澳洲坚果开花相关MiMADS-box基因克隆及表达载体构建

doi:10.7525/j.issn.1673-5102.2021.06.006

植物研究 ›› 2021, Vol. 41 ›› Issue (6): 888-895.doi: 10.7525/j.issn.1673-5102.2021.06.006

澳洲坚果开花相关MiMADS-box基因克隆及表达载体构建

谭秋锦, 张涛, 韦媛荣, 郑树芳, 汤秀华, 王文林()

广西南亚热带农业科学研究所，龙州 532400

收稿日期:2021-03-04 出版日期:2021-11-20 发布日期:2021-10-29
通讯作者: 王文林 E-mail:wwlin3949@163.com
作者简介:谭秋锦（1980—），女，高级农艺师，主要从事热带果树遗传育种研究。
基金资助:
广西农业科学院科技发展基金项目(桂农科2020YM141);广西农业科学院基本业务费项目(桂农科2020YM46);广西农业科学院科技发展基金自选项目(2019ZX17);广西农业科学院科技发展基金自选项目(桂农科2019ZX2015);广西科技重大专项（桂科AA17204058-4）;国家重点研发计划(2016YFC0502406)

Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia

Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG()

Guangxi South Subtropical Agricultural Science Research Institute，Longzhou 532400

Received:2021-03-04 Online:2021-11-20 Published:2021-10-29
Contact: Wen-Lin WANG E-mail:wwlin3949@163.com
About author:TAN Qiu-Jin（1980—），famale，senior agronomist，mainly engaged in the genetic breeding of tropical fruit trees.
Supported by:
Guangxi Academy of Agricultural Sciences Science and Technology Development Fund（Guinongke 2020YM141,Guinongke 2020YM46,Guinongke 2019ZX2015,2019ZX17）;Major Science and Technology Projects of Guangxi(Guike AA17204058-4);National Key Research and Development Project(2016YFC0502406)

摘要/Abstract

摘要：

为研究MADS-box转录因子家族成员在澳洲坚果开花及花器官发育过程的作用机制，利用PCR技术从澳洲坚果品种“桂热1号”叶片中克隆MiMADS-box基因并采用生物信息学对其结构及功能进行分析。结果表明，克隆获得的MiMADS-box基因cDNA序列全长1 179 bp，开放阅读框（ORF）729 bp，编码242个氨基酸。MiMADS-box编码一个无信号肽、无跨膜结构且定位于细胞核的不稳定亲水蛋白，存在MADS superfamily和K-box superfamily结构域，属于MIKCc型家族。系统进化分析将MiMADS-box与AGL7/AP1、AGL10/CAL1、AGL8/FUL、AGL79亲缘关系较近，聚类为FUL-AP1（SQUA）亚族。通过转录组数据分析MiMADS-box基因在“桂热1号”和“695”品种澳洲坚果枝条、花和叶片中的表达模式，表明MiMADS-box在“桂热1号”品种花中的表达量最低，“695”品种花中的表达量最高。利用同源重组的方法构建了绿色荧光蛋白融合表达载体pGREEN-MiMADS-box-ORF-GFP。推测MiMADS-box与澳洲坚果开花及花器官发育过程密切相关。本研究为阐明MiMADS-box基因在澳洲坚果开花调控机制提供理论基础和技术指导。

关键词: 澳洲坚果, MiMADS-box, 基因克隆, 载体构建

Abstract:

In order to clarify the mechanism of MADS-box transcription factor gene family member in flowering and floral organ development in Macadamia integrifolia， aMiMADS-box gene was cloned from the leaves of M. integrifolia “GUIRE1” cultivar by PCR technique and analyzed its structure and function by Bioinformatics method. The cloned cDNA sequence of MiMADS-box gene was 1 179 bp in length， 729 bp open reading frame （ORF）， encoded 242 amino acids. MiMADS-box， an unstable hydrophilic protein that had no transmembrane structure， no signal peptide and localized in the nucleus， had MADS superfamily and K-box superfamily conserved domains belonging to the MIKCc-type family. Phylogenetic analysis clustered MiMADS-box and AGL7/AP1， AGL10/CAL1， AGL8/FUL， AGL79 into FUL-AP1（SQUA） subfamily. The transcriptome data were used to analyze the expression patterns of MiMADS-box gene in branch， flower and leave of M. integrifolia “GUIRE1” and “695” cultivar. Results indicated that MiMADS-box had the lowest expression in the flower of “GUIRE1”， but the expression level of “695” was the highest. The green fluorescent protein fusion vector pGREEN-MiMADS-box-ORF-GFP was constructed by homologous recombination. It was speculated that MiMADS-box was closely related to flowering and floral organ development. The study provided a theoretical and technical guide for clarification the role MiMADS-box in flowering regulation mechanism of M. integrifolia.

Key words: Macadamia integrifolia, MiMADS-box, gene cloning, vector construction

中图分类号:

S664.9

谭秋锦, 张涛, 韦媛荣, 郑树芳, 汤秀华, 王文林. 澳洲坚果开花相关MiMADS-box基因克隆及表达载体构建[J]. 植物研究, 2021, 41(6): 888-895.

Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG. Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia[J]. Bulletin of Botanical Research, 2021, 41(6): 888-895.

图/表 6

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澳洲坚果开花相关MiMADS-box基因克隆及表达载体构建

Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia

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