摘要
【目的】解析涮辣与昆明皱皮椒苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)的差异,研究不同发育时期、不同环境和外源因子处理下PAL基因表达规律。【方法】基于基因克隆和生物信息学方法研究PAL基因和蛋白特性;利用荧光定量PCR及酶活性测定方法测定PAL表达量及酶活性。【结果】克隆得到涮辣PAL基因为2166 bp,昆明皱皮椒PAL基因为2154 bp,分别编码721和717个氨基酸。顺式元件预测结果显示:辣椒PAL可能受赤霉素、光与干旱的影响。PAL蛋白预测结果表明:PAL为脂溶性、亲水性的稳定蛋白,主要定位于质膜,无信号肽与跨膜结构,含58个磷酸化位点。在进化上PAL有保守性,但在亚组上存在一定差异。在不同发育阶段,2种辣椒PAL表达水平与酶活性的趋势都为先升高后急剧降低,且在大部分发育阶段均表现为露地栽培高于大棚栽培;同一环境条件下,涮辣PAL表达与酶活性整体高于昆明皱皮椒;且不同外源物质在一定时间内可影响涮辣PAL的表达。【结论】涮辣与昆明皱皮椒PAL基因和蛋白特性相似但存在差异,在辣椒生长发育、环境和外源物质响应方面有重要功能。
[Purpose]To analyze the differences between ShuanLa and Kunming wrinkled pepper phenylalanine ammonia-lyase(PAL), exploring the change rules of ShuanLa and Kunming wrinkled pepper PAL expression in different development stages, different environments and different exogenous factors. [Methods]The PAL gene and protein characteristics were studied based on gene cloning and bioinformatics methods. The relative expression level and enzyme activity of PAL were determined by qPCR and enzyme activity assay. [Results]The PAL gene of ShuanLa and Kunming wrinkled pepper was 2 166 bp and 2 154 bp, encoding 721 and 717 amino acids, respectively. The ciselement prediction results showed that the pepper’s PAL gene might be regulated by gibberellin, light and drought. The PAL protein prediction showed that it was fat soluble, hydrophilic and stable protein. PAL proteins had no signal peptide, no transmembrane structure, and both had 58 phosphorylation sites. It was evolutionarily conservative, but there were some differences in subgroups. At different developmental stages, the expression of PAL and enzyme activity increased first and then decreased sharply, and in most developmental stages, the expression of PAL and enzyme activity were higher in open field than in greenhouse. Under the same environmental conditions, the expression and enzyme activity of ShuanLa PAL were higher than those of Kunming wrinkled pepper. Different exogenous substances affected the expression of shabulipal in a certain time. [Conclusion]The PAL gene and protein characteristics of ShuanLa and Kunming wrinkled pepper were similar but different,which played an important role in pepper growth and development, environment and response to exogenous factor.
作者
张祥
刘雨婷
张婧柔
李平平
谢志和
张芮豪
邓明华
ZHANG Xiang;LIU Yuting;ZHANG Jingrou;LI Pingping;XIE Zhihe;ZHANG Ruihao;DENG Minghua(College of Landscape and Horticulture,Yunnan Agricultural University,Kunming 650201,China;DENG Minghua Expert Basic Scientific Research Workstation of Yuxi City,Economic Crop Workstation of Tonghai County,Tonghai 652700,China)
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第2期320-329,共10页
Journal of Yunnan Agricultural University:Natural Science
基金
云南省科技厅重大专项(2018BB020)
玉溪市专家基层科研工作站。
