Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Manipulation of cytokinin level in the ergot fungus Claviceps purpurea emphasizes its contribution to virulence

Manipulation of cytokinin level in the ergot fungus Claviceps purpurea emphasizes its... Pathogen-derived cytokinins (CKs) have been recognized as important virulence factor in several host–pathogen interactions and it was demonstrated multiple times that phytopathogenic fungi form CKs via the tRNA degradation pathway. In contrast to previous studies, the focus of this study is on the second step of CK formation and CK degradation to improve our understanding of the biosynthesis in fungi on the one hand, and to understand CK contribution to the infection process of Claviceps purpurea on the other hand. The ergot fungus Claviceps purpurea is a biotrophic phytopathogen with a broad host range including economically important crops causing harvest intoxication upon infection. Its infection process is restricted to unfertilized ovaries without causing macroscopic defense symptoms. Thus, sophisticated host manipulation strategies are implicated. The cytokinin (CK) plant hormones are known to regulate diverse plant cell processes, and several plant pathogens alter CK levels during infection. C. purpurea synthesizes CKs via two mechanisms, and fungus-derived CKs influence the host–pathogen interaction but not fungus itself. CK deficiency in fungi with impact on virulence has only been achieved to date by deletion of a tRNA-ipt gene that is also involved in a process of translation regulation. To obtain a better understanding of CK biosynthesis and CKs’ contribution to the plant–fungus interaction, we applied multiple approaches to generate strains with altered or depleted CK content. The first approach is based on deletion of the two CK phosphoribohydrolase (LOG)-encoding genes, which are believed to be essential for the release of active CKs. Single and double deletion strains were able to produce all types of CKs. Apparently, log gene products are dispensable for the formation of CKs and so alternative activation pathways must be present. The CK biosynthesis pathway remains unaffected in the second approach, because it is based on heterologous overexpression of CK-degrading enzymes from maize (ZmCKX1). Zmckx1 overexpressing C. purpurea strains shows strong CKX activity and drastically reduced CK levels. The strains are impaired in virulence, which reinforces the assumption that fungal-derived CKs are crucial for full virulence. Taken together, this study comprises the first analysis of a log depletion mutant that proved the presence of alternative cytokinin activation pathways in fungi and showed that heterologous CKX expression is a suitable approach for CK level reduction. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Genetics Springer Journals

Manipulation of cytokinin level in the ergot fungus Claviceps purpurea emphasizes its contribution to virulence

Download PDF
17 pages

Loading next page...
 
/lp/springer_journal/manipulation-of-cytokinin-level-in-the-ergot-fungus-claviceps-purpurea-uqUdVMzJZO
Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer-Verlag GmbH Germany, part of Springer Nature
Subject
Life Sciences; Microbial Genetics and Genomics; Microbiology; Biochemistry, general; Cell Biology; Plant Sciences; Proteomics
ISSN
0172-8083
eISSN
1432-0983
DOI
10.1007/s00294-018-0847-3
Publisher site
See Article on Publisher Site

Abstract

Pathogen-derived cytokinins (CKs) have been recognized as important virulence factor in several host–pathogen interactions and it was demonstrated multiple times that phytopathogenic fungi form CKs via the tRNA degradation pathway. In contrast to previous studies, the focus of this study is on the second step of CK formation and CK degradation to improve our understanding of the biosynthesis in fungi on the one hand, and to understand CK contribution to the infection process of Claviceps purpurea on the other hand. The ergot fungus Claviceps purpurea is a biotrophic phytopathogen with a broad host range including economically important crops causing harvest intoxication upon infection. Its infection process is restricted to unfertilized ovaries without causing macroscopic defense symptoms. Thus, sophisticated host manipulation strategies are implicated. The cytokinin (CK) plant hormones are known to regulate diverse plant cell processes, and several plant pathogens alter CK levels during infection. C. purpurea synthesizes CKs via two mechanisms, and fungus-derived CKs influence the host–pathogen interaction but not fungus itself. CK deficiency in fungi with impact on virulence has only been achieved to date by deletion of a tRNA-ipt gene that is also involved in a process of translation regulation. To obtain a better understanding of CK biosynthesis and CKs’ contribution to the plant–fungus interaction, we applied multiple approaches to generate strains with altered or depleted CK content. The first approach is based on deletion of the two CK phosphoribohydrolase (LOG)-encoding genes, which are believed to be essential for the release of active CKs. Single and double deletion strains were able to produce all types of CKs. Apparently, log gene products are dispensable for the formation of CKs and so alternative activation pathways must be present. The CK biosynthesis pathway remains unaffected in the second approach, because it is based on heterologous overexpression of CK-degrading enzymes from maize (ZmCKX1). Zmckx1 overexpressing C. purpurea strains shows strong CKX activity and drastically reduced CK levels. The strains are impaired in virulence, which reinforces the assumption that fungal-derived CKs are crucial for full virulence. Taken together, this study comprises the first analysis of a log depletion mutant that proved the presence of alternative cytokinin activation pathways in fungi and showed that heterologous CKX expression is a suitable approach for CK level reduction.

Journal

Current Genetics – Springer Journals

Published: May 30, 2018

References

  • T-DNA of Agrobacterium tumefaciens encodes an enzyme of cytokinin biosynthesis
    Akiyoshi, DE; Klee, H; Amasino, RM
  • Should I fight or should I grow now? The role of cytokinins in plant growth and immunity and in the growth–defence trade-off
    Albrecht, T; Argueso, CT
  • Gapped BLAST and PSI-BLAST: a new generation of protein database search programs
    Altschul, SF; Madden, TL; Schäffer, AA
  • Two-component elements mediate interactions between cytokinin and salicylic acid in plant immunity
    Argueso, CT; Ferreira, FJ; Epple, P; To, JPC; Hutchison, CE; Schaller, GE; Dangl, JL; Kieber, JJ; McDowell, JM
  • Current protocols in molecular biology
    Ausubel, FM
  • Conjugates of auxin and cytokinin
    Bajguz, A; Piotrowska, A
  • Remodeling of cytokinin metabolism at infection sites of Colletotrichum graminicola on maize leaves
    Behr, M; Motyka, V; Weihmann, F
  • Molecular and biochemical characterization of a cytokinin oxidase from maize
    Bilyeu, KD; Cole, JL; Laskey, JG
  • Subcellular locations of MOD5 proteins: mapping of sequences sufficient for targeting to mitochondria and demonstration that mitochondrial and nuclear isoforms commingle in the cytosol
    Boguta, M; Hunter, LA; Shen, WC
  • Ustilago maydis produces cytokinins and abscisic acid for potential regulation of tumor formation in maize
    Bruce, SA; Saville, BJ; Neil Emery, RJ
  • Cytokinins: new apoptotic inducers in plants
    Carimi, F; Zottini, M; Formentin, E
  • Rapid extraction of fungal DNA for PCR amplification
    Cenis, JL
  • The rice resistance protein pair RGA4/RGA5 recognizes the Magnaporthe oryzae effectors AVR-Pia and AVR1-CO39 by direct binding
    Cesari, S; Thilliez, G; Ribot, C
  • Plant hormones: a fungal point of view
    Chanclud, E; Morel, J-B
  • Cytokinin production by the rice blast fungus is a pivotal requirement for full virulence
    Chanclud, E; Kisiala, A; Emery, NRJ
  • Cytokinin biosynthesis and interconversion
    Chen, C
  • Multifunctional yeast high-copy-number shuttle vectors
    Christianson, TW; Sikorski, RS; Dante, M
  • A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors
    Colot, HV; Park, G; Turner, GE
  • Fasciation induction by the phytopathogen Rhodococcus fascians depends upon a linear plasmid encoding a cytokinin synthase gene
    Crespi, M; Messens, E; Caplan, AB
  • Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae
    Dihanich, ME; Najarian, D; Clark, R
  • The Gap1 general amino acid permease acts as an amino acid sensor for activation of protein kinase A targets in the yeast Saccharomyces cerevisiae
    Donaton, MCV; Holsbeeks, I; Lagatie, O
  • The three-dimensional structure of ‘Lonely Guy’ from Claviceps purpurea provides insights into the phosphoribohydrolase function of Rossmann fold-containing lysine decarboxylase-like proteins
    Dzurová, L; Forneris, F; Savino, S
  • Biosynthesis and function of posttranscriptional modifications of transfer RNAs
    Yacoubi, B; Bailly, M; Crécy-Lagard, V
  • Genetics of the ergot fungus Claviceps purpurea
    Esser, K; Tudzynski, P
  • Cytokinin oxidase/cytokinin dehydrogenase assay: optimized procedures and applications
    Frébort, I; Šebela, M; Galuszka, P
  • Studies in Natural Products Chemistry
    Galuszka, P; Spíchal, L; Kopečný, D
  • Inhibition of leaf senescence by autoregulated production of cytokinin
    Gan, S; Amasino, RM
  • The ABC transporter gene in the sirodesmin biosynthetic gene cluster of Leptosphaeria maculans is not essential for sirodesmin production but facilitates self-protection
    Gardiner, DM; Jarvis, RS; Howlett, BJ
  • Discovery of a cytokinin deaminase
    Goble, AM; Fan, H; Sali, A; Raushel, FM
  • Identification of a tRNA isopentenyltransferase gene from Arabidopsis thaliana
    Golovko, A; Sitbon, F; Tillberg, E; Nicander, B
  • The Pseudomonas type III effector HopQ1 activates cytokinin signaling and interferes with plant innate immunity
    Hann, DR; Domínguez-Ferreras, A; Motyka, V
  • The identification of phytohormone receptor homologs in early diverging fungi suggests a role for plant sensing in land colonization by fungi
    Hérivaux, A; Dugé de Bernonville, T; Roux, C
  • Molecular biology of food and water borne mycotoxigenic and mycotic fungi
    Hinsch, J; Tudzynski, P
  • De novo biosynthesis of cytokinins in the biotrophic fungus Claviceps purpurea
    Hinsch, J; Vrabka, J; Oeser, B
  • Functional characterization of the first filamentous fungal tRNA-isopentenyltransferase and its role in the virulence of Claviceps purpurea
    Hinsch, J; Galuszka, P; Tudzynski, P
  • Cytokinin metabolism in maize: Novel evidence of cytokinin abundance, interconversions and formation of a new trans-zeatin metabolic product with a weak anticytokinin activity
    Hluska, T; Dobrev, PI; Tarkowská, D
  • Transgenic studies on the involvement of cytokinin and gibberellin in male development
    Huang, S; Cerny, RE; Qi, Y
  • Variation in karyotype and ploidy level among field isolates of Claviceps purpurea
    Hüsgen, U; Buttner, P; Muller, U; Tudzynski, P
  • Gibepyrone biosynthesis in the rice pathogen Fusarium fujikuroi is facilitated by a small polyketide synthase gene cluster
    Janevska, S; Arndt, B; Niehaus, E-M
  • Cytokinins act synergistically with salicylic acid to activate defense gene expression in rice
    Jiang, C-J; Shimono, M; Sugano, S
  • A yield-associated gene TaCWI, in wheat: its function, selection and evolution in global breeding revealed by haplotype analysis
    Jiang, Y; Jiang, Q; Hao, C
  • The Claviceps purpurea glyceraldehyde-3-phosphate dehydrogenase gene: cloning, characterization, and use for the improvement of a dominant selection system
    Jungehülsing, U; Arntz, C; Smit, R; Tudzynski, P
  • TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions
    Kim, D; Pertea, G; Trapnell, C
  • The effect of modification of tRNA nucleotide-37 on the tRNA interaction with the P- and A-site of the 70S ribosome Escherichia coli
    Konevega, AL; Soboleva, NG; Makhno, VI
  • Structure and function of nucleoside hydrolases from Physcomitrella patens and maize catalyzing the hydrolysis of purine, pyrimidine, and cytokinin ribosides
    Kopečná, M; Blaschke, H; Kopečný, D
  • High-level expression and characterization of Zea mays cytokinin oxidase/dehydrogenase in Yarrowia lipolytica
    Kopečný, D; Pethe, C; Šebela, M
  • Direct control of shoot meristem activity by a cytokinin-activating enzyme
    Kurakawa, T; Ueda, N; Maekawa, M
  • Lack of tRNA modification isopentenyl-A37 alters mRNA decoding and causes metabolic deficiencies in fission yeast
    Lamichhane, TN; Blewett, NH; Crawford, AK
  • Extracellular invertase is an essential component of cytokinin-mediated delay of senescence
    Lara, MEB; Garcia, M-CG; Fatima, T
  • FeatureCounts: an efficient general purpose program for assigning sequence reads to genomic features
    Liao, Y; Smyth, GK; Shi, W
  • Analysis of relative gene expression data using real-time quantitative PCR and the 2∆∆CT method
    Livak, KJ; Schmittgen, TD
  • Plant membrane assays with cytokinin receptors underpin the unique role of free cytokinin bases as biologically active ligands
    Lomin, SN; Krivosheev, DM; Steklov, MY
  • Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2
    Love, MI; Huber, W; Anders, S
  • Differentiation of Claviceps purpurea in axenic culture
    Mantle, PG; Nisbet, LJ
  • Characteristic expression of twelve rice PR1 family genes in response to pathogen infection, wounding, and defense-related signal compounds
    Mitsuhara, I; Iwai, T; Seo, S
  • Activation of caspase-like proteases and induction of apoptosis by isopentenyladenosine in tobacco BY-2 cells
    Mlejnek, P; Procházka, S
  • The major Cu,Zn SOD of the phytopathogen Claviceps purpurea is not essential for pathogenicity
    Moore, S; Vries, OMH; Tudzynski, P
  • Detection of phytohormones in temperate forest fungi predicts consistent abscisic acid production and a common pathway for cytokinin biosynthesis
    Morrison, EN; Knowles, S; Hayward, A
  • Fungal derived cytokinins are necessary for normal Ustilago maydis infection of maize
    Morrison, EN; Emery, RJN; Saville, BJ
  • LONELY-GUY Knocks Every Door: Crosskingdom Microbial Pathogenesis
    Naseem, M; Sarukhanyan, E; Dandekar, T
  • Comparative ‘Omics’ of the Fusarium fujikuroi species complex highlights differences in genetic potential and metabolite synthesis
    Niehaus, E-M; Münsterkötter, M; Proctor, RH
  • Cross-talk of the biotrophic pathogen Claviceps purpurea and its host Secale cereale
    Oeser, B; Kind, S; Schurack, S
  • The ms 2io6A37 modification of tRNA in Salmonella typhimurium regulates growth on citric acid cycle intermediates
    Persson, BC; Ólafsson, Ó; Lundgren, HK; Björk, GR
  • Identification of Rhodococcus fascians cytokinins and their modus operandi to reshape the plant
    Pertry, I; Václavíková, K; Depuydt, S
  • tRNA biology charges to the front
    Phizicky, EM; Hopper, AK
  • Hydrolytic cleavage of N6-substituted adenine derivatives by eukaryotic adenine and adenosine deaminases
    Pospíšilová, H; Šebela, M; Novák, O; Frébort, I
  • Mod5 protein binds to tRNA gene complexes and affects local transcriptional silencing
    Pratt-Hyatt, M; Pai, DA; Haeusler, RA
  • Regulation of source/sink relations by cytokinins
    Roitsch, T; Ehneß, R
  • Cytokinins: activity, biosynthesis, and translocation
    Sakakibara, H
  • Proteasomal control of cytokinin synthesis protects Mycobacterium tuberculosis against nitric oxide
    Samanovic, MI; Tu, S; Novák, O
  • Molecular Cloning. A laboratory manual
    Sambrook, J; Fritsch, E; Maniatis, T
  • Plant-symbiotic fungi as chemical engineers: multi-genome analysis of the clavicipitaceae reveals dynamics of alkaloid loci
    Schardl, CL; Young, CA; Hesse, U
  • In vitro pathogenicity assay for the ergot fungus Claviceps purpurea
    Scheffer, J; Tudzynski, P
  • Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species
    Schmülling, T; Werner, T; Riefler, M
  • Tools for Botrytis cinerea: new expression vectors make the gray mold fungus more accessible to cell biology approaches
    Schumacher, J
  • No hormone to rule them all: interactions of plant hormones during the responses of plants to pathogens
    Shigenaga, AM; Argueso, CT
  • The cereal pathogen Fusarium pseudograminearum produces a new class of active cytokinins during infection
    Sørensen, JL; Benfield, AH; Wollenberg, RD
  • A yeast prion, Mod5, promotes acquired drug resistance and cell survival under environmental stress
    Suzuki, G; Shimazu, N; Tanaka, M
  • Arabidopsis CYP735A1 and CYP735A2 encode cytokinin hydroxylases that catalyze the biosynthesis of trans-zeatin
    Takei, K; Yamaya, T; Sakakibara, H
  • Structure and expression of two polygalacturonase genes of Claviceps purpurea oriented in tandem and cytological evidence for pectinolytic enzyme activity during infection of rye
    Tenberge, K; Homman, V; Oeser, B; Tudzynski, P
  • Cytokinin metabolism of pathogenic fungus Leptosphaeria maculans involves isopentenyltransferase, adenosine kinase and cytokinin oxidase/dehydrogenase
    Trdá, L; Barešová, M; Šašek, V
  • Plants and biotrophs: a pivotal role for cytokinins?
    Walters, DR; McRoberts, N
  • Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity
    Werner, T; Motyka, V; Laucou, V
  • Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C
    Winston, F; Dollard, C; Ricupero-Hovasse, SL