薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

doi:10.3969/j.issn.1000-2006.201910027

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南京林业大学学报（自然科学版） ›› 2020, Vol. 44 ›› Issue (4): 63-69.doi: 10.3969/j.issn.1000-2006.201910027

薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

王敏¹(), 席东², 莫正海¹, 陈于¹, 赵玉强¹, 朱灿灿¹()

^1.江苏省中国科学院植物研究所，江苏南京 210095
^2.上海市青浦区农业技术推广服务中心，上海 201700

收稿日期:2019-10-25 修回日期:2020-03-19 出版日期:2020-07-22 发布日期:2020-08-13
通讯作者: 朱灿灿
作者简介:王敏（wm120624@163.com）。
基金资助:
常州市科技支撑计划（农业）(CE20192012);江苏省林业科技创新与推广项目（LYKJ[2017]38）;中央财政林业科技推广示范资金项目（苏[2019]TG04）

Cloning, subcellular localization and expression analysis of CiAGL6 gene in pecan

WANG Min¹(), XI Dong², MO Zhenghai¹, CHEN Yu¹, ZHAO Yuqiang¹, ZHU Cancan¹()

^1.Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210095, China
^2.Shanghai Qingpu District Agricultural Technology Extension Service Center, Shanghai 201700, China

Received:2019-10-25 Revised:2020-03-19 Online:2020-07-22 Published:2020-08-13
Contact: ZHU Cancan

摘要/Abstract

摘要： 目的

研究薄壳山核桃MADS-box转录因子CiAGL6基因的特性、亚细胞定位及在雌花不同发育时期的表达水平，为揭示薄壳山核桃成花分子机理提供理论依据。

方法

根据薄壳山核桃转录组数据，采用RT-PCR方法获得CiAGL6基因，对其进行生物信息学分析和亚细胞定位鉴定。利用实时荧光定量PCR技术（qRT-PCR）分析CiAGL6基因在雌花发育过程中的表达模式。

结果

获得薄壳山核桃CiAGL6基因的完整开放阅读框（ORF）780 bp，共编码259个氨基酸，具有典型的MADS-box结构域和K-box区。在该蛋白的二级结构中，α-螺旋、延伸链和无规则卷曲分别占该蛋白质氨基酸残基总数的54.44%、17.37%和28.19%。系统进化分析表明，薄壳山核桃CiAGL6与壳斗科栓皮栎QsAGL6的亲缘关系最近。亚细胞定位显示，CiAGL6蛋白定位在细胞核内。qRT-PCR分析结果表明，CiAGL6基因在雌花中的表达量显著高于雄花中的。CiAGL6基因在雌花发育的各个阶段均有表达，其中在雌花盛开期的表达量最高。

结论

CiAGL6基因在薄壳山核桃雌花形成过程中发挥重要调控作用。

关键词: 薄壳山核桃, CiAGL6基因, 进化分析, 亚细胞定位, 基因表达

Abstract: Objective

The aim was to study the characteristics, subcellular localization and expression level in different development stages of female flowers of the MADS?box transcription factor CiAGL6 in pecan, which provides a theoretical basis for revealing the molecular mechanism of pecan flower formation.

Method

According to the transcriptome data of pecan, the CiAGL6 gene was cloned using RT?PCR. Next, the bioinformatic analysis and subcellular localization analysis were performed. And the expression pattern of CiAGL6 gene in female flower development was analyzed using real?time fluorescence quantitative PCR (qRT?PCR).

Result

The complete open reading frame (ORF) of the CiAGL6 gene was 780 bp, which encodes 259 amino acids and contains a conserved MADS?box domain and K?domain．In the secondary structure of the protein, a?helix, extension chain, and irregular curl accounted for 54.44%, 17.37%, and 28.19% of the total amino acid residues of the protein, respectively. The phylogenetic analysis showed that CiAGL6 has the closest relationship with QsAGL6 of Quercus suber in Fagaceae. The subcellular localization assay revealed CiAGL6 was located in the nucleus. The qRT?PCR analysis showed that the expression of CiAGL6 gene in female flowers was significantly higher than that in male flowers. CiAGL6 was expressed in all stages of female flower development, and the highest expression was found in female flowers at the blooming stage.

Conclusion

The CiAGL6 gene plays an important regulatory role in pecan female flower formation.

Key words: pecan, CiAGL6 gene, phylogenetic analysis, subcellular localization, gene expression

中图分类号:

S664.1

王敏,席东,莫正海,等. 薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达[J]. 南京林业大学学报（自然科学版）, 2020, 44(4): 63-69.

WANG Min, XI Dong, MO Zhenghai, CHEN Yu, ZHAO Yuqiang, ZHU Cancan. Cloning, subcellular localization and expression analysis of CiAGL6 gene in pecan[J].Journal of Nanjing Forestry University (Natural Science Edition), 2020, 44(4): 63-69.DOI: 10.3969/j.issn.1000-2006.201910027.

图/表 4

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图3

图4

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薄壳山核桃CiAGL6基因的克隆、亚细胞定位及表达

Cloning, subcellular localization and expression analysis of CiAGL6 gene in pecan

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