油用牡丹'丹凤'转录因子基因PoFUS3的克隆、表达及特征分析

张加强1,刘慧春1,王杰2,许雯婷1,周江华1,朱开元1,*
1浙江省农业科学院, 浙江省园林植物与花卉研究所, 杭州311251;2浙江省丽水市遂昌县农作物技术推广中心, 浙江丽水323300

通信作者:朱开元;E-mail: kyzhu1999@163.com

摘 要:

为了挖掘油用牡丹油脂合成相关转录因子, 以油用牡丹凤丹为试验材料, 采用RT-PCRRACE技术克隆PoFUS3基因全长, 并进行生物信息学和表达特征分析。研究结果显示, 该基因cDNA全长为1 217 bp, 开放阅读框长度为819 bp, 编码272个氨基酸, 相对分子量为31.18 kDa, 理论等电点为4.61, 无信号肽和跨膜域, 预测定位在细胞核, 属于含B3结构域的超家族。系统进化结果表明, PoFUS3与拟南芥AtFUS3亲缘关系最近。 RT-qPCR分析表明, PoFUS3凤丹中的表达具有组织特异性, 在荚果果皮中不表达, 在种子中的表达量最高。 PoFUS3在种子发育过程中呈现为先升高后降低的单峰型变化趋势, 在授粉后60 d时表达水平最高。

关键词:油用牡丹‘凤丹’; 转录因子; FUS3; 基因克隆; 表达分析

收稿:2020-11-17   修定:2021-03-04

资助:浙江省公益技术应用研究计划(LGN18C150007、 LGN19C150010)和2020年度浙江省农科院青年人才项目(2020R25R08E03)。

Cloning, expression and characterization of transcription factor gene PoFUS3 in Paeonia ostii

ZHANG Jiaqiang1, LIU Huichun1, WANG Jie2, XU Wenting1, ZHOU Jianghua1, ZHU Kaiyuan1,*
1Zhejiang Institute of Landscape Plants and Flowers, Zhejiang Academy of Agricultural Sciences, Hangzhou 311251, China; 2The Crop Technology Extension Center of Suichang County in Lishui City of Zhejiang Province, Lishui, Zhejiang 323300, China

Corresponding author: ZHU Kaiyuan; E-mail: kyzhu1999@163.com

Abstract:

In order to dig out the transcription factors related to the synthesis of oil from Paeonia ostii, the
full-length
PoFUS3 gene was cloned by RT-PCR and RACE technology, and the bioinformatics and expression characteristics were analyzed. The results showed that the full length cDNA of PoFUS3 was 1 217 bp, containing an 819 bp open reading frame which encoded 272 amino acids. The relative molecular weight of PoFUS3 was 31.18 kDa, and its isoelectric point was 4.61. There was no signal peptide or transmembrane domain in this protein, and it was predicted to be located in the nucleus, belonging to the superfamily containing the B3 domain. Phylogenetic results showed that PoFUS3 was closely related to AtFUS3. RT-qPCR analysis showed that the expression of PoFUS3 in P. ostii was tissue-specifc, it was not expressed in the pericarp of pod, and had the highest expression in seeds. The expression level of PoFUS3 was the highest at 60 days after pollination, and it showed a unimodal trend during seed development.

Key words: Paeonia ostii; transcription factor; FUS3; gene cloning; expression analysis

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