《植物生理学报》 2018, 54(8): 1316-1324
通信作者：陶俊；E-mail: taojun@yzu.edu.cn
摘 要：
研究钙离子传感器CIPK (calcineurin B-like protein-interacting protein kinase)基因在芍药(Paeonia lactiflora)不同组织及在正反向钙处理后花茎中的表达变化, 为通过采前喷施钙溶液调控芍药花茎挺直度提供理论依据。本文实验以芍药‘红艳争辉’花茎RNA为模板, 采用PCR技术对芍药CIPK基因进行克隆, 并通过定量实时PCR技术对PlCIPK基因表达进行分析。结果显示, 克隆获得的PlCIPK基因(GeaBank MH748106)长1 467 bp, 具有由1 314个碱基组成的完整开放阅读框, 共编码438个氨基酸。PlCIPK基因在N端具有一个蛋白激酶催化域, 在C端具有一个高度保守的NAF调控域, 属于CIPK家族基因。PlCIPK基因在芍药各组织中均有表达, 但在花茎中表达最高; 在芍药花茎中, PlCIPK基因的表达随着生长发育逐渐增加; 采前钙处理提高了芍药花茎中PlCIPK基因的表达, 而钙螯合剂处理降低了其表达, 推测钙能够调控芍药花茎中PlCIPK基因的表达, 影响其强度。本研究从一定程度上说明钙处理使钙调磷酸酶B蛋白(CBL)-CIPK调控增强, 花茎品质提高, 也为应用钙提高芍药切花花茎挺直度提供一定的理论参考。关键词：芍药; CIPK; 克隆; 表达分析; 钙调控
收稿：2018-05-16 修定：2018-08-15
资助：国家自然科学基金(31572148和31772341)、扬州市优秀青年基金(YZ2017097)和扬州大学优秀青年骨干教师项目。
Corresponding author: TAO Jun; E-mail: taojun@yzu.edu.cn
Abstract:
To provide a theoretical basis for regulating the straightness of the inflorescence stems in herbaceous peony (Paeonia lactiflora) by spraying calcium solution before harvesting, the expression of calcium ion sensor calcineurin B-like protein-interacting protein kinase gene (CIPK) in different tissues of P. lactiflora and the inflorescence stems after treatment with positive and negative calcium were studied. In this study, RNA extracted from P. lactiflora ‘Hongyan Zhenghui’ inflorescence stem were taking as templates by using PCR method to clone PlCIPK and quantitative real-time PCR method to examine the expression levels of PlCIPK. The results show that PlCIPK obtained by the clone was 1 467 bp in length and has a complete open reading frame consisting of 1 314 bases, encoding a total of 438 amino acids. PlCIPK had a protein kinase catalytic domain at the N-terminus and a highly conserved NAF regulatory domain at the C-terminus, belonging to the CIPK family. PlCIPK was expressed in all tissues of P. lactiflora with the expression being the highest in the inflorescence stem; besides, the expression of PlCIPK in P. lactiflora inflorescence stem increased with the development. Preharvest calcium treatment increased the expression of PlCIPK in the P. lactiflora in florescence stems, while the calcium chelator treatment reduced its expression, indicating that calcium would regulate the expression of PlCIPK in the P. lactiflora inflorescence stem and affect its mechanical strength as a result. To a certain extent, this study suggests that calcium treatment could enhance the regulation of calcineurin B-like protein (CBL)-CIPK and improve the quality of inflorescence stems. In addition, it also provides a theoretical basis for the application of calcium in improving the straightness of P. lactiflora inflorescence stems.Key words: herbaceous peony; CIPK; cloning; expression analysis; calcium regulation
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