No abstract

The mutualism between xyleborine beetles in the genus Euwallacea (Coleoptera: Curculionidae: Scolytinae) and members of the Ambrosia Fusarium Clade (AFC) represents one of 11 known evolutionary origins of fungiculture by ambrosia beetles. Female Euwallacea beetles transport fusarial symbionts in paired mandibular mycangia from their natal gallery to woody hosts where they are cultivated in galleries as a source of food. Native to Asia, several exotic Euwallacea species were introduced into the United States and Israel within the past two decades and they now threaten urban landscapes, forests and avocado production. To assess species limits and to date the evolutionary diversification of the mutualists, we reconstructed the evolutionary histories of key representatives of the Fusarium and Euwallacea clades using maximum parsimony and maximum likelihood methods. Twelve species-level lineages, termed AF 1-12, were identified within the monophyletic AFC and seven among the Fusarium-farming Euwallacea. Bayesian diversification-time estimates placed the origin of the Euwallacea-Fusarium mutualism near the Oligocene-Miocene boundary ∼19-24 Mya. Most Euwallacea spp. appear to be associated with one species of Fusarium, but two species farmed two closely related fusaria. Euwallacea sp. #2 in Miami-Dade County, Florida cultivated Fusarium spp. AF-6 and AF-8 on avocado, and Euwallacea sp. #4 farmed Fusarium ambrosium AF-1 and Fusarium sp. AF-11 on Chinese tea in Sri Lanka. Cophylogenetic analyses indicated that the Euwallacea and Fusarium phylogenies were largely incongruent, apparently due to the beetles switching fusarial symbionts (i.e., host shifts) at least five times during the evolution of this mutualism. Three cospeciation events between Euwallacea and their AFC symbionts were detected, but randomization tests failed to reject the null hypothesis that the putative parallel cladogenesis is a stochastic pattern. Lastly, two collections of Euwallacea sp. #2 from Miami-Dade County, Florida shared an identical cytochrome oxidase subunit 1 (CO1) allele with Euwallacea validus, suggesting introgressive hybridization between these species and/or pseudogenous nature of this marker. Results of the present study highlight the importance of understanding the potential for and frequency of host-switching between Euwallacea and members of the AFC, and that these shifts may bring together more aggressive and virulent combinations of these invasive mutualists.

Plant Dis. 98:694, 2014; published online as http:// dx.

Background Panama disease (aka Fusarium wilt) of banana (Musa spp.) has been a destructive problem for well over a century. Race 1 of the pathogen, Fusarium oxysporum f. sp. cubense (Foc), was responsible for the demise of the first export trades of banana that were based on the cultivar 'Gros Michel'. Currently, tropical race 4 (TR4) impacts the Cavendish cultivars, which are most important in both export and smallholder production. TR4 was confirmed in Jordan in 2013, but has probably been present in the country since at least 2005.

We summarize the information available on ambrosia beetle species that have been associated in Florida with Raffaelea lauricola T.C. Harr., Fraedrich & Aghayeva, the primary symbiont of Xyleborus glabratus Eichhoff and cause of laurel wilt. In total, 14 species in Ambrosiodmus, Euwallacea, Premnobius, Xyleborus, Xyleborinus, and Xylosandrus were either reared from laurel wilt-affected host trees or trapped in laurel wilt-affected stands of the same, and assayed for R. lauricola. In six collections from native species in the southeastern United States [Persea borbonia (L.), Persea palustris (Raf.) Sarg., and Persea humilis Nash] and four from avocado (Persea americana Mill.), extracted mycangia or heads (taxa with mandibular mycangia) or intact bodies (taxa with mycangia in other locations) were surface-disinfested before assays on a semi-selective medium for the isolation of Raffaelea (CSMA+). Raffaelea lauricola was identified based on its characteristic phenotype on CSMA+, and the identity of a random subset of isolates was confirmed with taxon-specific microsatellite markers. The pathogen was recovered from 34% (246 of 726) of the individuals that were associated with the native Persea spp., but only 6% (58 of 931) of those that were associated with avocado. Over all studies, R. lauricola was recovered from 10 of the ambrosia beetle species, but it was most prevalent in Xyleborus congeners. This is the first record of R. lauricola in Ambrosiodmus lecontei Hopkins, Xyleborinus andrewesi (Blandford), and Xyleborus bispinatus Eichhoff. The potential effects of R. lauricola's promiscuity are discussed.

Nutritional mutualisms that ambrosia beetles have with fungi are poorly understood. Although these interactions were initially thought to be specific associations with a primary symbiont, there is increasing evidence that some of these fungi are associated with, and move among, multiple beetle partners. We examined culturable fungi recovered from mycangia of ambrosia beetles associated with trees of Persea humilis (silk bay, one site) and P. americana (avocado, six commercial orchards) that were affected by laurel wilt, an invasive disease caused by a symbiont, Raffaelea lauricola, of an Asian ambrosia beetle, Xyleborus glabratus. Fungi were isolated from 20 adult females of X. glabratus from silk bay and 70 each of Xyleborus affinis, Xyleborus bispinatus, Xyleborus volvulus, Xyleborinus saxesenii, and Xylosandrus crassiusculus from avocado. With partial sequences of ribosomal (LSU and SSU) and nuclear (β-tubulin) genes, one to several operational taxonomic units (OTUs) of fungi were identified in assayed individuals. Distinct populations of fungi were recovered from each of the examined beetle species. Raffaelea lauricola was present in all beetles except X. saxesenii and X. crassiusculus, and Raffaelea spp. predominated in Xyleborus spp. Raffaelea arxii, R. subalba, and R. subfusca were present in more than a single species of Xyleborus, and R. arxii was the most abundant symbiont in both X. affinis and X. volvulus. Raffaelea aguacate was detected for the first time in an ambrosia beetle (X. bispinatus). Yeasts (Ascomycota, Saccharomycotina) were found consistently in the mycangia of the examined beetles, and distinct, putatively co-adapted populations of these fungi were associated with each beetle species. Greater understandings are needed for how mycangia in ambrosia beetles interact with fungi, including yeasts which play currently underresearched roles in these insects.

No abstract

Laurel wilt, caused by Raffaelea lauricola, is a destructive disease of avocado (Persea americana). The susceptibility of different cultivars and races was examined previously but more information is needed on how this host responds to the disease. In the present study, net CO2 assimilation (A), stomatal conductance of H2O (gs), transpiration (E), water use efficiency (WUE), and xylem sap flow rates were assessed in cultivars that differed in susceptibility. After artificial inoculation with R. lauricola, there was a close relationship between symptom development and reductions in A, gs, E, WUE, and mean daily sap flow in the most susceptible cultivar, 'Russell', and significantly greater disease and lower A, gs, E, WUE, and sap flow rates were usually detected after 15 days compared with the more tolerant 'Brogdon' and 'Marcus Pumpkin'. Significant differences in preinoculation A, gs, E, and WUE were generally not detected among the cultivars but preinoculation sap flow rates were greater in Russell than in Brogdon and Marcus Pumpkin. Preinoculation sap flow rates and symptom severity for individual trees were correlated at the end of an experiment (r=0.46), indicating that a plant's susceptibility to laurel wilt was related to its ability to conduct water. The potential management of this disease with clonal rootstocks that reduce sap flow rates is discussed.