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Insects
2012 Feb 10;31:141-60. doi: 10.3390/insects3010141.
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Phylogenetic Analysis of Fusarium solani Associated with the Asian Longhorned Beetle, Anoplophora glabripennis.
Geib SM
,
Scully ED
,
Jimenez-Gasco Mdel M
,
Carlson JE
,
Tien M
,
Hoover K
.
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Culture-independent analysis of the gut of a wood-boring insect, Anoplophora glabripennis (Coleoptera: Cerambycidae), revealed a consistent association between members of the fungal Fusarium solani species complex and the larval stage of both colony-derived and wild A. glabripennis populations. Using the translation elongation factor 1-alpha region for culture-independent phylogenetic and operational taxonomic unit (OTU)-based analyses, only two OTUs were detected, suggesting that genetic variance at this locus was low among A. glabripennis-associated isolates. To better survey the genetic variation of F. solani associated with A. glabripennis, and establish its phylogenetic relationship with other members of the F. solani species complex, single spore isolates were created from different populations and multi-locus phylogenetic analysis was performed using a combination of the translation elongation factor alpha-1, internal transcribed spacer, and large subunit rDNA regions. These analyses revealed that colony-derived larvae reared in three different tree species or on artificial diet, as well as larvae from wild populations collected from three additional tree species in New York City and from a single tree species in Worcester, MA, consistently harbored F. solani within their guts. While there is some genetic variation in the F. solani carried between populations, within-population variation is low. We speculate that F. solani is able to fill a broad niche in the A. glabripennis gut, providing it with fungal lignocellulases to allow the larvae to grow and develop on woody tissue. However, it is likely that many F. solani genotypes could potentially fill this niche, so the relationship may not be limited to a single member of the F. solani species complex. While little is known about the role of filamentous fungi and their symbiotic associations with insects, this report suggests that larval A. glabripennis has developed an intimate relationship with F. solani that is not limited by geographic location or host tree.
Figure 1. Phylogenetic analysis of ALB-derived and O’Donnell Fusarium solani isolates using the translation elongation factor 1-alpha locus. A rooted maximum likelihood tree was generated using TEF1-α sequences from all cultured representatives and a single representative from each OTU detected through culture-independent methods (n = 500 bootstrap replicates). These sequences were placed within the F. solani species complex, utilizing the dataset of O’Donnell [44]. Mating populations (abbreviated MP) are listed based off of the O’Donnell dataset. Nodes with bootstrap support values > 50 are reported, ML distance scale bar = 10 changes. ALB-derived isolates formed 3 strongly supported clusters, denoted as OTU1, OTU2A, and OTU2B. A single NYC isolate and all culture-independent sequences generated from this population were grouped into OTU1, while all MA-derived isolates were classified into cluster 2b. PSU-derived isolates can be observed in both OTU1 and OTU2a.
Figure 2. Multilocus phylogenetic analysis of ALB-derived and O’Donnell Fusarium solani isolates using the internal transcribed spacer region, the translation elongation factor 1-alpha locus, and the large ribosomal subunit locus. A rooted maximum likelihood tree was generated using a partitioned evolutionary model with sequences from three loci from all isolates cultured from ALB larvae (n = 500 bootstrap replicates). ALB-derived sequences were placed within the F. solani species complex, utilizing the dataset of O’Donnell [44]. Mating populations (abbreviated MP) are listed based off of the O’Donnell dataset. Nodes with bootstrap support values > 50 are reported, ML distance scale bar = 10 changes. ALB-derived isolates formed 3 strongly supported monophyletic clades, denoted as OTU1, OTU2a, and OTU2b. A single NYC isolate can be found in OTU1, while all MA-derived isolates were classified into OTU2b. PSU-derived isolates can be observed in both OTU1 and OTU2a.
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