Cloning and Functional Analysis of Flowering Regulation Gene MsCOL2 in Alfalfa

doi:10.11733/j.issn.1007-0435.2023.10.002

Acta Agrestia Sinica ›› 2023, Vol. 31 ›› Issue (10): 2905-2915.DOI: 10.11733/j.issn.1007-0435.2023.10.002

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Cloning and Functional Analysis of Flowering Regulation Gene MsCOL2 in Alfalfa

LU Dong-yu, WANG Xue, JIANG Xu, ZHANG Yun-xiu, ZHANG Li-li, LI Ya-jing, KANG Jun-mei

Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2023-04-15 Revised:2023-05-28 Online:2023-10-15 Published:2023-11-02

紫花苜蓿花期调控基因MsCOL2的克隆及功能研究

卢栋宇, 王雪, 蒋旭, 张云秀, 张丽丽, 栗亚静, 康俊梅

中国农业科学院北京畜牧兽医研究所, 北京 100193

通讯作者: 康俊梅,E-mail:kangjunmei@caas.cn
作者简介:卢栋宇(1995-),汉族,河北三河人,男,硕士研究生,主要从事饲草遗传育种研究,E-mail:1571687177@qq.com
基金资助:
国家自然科学基金（32071868）资助

Abstract

Abstract: Flowering is a turning point for plants from vegetative growth to reproductive growth. The yield and quality of alfalfa (Medicago sativa L.) are closely relevant to its flowering time. Photoperiod is one of the important factors affecting plant flowering. The CONSTANS-like (COL) gene family plays an important role in the regulation of photoperiod flowering. In order to reveal the function of COL family genes in alfalfa flowering regulation,MsCOL2 gene was cloned from alfalfa in this study. This gene encodes for 401 amino acids. Tissue differential expression analysis showed that MsCOL2 had the highest expression in flowers. Expression analysis at different developmental stages of flowering showed that MsCOL2 had the highest expression level at flower bud differentiation stage;under different photoperiod conditions,the expression level of MsCOL2 had different rhythmic expressions;After treatment with exogenous hormones 50 μmol·L^-1 Gibberellin 3 (GA3) and 100 μmol·L^-1Salicylic acid (SA),the expression level of MsCOL2 showed different trends. In order to verify the effect of overexpression of MsCOL2 on the expression of flowering-related genes,the expression levels of flowering-related genes under overexpression MsCOL2 lines were analyzed. The results showed that the expression level of the genes promoting flowering was down-regulated and the expression level of the genes inhibiting flowering was up-regulated. Flowering phenotype analysis showed that overexpression of MsCOL2 delayed flowering,indicating that MsCOL2 plays an important role in regulating of delayed flowering.

Key words: Alfalfa, MsCOL2, Flowering time, Expression analysis

摘要： 开花是植物由营养生长过渡到生殖生长的转折点，紫花苜蓿（Medicago sativa L.）的产量和品质与开花时间密切相关。光周期是影响植物开花的重要因素之一，CONSTANS-like（COL）基因家族在光周期开花调控途径中发挥重要的作用。为揭示COL家族基因在紫花苜蓿开花调控中的功能，本研究克隆了MsCOL2基因，该基因编码401个氨基酸；组织差异性表达分析显示，MsCOL2在花中表达量最高；花不同发育时期表达分析显示，MsCOL2在花芽分化期表达量最高；在不同光周期条件下，MsCOL2的表达水平具有不同的节律性表达；外源激素50 μmol·L^-1赤霉素和100 μmol·L^-1水杨酸处理后，MsCOL2表达水平呈现不同的变化趋势。为了验证过表达MsCOL2对开花相关基因表达的影响，分析了过表达MsCOL2株系中开花相关基因的表达水平，结果显示促进开花基因的表达水平下调，抑制开花基因的表达水平上调。表型观察显示，过表达株系延迟开花，表明MsCOL2对延迟开花有重要的调节作用。

关键词: 紫花苜蓿(Medicago sativa L.), MsCOL2, 开花时间, 表达分析

CLC Number:

S541.9

LU Dong-yu, WANG Xue, JIANG Xu, ZHANG Yun-xiu, ZHANG Li-li, LI Ya-jing, KANG Jun-mei. Cloning and Functional Analysis of Flowering Regulation Gene MsCOL2 in Alfalfa[J]. Acta Agrestia Sinica, 2023, 31(10): 2905-2915.

卢栋宇, 王雪, 蒋旭, 张云秀, 张丽丽, 栗亚静, 康俊梅. 紫花苜蓿花期调控基因MsCOL2的克隆及功能研究[J]. 草地学报, 2023, 31(10): 2905-2915.

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Cloning and Functional Analysis of Flowering Regulation Gene MsCOL2 in Alfalfa

紫花苜蓿花期调控基因MsCOL2的克隆及功能研究

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