High efficiency transformation and mutant screening of the laurel wilt pathogen, Raffaelea lauricola

Zhou, Yong‐Hong; Lü, Dingding; Joseph, Ross; Li, Tian; Keyhani, Nemat O.

doi:10.1007/s00253-020-10762-1

Cited by 9 publications

(5 citation statements)

References 35 publications

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“…Using a set of previously constructed green‐fluorescent and red‐fluorescent protein (GFP and RFP) expressing strains of H. lauricola (Zhou et al, 2020), a preliminary set of experiments were performed to determine whether the mycangia of the native (to the Southeastern United States) ambrosia beetle species X. affinis could be colonized by H. lauricola (Figure 1). X. affinis pupae were surface sterilized and newly emerged adults were fed on either GFP‐ or RFP‐expressing H. lauricola as detailed in Section 2.…”

Section: Resultsmentioning

confidence: 99%

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen

Joseph

Bansal

Keyhani

2023

Environmental Microbiology

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Ambrosia beetles require their fungal symbiotic partner as their cultivated (farmed) food source in tree galleries. While most fungal‐beetle partners do not kill the host trees they inhabit, since their introduction (invasion) into the United states around ~2002, the invasive beetle Xyleborus glabratus has vectored its mutualist partner (but plant pathogenic) fungus, Harringtonia lauricola, resulting in the deaths of over 300 million trees. Concerningly, indigenous beetles have been caught bearing H. lauricola. Here, we show colonization of the mycangia of the indigenous X. affinis ambrosia beetle by H. lauricola. Mycangial colonization occurred within 1 h of feeding, with similar levels seen for H. lauricola as found for the native X. affinis‐R. arxii fungal partner. Fungal mycangial occupancy was stable over time and after removal of the fungal source, but showed rapid turnover when additional fungal cells were available. Microscopic visualization revealed two pre‐oral mycangial pouches of ~100–200 × 25–50 μm/each, with narrow entry channels of 25–50 × 3–10 μm. Fungi within the mycangia underwent a dimorphic transition from filamentous/blastospore growth to yeast‐like budding with alterations to membrane structures. These data identify the characteristics of ambrosia beetle mycangial colonization, implicating turnover as a mechanism for host switching of H. lauricola to other ambrosia beetle species.

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Section: Resultsmentioning

confidence: 99%

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen

Joseph

Bansal

Keyhani

2023

Environmental Microbiology

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“…. Investigators have developed rapid field detection techniques(Abdulridha et al 2018, Hamilton et al 2021) and begun to increase the understanding of the pathogenicity of H. lauricola with molecular screening(Zhou et al 2020) and metabolic…”

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confidence: 99%

Symbiotic Fungi Associated With Xyleborine Ambrosia Beetles (Coleoptera: Curculionidae: Scolytinae) and the Imperative of Global Collaboration

Osborn

Castro

Duong

et al. 2022

Annals of the Entomological Society of America

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Ambrosia beetles from the tribe Xyleborini are part of nearly all forest ecosystems. Because of their small size, haplodiploid mating structure, and protected lives inside the sapwood of woody plants, they have a unique ability to expand into new regions via inadvertent human transport. A small number of invasive xyleborines cause significant damage to forests, lumber concerns, and agricultural systems. Most ambrosia pests damage or kill trees by the accumulation of beetle attacks, one is known to cause tree death through the introduction of pathogenic fungus into susceptible Lauraceae trees. The relationships between ambrosia fungi and their beetle vectors range from mutualistic symbiosis to facultative association, but most remain unstudied. Unresolved taxonomies, convergent morphologies, and the difficulty of sampling ambrosia fungi over their entire global ranges make comprehensive surveys of ambrosia fungi difficult to achieve. Ambrosia fungi from Europe and North America are moderately well documented, however, we have yet to sufficiently document those from Africa, Asia, Australia, and South America. Worldwide cooperation to improve and standardize scientific study of the ambrosia symbioses is needed to better understand these impactful organisms.

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“…These data can be used to complement existing genomic and transcriptomic datasets, allowing for genotype to phenotype observations of important metabolic and physiological traits such as substrate utilization, chemical sensitivities, and resistance to environmental stressors [ 51 , 70 ]. Coupled to more recent methods for the genetic manipulation of R. lauricola [ 41 ], specific hypotheses can be tested on the contributions of various physiological aspects of the fungus to virulence versus symbioses. Despite its varied lifestyles, our data show that the metabolic capacity of R. lauricola has certain restrictions, suggesting that specific mechanisms or pathways may be important for this fungus to persist in its disparate hosts.…”

Section: Discussionmentioning

confidence: 99%

“…A number of studies have examined the fungicide sensitivities of R. lauricola , and temperature and pH profiling indicated unique features of the fungus that include cold adaptation and poor growth at pH > 7.5 [ 39 , 40 , 41 ]. Here, using phenotyping arrays, we significantly expand characterization of the R. lauricola phenome to include a broad range of substrate utilization responses.…”

Section: Introductionmentioning

confidence: 99%

Unique Attributes of the Laurel Wilt Fungal Pathogen, Raffaelea lauricola, as Revealed by Metabolic Profiling

et al. 2021

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Raffaelea lauricola is the causative agent of laurel wilt, a devastating disease of lauraceous trees. R. lauricola is also an obligate nutritional symbiont of several ambrosia beetle species who act as vectors for the pathogen. Here, we sought to establish the baseline “phenome” of R. lauricola with knowledge concerning its metabolic capability, expanding our understanding of how these processes are impacted by environmental and host nutrients. Phenotypic screening using a microarray of over one thousand compounds was used to generate a detailed profile of R. lauricola substrate utilization and chemical sensitivity. These data revealed (i) relatively restricted carbon utilization, (ii) broad sulfur and phosphate utilization, and (iii) pH and osmotic sensitivities that could be rescued by specific compounds. Additional growth profiling on fatty acids revealed toxicity on C10 substrates and lower, with robust growth on C12–C18 fatty acids. Conditions for lipid droplet (LD) visualization and LD dynamics were examined using a series of lipid dyes. These data provide unique insights regarding R. lauricola metabolism and physiology, and identify distinct patterns of substrate usage and sensitivity which likely reflect important aspects of the host-microbe interface and can be exploited for the development of strategies for mitigating the spread of laurel wilt.

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High efficiency transformation and mutant screening of the laurel wilt pathogen, Raffaelea lauricola

Cited by 9 publications

References 35 publications

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen

Symbiotic Fungi Associated With Xyleborine Ambrosia Beetles (Coleoptera: Curculionidae: Scolytinae) and the Imperative of Global Collaboration

Unique Attributes of the Laurel Wilt Fungal Pathogen, Raffaelea lauricola, as Revealed by Metabolic Profiling

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