Huanghuaihai Semiarid Area,Ministry of Agriculture/National Cotton Improvement Center Hebei Branch,Shijiazhuang,Hebei 050051)

机构地区：[1]河北省农林科学院棉花研究所/农业农村部黄淮海半干旱棉花生物学与遗传育种重点实验室/国家棉花改良中心河北分中心,河北石家庄050051

出　　处：《核农学报》2022年第9期1713-1722,共10页Journal of Nuclear Agricultural Sciences

基　　金：河北省农林科学院现代农业科技创新工程基本科研业务费(2021070202);河北省农林科学院财政项目(F17R2017039379)。

摘　　要：9-顺式-环氧类胡萝卜素双加氧酶(NCED)是脱落酸(ABA)合成过程中的关键限速酶,被证实广泛参与植物的生长发育进程及对非生物胁迫的应答。为了挖掘、鉴定棉花抗旱相关的NCEDs基因,本研究克隆了亚洲棉(Gossypium arboreum)GaNCED3基因,利用实时荧光定量PCR(qRT-PCR)技术分析了干旱胁迫下该基因的表达特性;并遗传转化拟南芥(Arabidopsis thaliana),研究了该基因在干旱应答中的功能。结果显示,GaNCED3基因开放阅读框(ORF)全长为1794 bp,其编码的蛋白质包含597个氨基酸。该基因在干旱胁迫处理后上调表达,在处理3 h的根中表达量最高,是对照的27.6倍。在萌发期进行甘露醇模拟的干旱处理,超表达GaNCED3的转基因拟南芥种子萌发率和绿苗率均高于野生型。在幼苗期进行自然干旱处理后,转基因拟南芥植株叶片丙二醛含量显著低于对照(P<0.05),游离脯氨酸积累量显著高于对照(P<0.05)。本研究初步证明了外源超表达GaNCED3基因使植株抗旱能力增强,为进一步研究GaNCED3干旱应答的分子机制提供了理论依据,为抗旱育种提供了候选基因资源。Nine-cis-epoxy carotenoid dioxygenase(NCED),key rate-limiting enzymes responsible for biosynthesis of abscisic acid,are demonstrated to be essential for plant growth,development and abiotic stress response.In order to identify drought-related NCEDs in cotton and lay a foundation for breeding drought resistant cotton varieties,GaNCED3 gene was cloned and its expression profile under drought stress was analyzed using qRT-PCR.Moreover,function analysis of GaNCED3 was performed with the transgenic Arabidopsis under drought stress was performed in this study.The results showed that the ORF of GaNCED3 was 1794 bp in length and it coded 597 amino acids.The expression of GaNCED3 was up-regulated under drought stress,with a maximum 27.6-fold increase in roots after treated for 3 h.Under mannitol simulated drought stress,the seed germination rate and green plantlet rate of transgenic Arabidopsis were higher than those of wild type.Under natural drought stress at seedling stage,the malondialdehyde content in leaves of transgenic plants was significantly lower than that of control(P<0.05),while free proline content was significantly higher than that of control(P<0.05).This study preliminarily proved that exogenous overexpression of GaNCED3 gene enhanced the drought resistance of plants.The results laid a theoretical basis for further study on the molecular mechanism of GaNCED3 involved in drought response,and provided candidate gene resources for drought resistance breeding.

关 键 词：棉花 干旱 NCED3 9-顺式-环氧类胡萝卜素双加氧酶 基因功能 

分 类 号：S562[农业科学—作物学]