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      Pyrokinin β-Neuropeptide Affects Necrophoretic Behavior in Fire Ants ( S. invicta), and Expression of β-NP in a Mycoinsecticide Increases Its Virulence

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          Abstract

          Fire ants are one of the world's most damaging invasive pests, with few means for their effective control. Although ecologically friendly alternatives to chemical pesticides such as the insecticidal fungus Beauveria bassiana have been suggested for the control of fire ant populations, their use has been limited due to the low virulence of the fungus and the length of time it takes to kill its target. We present a means of increasing the virulence of the fungal agent by expressing a fire ant neuropeptide. Expression of the fire ant ( Solenopsis invicta) pyrokinin β -neuropeptide (β-NP) by B. bassiana increased fungal virulence six-fold towards fire ants, decreased the LT 50, but did not affect virulence towards the lepidopteran, Galleria mellonella. Intriguingly, ants killed by the β-NP expressing fungus were disrupted in the removal of dead colony members, i.e. necrophoretic behavior. Furthermore, synthetic C-terminal amidated β-NP but not the non-amidated peptide had a dramatic effect on necrophoretic behavior. These data link chemical sensing of a specific peptide to a complex social behavior. Our results also confirm a new approach to insect control in which expression of host molecules in an insect pathogen can by exploited for target specific augmentation of virulence. The minimization of the development of potential insect resistance by our approach is discussed.

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          Most cited references43

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          Global invasion history of the fire ant Solenopsis invicta.

          The fire ant Solenopsis invicta is a significant pest that was inadvertently introduced into the southern United States almost a century ago and more recently into California and other regions of the world. An assessment of genetic variation at a diverse set of molecular markers in 2144 fire ant colonies from 75 geographic sites worldwide revealed that at least nine separate introductions of S. invicta have occurred into newly invaded areas and that the main southern U.S. population is probably the source of all but one of these introductions. The sole exception involves a putative serial invasion from the southern United States to California to Taiwan. These results illustrate in stark fashion a severe negative consequence of an increasingly massive and interconnected global trade and travel system.
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            Two hydrophobins are involved in fungal spore coat rodlet layer assembly and each play distinct roles in surface interactions, development and pathogenesis in the entomopathogenic fungus, Beauveria bassiana.

            The entomogenous filamentous fungus, Beauveria bassiana expresses two hydrophobin genes, hyd1 and hyd2, hypothesized to be involved in cell surface hydrophobicity, adhesion, virulence, and to constitute the protective spore coat structure known as the rodlet layer. Targeted gene inactivation of hyd1 resulted in seemingly 'bald' conidia that contained significantly altered surface fascicles or bundles. These cells displayed decreased spore hydrophobicity, loss of water mediated dispersal, changes in surface carbohydrate epitopes and β-1,3-glucan distribution, lowered virulence in insect bioassays, but no effect on adhesion. In contrast, Δhyd2 mutants retained distorted surface bundles, but truncated/incomplete rodlets could be seen within the bundles. Δhyd2 conidia displayed both decreased cell surface hydrophobicity and adhesion, but the mutant was unaffected in virulence. The double Δhyd1Δhyd2 mutant was distinct from the single mutants, lacking both bundles and rodlets, and displaying additively decreased cell surface hydrophobicity, reduced cell attachment and lowered virulence than the Δhyd1 mutant. Epitope tagged constructs of the proteins were used to examine the expression and distribution of the proteins and to demonstrate the continued presence of Hyd2 in the Δhyd1 strain and vice versa. The implications of our results with respect to fascicle and rodlet assembly on the spore surface are discussed. © 2011 Blackwell Publishing Ltd.
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              Genetic engineering of fungal biocontrol agents to achieve greater efficacy against insect pests.

              Molecular biology methods have elucidated pathogenic processes in several fungal biocontrol agents including two of the most commonly applied entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana. In this review, we describe how a combination of molecular techniques has: (1) identified and characterized genes involved in infection; (2) manipulated the genes of the pathogen to improve biocontrol performance; and (3) allowed expression of a neurotoxin from the scorpion Androctonus australis. The complete sequencing of four exemplar species of entomopathogenic fungi including B. bassiana and M. anisopliae will be completed in 2010. Coverage of these genomes will help determine the identity, origin, and evolution of traits needed for diverse lifestyles and host switching. Such knowledge combined with the precision and malleability of molecular techniques will allow design of multiple pathogens with different strategies to be used for different ecosystems and avoid the possibility of the host developing resistance.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                4 January 2012
                : 7
                : 1
                : e26924
                Affiliations
                [1 ]Biotechnology Research Center, Southwest University, Beibei, Chongqing, People's Republic of China
                [2 ]Department of Entomology and Nematology, University of Florida, Gainesville, Florida, United States of America
                [3 ]Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida, United States of America
                University of Wisconsin – Madison, United States of America
                Author notes

                Conceived and designed the experiments: YF RMP NOK. Performed the experiments: YF RMP EK. Analyzed the data: YF RMP GC NOK. Contributed reagents/materials/analysis tools: GC. Wrote the paper: NOK. Contributed input to the writing of the manuscript: YF RMP EK GC.

                Article
                PONE-D-11-16736
                10.1371/journal.pone.0026924
                3251551
                22238569
                d5a51c11-2067-44fe-9361-0091a07f66b8
                Fan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 22 August 2011
                : 5 October 2011
                Page count
                Pages: 7
                Categories
                Research Article
                Agriculture
                Pest Control
                Biology
                Biotechnology
                Genetic Engineering
                Microbiology
                Mycology
                Plant Science
                Botany
                Mycology
                Zoology

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