杧果MiCO基因的克隆与表达模式分析  

Cloning and expression analysis of a MiCO gene in mango

作　　者：徐趁 罗聪[1] 余海霞[1] 陈锦文[1] 何新华[1] 王博[1] 方中斌[1] XU Chen LUO Cong YU Haixia CHEN Jinwen HE Xinhua WANG Bo FANG Zhongbin(College of Agriculture. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresourees, Guangxi University, Nanning 530004, Guangxi, Chin)

机构地区：[1]广西大学农学院.广西大学亚热带农业生物资源保护与利用国家重点实验室,南宁530004

出　　处：《果树学报》2017年第4期418-426,共9页Journal of Fruit Science

基　　金：国家自然科学基金(31660561);广西自然科学基金(2013 GXNSFDA019011;2014GXNSFBAII 8102);国家现代农业产业技术体系广西杧果创新团队栽培功能岗位项目(nycytxgxcxtd-06-02)

摘　　要：【目的】克隆杧果(Mangifera indica L.)MiCO基因并探讨其表达模式,为深入研究该基因的功能奠定基础。【方法】根据从杧果转录组测序数据中获得一个MiCO基因全长序列信息,设计基因全长引物,克隆具有不同开花习性的‘四季杧’和‘紫花杧’2个品种的MiCO基因全长序列,对其序列进行生物信息学分析,并利用荧光定量PCR技术对MiCO基因在杧果不同组织以及年周期的表达情况进行分析。【结果】序列分析显示,2个杧果品种的MiCO基因c DNA全长均为1 150 bp,都包含1个966 bp的开放阅读框,编码322个氨基酸,等电点为5.8,‘四季杧’品种的MiCO蛋白分子质量为35.30 ku,‘紫花杧’品种的MiCO蛋白分子质量为35.22 ku;MiCO基因编码的蛋白具有典型的CO同源蛋白结构,包括B-box1、B-box2和CCT结构域;系统进化树表明,杧果MiCO蛋白属于第一类CO蛋白,与拟南芥(Arabidopsis thaliana)At COL4相似性最高而聚类到一起。基因表达分析表明,MiCO基因在杧果不同组织中均表达,但不同组织的表达水平存在差异。年周期表达分析表明,MiCO基因在杧果成花转变期高度表达,而且在来年的5—6月还存在一个小的表达高峰期,但2个品种不同组织中MiCO基因表达存在差异。【结论】克隆获得杧果MiCO基因全长序列,发现2个具有不同开花习性的杧果品种的MiCO基因核苷酸和氨基酸序列高度同源,只存在少许差异。MiCO基因在杧果成花转变期高度表达,说明MiCO基因与杧果成花关系密切。[Objective] Floral induction is an important physiological process in higher plants from vegeta- tive stage to reproductive growth. Photoperiod is one of the important environmental triggers of flowering, and the CONSTANS （CO） gene is a key regulator of this pathway in the model plant Arabidopsis thaliana. Mango （Mangifera indica L.） is one of the world＇ s important woody plants, known as ＂the king of tropical fruits＂. Two mango cuhivars were used in present study with different flowering habits. ＇SiJi＇ blossoms several times a year and ＇ZiHua＇ blossoms once a year. Off-season production of mango has been achieved successful by using ＇SiJi＇ and much more income has been obtained by farmers. The molecular mechanism of different flowering habits in mango remains unclear. In present study, a MiCO gene of man- go was cloned and its bioinformatics and genetic evolutionary relationships as well as the expression pat- tern were analyzed. The results would provide scientists with a basis to further reveal the function of CO gene on flowering regulation in mango . [ Methods ] All materials were collected from the 8-year-old ＇SiJi＇ and ＇ZiHua＇ trees grown in the park of College of Agriculture, Guangxi University. The mature leaves were used for gene cloning. Young stems, young leaves, flowers, mature leaves and the fruits of the 7 d, 50 d and 90 d post full flowering of ＇ SiJi＇ were used for studying on the expression patterns of different orga- nizations. Mature stems, mature leaves and young stems of ＇SiJi＇ and ＇ZiHua＇ were used for analyzing the expression patterns of a year. All the materials collected were immediately stored at -80℃. Total RNA was isolated using hot-borate method, and quantified at OD260 and OD280. The RNA quality was veri- fied using agarose gel electrophoresis （1.2%）. The single- stranded cDNA was synthesized using a Reverse Transcriptase M-M kit （TaKaRa, Dalian, China） with the primer AUP1. In the previous studies, the full-length sequence of

关 键 词：杧果 克隆 CO基因 表达 

分 类 号：S667.7[农业科学—果树学]