在棉属中全基因组鉴定 GhRLCK-VII 亚家族基因及其在抗黄萎病中的功能研究。

Genome-wide identification of GhRLCK-VII subfamily genes in Gossypium hirsutum and investigation of their functions in resistance to Verticillium wilt.

机构信息

College of Agronomy, Northwest A&F University, No.3 Taicheng Road, Yangling, 712100, Shaanxi, China.

College of Life Sciences, Northwest A&F University, Yangling, 712100, Shaanxi, China.

出版信息

BMC Plant Biol. 2023 Sep 11;23(1):421. doi: 10.1186/s12870-023-04435-0.

DOI:10.1186/s12870-023-04435-0

PMID:37697254

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10494381/

Abstract

BACKGROUND

The receptor-like cytoplasmic kinases subfamily VII (RLCK-VII) is critical in regulating plant growth, development, and pattern-triggered immunity. However, a comprehensive exploration of these genes in the allotetraploid Gossypium hirsutum is still lacking. This study aimed to identify RLCK-VII genes in G. hirsutum and investigate their evolutionary history, structural features, expression patterns, and role in plant defense.

RESULTS

Seventy-two RLCK-VII genes in the G. hirsutum genome were unveiled and classified into nine groups following their phylogenetic analysis with Arabidopsis thaliana. Group VII-1 was the largest, accounting for 28%, while Groups VII-2 and VII-3 had only one member each. The analysis using MCScanX revealed that these 72 genes formed 166 collinear gene pairs and were resided on 26 chromosomes of G. hirsutum, suggesting that they were derived from whole genome segmental duplication events. Their calculated Ka/Ks values were below one, implying the occurrence of purification selection during the evolution and inhibition of gene function differentiation/loss. All members of the RLCK-VII subfamily possessed two conserved domains, PKinase-Tyr and PKinase, and several conserved PBS1 kinase subdomains, individually included in one of the ten motifs identified using MEME. The RNA-Seq results showed that RLCK-VII genes exhibited different spatiotemporal expression, indicating their involvement in cotton growth, development, and defense responses to Verticillium dahliae. The transcription patterns of RLCK-VII genes found by RNA-Seq were further validated using qRT-PCR assays after inoculating "20B12" (cotton cultivar) with "V991" (V. dahliae). The virus-induced gene silencing (VIGS) assays uncovered that two RLCK-VII genes (Gohir.A13G227248 and Gohir.A10G219900) were essential to G. hirsutum resistance to Verticillium wilt.

CONCLUSIONS

These observations offer valuable insight into the attributes and roles of RLCK-VII genes in G. hirsutum, potentially enable the breeding of new cotton cultivars with enhanced resistance to Verticillium wilt.

摘要

背景

受体样细胞质激酶亚家族 VII（RLCK-VII）在调控植物生长、发育和模式触发免疫中起着关键作用。然而，对异源四倍体棉属（Gossypium hirsutum）中这些基因的全面研究仍有待深入。本研究旨在鉴定 G. hirsutum 中的 RLCK-VII 基因，并研究其进化历史、结构特征、表达模式及其在植物防御中的作用。

结果

通过与拟南芥（Arabidopsis thaliana）的系统发育分析，揭示了 G. hirsutum 基因组中的 72 个 RLCK-VII 基因，并将其分为 9 组。第 VII-1 组最大，占 28%，而第 VII-2 和 VII-3 组各只有一个成员。使用 MCScanX 分析表明，这些 72 个基因形成了 166 个共线性基因对，分布在 G. hirsutum 的 26 条染色体上，表明它们是由全基因组片段重复事件产生的。这些基因的 Ka/Ks 值均低于 1，表明在进化过程中发生了纯化选择，抑制了基因功能的分化/丧失。RLCK-VII 亚家族的所有成员都具有两个保守结构域，PKinase-Tyr 和 Pkinase，以及几个保守的 PBS1 激酶亚结构域，分别包含在使用 MEME 鉴定的 10 个基序之一中。RNA-Seq 结果表明，RLCK-VII 基因表现出不同的时空表达，表明它们参与了棉花生长、发育以及对黄萎病菌（Verticillium dahliae）的防御反应。使用 qRT-PCR 检测在接种黄萎病菌（V. dahliae）后“20B12”（棉花品种）的转录模式，进一步验证了 RNA-Seq 发现的 RLCK-VII 基因的转录模式。病毒诱导的基因沉默（VIGS）试验表明，两个 RLCK-VII 基因（Gohir.A13G227248 和 Gohir.A10G219900）对棉花抗黄萎病至关重要。