Journal of Taiwan Agricultural Research

Author:Chao-Jung Wu, Sui-Li Hsu, Su-Yu Lai, Miao-Chun Lin, and Hui-Fang Ni*

Abstract:

In this study, healthy coffee leaves from seven coffee orchards were collected. The seven orchards were located in different areas, namely Ren-ai Township, Nantou County, Gukeng Township, Yunlin County, Chiayi Agricultural Experiment Station, Jhongpu, and Zhuqi Township, Chiayi County, Dongshan District, Tainan City, and Taiwu Township, Pingtung County. Coffee leaves were surface-disinfected and incubated on acidified potato dextrose agar to obtain endophytic fungi. Colletotrichum spp., Phyllosticta spp. and Xylariaceous fungi were isolated from these seven orchards with a high frequency, and Colletotrichum spp. were the most frequently isolated fungi in six of the seven orchards. According to the results of phylogenetic analysis based on actin, calmodulin, chitin synthase, glyceraldehyde 3-phosphate dehydrogenase, β-tubulin sequences constructed by Bayesian inference, the endophytic Colletotrichum spp. in coffee leaves contained nine species belonging to Colletotrichum gloeosporioides species complex, namely Colletotrichum cordylinicola, Colletotrichum fructicola, Colletotrichum gloeosporioides, Colletotrichum jiangxiense, Colletotrichum siamense, Colletotrichum theobromicola, Colletotrichum tropicale and an unknown new Colletotrichum sp., and two Colletotrichum boninense complex species, C. boninense and Colletotrichum karstii. Isolation frequency and species composition of endophytic Colletotrichum spp. from three orchards were compared. The Colletotrichum spp. composition of the orchard located in Ren-ai Township, Nantou County resembled that located in Jhongpu Township, Chiayi County. The isolation frequencies of C. cordylinicola were 83% and 27% in orchards of Ren-ai and Jhongpu, respectively. C. siamense was also isolated from both of the two orchards. In contrast, no C. cordylinicola or C. siamense was isolated from the orchard in Zhuqi Township, Chiayi County. The most frequently isolated Colletotrichum spp. in Zhuqi orchard were C. fructicola (45%) and C. karstii (45%). The result suggested that endophytic Colletotrichum spp. in coffee leaves are highly diverse, and the fungal composition is different among orchards. This is the first intensive study of endophytic Colletotrichum spp. in coffee leaves using multi-locus sequence analysis to resolve the species, and the result provides information for further study on coffee pathogen or endophyte.

Key words:Coffee, Endophyte, Colletotrichum spp.

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