油茶隐花色素基因CoCRY1的克隆及功能研究
作者:
作者单位:

中南林业科技大学林学院/经济林培育与保护教育部重点实验室/经济林育种与栽培国家林业和 草原局重点实验室/岳麓山实验室经济林品种创制中心,长沙 410004

作者简介:

研究方向为经济林栽培与利用,E-mail : rss1127@outlook.com

通讯作者:

李建安,研究方向为经济林栽培育种,E-mail : lja0731@126.com
阎晋东,研究方向为油茶开花分子机制研究,E-mail : yjd0731@outlook.com

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基金项目:

国家自然科学基金(32371930);中国博士后面上资助项目(2021M702653);湖南省自然科学青年基金(2023JJ41041);湖南省教育厅科学研究项目(22B0281);长沙市自然科学基金(kq2202281)


Cloning and Functional Study of Cryptochrome Gene CoCRY1 from Camellia oleifera Abe.
Author:
Affiliation:

College of Forestry, Central South University of Forestry and Technology / Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees of Ministry of Education / Key Laboratory of Breeding and Cultivation of Non-Wood Forest, National Forestry and Grassl and Administration / Yuelu Mountain Laboratory Non-Wood Forests Variety Innovation Center, Changsha 410004

Fund Project:

Foundation projects: National Natural Science Foundation of China (32371930); China Postdoctoral Science Foundation (2021M702653); Natural Science Foundation for Youth of Hunan Province (2023JJ41041); Research Foundation of Education Bureau of Hunan Province (22B0281); Changsha Natural Science Foundation (kq2202281)

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    摘要:

    油茶(Camellia oleifera Abe.)是我国重要木本油料树种,具有重要的经济价值和社会效益。隐花色素(Cryptochrome)是植物蓝光受体之一,参与植物开花调控、光形态建成等生长发育过程。本研究从油茶华硕中克隆到CoCRY1基因,通过生物信息学分析发现,其CDS序列长度为2262 bp,编码的蛋白质序列包含684个氨基酸,蛋白分子式为C3454H5281N971O1027S18,分子量77.42 kDa,是一种不稳定的疏水蛋白,进一步分析表明其蛋白序列包含3个结构域,分别为DNA_photolyase、FAD_binding_7和Cryptochrome_C,可证明CoCRY1蛋白属于隐花色素家族。同源序列及系统发育树分析发现CoCRY1蛋白与茶树(Camellia sinensis) CsCRY1序列相似性最高。组织表达分析显示,CoCRY1的表达量在茎中最高,在花中最低。通过农杆菌转化得到CoCRY1基因异源表达的拟南芥(Arabidopsis thaliana)植株,并且从DNA水平及RNA水平鉴定到阳性植株,对CoCRY1异源表达的拟南芥进行表型分析发现,CoCRY1基因的过量表达导致拟南芥提前开花,并对下胚轴伸长具有光特异性的抑制作用。本研究通过生物信息学、定量分析及异源表达发现,油茶CoCRY1基因在油茶开花等生物学过程中具有重要作用。

    Abstract:

    Camellia oleifera Abe. is an important woody oil tree in China, which has important economic and social benefits. Cryptochrome is one of the blue light receptors in plants, which is involved in the growth and development of plants such as flowering regulation and photomorphogenesis. In this study, CoCRY1 gene was cloned from C.oleifera 'Huashuo'. Bioinformatics analysis showed that CDS sequence length of CoCRY1 was 2262 bp encoding for 684 amino acids, the molecular formula of the CoCRY1 protein was C3454H5281N971O1027S18, and the molecular weight was 77.42 kDa, further analysis showed that this protein sequence contained three domains, namely DNA_photolyase, FAD_binding_7 and Cryptochrome_C, which proved that CoCRY1 protein belongs to cryptochrome family. Homologous sequence and phylogenetic tree analysis showed that CoCRY1 protein had the highest similarity with the homologous sequence of tea tree (Camellia sinensis). Tissue expression analysis showed that the transcripts of CoCRY1 were the highest in stem and the lowest in flower. Arabidopsis thaliana plants with heterogenous expression of CoCRY1 gene were obtained by Agrobacterium transformation, and transgenic plants were identified from DNA and RNA level. Phenotypic analysis of transgenic Arabidopsis showed that overexpression of CoCRY1 gene induced early flowering under long day londition and had photospectific inhibitory effect on hypocotyl elongation. Through bioinformatics, quantitative analysis and heterologous expression, it was found that CoCRY1 gene plays an important role in the biological process of C.oleifera flowering.

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任爽爽,隽乐梅,柳倩,等.油茶隐花色素基因CoCRY1的克隆及功能研究[J].植物遗传资源学报,2024,25(3):440-450.

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  • 收稿日期:2023-09-25
  • 最后修改日期:2023-10-24
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  • 在线发布日期: 2024-03-05
  • 出版日期: 2024-02-26
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