ORIGINAL ARTICLE
Sensitive and specific detection of Xanthomonas hortorum pv. pelargonii in geranium by real-time PCR
 
More details
Hide details
1
Department of Plant Protection, College of Agriculture, Shiraz University, 71441-65186 Shiraz, Iran
CORRESPONDING AUTHOR
Seyyed Mohsen Taghavi
Department of Plant Protection, College of Agriculture, Shiraz University, 71441-65186 Shiraz, Iran
Submission date: 2016-04-24
Acceptance date: 2016-08-04
 
Journal of Plant Protection Research 2016;56(3):265–269
KEYWORDS
TOPICS
ABSTRACT
Xanthomonas hortorum pv. pelargonii is the causal agent of bacterial blight of geranium. A specific and rapid real-time PCR assay for detecting the bacterium in plant was developed in this study. We compared sensitivity of conventional and real-time PCR for detection of the pathogen in inoculated plants. For application to disease management programs, PCR amplification must be able to detect latent infections of asymptomatic geraniums. Our results displayed that conventional PCR lacks sufficient sensitivity to be used as diagnostic tools for detection of X. hortorum pv. pelargonii in latent infections. On the other hand, real-time PCR is suitable method for detection of latent infection of the bacterium in planting materials and can be considered in management programs. The ability for accurate and reliable detection of X. hortorum pv. pelargonii in asymptomatic tissue is a significant step in setting up “pathogen free” certification programs for bacterial blight of geranium.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
REFERENCES (19)
1.
Anderson M.J., Nameth S.T. 1990. Development of a polyclonal antibody-based serodiagnostic assay for the detection of Xanthomonas campestris pv. pelargonii in geranium plants. Phytopathology 80: 357–360.
 
2.
Cullen D.W., Lees A.K., Toth I.K., Duncan J.M. 2002. Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR. Plant Pathology 51 (3): 281–292.
 
3.
Daughtrey M., Macksel M. 1993. Nursery-cultivated Geranium spp. as a possible inoculum source for Xanthomonas campestris pv. pelargonii causing bacterial blight disease of a greenhouse Pelargonium crop. Phytopathology 83: 242.
 
4.
Daughtrey M., Wick R.L. 1993. Vascular wilt diseases. p. 237–242. In: “Geraniums IV. The Grower’s Manual” (J.W. White, ed.). Ball Publishing, Geneva, Switzerland, 412 pp.
 
5.
Faghihi M., Taghavi S.M., Hamzezarghani H. 2014. Comparison of the efficiency of conventional PCR, nested-PCR, real-time PCR and loop-mediated isothermal amplification (LAMP) methods for detection of the causal agent of citrus huanglongbing in Iran. Iranian Journal of Plant Pathology 50: 237–255.
 
6.
Hodge N.C., Chase A.R., Stall R.E. 1992. Diversity of four species of Xanthomonas as determined by cellular fatty acid analysis. Phytopathology 82: 1153.
 
7.
Lees A.K., Cullen D.W., Sullivan L., Nicolson M.J. 2002. Development of conventional and quantitative real-time PCR assays for the detection and identification of Rhizoctonia solani AG-3 in potato and soil. Plant Pathology 51 (3): 293–302.
 
8.
Manulis S., Valinsky L., Lichter A., Gabriel D.W. 1994. Sensitive and specific detection of Xanthomonas campestris pv. pelargonii with DNA primers and probes identified by random amplified polymorphic DNA analysis. Applied and Environmental Microbiology 60 (11): 4094–4099.
 
9.
Nameth S.T., Daughtrey M.L., Moorman G.W., Sulzinski M.A. 1999. Bacterial blight of geranium: a history of diagnostic challenges. Plant Disease 83 (3): 204–212.
 
10.
NCBI (National Center for Biotechnology Information). Available on: http://www.ncbi.nlm.nih.gov/. [Accessed: December 21, 2015].
 
11.
Oglevee-O’Donovan W.A. 1986. Production of culture virus-indexed geraniums. p. 119–123. In: “Tissue Culture as a Plant Production System for Horticultural Crops” (R.H. Zimmerman, ed.). Springer, Netherlands, 371 pp.
 
12.
Safaie Farahani A., Taghavi S.M., Taher-khani K. 2014. Comparison of conventional, nested and real-time PCR for detection of the causal agent of ratoon stunt in Iran. Journal of Plant Pathology 97 (2): 259–263.
 
13.
Sasser M.J. 1990. Identification of bacteria by gas chromatography of cellular fatty acids. MIDI Technical Note # 101. Available on: http://natasha.eng.usf.edu/gil.... [Accessed: December 10, 2015].
 
14.
Schaad N.W., Opgenorth D., Gaush P. 2002. Real-time polymerase chain reaction for one-hour on-site diagnosis of Pierces disease of grape in early season asymptomatic vines. Phytopathology 92 (7): 721–728.
 
15.
Sulzinski M.A., Moorman G.W., Schlanhaufer B., Romaine C.P. 1996. Characteristics of a PCR-based assay for in planta detection of Xanthomonas campestris pv. pelargonii. Journal of Phytopathology 144 (7–8): 393–398.
 
16.
Sulzinski M.A., Schlagnhaufer B., Moorman G.W., Romaine C.P. 1998. PCR-Based detection of artificial latent infections of geranium by Xanthomonas campestris pv. pelargonii. Journal of Phytopathology 146 (2–3): 111–114.
 
17.
Tuinier J.E., Stephens C.T. 1989. Use of serology to detect Xanthomonas campestris pv. pelargonii in aqueous extracts of geranium plants. Plant Disease 73 (11): 875–878.
 
18.
Weller S.A., Elphinstone J.G., Smith N.C., Boonham N., Stead D.E. 2000. Detection of Ralstonia solanacearum strains with a quantitative, multiplex, real-time, fluorogenic PCR (TaqMan) assay. Applied and Environmental Microbiology 66 (7): 2853–2858.
 
19.
Weller S.A., Simpkins S.A., Stead D.E., Kurdziel A., Hird H., Weekes R.J. 2002. Identification of Agrobacterium spp. present within Brassica napus seed by TaqMan PCR – implications for GM screening procedures. Archives of Microbiology 178 (5): 338–343.
 
eISSN:1899-007X
ISSN:1427-4345