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Knockdown of the chitin-binding protein family gene CaChiIV1 increased sensitivity to Phytophthora capsici and drought stress in pepper plants

  • Muhammad Ali
  • Wen-Xian Gai
  • Abdul Mateen Khattak
  • Abid Khan
  • Saeed Ul Haq
  • Xiao Ma
  • Ai-Min Wei
  • Izhar Muhammad
  • Ibadullah Jan
  • Zhen-Hui GongEmail author
Original Article
  • 15 Downloads

Abstract

Phytophthora capsici has been the most destructive pathogen of pepper plants (Capsicum annuum L.), possessing the ability to quickly overcome the host defense system. In this context, the chitin-binding protein (CBP) family member CaChiIV1 regulates the response to P. capsici and abiotic stresses. The relevance of functional characterization and regulation of CaChiIV1 has not been explored in horticultural crops, especially pepper plants. The target gene (CaChiIV1) was isolated from pepper plants and cloned; the encoded protein carries a chitin-binding domain (CBD) that is rich in cysteine residues and has a hinge region with an abundance of proline and glycine residues. Additionally, the conserved regions in the promoter have a remarkable motif, “TTGACC”. The expression of CaChiIV1 was markedly regulated by methyl-jasmonate (MeJA), hydrogen peroxide (H2O2), melatonin, mannitol and P. capsici (PC and HX-9) infection. Knockdown of CaChiIV1 in pepper plants increased sensitivity to P. capsici (PC strain). Higher malondialdehyde (MDA) content and relative electrolyte leakage (REL) but lower antioxidant enzyme activities, chlorophyll content, root activity, and proline content were observed in CaChiIV1-silenced plants than in control plants. In conclusion, CaChiIV1-silenced pepper plants displayed increased susceptibility to P. capsici infection due to changes in expression of defense-related genes, thus showing its coregulation affect in particular conditions. Furthermore, antioxidant enzymes and proline content were largely diminished in CaChiIV1-silenced plants. Therefore, this evidence suggests that the CaChiIV1 gene plays a prominent role in the defense mechanism of pepper plants against P. capsici infection. In the future, the potential role of the CaChiIV1 gene in defense regulatory pathways and its coregulation with other pathogen-related genes should be identified.

Keywords

Biotic stress CaChiIV1 gene Chitin-binding protein Chitinase Pepper Phytophthora capsici Proline Resistance 

Abbreviations

APX

Ascorbate peroxidase

CBD

Chitin-binding domain

CBP

Chitin-binding protein

DAB

Diaminobenzidine

ETI

Induce effector-triggered immunity

GFP

Green fluorescent protein

HR

Hypersensitive response

MDA

Malondialdehyde

mg/g

Milligram per gram

NBT

Nitro-blue tetrazolium

ng/µL

Nano-gram per micro liter

ORF

Open reading frame

PAMPs

Pathogen-associated molecular patterns

PDA

Potato dextrose agar

PDS

Phytoene desaturase

qRT-PCR

Quantitative real-time polymerase chain reaction

REL

Relative electrolyte leakage

TBARS

Thiobarbituric acid-reactive substances

TRV

Tobacco rattle virus

TTC

Triphenyl-tetrazolium chloride

Ubi

Ubiquitin-conjugating protein

UTR

Untranslated region

VIGS

Virus-induced gene silencing

Notes

Author contributions

MA and ZHG conceived the study. MA collected the data and performed the experiment. AK and SUH performed the data analysis. WXG and XA performed the supplementary experiment during revision. MA drafted the manuscript. IM, AMK and IJ review and edited the manuscript. ZHG, WAM funding acquisition. All authors critically reviewed, read and approved the final manuscript.

Funding

This work was supported through funding from National Key R&D Program of China (No. 2016YFD0101900) and the National Natural Science Foundation of China (No. U1603102, No. 31272163).

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

438_2019_1583_MOESM1_ESM.docx (16 kb)
Supplementary material 1 (DOCX 16 kb)

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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of HorticultureNorthwest A&F UniversityYanglingChina
  2. 2.Department of HorticultureThe University of AgriculturePeshawarPakistan
  3. 3.College of Information and Electrical EngineeringChina Agricultural UniversityBeijingPeople’s Republic of China
  4. 4.Tianjin Vegetable Research CenterTianjinPeople’s Republic of China
  5. 5.State Key Laboratory of Crop Stress Biology in Arid Areas, College of AgronomyNorthwest A&F UniversityYanglingChina
  6. 6.Department of AgricultureUniversity of SwabiKhyber PakhtunkhwaPakistan

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