Identification, pathogenicity and abundance of Paracremonium pembeum sp nov and Graphium euwallaceae sp nov.-two newly discovered mycangial associates of the polyphagous shot hole borer (Euwallacea sp.) in California

WOS: 000375239600006PubMed ID: 26740544Fusarium euwallaceaeis a well-characterized fungal symbiont of the exotic ambrosia beetle Euwallacea sp. (polyphagous shot hole borer [PSHB]), together inciting Fusarium dieback on many host plants in Israel and California. Recent discoveries of additional fungal symbionts within ambrosia beetle mycangia suggest these fungi occur as communities. Colony-forming units of Graphium euwallaceae sp. nov. and Paracremonium pembeum sp. nov., two novel fungal associates of PSHB from California, grew from 36 macerated female heads and 36 gallery walls collected from Platanus racemosa, Acer negundo, Persea americana and Ricinus communis. Fungi were identified based on micromorphology and phylogenetic analyses of the combined internal transcribed spacer region (nuc rDNA ITS1- 5.8S-ITS2 [ITS barcode]), elongation factor (EF 1-alpha), small subunit (18S rDNA) sequences for Graphium spp., ITS, EF 1-alpha, calmodulin (cmdA), large subunit of the ATP citrate lyase (acl1), beta-tubulin (tub2), RNA polymerase II second largest subunit (rpb2) and large subunit (28S rDNA) sequences for Paracremonium spp. Other Graphium spp. recovered from PSHB in Vietnam, Euwallacea fornicates in Thailand, E. validus in Pennsylvania and Paracremonium sp. recovered from PSHB in Vietnam were identified. F. euwallaceae was recovered from mycangia at higher frequencies and abundances in all hosts except R. communis, in which those of F. euwallaceae and P. pembeum were equal. P. pembeum was relatively more abundant within gallery walls of A. negundo and R communis. In all hosts combined F. euwallaceae was relatively more abundant within PSHB heads than gallery walls. All three fungi grew at different rates and colonized inoculated excised stems of P. americana and A. negundo. P. pembeum produced longer lesions than F euwallaceae and G. euwallaceae on inoculated avocado shoots. Results indicate PSHB is associated with a dynamic assemblage of mycangial fungal associates that pose additional risk to native and nonnative hosts in California.California Avocado Commission; USDA Forest Service, Forest Health Protection and Forest Health Monitoring; USDA Farm Bill; National Institute of Food and AgricultureUnited States Department of Agriculture (USDA)National Institute of Food and AgricultureThis project was financially supported by the California Avocado Commission, USDA Forest Service, Forest Health Protection and Forest Health Monitoring, the USDA Farm Bill and the National Institute of Food and Agriculture. Special thanks to the California Avocado Commission, Huntington Botanical Gardens and Orange County Parks for permission to collect samples. Tim Thibault, curator at Huntington Botanical Gardens, advised on nomenclature for fungal descriptions. Dan Berry, nursery manager at Huntington Botanical Gardens, Dr John Kabashima, UCCE farm advisor, and Monica Dimson assisted with sample collections. Karen Xu provided statistical consulting. We also thank the anonymous reviewers for their comments and suggestions on this manuscript