Journal of General Plant Pathology

, Volume 71, Issue 4, pp 253–262 | Cite as

Possible roles and functions of LPL1 gene encoding lysophospholipase during early infection by Magnaporthe grisea

  • Masaki Kanamori
  • Ken-ichiro Saitoh
  • Tsutomu Arie
  • Takashi Kamakura
  • Tohru Teraoka
FUNGAL DISEASES
Received:
Accepted:

Abstract

The rice blast fungus Magnaporthe grisea differentiates appressoria, which are required to attack its rice plant host. Clone A26, tentatively named LPL1, was previously found to be homologous to the known lysophospholipase genes from our subtractive cDNA library. The LPL1 protein had a consensus motif (GxSxG) and a catalytic triad (S, D, H) of esterases in the deduced amino acid sequence, and the protein expressed in Escherichia coli had lysophospholipase activity. To clarify the functions and possible roles of LPL1, the gene was disrupted by targeted gene replacement. The ΔLPL1 mutants formed fewer appressoria on the hydrophobic surface of GelBond film, and the appressoria had reduced turgor pressure and penetration into cells of the leaf sheath. The ΔLPL1 mutants and wild-type differentiated normal appressoria on other artificial substrata such as polycarbonate plate and on rice leaf sheath. Cytological analysis of the appressoria indicated that ΔLPL1 mutants had a delay in the disappearance of lipid droplets. These findings imply that LPL1, phospholipid metabolism, or both are involved in glycerol biosynthesis and accumulation to generate turgor pressure in the appressorium. LPL1 was, however, dispensable for full pathogenicity, suggesting that other complementary pathways or similar genes related to phospholipid metabolism probably function in M. grisea.

Key words

Magnaporthe grisea Lipid droplets Penetration peg Appressorium Lysophospholipase Turgor pressure 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Akiyama, K, Watanabe, H, Tsukada, S, Sasai, H 2000A novel method for constructing gene-targeting vectorsNucleic Acids Res28E77CrossRefPubMedGoogle Scholar
  2. Altschul, SF, Madden, TL, Schaffer, AA, Zhang, J, Zhang, Z, Miller, W, Lipman, DJ 1997Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Res2533893402PubMedGoogle Scholar
  3. Bradford, MM 1976A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnal Biochem72248252PubMedGoogle Scholar
  4. Choi, W, Dean, RA 1997The adenylate cyclase gene MAC1 of Magnaporthe grisea controls appressorium formation and other aspects of growth and developmentPlant Cell919731983CrossRefPubMedGoogle Scholar
  5. Davis, DJ, Burlak, C, Money, NP 2000Biochemical and biomechanical aspects of appressorial development in Magnaporthe griseaTharreau, DLebrun, MHTalbot, NJNotteghem, JL eds. Advances in rice blast researchKluwer AcademicDordrecht248256Google Scholar
  6. De Jong, JC, McCormack, BJ, Smirnoff, N, Talbot, NJ 1997Glycerol generates turgor in rice blastNature389244245CrossRefGoogle Scholar
  7. Dixon, KP, Xu, JR, Smirnoff, N, Talbot, NJ 1999Independent signaling pathways regulate cellular turgor during hyperosmotic stress and appressorium-mediated plant infection by Magnaporthe griseaPlant Cell1120452058CrossRefPubMedGoogle Scholar
  8. Duncan, JA, Gilman, AG 1998A cytoplasmic acyl-protein thioesterase that removes palmitate from G protein α subunits and p21RASJ Biol Chem2731583015837CrossRefPubMedGoogle Scholar
  9. Duncan, JA, Gilman, AG 2002Characterization of Saccharomyces cerevisiae acyl-protein thioesterase 1, the enzyme responsible for G protein α subunit deacylation in vivoJ Biol Chem2773174031752CrossRefPubMedGoogle Scholar
  10. Gilbert, RD, Johnson, AM, Dean, RA 1996Chemical signals responsible for appressorium formation in the rice blast fungus Magnaporthe griseaPhysiol Mol Plant Pathol48335346CrossRefGoogle Scholar
  11. Gross, RW, Sobel, BE 1983Rabbit myocardial cytosolic lysophospholipase: purification, characterization, and competitive inhibition by l-palmitoyl carnitineJ Biol Chem25852215226PubMedGoogle Scholar
  12. Howard, RJ, Valent, B 1996Breaking and entering: host penetration by the fungal rice blast pathogen Magnaporthe griseaAnnu Rev Microbiol50491512CrossRefPubMedGoogle Scholar
  13. Howard, RJ, Ferrari, MA, Roach, DH, Money, NP 1991Penetration of hard substrates by a fungus employing enormous turgor pressuresProc Natl Acad Sci USA881128111284PubMedGoogle Scholar
  14. Jelitto, TC, Page, HA, Read, ND 1994Role of external signals in regulating the pre-penetration phase of infection by the rice blast fungus, Magnaporthe griseaPlanta194471477CrossRefGoogle Scholar
  15. Kamakura, T, Xiao, JZ, Choi, WB, Kochi, T, Yamaguchi, S, Teraoka, T, Yamaguchi, I 1999cDNA subtractive cloning of genes expressed during early stage of appressorium formation by Magnaporthe griseaBiosci Biotechnol Biochem6314071413CrossRefGoogle Scholar
  16. Kamakura, T, Yamaguchi, S, Saitoh, K, Teraoka, T, Yamaguchi, I 2002A novel gene, CBP1, encoding a putative extracellular chitin-binding protein, may play an important role in the hydrophobic surface sensing of Magnaporthe grisea during appressorium differentiationMol Plant Microbe Interact15437444PubMedGoogle Scholar
  17. Kimura, M, Izawa, K, Yoneyama, K, Arie, T, Kamakura, T, Yamaguchi, I 1995A novel transformation system for Pyricularia oryzae: adhesion of regenerating fungal protoplasts to collagen-coated dishesBiosci Biotechnol Biochem5911771180Google Scholar
  18. Lee, KS, Patton, JL, Fido, M, Hines, LK, Kohlwein, SD, Paltauf, F, Henry, SA, Levin, DE 1994The Saccharomyces cerevisiae PLB1 gene encodes a protein required for lysophospholipase and phospholipase B activityJ Biol Chem2691972519730PubMedGoogle Scholar
  19. Lee, YH, Dean, RA 1993cAMP regulates infection structure formation in the plant pathogenic fungus Magnaporthe griseaPlant Cell5693700CrossRefPubMedGoogle Scholar
  20. Lee, YH, Dean, RA 1994Hydrophobicity of contact surface induces appressorium formation in Magnaporthe griseaFEMS Microbiol Lett1157175CrossRefGoogle Scholar
  21. Liu, S, Dean, RA 1997G protein α subunit genes control growth, development, and pathogenicity of Magnaporthe griseaMol Plant Microbe Interact1010751086PubMedGoogle Scholar
  22. Mitchell, TK, Dean, RA 1995The cAMP-dependent protein kinase catalytic subunit is required for appressorium formation and pathogenesis by the rice blast pathogen Magnaporthe griseaPlant Cell718691878CrossRefPubMedGoogle Scholar
  23. Money, NP, Howard, RJ 1996Confirmation of link between fungal pigmentation, turgor pressure, and pathogenicity using a new method of tugor measurmentFungal Genet Biol20217227CrossRefGoogle Scholar
  24. Murphy, DJ 2001The biogenesis and functions of lipid bodies in animals, plants and microorganismsProg Lipid Res40325438CrossRefPubMedGoogle Scholar
  25. Nishimura, M, Park, G, Xu, JR 2003The G-beta subunit MGB1 is involved in regulating multiple steps of infection-related morphogenesis in Magnaporthe griseaMol Microbiol50231243CrossRefPubMedGoogle Scholar
  26. Saitoh, K, Arie, T, Teraoka, T, Yamaguchi, I, Kamakura, T 2003Targeted gene disruption of the neuronal calcium sensor 1 homologue in rice blast fungus, Magnaporthe griseaBiosci Biotechnol Biochem67651653CrossRefPubMedGoogle Scholar
  27. Sambrook, J, Fritsch, EF, Maniatis, T 1989Molecular cloning: a laboratory manual2nd edn.Cold Spring Harbor Laboratory PressCold Spring Harbor, NYGoogle Scholar
  28. Schlenk, H, Gellerman, J 1960Esterification of fatty acids with diazomethane on a small scaleAnal Chem3214121414CrossRefGoogle Scholar
  29. Sugimoto, H, Hayashi, H, Yamashita, S 1996Purification, cDNA cloning, and regulation of lysophospholipase from rat liverJ Biol Chem27177057711CrossRefPubMedGoogle Scholar
  30. Takahashi, Y 1958A method for disease forecast of rice blast: use of leaf sheath assayPlant Protection12339345 (in Japanese)Google Scholar
  31. Talbot, NJ 1995Having a blast: exploring the pathogenicity of Magnaporthe griseaTrends Microbiol3916CrossRefPubMedGoogle Scholar
  32. Talbot, NJ 2003On the trail of a cereal killer: exploring the biology of Magnaporthe griseaAnnu Rev Microbiol57177202CrossRefPubMedGoogle Scholar
  33. Talbot, NJ, McCafferty, HRK, Ma, M, Moore, K, Hamer, JE 1997Nitrogen starvation of the rice blast fungus Magnaporthe grisea may act as an environmental cue for disease symptom expressionPhysiol Mol Plant Pathol50179195CrossRefGoogle Scholar
  34. Thines, E, Weber, RW, Talbot, NJ 2000MAP kinase and protein kinase A-dependent mobilization of triacylglycerol and glycogen during appressorium turgor generation by Magnaporthe griseaPlant Cell1217031718CrossRefPubMedGoogle Scholar
  35. Thompson, JD, Higgins, DG, Gibson, TJ 1994CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucleic Acids Res2246734680PubMedGoogle Scholar
  36. Wang, A, Dennis, EA 1999Mammalian lysophospholipasesBiochim Biophys Acta1439116PubMedGoogle Scholar
  37. Wang, A, Deems, RA, Dennis, EA 1997aCloning, expression, and catalytic mechanism of murine lysophospholipase IJ Biol Chem2721272312729CrossRefGoogle Scholar
  38. Wang, A, Loo, R, Chen, Z, Dennis, EA 1997bRegiospecificity and catalytic triad of lysophospholipase IJ Biol Chem2722203022036CrossRefGoogle Scholar
  39. Wang, A, Yang, HC, Friedman, P, Johnson, CA, Dennis, EA 1999A specific human lysophospholipase: cDNA cloning, tissue distribution and kinetic characterizationBiochim Biophys Acta1437157169PubMedGoogle Scholar
  40. Wang, A, Johnson, CA, Jones, Y, Ellisman, MH, Dennis, EA 2000Subcellular localization and PKC-dependent regulation of the human lysophospholipase A/acyl-protein thioesterase in WISH cellsBiochim Biophys Acta1484207214PubMedGoogle Scholar
  41. Weber, RW, Wakley, GE, Thines, E, Talbot, NJ 2001The vacuole as central element of the lytic system and sink for lipid droplets in maturing appressoria of Magnaporthe griseaProtoplasma216101112PubMedGoogle Scholar
  42. Xiao, JZ, Watanabe, T, Kamakura, T, Ohshima, A, Yamaguchi, I 1994Studies on cellular differentiation of Magnaporthe grisea: physicochemical aspects of substratum surfaces in relation to appressorium formationPhysiol Mol Plant Pathol44227236CrossRefGoogle Scholar
  43. Xu, JR, Hamer, JE 1996MAP kinase and cAMP signaling regulate infection structure formation and pathogenic growth in the rice blast fungus Magnaporthe griseaGenes Dev1026962706PubMedGoogle Scholar
  44. Xu, JR, Urban, M, Sweigard, JA, Hamer, JE 1997The CPKA gene of Magnaporthe grisea is essential for appressorial penetrationMol Plant Microbe Interact10187194Google Scholar
  45. Zhang, YY, Dennis, EA 1988Purification and characterization of a lysophospholipase from a macrophage-like cell line P388D1J Biol Chem26399659972PubMedGoogle Scholar

Copyright information

© The Phytopathological Society of Japan and Springer-Verlag Tokyo 2005

Authors and Affiliations

  • Masaki Kanamori
    • 1
  • Ken-ichiro Saitoh
    • 1
  • Tsutomu Arie
    • 1
  • Takashi Kamakura
    • 2
  • Tohru Teraoka
    • 1
  1. 1.Graduate School of AgricultureTokyo University of Agriculture and TechnologyTokyoJapan
  2. 2.Faculty of Science and TechnologyTokyo University of ScienceChibaJapan

Personalised recommendations