藜麦LOC110717159基因克隆、生物信息学分析及其在拟南芥过表达分析

doi:10.11983/CBB25040

植物学报

藜麦LOC110717159基因克隆、生物信息学分析及其在拟南芥过表达分析

陶小所¹, 姚晓华^1,2, 姚有华^1,2

¹青海大学农林科学院, 西宁 810016; ²青藏高原种质资源研究与利用实验室, 西宁 810016

收稿日期:2025-03-19 修回日期:2025-05-20 出版日期:2025-06-11 发布日期:2025-06-11
通讯作者: 姚有华
基金资助:
青海省科技成果转化专项项目(No.2022-NK-112)

Cloning, Bioinformatics Analysis of LOC110717159 Gene from Chenopodium Quinoa and Overexpression in Transgenic Arabidopsis

Tao Xiaosuo¹, Yao Xiaohua^1,², Yao youhua^1,²

¹Academy of Agricultural and Forestry Sciences, Qinghai University, Xining 810016, China; ²Qinghai-Tibet Plateau Germplasm Resources Research and Utilization Laboratory, Qinghai Province, Xining 810016, China

Received:2025-03-19 Revised:2025-05-20 Online:2025-06-11 Published:2025-06-11
Contact: Yao Youhua

摘要/Abstract

摘要： 糖基水解酶家族在调控植物籽粒大小方面起关键作用。本研究通过克隆藜麦(Chenopodium quinoa)糖基水解酶家族LOC110717159基因, 并通过生物信息学、表达模式分析和拟南芥(Arabidopsis thaliana)异源表达实验, 解析其在籽粒充实过程中的生物学功能。生物信息学分析表明, 大、小粒藜麦种质LOC110717159编码序列全长为1 023 bp, 编码339个氨基酸, 存在1个碱基差异; 其启动子区 (~2 000 bp)含有119个顺式作用元件, 包括3个激素响应元件(茉莉酸甲酯、水杨酸和赤霉酸)。系统进化分析显示, 该基因与甜菜(Beta vulgaris)、菠菜(Spinacia oleracea)和苋(Amaranthus tricolor)的亲缘关系较近。qRT-PCR分析表明, 灌浆21天时, LOC110717159在大粒藜麦种质的籽粒中高表达。拟南芥过表达株系的千粒重显著低于野生型(P<0.05), 但粒长和粒宽无显著差异。单倍型分析发现, LOC110717159编码序列在17个藜麦种质中存在差异, 62.5%的千粒重大于4.0 g的种质与大粒藜麦种质碱基一致, 而77.8%的千粒重小于3.0 g的种质与小粒藜麦种质碱基一致。综上, LOC110717159可能负调控藜麦籽粒的充实过程。

Abstract: INTRODUCTION: The glycoside hydrolase (GH) gene family plays a crucial role in regulating plant grain size.

RATIONALE: To elucidate its biological role during grain filling, we cloning and functionally characterized LOC110717159, a GH gene in quinoa, through bioinformatics analysis, spatiotemporal expression profiling, and heterologous expression experiments in Arabidopsis.

RESULTS: LOC110717159 has a 1 023 bp coding sequence (encoding 339 amino acids) in both germplasms, differing by a single nucleotide. Analysis of the ~ 2 000 bp LOC110717159 promoter identified 119 cis-elements, including three hormone-responsive elements (MeJA, SA, and GA). Phylogenetically, it is closely related to Beta vulgaris, Spinacia oleracea and Amaranthus tricolor. qRT-PCR revealed high expression of LOC110717159 in large-grained quinoa at 21 days after anthesis (21 DAA). Arabidopsis overexpression lines showed significantly reduced 1 000-grain weight (P < 0.05) but no change in grain length and width. Haplotype analysis of 17 quinoa germplasms indicated that 62.5% of large germplasms (≥ 4.0 g/1 000 grains) carried the large-grained haplotype, while 77.8% of small germplasms (≤ 3.0 g) shared the small-grained haplotype.

CONCLUSION: In conclusion, LOC110717159 likely functions as a negative regulator in quinoa grain filling.

T3-generation transgenic Arabidopsis lines and seed morphological characteristics. (A)–(C) Wild-type plants; (D)–(F) AtOE1、AtOE2、AtOE3; ns: No significant difference; *P ≤ 0.05

陶小所, 姚晓华, 姚有华. 藜麦LOC110717159基因克隆、生物信息学分析及其在拟南芥过表达分析. 植物学报, DOI: 10.11983/CBB25040.

Tao Xiaosuo, Yao Xiaohua, Yao Youhua. Cloning, Bioinformatics Analysis of LOC110717159 Gene from Chenopodium Quinoa and Overexpression in Transgenic Arabidopsis. Chinese Bulletin of Botany, DOI: 10.11983/CBB25040.

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藜麦LOC110717159基因克隆、生物信息学分析及其在拟南芥过表达分析

Cloning, Bioinformatics Analysis of LOC110717159 Gene from Chenopodium Quinoa and Overexpression in Transgenic Arabidopsis

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