Department of Horticulture, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China.
BMC Plant Biol. 2021 Oct 30;21(1):499. doi: 10.1186/s12870-021-03279-w.
Papain-like cysteine proteases (PLCPs), a large group of cysteine proteases, are structurally related to papain. The members belonging to PLCPs family contribute to plant immunity, senescence, and defense responses in plants. The PLCP gene family has been identified in Arabidopsis, rice, soybean, and cotton. However, no systematic analysis of PLCP genes has been undertaken in grapevine. Since Plasmopara viticola as a destructive pathogen could affect immunity of grapes in the field, we considered that the members belonged to PLCPs family could play a crucial role in defensive mechanisms or programmed cell death. We aimed to evaluate the role of PLCPs in 2 different varieties of grapevines and compared the changes of their expressions with the transcriptional data in response to P. viticola.
In this study, 23 grapevine PLCP (VvPLCP) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Numerous cis-acting elements related to plant development, hormone, and stress responses were identified in the promoter of the VvPLCP genes. Phylogenetic analysis grouped the VvPLCP genes into nine subgroups. The transcription of VvPLCP in different inoculation time points and varieties indicated that VvPLCP may have vital functions in grapevine defense against Plasmopara viticola. According to transcriptome data and qPCR analysis, we observed the increasing expression levels of VvRD21-1 at 72 h after inoculation in resistant variety, inferring that it was related to grape downy mildew resistance. Meanwhile, 3 genes including VvXBCP1, VvSAG12-1, and VvALP1 showed higher expression at 24 h after pathogen inoculation in the susceptible variety and might be related to the downy mildew phenotype. We nominated these four genes to function during hypersensitive response (HR) process, inferring that these genes could be associated with downy mildew resistance in grapes.
Our results provide the reference for functional studies of PLCP gene family, and highlight its functions in grapevine defense against P. viticola. The results help us to better understand the complexity of the PLCP gene family in plant immunity and provide valuable information for future functional characterization of specific genes in grapevine.
木瓜蛋白酶样半胱氨酸蛋白酶(PLCPs)是一大类半胱氨酸蛋白酶,与木瓜蛋白酶在结构上相关。属于 PLCP 家族的成员有助于植物的免疫、衰老和防御反应。PLCP 基因家族已在拟南芥、水稻、大豆和棉花中被鉴定出来。然而,在葡萄中尚未进行 PLCP 基因的系统分析。由于葡萄霜霉病菌作为一种破坏性病原体,可能会影响田间葡萄的免疫力,因此我们认为属于 PLCPs 家族的成员可能在防御机制或程序性细胞死亡中发挥关键作用。我们旨在评估 PLCPs 在 2 个不同葡萄品种中的作用,并比较其表达变化与对 Plasmopara viticola 的转录数据。
在这项研究中,通过综合生物信息学分析,鉴定出 23 个葡萄 PLCP(VvPLCP)基因。随后,对这些基因的染色体定位、基因结构、保守结构域、系统进化关系、基因复制和顺式作用元件进行了分析。在 VvPLCP 基因的启动子中发现了许多与植物发育、激素和应激反应相关的顺式作用元件。系统进化分析将 VvPLCP 基因分为 9 个亚组。在不同接种时间点和品种中 VvPLCP 的转录表明,VvPLCP 可能在葡萄抵御 Plasmopara viticola 方面具有重要功能。根据转录组数据和 qPCR 分析,我们观察到在抗性品种中接种后 72 小时 VvRD21-1 的表达水平增加,推断其与葡萄霜霉病抗性有关。同时,在感病品种中,3 个基因(包括 VvXBCP1、VvSAG12-1 和 VvALP1)在接种病原菌后 24 小时表达水平较高,可能与霜霉病表型有关。我们将这 4 个基因命名为参与过敏反应(HR)过程的基因,推断它们可能与葡萄霜霉病抗性有关。
我们的结果为 PLCP 基因家族的功能研究提供了参考,并强调了其在葡萄抵御 P. viticola 中的作用。这些结果有助于我们更好地理解植物免疫中 PLCP 基因家族的复杂性,并为未来葡萄中特定基因的功能表征提供有价值的信息。