Rhodococcus fascians

Taxobox
name = "Rhodococcus fascians"
regnum = Bacteria
phylum = Actinobacteria
ordo = Actinomycetales
subordo = Corynebacterineae
familia = Nocardiaceae
genus = "Rhodococcus"
species = "R. fascians"
binomial = "Rhodococcus fascians"

"Rhodococcus fascians" (known as "Corynebacterium fascians" until 1984) is a Gram positive bacterial phytopathogen that causes leafy gall disease.K.Goethals, D. Vereecke, M.Jaziri, M.Van Montagu, M. Holster " [http://arjournals.annualreviews.org/doi/abs/10.1146/annurev.phyto.39.1.27 Leafy gall formation by Rhodococcus fascians] " Annual Review of Phytopathology, 2001. 39:27-52] "R. fascians" is the only phytopathogenic member of the "Rhodococcus" genus; its host range includes both dicotyledonous and monocotyledonous hosts. Because it commonly afflicts tobacco ("Nicotiana") plants, it is an agriculturally significant pathogen.

Physiology and morphology

"R. fascians" is an aerobic, pleiomorphic, actinomycete that is nonmotile and does not form spores.D.Vereeke, K. Cornelis, W.Temmerman, M.Jaziri, M. Van Montagu, M. Holsters, K.Goenthals " [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=134788 Chromosomal locus that affects pathogenicity of Rhodococcus fascians] " Journal of Bacteriology, Feb, 2002, 184(4):1112–1120] When grown on the surface of an agar plate, colonies are orange in color and appear both smooth or rough.

Virulence

"R. fascians" can be a pathogen of plants, both angiosperm or gymnosperm. Infected plants show typical symptoms, such as leaf deformation, witches broom and leaf gall, which development depends on the plant's cultivar, plant's age, and the bacterial strain.

Leaf deformation consists of widening of parenchyma and growth of vascular system, resulting in wrinkling of laminae and widening of veins. Leafy gall is a gall originated from a bud which would not develop under normal conditions. All effects coming from the infection of "R. fascians" do not depend on plant cells transformation (as for Agrobacterium tumefaciens or Agrobacterium rhizogenes), but on expression of virulence-related genes of bacterium and on the production of compounds that can interfere with normal plant growth and development. During the infection, "R. fascians" usually stays outside vegetal tissues, near a junction or cavity of a plant's cell walls, maybe to avoid environmental stresses. Presence of "R. fascians" was also observed in intercellular spaces inside tissues (in leaf or galls) and even inside cell walls. Presence of "R. fascians" on the infected plant is necessary, not only for the initiation of infection, but also for its maintenance.

Genes that control virulence

Virulence in "R. fascians" is controlled by genes on a plasmid (strains lacking that plasmid are not virulent) and on the chromosome. Using deletion mutations, it was possible to identify three loci on the plasmid: "fas", "att", and "hyp", and one locus on the chromosome, "vic".T.Maels, D.Veereke, T.Ritsema, K.Cornelis, H.N.Thi Thu, M. Van Montagu, M. Holsters, K.Goenthals " [http://www.blackwell-synergy.com/doi/full/10.1046/j.1365-2958.2001.02615.x?cookieSet=1 The att locus of Rhodococcus fascians strain D188 is essential for full virulence on tobacco through the production of an autoregulatory compound] " Molecular Bacteriology, 2001, 42(1), 13-28]

The "fas" is an operon made of six genes (orf 1-6) and a regulatory gene, "fasR". Because deletions of some "fas" genes give a non-virulent phenotype, for fas a main role in virulence was proposed .W.Temmermann, D.Vereeke, R.Dreesen, M.Van Montagu, M.Holsters, K.Goethals " [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=94707 Leafy gall formation is controlled by fasR, an AraC-Type regulatory gene in Rhodococcus fascians] " Journal of Bacteriology, 2000 October; 182(20): 5832–5840] Gene "fasR" is an araC-like transcriptional regulator. Its transcription can be induced "in vitro" in cultures containing certain carbon sources (such as glucose, sucrose, arabinose, glycerol, pyruvate, mannitol, mannose) or nitrogen sources (such as histidine), and is influenced by culture pH and optical density. Also, fasR can be induced by gall extract created by virulent strain.The operon codifies for genes involved in cytokinin synthesis and degradation (orf 4,5,6), in particular for an isopentenyl transferase, a cytokinin oxidase and a glutation-s transferase.The orf1,2,3 transcribe for a cytochrome 450, a ferridoxine containing also a pyruvate dehydrogenase alfa-like domain and a pyruvate dehydrogenase beta subunit. It was suppposed that the first three genes supply energy for the synthesis and degradation of cytokinin, performed by the last three genes of the operon: "R. fascians" can actually produce and degradate zeatin and isopentenil adenine.The compound cytokin oxidase(orf4) can also create adenine with a reactive nitrogen in position 6, which can react with other lateral chains, to form cytokininn-like compounds, more efficient in inducing plant tissue growth.

The "att" is an operon composed of nine genes: "attR", a transcriptional regulator, "attX", a gene including domains for transmembrane localization (perhaps needed for exportation of compounds made by other "att" genes), and several genes "attA"-"attH". Many point and Δ"att" mutants show an attenuated virulence.

Gene "attR" is a transcriptional factor including a helix-turn-helix motif. Its transcription is regulated by the same factors that regulate "fasR" transcription, but with a higher intensity, suggesting, with the attenuation of virulence in "att" mutants, that "att" may regulate fas transcription. Transcription of "att" operon is regulated with a quorum-sensing mechanism: indeed, density of cultures can influence transcription of "attR", and leafy gall extracts coming from galls made by "att" mutant strains are less effective on transcription of "attR".

Genes "attA"-"attH" may be involved in synthesis of compounds needed for transcription of "attR" and "attX". In fact, "attA", "attD" & "attH" are involved in betalactamase synthesis, but no traces of those compounds were found in culture supernatants.

The "hyp" codifies for an RNA-helicase; mutants for this gene are hypervirulent. Also, "hyp" is involved in post transcriptional control of virulence-related genes, maybe on "fas" products.

Operon "vic" is an operon made of five genes, located on the bacterial chromosome. The only known gene is "vicA", the fourth gene in the operon, whose product is a Mas homologue, a protein needed for the switch from citric acid cycle to glyoxylate cycle, both for metabolic reasons and to avoid glyoxylate accumulation, which is toxic for the bacteria. Mutations in "vicA" reduce virulence due to incapacity of "R. fascians" to resist glyoxylate accumulation.

Induction of transcription in infected plant

In tobacco, infection of "R. fascians" leads to hyperexpression of a cytochrome P450, homologue to a gene involved in inactivation of abscissic acid in "Arabidopsis thaliana", of a gibberellic acid oxidase, which inactivates this hormone and its precursors, a proline dehidrogenase, which has its trascription induced by cytokinin and turns proline into glutamic acid, and a factor involved in molybdenum cofactor, needed for sulfur, carbon and nitrogen metabolism control and for abscissic acid synthesis.C.Simon-Mateo, S. Depuydt, C.L. De Oliveira Manes, F.Cnudde, M.Holsters, K.Goethals, D.Vereeke " [http://www.ingentaconnect.com/content/bsc/mpp/2006/00000007/00000002/art00003 The phytopathogen Rhodococcus fascians breaks apical dominance and activate auxiliary meristems by inducing plant genes involved in hormones metabolism] " Molecular Plant Pathology, 2006, 7(2), 103-112]

Role of phytohormones during infection

All the effects of "R. fascians" infection can be attributed to hormone hyperdosage. In particular, most of the effects are connected to auxin and cytokinin, such as: formation of green islands on leaves, wrinkling of laminae, bud proliferation, delay of senescence, and inhibition of lateral roots. In fact, "R. fascians" can produce itself cytokinin, or citokinin-like compounds: using "orf4" and "orf5" in "fas" operon, it can stimulate infected plants to produce cytokinin, and it can produce itself indole-3-acetic acid, using a pathway starting from tryptophane and passing through production of 3-indol-piruvic acid and 3-indol-acetaldeid.O.Vandeputte, S. Oden, A. Mol, D.Vereeke, K.Goethals, M.Jaziri, E.Prinsen " [http://aem.asm.org/cgi/content/full/71/3/1169 Biosynthesis of auxin by the gram positive phytopathogen Rhodococcus fascians is controlled by compounds specific to plant infected tissues] " Applied and Environmental Microbiology, Mar.2005, 71(3):1169-1177] "R.fascians" can also degradate cytokinin to influence the cytokinin/auxin ratio.

Beside cytokinin and auxin, "R. fascians" acts on other hormones: in particular, it can block abscissic acid and gibberellic acid synthesis in infected plants. Abscissic acid represses growth, so a block of production is needed to allow proliferation of cells in leafy galls. Giberellic acid controls cellular differentiation, so its block is needed for maintenance of meristematics cells and for their proliferation.

References