李乾玉 王秀美 倪珊珊 王乐 张舒婷 夏朝水 叶炜 陈裕坤 赖钟雄

摘  要：以非洲菊‘G088為材料，基于本研究所测序得到的非洲菊转录组数据（SRA： SRR9937065）的基础上，采用RT-PCR技术克隆获得2个病程相关蛋白1（PR-1）基因，分别命名为GjPR-1-like1和GjPR-1-like2（GenBank登录号：MW057342和MW057343），GjPR-1-like1和GjPR-1-like2编码序列长度分别为534 bp和489 bp，可编码177和162个氨基酸。生物信息学分析表明，GjPR-1-like1属于碱性不稳定性蛋白，GjPR-1-like2属于弱酸性稳定蛋白;它们编码的蛋白质均含有CAP_PR-1和CAP superfamily保守结构域，属于富含半胱氨酸超家族蛋白，含有信号肽、跨膜结构和磷酸化位点;亚细胞定位表明均位于液泡中。PR-1基因在不同组织（叶、叶柄和根）、不同激素与逆境（SA、MeJA、ABA、NaCl）以及根腐病病原菌处理非洲菊的表达模式分析结果表明，GjPR-1-like1和GjPR-1-like2均在叶片中表达水平最高，在SA、MeJA、ABA和NaCl胁迫以及根腐病病原菌处理下，均显著影响GjPR-1-like1和GjPR-1-like2的表达水平。研究表明，GjPR-1-like1和GjPR-1-like2可能参与调控非洲菊生长发育过程，尤其是叶片的发育过程;且可能参与了非洲菊生物及非生物胁迫过程，尤其在响应非洲菊根腐病的抗病防御过程中发挥了一定的作用。

关键词：非洲菊;病程相关蛋白1;基因克隆;激素;根腐病

中图分类号：Q949.783.5      文献标识码：A

Cloning and Expression Analysis of PR-1 Genes in Gerbera jamesonii

LI Qianyu1， WANG Xiumei1， NI Shanshan1， WANG Le1， ZHANG Shuting1， XIA Chaoshui2， YE Wei2， CHEN Yukun1*， LAI Zhongxiong1*

1. Institute of Horticultural Biotechnology， Fujian Agriculture and Forestry University， Fuzhou， Fujian 350002， China; 2. Sanming Academy of Agricultural Sciences， Sanming， Fujian 365000， China

Abstract： In this study， based on the transcriptome database of Gerbera jamesonii （SRA： SRR9937065）， we successfully cloned two PR-1 genes， named as GjPR-1-like1 and GjPR-1-like2 （GenBank accession number： MW057342 and MW057343）， from G. jamesonii cv.‘G088 using the RT-PCR method. The coding sequence of GjPR-1-like1 and GjPR-1-like2 was 534 bp and 489 bp， encoding 177 and 162 amino acids. Bioinformatics analysis indicated that GjPR-1-like1 and GjPR-1-like2 were belonged to the basic unstable protein and acid stable protein; all the two GjPR-1s possessed CAP_PR-1 and CAP superfamily conserved domain， belonged to cysteine-rich secretory super family proteins and contained signal peptide， transmembrane structure and phosphorylation sites; subcellular localization indicated that they both located in the vacuole. The expression patterns of PR-1 in different tissues （including leaf， petiole and root）， and under different adversity stresses （including SA， MeJA， ABA， NaCl treatments） and Phytophthora cryptogea treatments in Gerbera transplant seedlings， the results showed that both GjPR-1-like1 and GjPR-1-like2 showed the highest expression level in the leaf. Under stress conditions of SA， MeJA， ABA， NaCl treatments and P. cryptogea treatments， the expression level of GjPR-1-like1 and GjPR-1-like2 were significantly changed. GjPR-1-like1 and GjPR-1-like2 may participate in the regulation of the growth and development process of G. jamesonii particularly in leaf development， and were concerned with biotic stresses and abiotic stresses， especially play a certain role in the process of disease resistance and defense in response to the gerbera P. cryptogea interaction.

Keywords： Gerbera jamesonii; pathogenesis-related protein 1; gene clone; hormone; Phytophthora cryptogea

DOI： 10.3969/j.issn.1000-2561.2021.07.006

植物生长发育的过程中会面临多种病毒、真菌以及外界胁迫的侵袭。为了保护自己免受侵袭，植物细胞壁硬化并产生抗菌化合物（植物抗毒素）和抗菌蛋白[1-2]，启动体内的防御机制。植物病程相关蛋白（pathogenesis-related proteins，PRs）是植物在受病原菌侵染过程中诱导产生的[3]，在植物抗病过程中发挥重要作用。根据PRs蛋白家族的类型及特性主要归纳为17个家族[4-5]，其中PR-1家族是PRs家族中重要的多基因家族，属于富含半胱氨酸的分泌蛋白、抗原5和病程相关蛋白1超家族（CAP superfamily）[6]，并且进化上高度保守。……