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Functional analysis of MeCIPK23 and MeCBL1/9 in cassava defense response against Xanthomonas axonopodis pv. manihotis

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Abstract

Key message

MeCIPK23 interacts with MeCBL1/9, and they confer improved defense response, providing potential genes for further genetic breeding in cassava.

Abstract

Cassava (Manihot esculenta) is an important food crop in tropical area, but its production is largely affected by cassava bacterial blight. However, the information of defense-related genes in cassava is very limited. Calcium ions play essential roles in plant development and stress signaling pathways. Calcineurin B-like proteins (CBLs) and CBL-interacting protein kinases (CIPKs) are crucial components of calcium signals. In this study, systematic expression profile of 25MeCIPKs in response to Xanthomonas axonopodis pv. manihotis (Xam) infection was examined, by which seven candidate MeCIPKs were chosen for functional investigation. Through transient expression in Nicotiana benthamiana leaves, we found that six MeCIPKs (MeCIPK5, MeCIPK8, MeCIPK12, MeCIPK22, MeCIPK23 and MeCIPK24) conferred improved defense response, via regulating the transcripts of several defense-related genes. Notably, we found that MeCIPK23 interacted with MeCBL1 and MeCBL9, and overexpression of these genes conferred improved defense response. On the contrary, virus-induced gene silencing of either MeCIPK23 or MeCBL1/9 or both genes resulted in disease sensitive in cassava. To our knowledge, this is the first study identifying MeCIPK23 as well as MeCBL1 and MeCBL9 that confer enhanced defense response against Xam.

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Abbreviations

ABA:

Abscisic acid

BiFC:

Biomolecular fluorescence complementation

CaM:

Calmodulin

CBL:

Calcineurin B-like protein

CIPK:

CBL-interacting protein kinase

CML:

Calmodulin-like protein

CPK:

Calcium-dependent protein kinase

DAPI:

4′,6-Diamidino-2-phenylindole

dpi:

Day post infiltration

EL:

Electrolyte leakage

GFP:

Green fluorescent protein

H+ :

Hydrogen ion

H2O2 :

Hydrogen peroxide

MAMPs/PAMPs:

Microbe/pathogen-associated molecular patterns

NOX:

NADPH oxidase

NPR1:

Nonexpresser of PR genes 1

O2 ·− :

Superoxide radical

PM:

Plasma membrane

PP2C:

2C-type protein phosphatase

PPI:

Protein–phosphatase interaction

PR:

Pathogensis-related gene

Rboh:

Respiratory burst oxidase homologues

ROS:

Reactive oxygen species

VIGS:

Virus-induced gene silencing

Xam :

Xanthomonas axonopodis pv. manihotis

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Acknowledgements

We thank Dr. Chris R. Somerville, Dr. Jie Zhou, Dr. Yanru Hu, and Dr. Jiang Tian for sharing their vector plasmids. This research was supported by the National Natural Science Foundation of China (No. 31760067), the startup funding and the scientific research foundation of Hainan University (No. kyqd1531) to Haitao Shi.

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  1. Yu Yan, Xinyi He and Wei Hu have contributed equally to this work.

Authors and Affiliations

  1. Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources and College of Biology, Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, 570228, China

    Yu Yan, Xinyi He, Guoyin Liu, Peng Wang, Chaozu He & Haitao Shi

  2. Key Laboratory of Biology and Genetic Resources of Tropical Crops, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Xueyuan Road 4, Haikou, 571101, Hainan Province, China

    Wei Hu

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  1. Yu Yan
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  2. Xinyi He
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  4. Guoyin Liu
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Correspondence to Haitao Shi.

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Communicated by Ying-Tang Lu.

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Yan, Y., He, X., Hu, W. et al. Functional analysis of MeCIPK23 and MeCBL1/9 in cassava defense response against Xanthomonas axonopodis pv. manihotis. Plant Cell Rep 37, 887–900 (2018). https://doi.org/10.1007/s00299-018-2276-7

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