Tracking and assessment of Puccinia graminis f. sp. tritici colonization on rice phyllosphere by integrated fluorescence imaging and qPCR for nonhost resistance phenotyping
Tóm tắt
Fluorescence microscopy and qPCR-based pathogen tracking tools were developed to elucidate the growth and development of Puccinia graminis f. sp. tritici on leaves of nonhost plant (rice: Oryza sativa L.). Stem rust race-Puccinia graminis f. sp. tritici 40A was identified using Internal Transcribed Spacer (ITS) sequences and gene sequences coding for hypothetical protein (PGTG_08233_hypothetical protein, HP). A reliable qPCR-based quantitation assay was developed for stem rust fungus exploiting HP primers that yielded 377-bp amplicon in conventional PCR and a forma specialis-specific 146 bp in qPCR with a detection sensitivity of 250-fg genomic DNA. The fungal cell wall N-acetyl-glucosamine was green fluorescence labelled and visualized on propidium iodide-stained nonhost leaf. This technique along with scanning electron microscopy allowed imaging of various developmental structures of Puccinia graminis f. sp. tritici 40A on plant epiphytic and endophytic niches. The microscopy coupled with qPCR-based pathogen load estimation revealed that the nonhost (rice) and host (wheat) phyllosphere surface supported uredospore germination, germ tube formation, hyphal elongation, epiphytic growth, stomatal entry and endophytic growth of fungus in an identical manner. However, the nonhost plant did not show any sign of rusting caused by uredospore production, which instead displayed induced H2O2 accumulation, on leaf. Development of qPCR-based Puccinia quantitation and green fluorescent tag-based qualitative assessment of Puccini proliferation will facilitate nonhost resistance phenotyping not only in rice but also in other nonhost plants of stem rust pathogen.
Tài liệu tham khảo
Ayliffe M, Devilla R, Mago R, White R, Talbot M, Pryor A et al (2011) Nonhost resistance of rice to rust pathogens. Mol Plant Microbe In 24:1143–1155. https://doi.org/10.1094/mpmi-04-11-0100
Ayliffe M, Jin Y, Kang Z, Persson M, Steffenson B, Wang S et al (2011) Determining the basis of nonhost resistance in rice to cereal rusts. Euphytica 179:33–40. https://doi.org/10.1007/s10681-010-0280-2
Barnes CW, Szabo LJ (2007) Detection and identification of four common rust pathogens of cereals and grasses using Real-Time polymerase chain reaction. Phytopathology 97(6):717–727. https://doi.org/10.1094/PHYTO-97-6-0717
Baskarathevan J, Taylor RK, Ho W, McDougal RL, Shivas RG, Alexander BJR (2016) Real-Time PCR assays for the detection of Puccinia psidii. Plant Dis 100(3):617–624. https://doi.org/10.1094/PDIS-08-15-0851-RE
Bettgenhaeuser J, Gilbert B, Ayliffe M, Moscou MJ (2014) Nonhost resistance to rust pathogens—a continuation of continua. Front Plant Sci 5:664. https://doi.org/10.3389/fpls.2014.00664
Bini AP, Quecine MC, da Silva TM, Silva LD, Labate CA (2018) Development of a quantitative real-time PCR assay using SYBR Green for early detection and quantification of Austropuccinia psidii in Eucalyptus grandis. Eur J Plant Pathol 150:735. https://doi.org/10.1007/s10658-017-1321-7
Chen XM (2005) Epidemiology and control of stripe rust Puccinia striiformis f. sp. tritici on wheat. Can J Plant Pathol 27:314–337. https://doi.org/10.1080/07060660509507230
Chen WQ, Wellings C, Chen X, Kang ZS, Liu TG (2014) Wheat stripe (yellow) rust caused by Puccinia striiformis f. sp. tritici. Mol Plant Pathol 15:433–446. https://doi.org/10.1111/mpp.12116
Dawson AM, Bettgenhaeuser J, Gardiner M, Green P, Hernandez-Pinzon I, Hubbard A et al (2015) The development of quick, robust, quantitative phenotypic assays for describing the host-nonhost landscape to stripe rust. Front Plant Sci 6:876. https://doi.org/10.3389/fpls.2015.00876
Demers JE, Crouch JA, Lisa A, Castlebury LA (2015) A multiplex Real-Time PCR assay for the detection of Puccinia horiana and P. chrysanthemi on Chrysanthemum. Plant Dis 99:691–698. https://doi.org/10.1094/PDIS-06-14-0632-RE
Gao L, Yu HX, Kang XH, Shen HM, Li C, Liu TG, Liu B, Chen WQ (2016) Development of SCAR markers and an SYBR green assay to detect Puccinia striiformis f. sp. tritici in infected wheat leaves. Plant Dis 100(9):1840–1847. https://doi.org/10.1094/PDIS-06-15-0693-RE
Gill US, Lee S, Mysore KS (2015) Host versus nonhost resistance: distinct wars with similar arsenals. Phytopathology 105:580–587. https://doi.org/10.1094/phyto-11-14-0298-rvw
Ginzinger DG (2002) Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Exp Hematol 30:503–512. https://doi.org/10.1016/S0301-472X(02)00806-8
Glynn NC, Dixon LJ, Castlebury LA, Szabo LJ, Comstock JC (2010) PCR assays for the sugarcane rust pathogens Puccinia kuehnii and P. melanocephala and detection of a SNP associated with geographical distribution in P. kuehnii. Plant Pathol 59:703–711. https://doi.org/10.1111/j.1365-3059.2010.02299.x
Heath MC (1985) Implications of nonhost resistance for understanding host–parasite interactions. In: Groth JV, Bushnell WR (eds) Genetic basis of biochemical mechanisms of plant disease. APS Press, St Paul, pp 25–42. http://agris.fao.org/agris-search/search.do?recordID=US8733484
Heath MC (1991) The role of gene-for-gene interactions in the determination of host species specificity. Phytopathology 81:127–130
Kutschera U, Hossfeld U (2012) Physiological phytopathology: origin and evolution of a scientific discipline. J Appl Bot Food Qual 85:1–5
Kuzdraliński A, Kot A, Szczerba H, Ostrowska A, Nowak M, Muszyńska M, Lechowski M, Muzyka P (2017) Novel PCR assays for the detection of biological agents responsible for wheat rust diseases: Puccinia triticina and Puccinia striiformis f. sp. tritici. J Mol Microbiol Biotechnol 27:299–305. https://doi.org/10.1159/000481799
Langenbach C, Schultheiss H, Rosendahl M, Tresch N, Conrath U, Goellner K (2016) Interspecies gene transfer provides soybean resistance to a fungal pathogen. Plant Biotechnol J 14:699–708. https://doi.org/10.1111/pbi.12418
Lipka V et al (2005) Pre- and post invasion defenses both contribute to nonhost resistance in Arabidopsis. Science 310:1180–1183
Liu M, McCabe E, Chapados JT et al (2015) Detection and identification of selected cereal rust pathogens by TaqMan® real-time PCR. Can J Plant Path 37:92–105
Mellersh DG, Heath MC (2003) An investigation into the involvement of defense signaling pathways in components of the nonhost resistance of Arabidopsis thaliana to rust fungi also reveals a model system for studying rust fungal compatibility. Mol Plant-Microbe Interact 16:398–404. https://doi.org/10.1094/MPMI.2003.16.5.398
Mishina TE, Zeier J (2007) Bacterial non-host resistance: interactions of Arabidopsis with non-adapted Pseudomonas syringae strains. Physiol Plantarum 131:448–461. https://doi.org/10.1111/j.1399-3054.2007.00977.x
Mojerlou Sh, Safaie N, Abasi-Moghadam A, Shams-Bakhsh M (2013) Evaluation of some Iranian wheat landraces resistance against stem rust disease at seedling stage in the greenhouse. Plant Prot J 35:69–82
Nampally M, Moerschbacher BM, Kolkenbrock S (2012) Fusion of a novel genetically engineered chitosan affinity protein and green fluorescent protein for specific detection of chitosan in vitro and in situ. Appl Environ Microbiol 78(9):3114–3119. https://doi.org/10.1128/AEM.07506-11
Niks RE, Marcel TC (2009) Nonhost and basal resistance: how to explain specificity? New Phytol 182:817–828
Niks RE, Alemu SK, Marcel TC, Van Heyzen S (2015) Mapping genes in barley for resistance to Puccinia coronata from couch grass and to P. striiformis from brome, wheat and barley. Euphytica 206:487–499. https://doi.org/10.1007/s10681-015-1516-y
Nurnberger T, Lipka V (2005) Non-host resistance in plants: new insights into an old phenomenon. Mol Plant Pathol 6:335–345. https://doi.org/10.1111/j.1364-3703.2005.00279.x
Pan JJ, Luo Y, Huang C, Sun ZY, Zhao L, Yan JH, Ma ZH (2010) Application of real-time PCR to establish the quantitative detection of latent infection of wheat stripe rust quantity method. Acta Phytopathol Sin 40:504–510
Schulze-Lefert P, Panstruga R (2011) A molecular evolutionary concept connecting nonhost resistance, pathogen host range, and pathogen speciation. Trends Plant Sci 16:117–125. https://doi.org/10.1016/j.tplants.2011.01.001
Senthil-Kumar M, Mysore KS (2013) Nonhost resistance against bacterial pathogens: retrospectives and prospects. Annu Rev Phytopathol 51:407–427
Shafiei R, Hang C, Kang JG, Loake GJ (2007) Identification of loci controlling non-host disease resistance in Arabidopsis against the leaf rust pathogen Puccinia triticina. Mol Plant Pathol 8:773–784. https://doi.org/10.1111/J.1364-3703.2007.00431.X
Singh RP, Hodson DP, Jin Y, Lagudah ES, Ayliffe MA, Bhavani S et al (2015) Emergence and spread of new races of wheat stem rust fungus: continued threat to food security and prospects of genetic control. Phytopathology 105:872–884
Tadesse K, Ayalew A, Badebo A (2010) Effect of fungicide on the development of wheat stem rust and yield of wheat varieties in highlands of Ethiopia. Afr Crop Sci J 18(1):23–33
Thordal-Christensen H (2003) Fresh insights into processes of nonhost resistance. Curr Opin Plant Biol 6(4):351–357. https://doi.org/10.1016/s1369-5266(03)00063-3 (PMID: 12873530)
Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localisation of H2O2 in plants. H202 accumulation in papillae and hypersensitive response during the barley powdery mildew interaction. Plant J 11:187–1194. https://doi.org/10.1046/j.1365-313X.1997.11061187.x
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Gelfand MA, Sninsky DH, Thomas JJ, San Diego J (eds) PCR protocols: a guide to methods and applications. Academic Press Inc, New York, p 315
Yang Y, Zhao J, Xing H, Wang J, Zhou K, Zhan G et al (2014) Different nonhost resistance responses of two rice subspecies, japonica and indica, to Puccinia striiformis f. sp. tritici. Plant Cell Rep 33:423–433. https://doi.org/10.1007/s00299-013-1542-y
Zellerhoff N, Himmelbach A, Dong W, Bieri S, Schaffrath U, Schweizer P (2010) Nonhost resistance of barley to different fungal pathogens is associated with largely distinct, quantitative transcriptional responses. Plant Physiol 152:2053–2066. https://doi.org/10.1104/pp.109.151829
Zhao J, Yang Y, Yang D, Cheng Y, Jiao M, Zhan G, Zhang H, Wang J, Zhou K, Huang L, Kang Z (2016) Characterization and genetic analysis of Rice Mutant crr1 exhibiting compromised non-host resistance to Puccinia striiformis f. sp. tritici (Pst). Front Plant Sci 7:1822. https://doi.org/10.3389/fpls.2