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


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.


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



Ascorbate peroxidase


Chitin-binding domain


Chitin-binding protein




Induce effector-triggered immunity


Green fluorescent protein


Hypersensitive response




Milligram per gram


Nitro-blue tetrazolium


Nano-gram per micro liter


Open reading frame


Pathogen-associated molecular patterns


Potato dextrose agar


Phytoene desaturase


Quantitative real-time polymerase chain reaction


Relative electrolyte leakage


Thiobarbituric acid-reactive substances


Tobacco rattle virus


Triphenyl-tetrazolium chloride


Ubiquitin-conjugating protein


Untranslated region


Virus-induced gene silencing


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.


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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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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