Functional analysis of the cotton CLE polypeptide signaling gene family in plant growth and development

Ke Wan,Yuxin He,Dong-Lei Yang,Xueying Guan,Guosheng Xiong,Ting Zhao,Xiaoyuan Tao,Kening Lu,Mengtao Gao

doi:10.1038/s41598-021-84312-8

Ke Wan, Yuxin He + Show 7 more

Open Access

https://doi.org/10.1038/s41598-021-84312-8

Copy DOI

Abstract

The CLAVATA3 (CLV3)/EMBRYO SURROUNDING REGION (ESR)–RELATED (CLE) gene family encodes a large number of polypeptide signaling molecules involved in the regulation of shoot apical meristem division and root and vascular bundle development in a variety of plants. CLE family genes encode important short peptide hormones; however, the functions of these signaling polypeptides in cotton remain largely unknown. In the current work, we studied the effects of the CLE family genes on growth and development in cotton. Based on the presence of a conserved CLE motif of 13 amino acids, 93 genes were characterized as GhCLE gene family members, and these were subcategorized into 7 groups. A preliminary analysis of the cotton CLE gene family indicated that the activity of its members tends to be conserved in terms of both the 13-residue conserved domain at the C-terminus and their subcellular localization pattern. Among the 14 tested genes, the ectopic overexpression of GhCLE5::GFP partially mimicked the phenotype of the clv3 mutant in Arabidopsis. GhCLE5 could affect the endogenous CLV3 in binding to the receptor complex, comprised of CLV1, CLV2, and CRN, in the yeast two-hybrid assay and split-luciferase assay. Silencing GhCLE5 in cotton caused a short seedling phenotype. Therefore, we concluded that the cotton GhCLE gene family is functionally conserved in apical shoot development regulation. These results indicate that CLE also plays roles in cotton development as a short peptide hormone.

Highlights

Intercellular communication is essential for the development of tissues and organs
To determine how the ectopic overexpression of GhCLE5 in Arabidopsis led to a phenotype partially resembling the Atclv[3] phenotype, we further examined the binding capacity of GhCLE5 in each of the receptor complexes of CLV1, CLV2, and CRN/CLV2
Based on the presented data, we propose that GhCLE5 could occupy the CLV3 motif binding sites with CLV1, CLV2, and CRN in the Arabidopsis ectopic overexpression line (Fig. 8)

Summary

Introduction

Peptide-receptor signaling modules play an important role in mediating intercellular communication and interactions during development, as well as responding to environmental stimuli[1]. The WUSCHEL-CLAVATA (WUS-CLV) feedback loop of Arabidopsis thaliana maintains stem cell homeostasis in shoot apical meristems (SAMs)[8,9,10]. In addition to maintain the homeostats of SAM, other CLE gene family members play important roles in multiple plant organ development control[25]. We carried out a functional analysis of CLE genes in the upland cotton (Gossypium hirsutum) genome. By means of a genome-wide study, translational analysis, protein interaction assay, and virus-induced gene silencing (VIGS) technology, we identified members of the cotton CLE gene and screened out the cotton GhCLE5 with ectopic effects in the reassembly of the Atclv[3] mutant

Methods

Results

Conclusion