Isolation and identification of Colletotrichum as fungal pathogen from tea and preliminary fungicide screening

Meixia Chen; Lingkun Zhong; Zhenxiong Zhang; Chengbin Peng; Dongxian Ke; Peng Gan; Zerong Wang; Rifeng Wei; Wei Liu; Jiangfan Yang

doi:10.15586/qas.v14i1.1000

Meixia Chen

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, China; College of Life Science, Ningde Normal University, Ningde, China.

Lingkun Zhong

The Government of Yantian, Ningde, China.

Zhenxiong Zhang

College of Life Science, Ningde Normal University, Ningde, China; Industry and University Research Cooperation Demonstration Base in Fujian Province, Ningde Normal University, Ningde, China.

Chengbin Peng

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, China; College of Life Science, Ningde Normal University, Ningde, China.

Dongxian Ke

College of Life Science, Ningde Normal University, Ningde, China; Industry and University Research Cooperation Demonstration Base in Fujian Province, Ningde Normal University, Ningde, China.

Peng Gan

College of Life Science, Ningde Normal University, Ningde, China; Industry and University Research Cooperation Demonstration Base in Fujian Province, Ningde Normal University, Ningde, China.

Zerong Wang

College of Life Science, Ningde Normal University, Ningde, China; Industry and University Research Cooperation Demonstration Base in Fujian Province, Ningde Normal University, Ningde, China.

Rifeng Wei

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, China.

Wei Liu

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, China; College of Life Science, Ningde Normal University, Ningde, China.

Jiangfan Yang

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, China.

Keywords

anthracnose, Colletotrichum, fungicide screening, isolation and identification, tea

Abstract

The increasing incidence of anthracnose disease in tea plant caused by Colletotrichum has become an important global concern. It is the cause of withered leaves of the tea plant, leading to a considerable decrease in the economic yield of tea, and thereby threatening the sustainable development of tea industries. In this study, Colletotrichum infected tea leaves were collected from three separate regions: Zhouning, Longyan, and Ningde in Fujian province, China. The pathogen was isolated from the leaves and identified based on morphology following Koch’s postulates and DNA sequencing of the nuclear rDNA internal transcribed spacer (ITS) region, beta-tubulin 2 (β-Tub2), the large subunit of the nuclear ribosomal RNA gene (LSU), Glutamine synthetase (GS), and an oligonucleotide primer (CgInt). A total of six strains were identified with different cultural, morphological, and molecular characteristics. Furthermore, multi-locus phylogenetic analysis showed the six strains that belong to C.fructicola. Finally, a preliminary screening of nine chemical fungicides to inhibit the strain N by toxicity test identified 40% prochloraz at 0.1 μg mL^–1 as the most effective method. The phylogenetic tree analysis revealed a close relationship between the identified strains, and the strains were classified based on cultural, morphological, and molecular characteristics. The findings of this study add to our understanding of C.fructicola, which will aid in the development of preventive measures, the improvement of tea quality, and the assurance of safe production.

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References

Cai, L., Hyde, K.D., Taylor, P.W.J., Weir, B.S., Waller, J.M., Abang, M.M., Zhang, J.Z., Yang, Y.L., Phoulivong, S., Liu, Z.Y., Prihastuti, H., Shivas, R.G., Mc Kenzie, E.H.C. and Johnston, P.R., 2009. A polyphasic approach for studying Colletotrichum. Fungal Diverisity 39: 183–204. 10.1016/j.riam.2009.11.001

Diao, Y.Z., Zhang, C., Liu, F., Wang, W.Z., Liu, L., Cai, L. and Liu, X.L., 2017. Colletotrichum species causing anthracnose disease of chili in China. Persoonia: Molecular Phylogeny and Evolution of Fungi 38: 20–37. 10.3767/003158517X692788

Fang, Z.D., 2001. The research method of plant disease, 3rd ed. China Agriculture Press, Beijing.

Guo, M., Pan, Y.M., Dai, Y.L. and Gao, Z.M., 2014. First report of brown blight disease caused by Colletotrichum gloeosporioides on camellia sinensis in Anhui province, China. Plant Disease 98: 284. 10.1094/pdis-08-13-0896-pdn

Gutiérrez-Alonso, J.G., Gutiérrez-Alonso, O., Nieto-Angel, D., Téliz-Ortiz, D., Zavaleta-Mejía, E., Delgadillo-Sánchez, F. and Vaquera-Huerta, H., 2003. Evaluación de resistencia a imazalil, prochloraz y azoxystrobin en aislamientos de Colletotrichum loeosporioides (Penz.) Penz. y Sacc. y control de la antracnosis del mango (Mangifera indica L.) en postcosecha. Revista Mexicana De Fitopatología 121: 228–232.

Hirayama, Y., Asano, S., Okayam, K.O., Ohki, S.T. and Tojo, M., 2018. Weeds as the potential inoculum source of Colletotrichum fructicola responsible for strawberry anthracnose in Nara, Japan. Journal of General Plant Pathology 84: 12–19. 10.1007/s10327-017-0753-4

Kumita, K., Kitazawa, Y., Tokuda, R., Miyazaki, A., Maejima, K., Namba, S. and Yamaji, Y., 2021. First report of anthracnose on tillandsia caused by Colletotrichum sp. in Japan. Journal of General Plant Pathology 87: 254–258. 10.1007/s10327-021-00995-x

Liu, F., Damm, U., Cai, L. and Crous, P.W., 2013. Species of the Colletotrichum gloeosporioides complex associated with anthracnose diseases of Proteaceae. Fungal Diversity 61: 89–105. 10.1007/s13225-013-0249-2

Liu, F., Weir, B.S., Damm, U., Crous, P.W., Wang, Y., Liu, B., Wang, M., Zhang, M. and Cai, L., 2015. Unravelling Colletotrichum species associated with Camellia: employing ApMat and GS loci to resolve species in the C.gloeosporioides complex. Persoonia: Molecular Phylogeny and Evolution of Fungi 35: 863–866. 10.3767/003158515X687597

Liu, W., 2013. Anthracnose pathogen identification and the genetic diversity of tea plant. Fujian Agriculture and Forestry University, Fuzhou, China.

Lu, Q.H., Wang, Y.C., Li, N.N., Ni, D.J., Yang, Y.J. and Wang, X.C., 2018. Differences in the characteristics and pathogenicity of Colletotrichum camelliae and C. fructicola isolated from the tea plant (Camellia sinensis (L.) O. Kuntze). Fronties in Microbiology 9: 3060. 10.3389/fmicb.2018.03060

Marais, L., 1990. Efficacy of fungicides against Colletotrichum coccodes on potato tubers. Potato Research 33: 275–281. 10.1007/bf02358457

Mills, P.R., Sreenivasaprasad, S. and Brown, A.E., 1992. Detection and differentiation of Colletotrichum gloeosporioides isolates using PCR. Federation of European Microbiology Societies Microbiology Letters 98: 137–144. 10.1111/j.1574-6968.1992.tb05503.x

Nilsson, R.H., Kristiansson, E., Ryberg, M., Hallenberg, N. and Larsson, K.H., 2008. Intraspecific ITS variability in the kingdom fungi as expressed in the international sequence databases and its implications for molecular species identification. Evolutionary Bioinformatics 4: 193–201. 10.4137/ebo.s653

Noireung, P., Phoulivong, S., Liu, F., Cai, L., Mckenzie, E., Chukeatirote, E., Jones, E.B.G., Bahkali, A. and Hyde, K.D., 2012. Novel species of Colletotrichum revealed by morphology and molecular analysis. Cryptogamie Mycologie 33: 347–362. 10.7872/crym.v33.iss3.2012.347

Oliveira, P.D.L., de Oliveira, K.Á.R., dos Santos Vieira, W.A., Câmara, M.P.S. and de Souza, E.L., 2018. Control of anthracnose caused by Colletotrichum species in guava, mango and papaya using synergistic combinations of chitosan and Cymbopogon citratus (D.C. ex Nees) Stapf. essential oil. International Journal of Food Microbiology 266: 87–94. 10.1016/j.ijfoodmicro.2017.11.018

Rahmani, A.H., Al Shabrmi, F.M., Allemailem, K., Aly, S.M. and Khan, M.A., 2015. Implications of green tea and its constituents in the prevention of cancer via the modulation of cell signalling pathway. BioMed Research International 2015:1–12. 10.1155/2015/925640

Reyes-Estebanez, M., Sanmartín, P., Carlos Camacho-Chab, J., De la Rosa-García, S.C., Jesús Chan Bacab, M., Noemí Águila Ramírez, R., Carrillo-Villanueva, F., De la Rosa-Escalante, E., Luis Arteaga-Garma, J., Serrano, M. and Otto Ortega-Morales, B., 2020. Characterization of a native Bacillus velezensis-like strain for the potential biocontrol of tropical fruit pathogens. Biological Control 141:1–12. 10.1016/j.biocontrol.2020.104127

Taylor, J.W., Jacobson, D.J., Kroken, S., Kasuga, T., Geiser, D.M., Hibbett, D.S. and Fisher, M.C., 2000. Phylogenetic species recognition and species concepts in fungi. Fungal Genetics and Biology 31: 21–32. 10.1006/fgbi.2000.1228

Vingaard, A.M., Hass, U., Dalgaard, M., Andersen, H.R., Bonefeld-Jørgensen, E., Christiansen, S., Laier, P. and Poulsen, M.E., 2006. Prochloraz: an imidazole fungicide with multiple mechanisms of action. International Journal of Andrology 29: 186–192. 10.1111/j.1365-2605.2005.00604.x

Walsh, T.J., Groll, A., Hiemenz, J., Fleming, R., Roilides, E. and Anaissie, E., 2004. Infections due to emerging and uncommon medically important fungal pathogens. Clinical Microbiology and Infection 10: 48–66. 10.1111/j.1470-9465.2004.00839.x

Wang, Y.C., Hao, X.Y., Wang, L., Bin, X., Wang, X.C. and Yang Y.J., 2016. Diverse Colletotrichum species cause anthracnose of tea plants (Camellia sinensis (L.) O. Kuntze) in China. Scientific Reports 6: 35287. 10.1038/srep35287

Wei, J.C., 1979. Fungus identification manual. Shanghai Science and Technology Publishing House. Shanghai, China.

Weir, B.S., Johnston, P.R. and Damm, U., 2012. The Colletotrichum gloeosporioides species complex. Studies in Mycology 73: 115–180. 10.3114/sim0011

Yoshida, K., Ogino, A., Yamada, K. and Sonoda, R., 2010. Induction of disease resistance in tea (Camellia sinensis L.) by plant activators. Japan Agricultural Research Quarterly 44: 391–398. 10.6090/jarq.44.391