拟南芥转录因子AtOFP19功能的初步研究

doi:10.7668/hbnxb.20191995

摘要/Abstract

摘要： AtOFP19是植物中特有的蛋白质，是拟南芥卵形家族蛋白（AtOFPs）新发现的成员之一。为确定AtOFP19在植物体中的特定作用，采取PCR克隆方法，获得了AtOFP19基因，并利用生物信息学的方法对AtOFP19基因进行分析。采取农杆菌介导法浸染野生型拟南芥，获得35S∶HA-AtOFP19转基因植株。对比35S∶HA-AtOFP19转基因植株、atofp19突变体植株与野生型植株的形态差异，结果表明，atofp19表型与WT无明显差别，OE与同时期WT相比较，其植株莲座叶较小，茎细，植株整体较细小。此外还成功构建了PUC-GD-AtOFP19载体，应用原生质体瞬时转染技术，利用酶标仪测定GUS相对表达量，证明AtOFP19是转录抑制子。qRT-PCR测定分析表明，AtOFP19基因在拟南芥各个部位均有表达，在花中表达量最高。在拟南芥不同发育阶段中，AtOFP19基因表达量在拟南芥发育初期较高，随着拟南芥生长发育AtOFP19基因表达量降低，在开花期又升高。进一步探究AtOFP19基因在植物体内的功能，对比OE、atofp19与WT的种子萌发率，发现AtOFP19对种子萌发具有抑制作用。为探究AtOFP19基因对激素的反应，外源添加5种激素于1/2MS培养基中，挑选生长状况良好的幼苗于土壤中培养，进行qRT-PCR分析，结果表明AtOFP19基因的表达主要受到6-BA激素的调控。以上结果表明，AtOFP19是转录抑制子，对种子萌发有抑制作用，且在植株体内主要受到6-BA调控。

关键词: 拟南芥, AtOFP19基因, 种子萌发率, 植物激素

Abstract: AtOFP19 is a unique protein in plants and it is a new member of Arabidopsis ovate family proteins(AtOFPs).To determine the specific function of AtOFP19, PCR was used to obtain AtOFP19 gene, and bioinformatics analysis was conducted.Using the method of Agrobacterium-mediated constructed 35S:HA-AtOFP19 transgenic plants. Comparing the morphological diffenerces between 35S:HA-AtoFP19 transgenic, atofp19 mutant, and wild-type plants, there is no significant difference in atofp19 and WT. Comparing with WT in the same period, OE had small leaflets, thin stem and the whole plant was small. Successfully constructing the PUC-GD-AtOFP19 vector, the relative expression of GUS was determined by enzyme-labelled meter with protoplast transient transfection technique, which proved that AtOFP19 was a transcriptional suppressor.qRT-PCR analysis showed that the AtOFP19 gene was expressing in all plant and the expression in flowers was highest. In the early stage of development, the expression of AtOFP19 gene decreased and increased in flowering stage.To explore the function of AtOFP19 gene in plants, the seed germination rate of OE, atofp19 and WT were compared, and it showed that AtOFP19 had inhibitory effect.To explore the response of AtOFP19 gene to plant hormones, five hormones were applied in 1/2 MS culture medium. Seedlings with good growth condition were cultured in soil. qRT-PCR analysis showed that AtOFP19 gene was regulated by 6-BA. The results showed that AtOFP19 gene was a transcriptional repressor, which inhibited seed germination and was mainly regulated by 6-BA.

Key words: Arabidopsis thaliana, AtOFP19 gene, Seed germination rate, Plant hormone

中图分类号:

Q946.2
S188

韩佳良, 王鹤萌, 张冬瑞, 常缨. 拟南芥转录因子AtOFP19功能的初步研究[J]. 华北农学报, 2021, 36(4): 56-63. doi: 10.7668/hbnxb.20191995.

HAN Jialiang, WANG Hemeng, ZHANG Dongrui, CHANG Ying. Preliminary Study on the Function of Transcription Factor AtOFP19 in Arabidopsis thaliana[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 56-63. doi: 10.7668/hbnxb.20191995.

http://www.hbnxb.net/CN/Y2021/V36/I4/56

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