Screening and Functional Identification of Proteins Specifically Expressed in Terminal Buds of Different Fall Dormancy Alfalfa

doi:10.11733/j.issn.1007-0435.2022.01.007

Acta Agrestia Sinica ›› 2022, Vol. 30 ›› Issue (1): 55-61.DOI: 10.11733/j.issn.1007-0435.2022.01.007

Screening and Functional Identification of Proteins Specifically Expressed in Terminal Buds of Different Fall Dormancy Alfalfa

DU Hong-qi¹, LOU Zhi-guo¹, ZOU Jing³, FANG Wei-ping², FENG Chang-song¹

1. Institute of Animal Husbandry and Veterinary Research, Henan Academy of Agricultural Sciences, Zhengzhou, Henan Province 450002, China;
2. Economic Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou, Henan Province 450002, China;
3. Henan Xinyang animal husbandry workstation, Xinyang, Henan Province 464000, China

Received:2021-05-20 Revised:2021-08-05 Published:2022-01-27

不同秋眠型苜蓿顶芽中特异表达蛋白筛选及其功能鉴定

杜红旗¹, 娄治国¹, 邹靖³, 房卫平², 冯长松¹

1. 河南省农业科学院畜牧兽医研究所，河南郑州 450002;
2. 河南省农业科学院经济作物研究所，河南郑州 450002;
3. 河南省信阳市畜牧工作站，河南信阳 464000

通讯作者: 冯长松, E-mail: fengchangsong72@163.com
作者简介:杜红旗(1981-)，男，汉族，河南睢县人，助理研究员，博士，主要从事牧草分子生理研究和育种工作，E-mail: biodhq@126.com
基金资助:
河南省科技攻关项目(202102110084)；河南省博士后科研项目启动经费(19030086)；河南省农业科学院新兴学科发展专项项目(2020XK03、2021XK06)资助

Abstract

Abstract: To obtain proteins specifically expressed in the terminal buds of fall dormant and non-fall dormant alfalfa during the fall dormant period, and identify their function in growth and fall dormancy of alfalfa, prove that the proteins xpecifically expressed were not included in differential proteins obtained by the conventional high-throughput screening, We analyzed the presence or absence of proteins in the proteome results of terminal bud of fall dormancy Ⅰ standard variety 'Maverick' and fall dormancy Ⅸ alfalfa standard variety 'Cuf101' at fall dormant stage.Proteins specifically expressed in the apical buds of both varieties were screened. The mRNA relative expression level in the apical buds in spring, summer and autumn were detected by qRT-PCR. Transgenic plants were established by Agrobacterium tumefaciens transformation of seedlings injured cotyledon node. Phenotypic changes of transgenic plants were observed. The results showed that RBP47C protein was expressed specifically in the terminal buds of 'Maverick' alfalfa in autumn. Its mRNA was significantly highly expressed in the apical buds in autumn. The high expression of RBP47C inhibited growth of non-fall dormant alfalfa, while silencing RBP47C promoted the growth of fall dormant alfalfa. The hypothetical protein AES61470 was expressed specifically in the terminal buds of 'Cuf101' alfalfa. AES61470 mRNA level in apical buds of 'Cuf101'alfalfa was significant higher than that in apical buds of 'Maverick' alfalfa. Therefore, different fall dormancy alfalfa have specific expression proteins during fall dormant period, the highly specific expression of RBP47C protein promotes the fall dormancy of alfalfa, while its low expression accelerates the growth of alfalfa.

Key words: Alfalfa, Fall dormancy, High-Throughput, RBP47C, Specific expression

摘要： 为了获得秋眠时期秋眠型和非秋眠型苜蓿(Medicago sativa L.)顶芽中各自特异表达蛋白，鉴定其在苜蓿生长和秋眠中的功能，同时证明常规高通量筛选差异蛋白不包括特异表达蛋白，本研究拟分析秋眠时期秋眠1级苜蓿标准品种'Maverick’和9级苜蓿标准品种'Cuf101’顶芽蛋白质组结果中各蛋白在两品种顶芽中是否存在筛选各自特异表达蛋白，并应用qRT-PCR检测特异蛋白mRNA从春季至秋季在两品种苜蓿顶芽中相对表达量，应用农杆菌侵染子叶节点受伤幼苗建立各自的转基因植株并观察其表型变化。结果表明：在秋季，聚腺苷结合蛋白RBP47C在'Maverick’苜蓿顶芽中特异表达，其mRNA在'Maverick’苜蓿顶芽中显著特异高表达，其过表达抑制非秋眠苜蓿的生长，其沉默促进了秋眠型苜蓿的生长；假设蛋白AES61470在'Cuf101’苜蓿顶芽中特异表达, 其mRNA在'Cuf101’苜蓿顶芽中较'Maverick’苜蓿显著特异高表达。因此，不同秋眠型苜蓿在秋眠时期具有特异表达的蛋白，且RBP47C蛋白特异高表达促进了苜蓿的秋眠、低表达促进了苜蓿的生长。

关键词: 苜蓿, 秋眠, 特异表达, RBP47C, 高通量

CLC Number:

S551+.7

DU Hong-qi, LOU Zhi-guo, ZOU Jing, FANG Wei-ping, FENG Chang-song. Screening and Functional Identification of Proteins Specifically Expressed in Terminal Buds of Different Fall Dormancy Alfalfa[J]. Acta Agrestia Sinica, 2022, 30(1): 55-61.

杜红旗, 娄治国, 邹靖, 房卫平, 冯长松. 不同秋眠型苜蓿顶芽中特异表达蛋白筛选及其功能鉴定[J]. 草地学报, 2022, 30(1): 55-61.

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Screening and Functional Identification of Proteins Specifically Expressed in Terminal Buds of Different Fall Dormancy Alfalfa

不同秋眠型苜蓿顶芽中特异表达蛋白筛选及其功能鉴定

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