Colonization by the arbuscular mycorrhizal fungus Glomus versiforme induces a defense response against the root-knot nematode Meloidogyne incognita in the grapevine (Vitis amurensis Rupr.), which includes transcriptional activation of the class III chitinase gene VCH3.

Inoculation of the grapevine (Vitis amurensis Rupr.) with the arbuscular mycorrhizal (AM) fungus Glomus versiforme significantly increased resistance against the root-knot nematode (RKN) Meloidogyne incognita. Studies using relative quantitative reverse transcription-PCR (RQRT-PCR) analysis of grapevine root inoculation with the AM fungus revealed an up-regulation of VCH3 transcripts. This increase was greater than that observed following infection with RKN. However, inoculation of the mycorrhizal grapevine roots with RKN was able to enhance VCH3 transcript expression further. Moreover, the increase in VCH3 transcripts appeared to result in a higher level of resistance against subsequent RKN infection. Constitutive expression of VCH3 cDNA in transgenic tobacco under the control of the cauliflower mosaic virus 35S promoter also conferred resistance against RKN, but had no significant effect on the growth of the AM fungus. We analyzed beta-glucuronidase (GUS) activity directed by a 1,216 bp VCH3 promoter in transgenic tobacco following inoculation with both the AM fungus and RKN. GUS activity was negligible in the root tissues before inoculation, and was more effectively induced after inoculation with the AM fungus than with RKN. Moreover, GUS staining in the mycorrhizal transgenic tobacco roots was enhanced by subsequent RKN infection, and was found ubiquitously throughout the whole root tissue. Together, these results suggest that AM fungus induced a defense response against RKN in the mycorrhizal grapevine roots, which appeared to involve transcriptional control of VCH3 expression throughout the whole root tissue.

[1]  T. Boller,et al.  Sinorhizobium meliloti-induced chitinase gene expression in Medicago truncatula ecotype R108-1: a comparison between symbiosis-specific class V and defence-related class IV chitinases , 2004, Planta.

[2]  C. Town,et al.  Transcript Profiling Coupled with Spatial Expression Analyses Reveals Genes Involved in Distinct Developmental Stages of an Arbuscular Mycorrhizal Symbiosis Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tp , 2003, The Plant Cell Online.

[3]  E. Oerke,et al.  Effects of arbuscular mycorrhizal fungi and a non-pathogenic Fusarium oxysporum on Meloidogyne incognita infestation of tomato , 2003, Mycorrhiza.

[4]  J. Barea,et al.  Localized versus systemic effect of arbuscular mycorrhizal fungi on defence responses to Phytophthora infection in tomato plants. , 2002, Journal of experimental botany.

[5]  R. Kjøller,et al.  Interactions between indigenous arbuscular mycorrhizal fungi and Aphanomyces euteiches in field-grown pea , 2002, Mycorrhiza.

[6]  S. Burleigh Relative quantitative RT-PCR to study the expression of plant nutrient transporters in arbuscular mycorrhizas. , 2001, Plant science : an international journal of experimental plant biology.

[7]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[8]  T. Boller,et al.  Differential expression of eight chitinase genes in Medicago truncatula roots during mycorrhiza formation, nodulation, and pathogen infection. , 2000, Molecular plant-microbe interactions : MPMI.

[9]  J. Barea,et al.  Chitosanase and chitinase activities in tomato roots during interactions with arbuscular mycorrhizal fungi or Phytophthora parasitica , 1998 .

[10]  H. Shinshi,et al.  A tobacco gene encoding a novel basic class II chitinase: a putative ancestor of basic class I and acidic class II chitinase genes , 1998, Molecular and General Genetics MGG.

[11]  G. Galili,et al.  Suppression of tobacco basic chitinase gene expression in response to colonization by the arbuscular mycorrhizal fungus Glomus intraradices. , 1998, Molecular plant-microbe interactions : MPMI.

[12]  T. Boller,et al.  Plant defence genes are induced in the pathogenic interaction between bean roots and Fusarium solani, but not in the symbiotic interaction with the arbuscular mycorrhizal fungus Glomus mosseae , 1998 .

[13]  R. Tenhaken,et al.  Chitinase in cucumber hypocotyls is induced by germinating fungal spores and by fungal elicitor in synergism with inducers of acquired resistance , 1998 .

[14]  C. Chlan,et al.  Plant chitinase consensus sequences , 1997, Plant Molecular Biology Reporter.

[15]  J. Barea,et al.  Arbuscular mycorrhizas and biological control of soil-borne plant pathogens – an overview of the mechanisms involved , 1997, Mycorrhiza.

[16]  M. Lambais,et al.  Soybean roots infected by Glomus intraradices strains differing in infectivity exhibit differential chitinase and β-1,3-glucanase expression , 1996 .

[17]  M. Pozo,et al.  Plant hydrolytic enzymes (chitinases and β-1,3-glucanases) in root reactions to pathogenic and symbiotic microorganisms , 1996, Plant and Soil.

[18]  E. Dumas‐Gaudot,et al.  Cellular and molecular defence‐related root responses to invasion by arbuscular mycorrhizal fungi , 1996 .

[19]  M. Lambais,et al.  DIFFERENTIAL EXPRESSION OF DEFENSE-RELATED GENES IN ARBUSCULAR MYCORRHIZA , 1995 .

[20]  T. Boller,et al.  Colonization of Transgenic Tobacco Constitutively Expressing Pathogenesis-Related Proteins by the Vesicular-Arbuscular Mycorrhizal Fungus Glomus mosseae , 1995, Applied and environmental microbiology.

[21]  A. Bleecker,et al.  Analysis of Ethylene Signal-Transduction Kinetics Associated with Seedling-Growth Response and Chitinase Induction in Wild-Type and Mutant Arabidopsis , 1995, Plant physiology.

[22]  R.-J. Liu,et al.  A new method to quantify the inoculum potential of arbuscular mycorrhizal fungi. , 1994, The New phytologist.

[23]  J. Mikkelsen,et al.  Plant chitinase genes , 1994, Plant Molecular Biology Reporter.

[24]  T. Boller,et al.  Ethylene Biosynthesis and Activities of Chitinase and ß-1,3-Glucanase in the Roots of Host and Non-Host Plants of Vesicular-Arbuscular Mycorrhizal Fungi after Inoculation with Glomus mosseae , 1994 .

[25]  Y. Elkind,et al.  A Vesicular Arbuscular Mycorrhizal Fungus (Glomus intraradix) Induces a Defense Response in Alfalfa Roots , 1994, Plant physiology.

[26]  E. Dumas‐Gaudot,et al.  New acidic chitinase isoforms induced in tobacco roots by vesicular-arbuscular mycorrhizal fungi , 1992, Mycorrhiza.

[27]  S. Gianinazzi Vesicular-arbuscular (endo-) mycorrhizas: cellular, biochemical and genetic aspects , 1991 .

[28]  T. Boller,et al.  Chitinase in roots of mycorrhizal Allium porrum: regulation and localization , 1989, Planta.

[29]  R. Jefferson Assaying chimeric genes in plants: The GUS gene fusion system , 1987, Plant Molecular Biology Reporter.

[30]  R. Hussey,et al.  Interaction of Endomycorrhizal Fungi, Superphosphate, and Meloidogyne incognita on Cotton in Microplot and Field Studies. , 1986, Journal of nematology.

[31]  J. Fry,et al.  A simple and general method for transferring genes into plants. , 1985, Science.

[32]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[33]  L. Hai A PRELIMINARY REPORT ON INTERACTIONS BETWEEN ARBUSCULAR MYCORRHIZAL FUNGI AND SOYBEAN CYST NEMATODE , 2002 .

[34]  C. Zheng,et al.  PNZIP is a novel mesophyll-specific cDNA that is regulated by phytochrome and the circadian rhythm and encodes a protein with a leucine zipper motif. , 1998, Plant physiology.

[35]  R. Sundarababu,et al.  Interaction of vesicular-arbuscular mycorrhizae with reniform nematode, rotylenchulus reniformis on Ragi , 1998 .

[36]  J. Barea,et al.  agronomie : plant genetics and breeding Induction of new chitinase isoforms in tomato roots during interactions with Glomus mosseae and / or Phytophthora nicotianae var parasitica , 2007 .

[37]  E. Dumas‐Gaudot,et al.  Gene Expression and Molecular Modifications Associated with Plant Responses to Infection by Arbuscular Mycorrhizal Fungi , 1994 .

[38]  M. St-Arnaud,et al.  Resistance responses of mycorrhizal Ri T-DNA-transformed carrot roots to infection by Fusarium oxysporum s. sp. chrysanthemi , 1994 .

[39]  M. Margis-Pinheiro,et al.  Bean class IV chitinase gene: structure, developmental expression and induction by heat stress , 1994 .

[40]  A. Osbourn,et al.  Advances in Molecular Genetics of Plant-Microbe Interactions , 1994, Current Plant Science and Biotechnology in Agriculture.

[41]  M. Lambais,et al.  Suppression of endochitinase, β-1,3-endoglucanase, and chalcone isomerase expression in bean vesicular-arbuscular mycorrhizal roots under different soil phosphate conditions , 1993 .

[42]  A. Stomp Histochemical localization of β-glucuronidase. , 1992 .

[43]  S. Gallagher GUS protocols: using the GUS gene as a reporter of gene expression. , 1992 .

[44]  R. Hussey Vesicular-Arbuscular Mycorrhizae May Limit Nematode Activity and Improve Plant Growth , 1982 .

[45]  B. Biermann,et al.  QUANTIFYING VESICULAR-ARBUSCULAR MYCORRHIZAE: A PROPOSED METHOD TOWARDS STANDARDIZATION * , 1981 .