HvGCN2 silencing in barley displays enhanced Blumeria graminis f. sp. hordei susceptibility
Keywords:general control non-depressible 2 (GCN2), barley, Blumeria graminis f. sp. hordei, powdery mildew, virus-induced gene silencing (VIGS), plant disease resistance
- Arabidopsis thaliana GCN2 (AtGCN2) was shown to be involved in disease resistance against biotrophic and necrotrophic pathogens, suggesting a similar role for Hordeum vulgare GCN2 (HvGCN2).
- This is the first study showing the potential importance of HvGCN2 in powdery mildew disease of barley. Under susceptible condition, powdery mildew development was increased in HvGCN2 silenced plants compared to control plants, supported both by hyphal length and number of germinated spores. However, no difference was observed between HvGCN2-silenced and control plants under resistant condition.
Abstract: Powdery mildew disease, caused by Blumeria graminis f. sp. hordei (Bgh), which belongs to the order Erysiphales, is a major crop disease. The general control nondepressible-2 (GCN2) gene of barley was previously found to be overexpressed during the powdery mildew resistance response. Recently, Arabidopsis thaliana GCN2 (AtGCN2) was shown to be involved in disease resistance against biotrophic and necrotrophic pathogens. In order to understand the function of Hordeum vulgare GCN2 (HvGCN2) in the barley powdery mildew resistance response, this gene was silenced by barley stripe mosaic virus (BSMV), mediated by virus-induced gene silencing (VIGS). This is the first study showing the potential importance of HvGCN2 in powdery mildew disease of barley. Based on our observations, when HvGCN2 was silenced on average by 53.5%, Bgh development was increased by 18.7 to 32.1%, which was determined by primary, secondary and longest hyphae measurements. The number of germinated spores also increased 2.8-fold in HvGCN2 silenced plants compared to control plants (BSMV:00). On the other hand, under the resistant condition, no difference was observed in HvGCN2-silenced plants compared to non-silenced lines although the gene was found to be overexpressed in incompatible interaction.
Received: October 17, 2017; Revised: February 20, 2018; Accepted: March 20, 2018; Published onlione: March 29, 2018
How to cite this article: Ozturk IK, Ersoy F, Akkaya MS. HvGCN2 silencing in barley displays enhanced Blumeria graminis f. sp. hordei susceptibility. Arch Biol Sci. 2018;70(3):…
Dean R, Van Kan JAL, Pretorius ZA, Hammond-Kosack KE, Di Pietro A, Spanu PD, Rudd JJ, Dickman M, Kahmann R, Ellis J, Foster GD. The Top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol. 2012;13(4):414-30.
Glawe DA. The powdery mildews: a review of the world’s most familiar (yet poorly known) plant pathogens. Annu Rev Phytopathol. 2008;46:27-51.
Wyand RA, Brown JKM. Genetic and forma specialis diversity in Blumeria graminis of cereals and its implications for host-pathogen co-evolution. Mol Plant Pathol. 2003;4(3):187-98.
Inuma T, Khodaparast SA, Takamatsu S. Multilocus phylogenetic analyses within Blumeria graminis, a powdery mildew fungus of cereals. Mol Phylogenet Evol. 2007;44(2):741-51.
Spanu PD, Abbott JC, Amselem J, Burgis TA, Soanes DM, Stüber K, Ver Loren van Themaat E, Brown JK, Butcher SA, Gurr SJ, Lebrun MH, Ridout CJ, Schulze-Lefert P, Talbot NJ, Ahmadinejad N, Ametz C, Barton GR, Benjdia M, Bidzinski P, Bindschedler LV, Both M, Brewer MT, Cadle-Davidson L, Cadle-Davidson MM, Collemare J, Cramer R, Frenkel O, Godfrey D, Harriman J, Hoede C, King BC, Klages S, Kleemann J, Knoll D, Koti PS, Kreplak J, López-Ruiz FJ, Lu X, Maekawa T, Mahanil S, Micali C, Milgroom MG, Montana G, Noir S, O'Connell RJ, Oberhaensli S, Parlange F, Pedersen C, Quesneville H, Reinhardt R, Rott M, Sacristán S, Schmidt SM, Schön M, Skamnioti P, Sommer H, Stephens A, Takahara H, Thordal-Christensen H, Vigouroux M, Wessling R, Wicker T, Panstruga R. Genome expansion and gene loss in powdery mildew fungi reveal tradeoffs in extreme parasitism. Science. 2012;330(6010):1543-6.
Dong X .Genetic dissection of systemic acquired resistance. Curr Opin Plant Biol 2001;4(4):309-14.
Spoel SH, Dong X. How do plants achieve immunity? Defense without specialized immune cells. Nature Rev Immunol. 2012;12(2):89-100.
Glazebrook J. Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Annu Rev Phytopathol. 2005;43:205-27.
Wek SA, Zhu S, Wek RC. The histidyl-tRNA synthetase-related sequence in the eIF-2 alpha protein kinase GCN2 interacts with tRNA and is required for activation in response to starvation for different amino acids. Mol Cell Biol. 1995;15:4497-506.
Hinnebusch AG. Translational regulation of GCN4 and the general amino acid control of yeast. Annu Rev Microbiol. 2005;59:407-50.
Halford NG. Regulation of carbon and amino acid metabolism, roles of sucrose nonfermenting-1-related protein kinase-1 and general control nonderepressible-2-related protein kinase. Adv Bot Res. 2006;43:93-142.
Lageix S, Lanet E, Pouch-Pélissier, MN, Espagnol MC, Robaglia C, Deragon JM, Pélissier T. Arabidopsis eIF2alpha kinase GCN2 is essential for growth in stress conditions and is activated by wounding. BMC Plant Biol. 2008;8:134.
Zhang Y, Wang Y, Kanyuka K, Parry MAJ, Powers SJ, Halford NG. GCN2-dependent phosphorylation of eukaryotic translation initiation factor-2α in Arabidopsis. J Exp Bot. 2008;59:3131-41.
Chen JJ, London IM .Regulation of protein synthesis by heme-regulated eIF-2 alpha kinase. Trends Biochem Sci. 1995;20(3):105-8.
Feng GS, Chong K, Kumar A, Williams BR. Identification of double-stranded RNA-binding domains in the interferon-induced double-stranded RNA-activated p68 kinase. Proc Natl Acad Sci USA. 1992;89(12):5447-51.
Shi Y, Vattem KM, Sood R, An J, Liang J, Stramm L, Wek RC. Identification and characterization of pancreatic eukaryotic initiation factor 2 alpha-subunit kinase, PEK, involved in translational control. Mol Cell Biol. 1998;18(12):7499-509.
Terry BC, Liu X, Muphey AM, Pajerowska-Mukhtar KM. Arabidopsis thaliana GCN2 is Involved in Responses to Osmotic and Heat Stresses. Int J Plant Res. 2015;5(4):87-95.
Liu X, Kørner CJ, Hajdu D, Guo T, Ramonell K, Argueso C, Pajerowska-Mukhtar KM. Arabidopsis thaliana AtGCN2 Kinase is Involved in Disease Resistance Against Pathogens with Diverse Life Styles. Int J Phytopathol. 2015;04(02):93-104.
Merchant A, Pajerowska-Mukhtar K. Arabidopsis thaliana Dynamic Phenotypic Plasticity in Response to Environmental Conditions. Int J Mod Bot. 2015;5:23-8.
Faus I, Zabalza A, Santiago J, Nebauer SG, Royuela M, Serrano R, Gadea J. Protein kinase GCN2 mediates responses to glyphosate in Arabidopsis. BMC Plant Biol, 2015;15:14.
Luna E, van Hulten M, Zhang Y, Berkowitz O, López A, Pétriacq P, Sellwood MA, Chen B, Burrell M, van de Meene A. Plant perception of β-aminobutyric acid is mediated by an aspartyl-tRNA synthetase. Nat Chem Biol. 2014;10:450-6.
Yildirim-Ersoy F, Ridout CJ, Akkaya MS. Detection of physically interacting proteins with the CC and NB-ARC domains of a putative yellow rust resistance protein, Yr10, in wheat. J Plant Dis Prot. 2011;118(3/4):119-26.
Lu R, Martin-Hernandezn AM, Peart JR, Malcuit I, Baulcombe DC. Virus-induced gene silencing in plants. Methods. 2003;30(4):296-303.
Gould B, Kramer EM. Virus-induced gene silencing as a tool for functional analyses in the emerging model plant Aquilegia (columbine, Ranunculaceae). Plant Methods. 2007;3:6.
Velásquez AC, Chakravarthy S, Martin GB. Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. J Vis Exp. 2009;(10):1292.
Lee WS, Rudd JJ, Kanyuka K. Virus induced gene silencing (VIGS) for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance. Fungal Genet Biol. 2015;79:84-8.
Holzberg S, Brosio P, Gross C, Pogue GP. Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J. 2002;30(3):315-27.
Dagdas YF, Dagdas G, Unver T, Akkaya MS. A new ZTL-type F-box functions as a positive regulator in disease resistance: VIGS analysis in barley against powdery mildew. Physiol Mol Plant Pathol. 2009;74(1):41-4.
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;18,3(7):research0034.1–0034.11.
Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29(9):e45.
Hein I, Barciszewska-Pacak M, Hrubikova K, Williamson S, Dinesen M, Soenderby IE, Sundar S, Jarmolowski A, Shirasu K, Lacomme C. Virus-induced gene silencing-based functional characterization of genes associated with powdery mildew resistance in barley. Plant Physiol. 2005;138(4):2155-64.
Sattlegger E, Hinnebusch AG, Barthelmess IB. Cpc-3, the Neurospora crassa homologue of yeast GCN2, encodes a polypeptide with juxtaposed eIF2 alpha kinase and histidyl-tRNA synthetase-related domains required for general amino acid control. J Biol Chem. 1998;273:20404-16.
Baena-González E. Energy signaling in the regulation of gene expression during stress. Mol Plant. 2010;3(2),300-13.
Meng Y, Moscou MJ, Wise RP. Blufensin1 negatively impacts basal defense in response to barley powdery mildew. Plant Physiol. 2009;149,271-85.
Yin C, Jurgenson JE, Hulbert SH. Development of a Host-Induced RNAi System in the Wheat Stripe Rust Fungus Puccinia striiformis f. sp. tritici. Mol Plant Microbe Interact. 2011;24(5):554-61.