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Insertion of an Esterase Gene into a Specific Locust Pathogen () Enables It to Infect Caterpillars


An enduring theme in pathogenic microbiology is poor understanding of the mechanisms of host specificity. Metarhizium is a cosmopolitan genus of invertebrate pathogens that contains generalist species with broad host ranges such as M. robertsii (formerly known as M. anisopliae var. anisopliae) as well as specialists such as the acridid-specific grasshopper pathogen M. acridum. During growth on caterpillar (Manduca sexta) cuticle, M. robertsii up-regulates a gene (Mest1) that is absent in M. acridum and most other fungi. Disrupting M. robertsii Mest1 reduced virulence and overexpression increased virulence to caterpillars (Galleria mellonella and M. sexta), while virulence to grasshoppers (Melanoplus femurrubrum) was unaffected. When Mest1 was transferred to M. acridum under control of its native M. robertsii promoter, the transformants killed and colonized caterpillars in a similar fashion to M. robertsii. MEST1 localized exclusively to lipid droplets in M. robertsii conidia and infection structures was up-regulated during nutrient deprivation and had esterase activity against lipids with short chain fatty acids. The mobilization of stored lipids was delayed in the Mest1 disruptant mutant. Overall, our results suggest that expression of Mest1 allows rapid hydrolysis of stored lipids, and promotes germination and infection structure formation by M. robertsii during nutrient deprivation and invasion, while Mest1 expression in M. acridum broadens its host range by bypassing the regulatory signals found on natural hosts that trigger the mobilization of endogenous nutrient reserves. This study suggests that speciation in an insect pathogen could potentially be driven by host shifts resulting from changes in a single gene.


Vyšlo v časopise: Insertion of an Esterase Gene into a Specific Locust Pathogen () Enables It to Infect Caterpillars. PLoS Pathog 7(6): e32767. doi:10.1371/journal.ppat.1002097
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002097

Souhrn

An enduring theme in pathogenic microbiology is poor understanding of the mechanisms of host specificity. Metarhizium is a cosmopolitan genus of invertebrate pathogens that contains generalist species with broad host ranges such as M. robertsii (formerly known as M. anisopliae var. anisopliae) as well as specialists such as the acridid-specific grasshopper pathogen M. acridum. During growth on caterpillar (Manduca sexta) cuticle, M. robertsii up-regulates a gene (Mest1) that is absent in M. acridum and most other fungi. Disrupting M. robertsii Mest1 reduced virulence and overexpression increased virulence to caterpillars (Galleria mellonella and M. sexta), while virulence to grasshoppers (Melanoplus femurrubrum) was unaffected. When Mest1 was transferred to M. acridum under control of its native M. robertsii promoter, the transformants killed and colonized caterpillars in a similar fashion to M. robertsii. MEST1 localized exclusively to lipid droplets in M. robertsii conidia and infection structures was up-regulated during nutrient deprivation and had esterase activity against lipids with short chain fatty acids. The mobilization of stored lipids was delayed in the Mest1 disruptant mutant. Overall, our results suggest that expression of Mest1 allows rapid hydrolysis of stored lipids, and promotes germination and infection structure formation by M. robertsii during nutrient deprivation and invasion, while Mest1 expression in M. acridum broadens its host range by bypassing the regulatory signals found on natural hosts that trigger the mobilization of endogenous nutrient reserves. This study suggests that speciation in an insect pathogen could potentially be driven by host shifts resulting from changes in a single gene.


Zdroje

1. WaltonJD 2006 HC-toxin. Phytochemistry 67 1406 1413

2. WolpertTJDunkleLDCiuffettiLM 2002 Host-selective toxins and avirulence determinants: What's in a name? Annu Rev Phytopathol 40 251 285

3. van der DoesHCRepM 2007 Virulence genes and the evolution of host specificity in plant-pathogenic fungi. Mol Plant Microbe Interact 20 1175 1182

4. MylonakisECasadevallAAusubelFM 2007 Exploiting amoeboid and non-vertebrate animal model systems to study the virulence of human pathogenic fungi. PLoS Pathog 3 e101

5. BischoffJFRehnerSAHumberRA 2009 A multilocus phylogeny of the Metarhizium anisopliae lineage. Mycologia 101 512 530

6. BlanfordSChanBHJenkinsNSimDTurnerRJ 2005 Fungal pathogen reduces potential for malaria transmission. Science 308 1638 1641

7. de FariaMRWraightSP 2007 Mycoinsecticides and Mycoacaricides: A comprehensive list with worldwide coverage and international classification of formulation types. Biol Control 43 237 256

8. KanzokSMJacobs-LorenaM 2006 Entomopathogenic fungi as biological insecticides to control malaria. Trends Parasitol 22 49 51

9. GottarMGobertVMatskevichAAReichhartJMWangC 2006 Dual detection of fungal infections in Drosophila via recognition of glucans and sensing of virulence factors. Cell 127 1425 1437

10. DriverFMilnerRJTruemanJWH 2000 A taxonomic revision of Metarhizium based on a phylogenetic analysis of rDNA sequence data. Mycol Res 104 134 150

11. PengGXWangZKYinYPZengDYXiaYX 2008 Field trials of Metarhizium anisopliae var. acridum (Ascomycota: Hypocreales) against oriental migratory locusts, Locusta migratoria manilensis (Meyen) in Northern China. Crop Prot 27 1244 1250

12. ThomasMBReadAF 2007 Can fungal biopesticides control malaria? Nat Rev Microbiol 5 377 383

13. St LegerRJButtTMGoettelMSStaplesRSRobertsDW 1989 Production in vitro of appressoria by the entomopathogenic fungus Metarhizium anisopliae. Exp Mycol 13 274 288

14. St LegerRJJoshiLBidochkaMJRizzoNWRobertsDW 1996 Characterization and ultrastructural localization of chitinases from Metarhizium anisopliae, M. flavoviride, and Beauveria bassiana during fungal invasion of host (Manduca sexta) cuticle. Appl Environ Microbiol 62 907 912

15. WangCSt LegerRJ 2007 The Metarhizium anisopliae perilipin homolog MPL1 regulates lipid metabolism, appressorial turgor pressure, and virulence. J Biol Chem 282 21110 21115

16. WangCSt LegerRJ 2005 Developmental and transcriptional responses to host and nonhost cuticles by the specific locust pathogen Metarhizium anisopliae var. acridum. Eukaryot Cell 4 937 947

17. FreimoserFMScreenSBaggaSHuGSt LegerRJ 2003 Expressed sequence tag (EST) analysis of two subspecies of Metarhizium anisopliae reveals a plethora of secreted proteins with potential activity in insect hosts. Microbiology 149 239 247

18. WangSLeclerqueAPava-RipollMFangWSt LegerRJ 2009 Comparative genomics using microarrays reveals divergence and loss of virulence-associated genes in host-specific strains of the insect pathogen Metarhizium anisopliae. Eukaryot Cell 8 888 898

19. BergerRHoffmannMKellerU 1998 Molecular analysis of a gene encoding a cell-bound esterase from Streptomyces chrysomallus. J Bacteriol 180 6396 6399

20. BornscheuerUT 2002 Microbial carboxyl esterases: classification, properties and application in biocatalysis. FEMS Microbiol Rev 26 73 81

21. KugimiyaWOtaniYHashimotoYTakagiY 1986 Molecular cloning and nucleotide sequence of the lipase gene from Pseudomonas fragi. Biochem Biophys Res Commun 141 185 190

22. GaoQJinKYingSHZhangYXiaoG 2011 Genome sequencing and comparative transcriptomics of the model entomopathogenic fungi Metarhizium anisopliae and M. acridum. PLoS Genet 7 e1001264

23. EbboleDJ 1998 Carbon catabolite repression of gene expression and conidiation in Neurospora crassa. Fungal Genet Biol 25 15 21

24. GancedoJM 1998 Yeast carbon catabolite repression. Microbiol Mol Biol Rev 62 334 361

25. RequenaNFullerPFrankenP 1999 Molecular characterization of GmFOX2, an evolutionarily highly conserved gene from the mycorrhizal fungus Glomus mosseae, down-regulated during interaction with rhizobacteria. Mol Plant Microbe Interact 12 934 942

26. VergerR 1997 Interfacial activation of lipases: Facts and artifacts. Trends Biotechnol 15 32 38

27. St LegerRJMayBAlleeLLFrankDCStaplesRC 1992 Genetic differences in allozymes and information of infection structures among isolates of the entomopathogenic fungus Metarhizium anisopliae. J Invertebr Pathol 60 89 101

28. St LegerRJBidochkaMJRobertsDW 1994 Germination triggers of Metarhizium anisopliae conidia are related to host species. Microbiology 140 1651 1660

29. JandrositzaAPetschniggaJZimmermannaRNatteraKScholzebH 2005 The lipid droplet enzyme Tgl1p hydrolyzes both steryl esters and triglycerides in the yeast, Saccharomyces cerevisiae. Biochimica et Biophysica Acta 1735 50 58

30. ChapmanRF 1998 The Insects: structure and function New York Cambridge University Press 788

31. JarroldSLMooreDPotterUCharnleyAK 2007 The contribution of surface waxes to pre-penetration growth of an entomopathogenic fungus on host cuticle. Mycol Res 111 240 249

32. St LegerRJGoettelMRobertsDWStaplesRC 1991 Pre-penetration events during infection of host cuticle by Metarhizium anisopliae. J Invertebr Pathol 58 168 179

33. MoonYSDonzelliBGKrasnoffSBMcLaneHGriggsMH 2008 Agrobacterium-mediated disruption of a nonribosomal peptide synthetase gene in the invertebrate pathogen Metarhizium anisopliae reveals a peptide spore factor. Appl Environ Microbiol 74 4366 4380

34. DillonRJCharnleyAK 1991 The fate of fungal spores in the insect gut. ColeGTHochHC The fungal spore and disease initiation in plants and animals New York Plenum Press 129 156

35. FangWPava-RipollMWangSSt LegerR 2009 Protein kinase A regulates production of virulence determinants by the entomopathogenic fungus, Metarhizium anisopliae. Fungal Genet Biol 46 277 285

36. FangWPeiYBidochkaMJ 2006 Transformation of Metarhizium anisopliae mediated by Agrobacterium tumefaciens. Can J Microbiol 52 623 626

37. McCluskeyK 2003 The Fungal Genetics Stock center: from molds to molecules. Adv Appl Microbiol 52 245 262

38. WangCSt LegerRJ 2006 A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses. Proc Natl Acad Sci U S A 103 6647 6652

39. CovertSFKapoorPLeeMBrileyANairnCJ 2001 Agrobacterium tumefaciens mediated transformation of Fusarium circinatum. Mycol Res 105 259 264

40. WelteMACermelliSGrinerJVieraAGuoY 2005 Regulation of lipid-droplet transport by the perilipin homolog LSD2. Curr Biol 15 1266 1275

41. PhilipsJHerskowitzI 1997 Osmotic balance regulates cell fusion during mating in Saccharomyces cerevisiae. J Cell Biol 138 961 974

42. PrimNBlancoAMartinezJPastorFIJDiazP 2000 EstA, a gene coding for a cell-bound esterase from Paenibacillus sp. BP-23, is a new member of the bacterial subclass of type B carboxylesterases. Res Microbiol 151 303 312

43. NowierskiRMZengZJaronskiSDelgadoFSwearingenW 1996 Analysis and modeling of time-dose-mortality of Melanoplus sanguinipes, Locusta migratoria migratorioides, and Schistocerca gregaria (Orthoptera: Acrididae) from Beauveria, Metarhizium, and Paecilomyces isolates from Madagascar. J Invertebr Pathol 67 236 252

44. SPSS Inc 2001 SPSS for Windows, Rel. 11.0.1 Chicago SPSS Inc

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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