Target-specific PCR primers can detect and differentiate ophiostomatoid fungi from microbial communities associated with the mountain pine beetle Dendroctonus ponderosae

Khadempour, Lily; Alamouti, Sepideh Massoumi; Hamelin, Richard; Bohlmann, Jöerg; Breuil, Colette

doi:10.1016/j.funbio.2010.08.001

Cited by 12 publications

(18 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Samples were initially identified to species based on morphology and corroborated by PCR amplification of rDNA and other protein coding gene fragments using species‐specific primers coupled with Sanger sequencing (Khadempour et al . ) (data not shown).…”

Section: Methodsmentioning

confidence: 84%

Genetic and genomic evidence of niche partitioning and adaptive radiation in mountain pine beetle fungal symbionts

et al. 2017

Self Cite

View full text Add to dashboard Cite

Bark beetles form multipartite symbiotic associations with blue stain fungi (Ophiostomatales, Ascomycota). These fungal symbionts play an important role during the beetle's life cycle by providing nutritional supplementation, overcoming tree defences and modifying host tissues to favour brood development. The maintenance of stable multipartite symbioses with seemingly less competitive symbionts in similar habitats is of fundamental interest to ecology and evolution. We tested the hypothesis that the coexistence of three fungal species associated with the mountain pine beetle is the result of niche partitioning and adaptive radiation using SNP genotyping coupled with genotype-environment association analysis and phenotypic characterization of growth rate under different temperatures. We found that genetic variation and population structure within each species is best explained by distinct spatial and environmental variables. We observed both common (temperature seasonality and the host species) and distinct (drought, cold stress, precipitation) environmental and spatial factors that shaped the genomes of these fungi resulting in contrasting outcomes. Phenotypic intraspecific variations in Grosmannia clavigera and Leptographium longiclavatum, together with high heritability, suggest potential for adaptive selection in these species. By contrast, Ophiostoma montium displayed narrower intraspecific variation but greater tolerance to extreme high temperatures. Our study highlights unique phenotypic and genotypic characteristics in these symbionts that are consistent with our hypothesis. By maintaining this multipartite relationship, the bark beetles have a greater likelihood of obtaining the benefits afforded by the fungi and reduce the risk of being left aposymbiotic. Complementarity among species could facilitate colonization of new habitats and survival under adverse conditions.

show abstract

Section: Methodsmentioning

confidence: 84%

Genetic and genomic evidence of niche partitioning and adaptive radiation in mountain pine beetle fungal symbionts

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Their inclusion, had they been reliably distinguishable by morphological characteristics likely would not affect our inferences regarding the relationships between geographical location and fungal community composition. Together, these observations point to the utility of applying DNA-based identification techniques [25, 42–44] to the mixed-species environmental cultures that were obtained to confirm species identifications and to test for cryptic species assemblages.…”

Section: Discussionmentioning

confidence: 99%

Spatial Community Structure of Mountain Pine Beetle Fungal Symbionts Across a Latitudinal Gradient

et al. 2011

View full text Add to dashboard Cite

Symbiont redundancy in obligate insect–fungal systems is thought to buffer the insect host against symbiont loss and to extend the environmental conditions under which the insect can persist. The mountain pine beetle is associated with at least three well-known and putatively obligate ophiostomatoid fungal symbionts that vary in their environmental tolerances. To better understand the spatial variation in beetle–fungal symbiotic associations, we examined the community composition of ophiostomatoid fungi associated with the mountain pine beetle as a function of latitude and elevation. The region investigated represents the leading edge of a recent outbreak of mountain pine beetle in western Canada. Using regression and principal components analysis, we identified significant spatial patterns in fungal species abundances that indicate symmetrical replacement between two of the three fungi along a latitudinal gradient and little variation in response to elevation. We also identified significant variation in the prevalence of pair-wise species combinations that occur within beetle galleries. Frequencies of pair-wise combinations were significantly different from what was expected given overall species abundances. These results suggest that complex processes of competitive exclusion and coexistence help determine fungal community composition and that the consequences of these processes vary spatially. The presence of three fungal symbionts in different proportions and combinations across a wide range of environmental conditions may help explain the success of mountain pine beetle attacks across a broad geographic range.Electronic supplementary materialThe online version of this article (doi:10.1007/s00248-011-9841-8) contains supplementary material, which is available to authorized users.

show abstract

“…The presence of a specific band in gel electrophoresis indicates the presence of a plant pathogen. The choice of the target DNA region is a crucial parameter when designing molecular markers to detect and discriminate fungi (Khadempour et al 2010), with the ITS rDNA region being until recently the most common target region employed for this purpose (Zambounis et al 2007). However, the capacity of this region to differentiate variant strains of the same fungus or closely related taxa is quite limited (Wang et al 2010).…”

Section: Introductionmentioning

confidence: 99%

High-Resolution Melting approaches towards plant fungal molecular diagnostics

et al. 2014

View full text Add to dashboard Cite

Reliable and early molecular detection of phytopathogenic fungi is crucial in an era where the expansion of global trade in plant material is undoubtedly increasing the risk of invasive outbreaks, with devastating effects in crop production. Genetic variation within and between fungal species or strains is also important for screening isolates regarding various resistance attributes. Until today many approaches have been employed in fungal diagnostics which are either laborand time-consuming or costly and of reduced sensitivity. Here, we demonstrate and review recent advances on high-resolution melting (HRM) analysis as a rapid, accurate and powerful tool, capable of differentiating even closely related fungal isolates. HRM technique is based on monitoring the melting of PCR amplicons, using saturating concentrations of a fluorescent intercalating dye that binds to double-stranded DNA. Additionally, we discuss the four case studies inferring applications of HRM analysis towards either genotyping of closely related fungal species or screening for fungicide resistance. We focus on the promising results of these studies, giving some technical considerations and describing the advantages of the application of this approach. Finally, we discuss current prospects and applications for research and development related to this innovative HRM technique in plant fungal diagnostics.

show abstract

Target-specific PCR primers can detect and differentiate ophiostomatoid fungi from microbial communities associated with the mountain pine beetle Dendroctonus ponderosae

Cited by 12 publications

References 39 publications

Genetic and genomic evidence of niche partitioning and adaptive radiation in mountain pine beetle fungal symbionts

Genetic and genomic evidence of niche partitioning and adaptive radiation in mountain pine beetle fungal symbionts

Spatial Community Structure of Mountain Pine Beetle Fungal Symbionts Across a Latitudinal Gradient

High-Resolution Melting approaches towards plant fungal molecular diagnostics

Contact Info

Product

Resources

About