Sugarcane calcineurin B-like (CBL) genes play important but versatile roles in regulation of responses to biotic and abiotic stresses

Weihua Su,Hui Ling,Liping Xu,Huaying Mao,Long Huang,Ning Huang,Yongjuan Ren,Youxiong Que,Khushi Muhammad,Yachun Su,Dongjiao Wang

doi:10.1038/s41598-019-57058-7

Weihua Su, Hui Ling + Show 9 more

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https://doi.org/10.1038/s41598-019-57058-7

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Abstract

Free calcium ions are common second messengers in plant cells. The calcineurin B-like protein (CBL) is a special calcium sensor that plays an important role in plant growth and stress response. In this study, we obtained three CBL genes (GenBank accession nos. KX013374, KX013375, and KX013376) from sugarcane variety ROC22. The open reading frames of ScCBL genes ranged from 642 to 678 base pairs in length and encoded polypeptides from 213 to 225 amino acids in length. ScCBL2-1, ScCBL3-1, and ScCBL4 were all located in the plasma membrane and cytoplasm. ScCBL2-1 and ScCBL3-1 expression was up-regulated by treatment with salicylic acid (SA), methyl jasmonate (MeJA), hydrogen peroxide (H2O2), polyethylene glycol (PEG), sodium chloride (NaCl), or copper chloride (CuCl2). ScCBL4 expression was down-regulated in response to all of these stresses (abscisic acid (ABA), SA, MeJA, and NaCl) except for H2O2, calcium chloride (CaCl2), PEG, and CuCl2. Expression in Escherichia coli BL21 cells showed that ScCBLs can enhance tolerance to NaCl or copper stress. Overexpression of ScCBLs in Nicotiana benthamiana leaves promoted their resistance to infection with the tobacco pathogen Ralstonia solanacearum. The results from the present study facilitate further research regarding ScCBL genes, and in particular, their roles in the response to various stresses in sugarcane.

Highlights

Calcium (Ca2+) is an important inorganic nutritive element and a ubiquitous second messenger[1]
calcineurin B-like protein (CBL), a Ca2+ sensor relay protein that is expressed in a wide range of plants, can interact with a family of serine-threonine protein kinases known as CBL-interacting protein kinases (CIPKs)[7]
We performed the sampling after abiotic stress treatment as follows[33]: uniform four-month-old cultured ROC22 plantlets were transferred to water for one week and treated with eight exogenous treatments, including 100 μM Abscisic acid (ABA), 5 mM salicylic acid (SA), 100 μM methyl jasmonate (MeJA), 50 μM calcium chloride (CaCl2), 10 μM hydrogen peroxide (H2O2), 25% polyethylene glycol (PEG) 8000, 250 mM NaCl, or 100 mM copper chloride (CuCl2), by root dipping at 28 °C with 16 h light and 8 h darkness[34]

Summary

Introduction

Calcium (Ca2+) is an important inorganic nutritive element and a ubiquitous second messenger[1]. S. bicolor CBL genes are thought to regulate sodium carbonate stress-specific cellular adaptation responses and influence the plant growth and developmental patterns[14]. Analysis of CBL transcripts in Populus euphratica under abiotic stress suggested that seven CBL (PeCBL1, 2, 3, 4, 5, 9, and 10) members may play important roles in responding to specific external stimuli[12]. This study aims to provide useful information about the sequence characteristics of these three ScCBL proteins as well as their expression patterns in response to phytohormones and various stresses. This increased knowledge of ScCBL genes could be applied by sugarcane breeder to develop resistant variety

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