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Colletotrichum siamense was demonstrated as the dominant species among Colletotrichum spp. that infected rubber tree, areca palm, and coffee in Hainan, China. However, the extent of genetic differentiation within the species C. siamense in relation to geographical regions and host species is not known. In this study, 112 C. siamense isolates were genotyped with 12 microsatellite markers. In total, there were 99 multilocus genotypes. Results from permutational multivariate analysis of variance and analysis of molecular variance indicated that there was no significant genetic differentiation between fungal populations with respect to host, location (county), and year. Discriminant analysis of principal components and STRUCTURE analysis showed that C. siamense isolates grouped into three clusters; further analysis confirmed that there were significant ( P < 0.001) genetic differences among the three clusters. However, each cluster had isolates from different hosts, counties, or years, supporting the lack of genetic differentiation with respect to host, county, and year. Statistical analyses of allelic associations indicated some evidence for recombination within the populations defined on the basis of host or county. The present findings provide insights into the genetic structure of C. siamense on the three perennial host species in Hainan and suggest that the disease on these three crops can be effectively considered as one disease and, hence, needs to be controlled simultaneously in mixed plantations.

Genus Colletotrichum is one of the most important genera of plant-pathogenic fungi affecting numerous species, particularly tropical and subtropical crops and fruit trees. In this study, 43 Colletotrichum strains were isolated from areca palm leaves with anthracnose symptoms in 11 areca palm plantations in eight counties of Hainan, China. Based on the morphology, phylogenetic analysis of six loci (internal transcribed spacer, actin, chitin synthase 1, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, and mating type locus MAT1-2), and pathogenicity tests, eight Colletotrichum species were distinguished, comprising five previously known species ( C. cordylinicola , C. fructicola , C. gloeosporioides , C. siamense , and C. tropicale ), one unidentified Colletotrichum species, a new species ( C. arecicola ) in the gloeosporioides species complex, and C. karstii in the boninense species complex. C. siamense was the most common species found in areca palm in Hainan, followed by C. arecicola . Pathogenicity tests showed that all eight species could cause anthracnose symptoms on areca palm leaves using a wound inoculation method and that the isolates from the gloeosporioides species complex caused larger lesions than the isolates from the boninense species complex. Further research is needed to understand the epidemiology of these pathogenic species on areca palm in order to develop management strategies.

Colletotrichum gloeosporioides and C. acutatum have been reported to be causal agents of anthracnose disease of rubber tree. Recent investigations have shown that both C. gloeosporioides and C. acutatum are species complexes. The identities of Colletotrichum species causing anthracnose disease of rubber tree in Hainan, China, are unknown. In this study, 106 isolates obtained from rubber tree with symptoms of anthracnose were collected from 12 counties of Hainan and identified at the species complex level based on the ITS sequences and colony morphologies. Seventy-four isolates were identified as C. gloeosporioides species complex and the other 32 isolates as C. acutatum species complex. Forty-two isolates were selected for further multilocus phylogenetic analyses in order to identify the isolates to the species level. Twenty-six isolates from the C. gloeosporioides species complex were characterized for partial sequences of seven gene regions (ACT, TUB2, CHS-1, GAPDH, ITS, ApMat, and GS), and the other 16 isolates from the C. acutatum species complex for five gene regions (ACT, TUB2, CHS-1, GAPDH, and ITS). Three species were identified: C. siamense and C. fructicola from the C. gloeosporioides species complex, and a new species C. wanningense from the C. acutatum species complex. Artificial inoculation of rubber tree leaves confirmed the pathogenicity of the three species. The present study improves the understanding of species causing anthracnose on rubber tree and provides useful information for the effective control of the disease.

Colletotrichum gloeosporioides and C. acutatum species complexes are causal agents of Colletotrichum leaf disease (CLD) of rubber trees worldwide. To determine the geographic distribution of Colletotrichum species complexes associated with CLD of rubber trees in Hainan, China, and their sensitivity to fungicides used in the region, a total of 275 Colletotrichum isolates were collected from 52 rubber tree plantations in 11 counties. These isolates were identified based jointly on morphological characteristics and PCR-based methodology. Of these isolates, 78 and 22% belonged to the C. gloeosporioides species complex (CGSC) and the C. acutatum complex (CASC), respectively. The incidence of CGSC isolates was greater than the CASC in all counties sampled. The incidence of CASC isolates appeared to be lower in the western and central south of Hainan than in other regions. There was no association in their presence at a given plantation between the two species complexes. The in vitro sensitivity of these two species complexes to carbendazim, chlorothalonil, and four demethylation inhibitor (DMI) fungicides (difenoconazole, propiconazole, myclobutanil, and prochloraz) was determined. Carbendazim was effective against CGSC but not against CASC with mean ED 50 values of 0.176 and 2.182 µg/ml, respectively. CASC isolates were more sensitive to difenoconazole, propiconazole, and myclobutanil (mean ED 50 values of 0.177, 0.129, and 1.424 µg/ml, respectively) than CGSC isolates (mean ED 50 values of 0.710, 0.348, and 3.496 µg/ml, respectively). Mean ED 50 values of CGSC against chlorothalonil and prochloraz were 173.341 and 0.035 µg/ml, respectively; corresponding values for CASC were 151.441 and 0.040 µg/ml. These results suggest that prochloraz, propiconazole, and difenoconazole are effective against both species complexes.

Areca palm, rubber tree, and coffee are always planted adjacent to each other or intercropped with each other, and Colletotrichum siamense was found to be the dominant species of Colletotrichum from these crops in Hainan, China. To study the population structure, pathogenicity, and fungicide sensitivity of C. siamense from these three crops in Hainan, isolates were first identified by their morphological characteristics and multilocus phylogeny. Then both permutational multivariate analysis of variance (PERMANOVA) and pairwise FST analysis showed that the original host and geographical origin (counties) had significant effects on genetic variation in the C. siamense population, explaining 5.6% and 16.3% of genetic variation, respectively. There were significant genetic differentiations between coffee and rubber tree populations but the genetic differentiation was small (FST = 0.04), while significant differentiations were observed among all populations from different counties except those between Haikou and Chengmai. Pathogenicity analysis using artificial inoculation showed that isolates had significantly higher disease incidence and diseased lesion diameter on wounded leaves than on nonwounded leaves. However, the original host of an isolate and geographical origin did not significantly affect the pathogenicity of the C. siamense populations from these three hosts. In vitro tests showed that there were no significant differences in the sensitivity of C. siamense populations to carbendazim, prochloraz, difenoconazole, and propiconazole from different original hosts and geographical origins. The present study suggests that host specialization of C. siamense has not occurred yet in coffee, rubber tree, and areca palm. [ABSTRACT FROM AUTHOR]