Transcriptome analysis of root-knot nematode functions induced in the early stages of parasitism.
Root-knot nematodes of the genus Meloidogyne are obligate biotrophic parasites able to infest > 2000 plant species. The nematode effectors responsible for disease development are involved in the adaptation of the parasite to its host environment and host response modulation. Here, the differences between the transcriptomes of preparasitic exophytic second-stage juveniles (J2) and parasitic endophytic third-stage juveniles (J3) of Meloidogyne incognita were investigated. Genes up-regulated at the endophytic stage were isolated by suppression subtractive hybridization and validated by dot blots and real-time quantitative polymerase chain reaction (PCR). Up-regulation was demonstrated for genes involved in detoxification and protein degradation, for a gene encoding a putative secreted protein and for genes of unknown function. Transcripts of the glutathione S-transferase gene Mi-gsts-1 were 27 times more abundant in J3 than in J2. The observed Mi-gsts-1 expression in the oesophageal secretory glands and the results of functional analyses based on RNA interference suggest that glutathione S-transferases are secreted during parasitism and are required for completion of the nematode life cycle in its host. Secreted glutathione S-transferases may protect the parasite against reactive oxygen species or modulate the plant responses triggered by pathogen attack.
Dubreuil
G
G
INRA-UNSA-CNRS, UMR 1064, Plant-Microbe Interactions and Plant Health, 400, Route des Chappes, BP 167, F-06903 Sophia-Antipolis, France.
Magliano
M
M
INRA-UNSA-CNRS, UMR 1064, Plant-Microbe Interactions and Plant Health, 400, Route des Chappes, BP 167, F-06903 Sophia-Antipolis, France.
Deleury
E
E
INRA-UNSA-CNRS, UMR 1064, Plant-Microbe Interactions and Plant Health, 400, Route des Chappes, BP 167, F-06903 Sophia-Antipolis, France.
Abad
P
P
INRA-UNSA-CNRS, UMR 1064, Plant-Microbe Interactions and Plant Health, 400, Route des Chappes, BP 167, F-06903 Sophia-Antipolis, France.
Rosso
M N
MN
INRA-UNSA-CNRS, UMR 1064, Plant-Microbe Interactions and Plant Health, 400, Route des Chappes, BP 167, F-06903 Sophia-Antipolis, France.
D000595:Amino Acid Sequence; D000818:Animals; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015723:Gene Library; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D015801:Helminth Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D006790:Host-Parasite Interactions / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010944:Plants / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D016415:Sequence Alignment; D014431:Tylenchoidea / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D015801:Helminth Proteins; D012333:RNA, Messenger; D005982:Glutathione Transferase
De Almeida J, Favery B, Engler G, Abad P. 2005. Loss of susceptibility as an alternative for nematode resistance. Current Opinion in Biotechnology 16: 112-117.
Atkinson HJ, Urwin PE, McPherson MJ. 2003. Engineering plants for nematode resistance. Annual Review of Phytopathology 41: 615-639.
Bendtsen JD, Jensen LJ, Blom N, Von Heijne G, Brunak S. 2004. Feature-based prediction of non-classical and leaderless protein secretion. Protein Engineering Design and Selection 17: 349-356.
Cha CJ, Kim SJ, Kim YH, Stingley R, Cerniglia CE. 2002. Molecular cloning, expression and characterization of a novel class glutathione S-transferase from the fungus Cunninghamella elegans. Biochemistry Journal 368: 589-595.
Davis EL, Allen R, Pratt LH, Hussey RS. 1994. Novel immunization procedures used to develop monoclonal antibodies that bind to specific structures in Meloidogyne spp. Phytopathology 82: 1244-1250.
Davis EL, Hussey RS, Baum TJ, Bakker J, Schots A, Rosso MN, Abad P. 2000. Nematode parasitism genes. Annual Review of Phytopathology 38: 365-396.
Diatchenko L, Lau YFC, Campbell AP, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov EDSiebert PD. 1996. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proceedings of the National Academy of Sciences, USA 93: 6025-6030.
Dixon DP, Lapthorn A, Edwards R. 2002. Plant glutathione transferases. Genome Biology 3: 3004.1-3004.10.
Doyle EA, Lambert KN. 2003. Meloidogyne javanica chorismate mutase 1 alters plant cell development. Molecular Plant-Microbe Interactions 16: 123-131.
Ewing B, Hillier L, Wendl MC, Green P. 1998. Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Research 3: 175-185.
Ferreira de Almeida V, Campos VP. 1993. Survival of Meloidogyne exigua in soil and coffee roots in field. Fitopatologia Brasileira 18: 147-150.
Fioretti L, Warry A, Porter A, Haydock P, Curtis R. 2001. Isolation and localization of an annexin gene (gp-nex) from the potato cyst nematode, Globodera pallida. Nematology 3: 45-54.
Fragoso R, Batista JA, Neto OB, Grossi de SaMF. 2005. Isolation and characterization of a cDNA encoding a serine proteinase from the root-knot nematode Meloidogyne incognita. Experimental Parasitology 110: 123-133.
Gheysen G, Fenoll C. 2002. Gene expression in nematode feeding sites. Annual Review of Phytopathology 40: 191-219.
Gürlebeck D, Thieme F, Bonas U. 2006. Type III effector proteins from the plant pathogen Xanthomonas and their role in the interaction with the host plant. Journal of Plant Physiology 163: 233-255.
Huang CS. 1985. Anatomy and physiology of giant cells induced by root-knot nematodes. In: Sasser JN, Carter CC, eds. Advanced treatise on Meloidogyne, Vol. I. Raleigh, NC, USA: North Carolina State University Graphics, 155-164.
Huang G, Dong R, Allen R, Davis EL, Baum TJ, Hussey RS. 2006. A root-knot nematode secretory peptide functions as a ligand for a plant transcription factor. Molecular Plant-Microbe Interactions 19: 463-470.
Huang G, Gao B, Maier T, Allen R, Davis EL, Baum TJ, Hussey RS. 2003. A profile of putative parasitism genes expressed in the oesophageal gland cells of the root-knot nematode Meloidogyne incognita. Molecular Plant-Microbe Interactions 16: 376-381.
Jammes F, Lecomte P, De Almeida-Engler J, Bitton F, Martin-Magniette ML, Renou JP, Abad P, Favery B. 2005. Genome-wide expression profiling of the host response to root-knot nematode infection in Arabidopsis. Plant Journal 44: 447-458.
Jasmer DP, Goverse A, Smant G. 2003. Parasitic nematode interactions with mammals and plants. Annual Review of Phytopathology 41: 245-270.
Jaubert S, Laffaire JB, Ledger TN, Escoubas P, Amri EZ, Abad P, Rosso MN. 2002a. Comparative analysis of two 14-3-3 homologues and their expression pattern in the root-knot nematode Meloidogyne incognita. International Journal for Parasitology 34: 873-880.
Jaubert S, Ledger TN, Laffaire JB, Piotte C, Abad P, Rosso MN. 2002b. Direct identification of stylet secreted proteins from root-knot nematodes by a proteomic approach. Molecular and Biochemical Parasitology 121: 205-211.
Jaubert S, Milac AL, Petrescu AJ, De Almeida-Engler J, Abad P, Rosso MN. 2005. In planta secretion of a calreticulin by migratory and sedentary stages of root-knot nematode. Molecular Plant-Microbe Interactions 18: 1277-1284.
Jones JT, Reavy B, Smant G, Prior AE. 2004. Glutathione peroxidases of the potato cyst nematode Globodera rostochiensis. Gene 324: 47-54.
Klion AD, Donelson JE. 1994. OvGalBP, a filarial antigen with homology to vertebrate galactoside-binding proteins. Molecular and Biochemical Parasitology 65: 305-315.
Kriksunov IA, Schuller DJ, Campbell AM, Barrett J, Brophy PM, Hao Q. 2003. Crystallization and preliminary crystallographic analysis of a new class of glutathione transferase from nematodes. Acta Crystallographica Section D, Biologica Crystallography 59: 262-264.
Lambert C, Leonard N, De Bolle X, Depiereux E. 2002. ESyPred3D: Prediction of protein 3D structures. Bioinformatics 18: 1250-1256.
Marrs KA. 1996. The functions and regulation of glutathione S-transferases in plants. Annual Review of Plant Physiology and Plant Molecular Biology 47: 127-158.
Melillo MT, Leonetti P, Bongiovanni M, Castagnone-Sereno P, Bleve-Zacheo T. 2006. Modulation of reactive oxygen species activities and H2O2 accumulation during compatible and incompatible tomato-root-knot nematode interactions. New Phytologist 170: 501-512.
Molinari S, Miacola C. 1997. Antioxidant enzymes in phytoparasitic nematodes. Journal of Nematology 29: 153-159.
Moons A. 2005. Regulatory and functional interactions of plant growth regulators and plant glutathione S-transferases (GSTs). Vitamins and Hormones 72: 155-202.
Neveu C, Abad P, Castagnone-Sereno P. 2003. Molecular cloning and characterization of an intestinal cathepsin L protease from the plant-parasitic nematode Meloidogyne incognita. Physiological and Molecular Pathology 63: 159-165.
Newton SE, Monti JR, Greenhalgh CJ, Ashman K, Meeusen EN. 1997. cDNA cloning of galectins from third stage larvae of the parasitic nematode Teladorsagia circumcincta. Molecular and Biochemical Parasitology 86: 143-153.
Pertea G, Huang X, Liang F, Antonescu V, Sultana R, Karamycheva S, Lee Y, White J, Cheung F, Parvizi B, Tsai J, Quackenbush J. 2003. TIGR gene indices clustering tools (TGICL): a software system for fast clustering of large EST datasets. Bioinformatics 19: 651-652.
Rao UR, Salinas G, Mehta K, Klei TR. 2000. Identification and localization of glutathione S-transferase as a potential target enzyme in Brugia species. Parasitology Research 86: 908-915.
Reinhardt A, Hubbard T. 1998. Using neural networks for prediction of the subcellular location of proteins. Nucleic Acids Research 9: 2230-2236.
Robertson L, Robertson WM, Sobczak JH, Tetaud E, Ariyanayagam MR, Ferguson MAJ, Fairlamb A, Jones JT. 2000. Cloning, expression and functional characterization of a peroxiredoxin from the potato cyst nematode Globodera rostochiensis. Molecular and Biochemical Parasitology 111: 41-49.
Rosso MN, Dubrana MP, Cimbolini N, Jaubert S, Abad P. 2005. Application of RNA interference to root-knot nematode genes encoding esophageal gland proteins. Molecular Plant-Microbe Interactions 18: 615-620.
Rosso MN, Favery B, Piotte C, Arthaud L, De Boer JM, Hussey RS, Bakker J, Baum TJ, Abad P. 1999. Isolation of a cDNA encoding a beta-1,4-endoglucanase in the root-knot nematode Meloidogyne incognita and expression analysis during plant parasitism. Molecular Plant-Microbe Interactions 12: 585-591.
Rubartelli A, Sitia R. 1991. Interleukin 1 beta and thioredoxin are secreted through a novel pathway of secretion. Biochemical Society Transactions 2: 255-259.
Scholl EH, Thorne JL, McCarter JP, Bird DM. 2003. Horizontally transferred genes in plant-parasitic nematodes: a high-throughput genomic approach. Genome Biology. 4: R39.
Schuller DJ, Liu Q, Kriksunov IA, Campbell AM, Barrett J, Brophy PM, Hao Q. 2005. Crystal structure of a new class of glutathione transferase from the model human hookworm nematode Heligmosoides polygorus. Proteins 61: 1024-1031.
Sheehan D, Maede G, Foley VM, Dowd CA. 2001. Structure, function and evolution of glutathione transferases: implications for classification non-mammalian members of an ancient enzyme superfamily. Biochemical Journal 360: 1-16.
Shingles J, Lilley CJ, Atkinson HJ, Urwin PE. 2007. Meloidogyne incognita: molecular and biochemical characterisation of a cathepsin L cysteine proteinase and the effect on parasitism following RNAi. Experimental Parasitology 115: 114-120.
Siemering KR, Golbik R, Sever R, Haseloff J. 1996. Mutations that suppress the thermosensitivity of green fluorescent protein. Current Biology 12: 1653-1663.
Sijmons PC, Grundler FMW, Von Mende N, Burrows PR, Wyss U. 1991. Arabidopsis thaliana as a new model host for plant-parasitic nematodes. Plant Journal 1: 245-254.
Sommer A, Nimtz M, Conradt HS, Brattig N, Boettcher K, Fischer P, Walter RD, Liebau E. 2001. Structural analysis and antibody response to the extracellular glutathione S-transferases from Onchocerca volvulus. Infection and Immunity 69: 7718-7728.
Van Gundy SD. 1965. Factors in survival of nematodes. Annual Review of Phytopathology 3: 43-68.
Vanholme B, De Meutter J, Tytgat T, Van Montagu M, Coomans A, Gheysen G. 2004. Secretions of plant-parasitic nematodes: a molecular update. Gene 332: 13-27.
Wang X, Meyers D, Yan Y, Baum T, Smant G, Hussey R, Davis E. 1999. In planta localization of a beta-1,4-endoglucanase secreted by Heterodera glycines. Molecular Plant-Microbe Interactions 12: 64-67.
Wilce MCJ, Parker MW. 1994. Structure and function of glutathione- S-transferases. Biochimica et Biophysica Acta 1205: 1-18.
Young AR, Meeusen EN. 2004. Galectins in parasite infection and allergic inflammation. Glycoconjugate Journal 19: 601-606.
Zdobnov EM, Apweiler R. 2001. InterProScan: an integration platform for the signature-recognition methods in InterPro. Bioinformatics 17: 847-848.
Zhan B, Liu S, Perally S, Xue J, Fujiwara R, Brophy P, Xiao S, Liu Y, Feng J, Williamson A, Wang Y, Bueno LL, Mendez S, Goud G, Bethony JM, Hawdon JM, Loukas A, Jones K, Hotez PJ. 2005. Biochemical characterization and vaccine potential of a heme-binding glutathione transferase from the adult hookworm Ancylostoma caninum. Infection and Immunity 73: 6903-6911.