Suppression of CaCYP1, a novel cytochrome P450 gene, compromises the basal pathogen defense response of pepper plants

https://doi.org/10.1016/j.bbrc.2006.04.124Get rights and content

Abstract

A putative cytochrome P450 gene from chili pepper, Capsicum annuum L. Bukang cytochrome P450 (CaCYP1), was identified using cDNA microarray analysis of gene expression following induction of the leaf hypersensitive response by inoculation of pepper plants with the non-host pathogen Xanthomonas axonopodis pv. glycines 8ra. The full-length cDNA of CaCYP1 encoded a protein of 514 amino acid residues, which contained a putative hydrophobic membrane anchoring domain in the N-terminal region, and a heme-binding motif in the C-terminal region. Analysis of the deduced amino acid sequence of CaCYP1 revealed that it has high homology to Arabidopsis CYP89A5, the function of which is unknown. Expression of CaCYP1 was preferentially increased in pepper plants in response to non-host pathogen inoculation and also during the host resistance response. CaCYP1 expression also increased following treatment with salicylic acid and abscisic acid, while treatment with ethylene had a mild effect. Using a virus-induced gene silencing-based reverse genetics approach, we demonstrated that suppression of CaCYP1 results in enhanced susceptibility to bacterial pathogens. Interestingly, gene silencing of CaCYP1 in pepper plants resulted in the reduced expression of the defense-related genes CaLTP1, CaSIG4, and Cadhn. Our results indicated that CaCYP1, a novel cytochrome P450 in pepper plants, may play a role in plant defense response pathways that involve salicylic acid and abscisic acid signaling pathways.

Section snippets

Materials and methods

Plant materials and pathogen inoculation. Chili pepper (Capsicum annuum L. cv. Bukang) and the bell pepper (C. annum L. cv. Early Calwonder 30R;bs1/bs1, bs2/bs2, Bs3/Bs3) were used in the current study. Peppers were grown in a plant growth room at 24 ± 1 °C with a photoperiod of 16 h light. Six weeks after seed germination, pepper plants were treated with bacterial pathogens or various chemicals. The bacterial pathogens used for inoculation were X. axonopodis pv. glycines 8ra (Xag 8ra), a soy bean

Isolation of pepper CaCYP1 using cDNA microarray analysis following non-host pathogen infection

To isolate pepper genes induced during the non-host bacterial pathogen HR, a pepper cDNA microarray was probed with RNA extracted from Xag8ra-infected hot pepper leaves. Xag8ra is not a pathogen of pepper, but does elicit an HR in pepper leaves, as well as induce the expression of a number of PR genes [30]. Microarray analysis indicated that 40 (out of 350 total ESTs) gene sequences were up-regulated more than 2-fold following Xag infiltration. This result was subsequently confirmed by reverse

Acknowledgments

This work was supported by the 21st frontier research programs of MOST, Crop Functional Genomics Center, Plant Diversity Research Center and in part by PMGBRC, one of the KOSEF science research Center.

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