Reaction of Chilli (Capsicum annuum L.) Accessions for Mixed Infection of Colletotrichum capsici and Colletotrichum gloeosporioides and Molecular Analysis of Associated NBS-LLR Disease Resistant Gene Analogues


Published: 2022-02-05

Page: 111-124

K. A. S. I. Kumari

Field Crops Research and Development Institute, Mahailluppallama, Sri Lanka.

D. M. J. B. Senanayake *

Field Crops Research and Development Institute, Mahailluppallama, Sri Lanka.

W. A. R. Dhammika

Field Crops Research and Development Institute, Mahailluppallama, Sri Lanka.

W. M. K. Fernando

Field Crops Research and Development Institute, Mahailluppallama, Sri Lanka.

P. J. K. Dasanayake

Field Crops Research and Development Institute, Mahailluppallama, Sri Lanka.

C. M. Nanayakkara

University of Colombo, Colombo, Sri Lanka.

A. Balasuriya

Rajarata University of Sri Lanka, Mihinthale, Sri Lanka.

*Author to whom correspondence should be addressed.


This study was carried out to investigate the phenotypic reaction of 21 selected chilli accessions available in Sri Lanka for chilli anthracnose caused by mixed infection of Colletotrichum capsici and Colletotrichum gloeosporioides and protein level changes underlying with the plant response. A mixed inoculum of 105 conidia ml-1 of fungus was used and disease severity index (DSI) were calculated. A set of primers, designed based on resistant gene analogues (RGA) was used for PCR amplification of selected possible resistant gene sequences. Amino acid profiles were generated and were compared with similar amino acid profiles available in the NCBI. The results revealed that chilli accessions show varying degrees of susceptibility from low to high for anthracnose and none were resistant to anthracnose. The lowest DSI values reported from MI Hot (19.47), Hen miris (24.73), Galkiriyagama selection (26.74), C. baccatum (26.95) and MICH3 (27.03). The amino acid profiles showed that RGAs of chilli accessions, except MICH3 belonged to NB-ARC domain. Homology analysis on amino acid profiles showed a high diversity among the studied RGAs and varying susceptible reactions observed among 21 chilli accessions to the anthracnose disease caused by C. capcisi and C. gloeosporioides may be due to non-formation of required proteins as a result. MI Hot, Hen Miris, Galkiriyagama selection, C. baccatum and MICH3, which showed lower susceptibility to anthracnose disease, can be used in different combinations in breeding programmes to improve anthracnose resistance in chilli.  Molecular and protein level findings of this study may be useful in creating genetic resistance for anthracnose caused by C. capsici and C. gloeosporioides.

Keywords: Anthracnose resistance, Colletotrichum capsici, Colletotrichum gloeosporioides, Capsicum annuum, resistant gene analogues

How to Cite

Kumari, K. A. S. I., Senanayake, D. M. J. B., Dhammika, W. A. R., Fernando, W. M. K., Dasanayake, P. J. K., Nanayakkara, C. M., & Balasuriya, A. (2022). Reaction of Chilli (Capsicum annuum L.) Accessions for Mixed Infection of Colletotrichum capsici and Colletotrichum gloeosporioides and Molecular Analysis of Associated NBS-LLR Disease Resistant Gene Analogues. Asian Research Journal of Current Science, 4(1), 111–124. Retrieved from


Download data is not yet available.


Than PP, Prihastuti H, Phoulivong S, Taylor PW, Hyde KD. Chilli anthracnose disease caused by Colletotrichum species. Journal of Zhejiang University Science B. 2008;9(10):764.

Agstat. Pocket book of Agricultural statistics Socio- Economic and Planning Center. Department of Agriculture, Peradeniya, Sri Lanka. 2019;15:23.

Senanayake DMJB, Jayasinghe JEARM, Shilpi S, Wasala SK, Mandal B. A new begomovirus–betasatellite complex is associated with chilli leaf curl disease in Sri Lanka. Virus Genes. 2013;46(1):128-139.

Mongkolporn O, Taylor PWJ. Chili anthracnose: Colletotrichum taxonomy and pathogenicity. Plant Pathology. 2018; 67(6):1255-1263.

Rajapakse RGAS, Ranasinghe JADAR. Development of variety screening method for anthracnose disease of chilli (Capsicum annum L.) under field conditions. Tropical Agricultural Research and Extension. 2002;5(1-2):7-11.

Rajapakse, R.G.A.S. 1998. Observations on anthracnose of chilli pepper (Capsicum annuum L.) caused by Colletotrichum species in Sri Lanka. Doctoral dissertation. Imperial College London University of London. London.

Don LD, Van Phuong TT, Vy TT, Kieu PTM. Colletotrichum spp. attacking on chilli pepper growing in Vietnam. Country Report. In Abstracts of the First International Symposium on Chilli Anthracnose. National Horticultural Research Institute, Rural Development of Administration, Republic of Korea. 2007;24.

Byung SK. Country report of Anthracnose research in Korea. In First International Symposium on Chili Anthracnose, Hoam Faculty House, Seoul National University (Seoul). 2007;24.

Ramachandran N, Rathnamma K. Colletotrichum acutatum a new addition to the species of chilli anthracnose pathogen in India. In Annual Meeting and Symposium of Indian Phytopathological Society, Central Plantation Crops Research Institute (Kasarago).

Flor HH. Inheritance of reaction to rust in flax. J. agric. Res. 1947;74(9):41.

Ravensdale M, Nemri A, Thrall PH, Ellis JG, Dodds PN. Co‐evolutionary interactions between host resistance and pathogen effector genes in flax rust disease. Molecular Plant Pathology. 2011;12(1):93-102.

Leister D, Ballvora A, Salamini F, Gebhardt C. A PCR–based approach for isolating pathogen resistance genes from potato with potential for wide application in plants. Nature genetics. 1996;14(4):421-429

He CY, Tian AG, Zhang JS, Zhang ZY, Gai JY, Chen SY. Isolation and characterization of a full-length resistance gene homolog from soybean. Theoretical and Applied Genetics. 2003; 106(5):786-793.

Collins NC, Webb CA, Seah S, Ellis JG, Hulbert SH, Pryor A. The isolation and mapping of disease resistance gene analogs in maize. Molecular plant-microbe interactions. 1998;11(10): 968-978.

Kastoori RR, Talluri KB, Subramonian S, Kodeboina VS, Pandravada SR, Ganta A, Khagendra MM. Isolation of Resistance Gene Candidates in Chilli and Use of Molecular Markers for Root Knot Nematode Resistance. International Journal of Plant Breeding. 2012;6(1):47-52

Mahasuk P, Taylor PWJ, Mongkolporn O. Identification of two new genes conferring resistance to Colletotrichum acutatum in Capsicum baccatum. Phytopathology. 2009;99(9):1100-1104.

Park HG. Problems of anthracnose in pepper and prospects for its management. In The First International Symposium on Chili Anthracnose, Convention Center, Seoul National University, Korea. 2007;19.

Sharma PN, Kaur M, Sharma OP, Sharma P, Pathania A. Morphological, pathological and molecular variability in Colletotrichum capsici, the cause of fruit rot of chillies in the subtropical region of north western India J. Phytopathol. 2005;153:232–237.

Montri P, Taylor PWJ, Mongkolporn O. Pathotypes of Colletotrichum capsici, the causal agent of chili anthracnose, in Thailand. Plant Disease. 2009;93(1):17-20.

MacQueen A, Bergelson J. Modulation of R-gene expression across environments. Journal of experimental botany. 2016;67(7):2093-2105.

Smith BJ, Black LL. Morphological, cultural, and pathogenic variation among Colletotrichum species isolated from strawberry. Plant Dis. 1990;74:69-76

Simmonds JH. A study of the species of Colletotrichum causing ripe fruit rots in Queensland. Queensland Journal Agriculture and Animal Science. 1965;22(4):437-459.

Yoon JB, Park HG. Screening method for resistance to pepper fruits anthracnose: Pathogen sporulation, inoculation methods related to inoculum concentrations, post-inoculation environments. Journal of Korean Society and Horticultural Science. 2001;42:382-393.

Yoon JB. Identification of Genetic Resource, Interspecific Hybridization and Inheritance Analysis for Breeding Pepper (Capsicum annuum) Resistant to Anthracnose. Ph.D. Thesis, Seoul National University, Korea; 2003.

Martin GB, Bogdanove AJ, Sessa G. Understanding the functions of plant disease resistance proteins. Annual Review of Plant Biology. 2003;54(1):23-61.

Sekhwal MK, Li P, Lam I, Wang X, Cloutier S, You FM. Disease resistance gene analogs (RGAs) in plants. International Journal of Molecular Sciences. 2015;16(8):19248-19290.

Totad AS, Fakrudin B, Kuruvinashetti MS. Isolation and characterization of resistance gene analogs (RGAs) from sorghum (Sorghum bicolor L. Moench). Euphytica. 2005;143(1-2):179-188.

Van der Biezen EA, Jones JD. The NB-ARC domain: A novel signaling motif shared by plant resistance gene products and regulators of cell death in animals. Current Biology. 1998;8(7): R226-R228.

Van Ooijen G, Mayr G, Kasiem MM, Albrecht M, Cornelissen BJ, Takken FL. Structure–function analysis of the NB-ARC domain of plant disease resistance proteins. Journal of Experimental Botany. 2008;59(6):1383-1397.

Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution. 2018;35:1547-1549.

Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution. 1987;4:406-425.

Dopazo J. Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach. Journal of Molecular Evolution. 1994;38:300-304.

Rzhetsky A. and M. Nei. 1992. A simple method for estimating and testing minimum evolution trees. Molecular Biology and Evolution. 2006;9:945-967.

Tajima F, Nei M. Estimation of evolutionary distance between nucleotide sequences. Molecular Biology and Evolution. 1984;1:269-285

Tajima F, Nei M. Estimation of evolutionary distance between nucleotide sequences. Molecular Biology and Evolution. 1984;1: 269-285.

Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl. Acids. Symp. Ser. 1999;41:95-98.

Coll NS, Epple P, Dangl JL. Programmed cell death in the plant immune system. Cell Death & Differentiation. 2011;18(8):1247-1256

Hwang CF, Williamson de Ilarduya OM, Nombela GVM, Muñiz M, Kaloshian I. Rme1 is necessary for Mi-1-mediated resistance and acts early in the resistance pathway. Molecular Plant-Microbe Interactions. 2004;17(1):55-61.

Kumari KASI, Senanayake DMJB, Nanayakkara CM, Balasuriya A, Dhammika WAR, Fernando WMK, Wijerathne WMSBK, Dasanayake PJK. Assessment of Colletotrichum capsici and C. gloeosporioides Resistance among Chilli Accessions Available in Sri Lanka. Sri Lankan Journal of Agriculture and Ecosystems. 2019;1(1):117-128.

ISSN: 2673-1401:

Sariah M. Incidence of Colletotrichum spp on chili in Malaysia and pathogenicity of C. gloeosporioides. Biotrop Special Publication. 1994;54:103-120.

Tameling WIL, Elzinga SD, Darmin PS, Vossen JH, Takken FLW, Haring MA, Cornelissen BJC. The tomato R gene products I-2 and Mi-1 are functional ATP binding proteins with ATPase activity. Plant Cell. 2002;14:2929–2939.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution. 2013;30:2725-2729