Bacterial leaf blight (BLB) of rice is a very destructive disease worldwide and is caused by Xanthomonas oryzae pv. oryzae (Xoo). The aim of the present study was to examine if the Xoo virulence pathotypes obtained using phenotypic pathotyping could be confirmed using molecular approach. After screening of 60 Operon primers with genomic DNA of two Xoo isolates (virulent pathotype, Vr, and mildly virulent pathotype, MVr), 12 Operon primers that gave reproducible and useful genetic information were selected and used to analyze 50 Xoo isolates from 7 West African countries. Genetic analysis revealed two major Xoo virulence genotypes (Mta and Mtb) with Mta having two subgroups (Mta1 and Mta2). Mta1 (Vr1) subgroup genotype has occurrence in six countries and Mta2 (Vr2) in three countries while Mtb genotype characterized mildly virulence (MVr) Xoo isolates present in five countries. The study revealed possible linkage and correlation between phenotypic pathotyping and molecular typing of Xoo virulence. Xoo virulence genotypes were known to exist within country and there was evidence of Xoo pathogen migration between countries. Durable resistance rice cultivars would need to overcome both Mta and Mtb Xoo virulence genotypes in order to survive after their deployment into different rice ecologies in West Africa. 1. Introduction Rice is perhaps the most widely cultivated food crop world over, but its production is constrained by diseases of fungal, bacterial, and viral origins. Bacterial leaf blight (BLB) of rice, caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very destructive disease and its incidence has been reported from different parts of Asia, northern Australia, Africa, and USA [1–3]. The disease is known to occur in epidemic proportions in many parts of the world, incurring severe crop loss of up to 50% [1, 2, 4]. In West Africa, disease incidence ranged from 70 to 85% and yield loss ranged from 50 to 90%, indicating a wide spread of BLB disease in farmers’ fields [2, 4]. Some selected Xoo isolates have shown high level of pathogenicity and virulence on the cultivated rice varieties [4]. Research studies have also revealed that BLB is an important rice disease in irrigated rice ecosystems in West Africa, mainly in Sahelian and Sudano-Sahelian countries [2, 5]. Crop loss assessment studies have revealed that this disease reduces grain yield to varying levels, depending on the stage of the crop, degree of cultivar susceptibility, and, to a great extent, the conduciveness of the environment in which it occurs [6]. The severity and significance of damage
References
[1]
T. B. Adhikari, C. M. Vera Cruz, Q. Zhang et al., “Genetic diversity of Xanthomonas oryzae pv. oryzae in Asia,” Applied and Environmental Microbiology, vol. 61, no. 3, pp. 966–971, 1995.
[2]
Y. Sere, A. Onasanya, V. Verdier et al., “Rice bacterial leaf blight in West Africa: preliminary studies on disease in farmers’ fields and screening released varieties for resistance to the bacteria,” Asian Journal of Plant Sciences, vol. 4, pp. 577–579, 2005.
[3]
G. H. Jiang, Z. H. Xia, Y. L. Zhou et al., “Testifying the rice bacterial blight resistance gene xa5 by genetic complementation and further analyzing xa5 (Xa5) in comparison with its homolog TFIIAγ1,” Molecular Genetics and Genomics, vol. 275, no. 4, pp. 354–366, 2006.
[4]
A. Onasanya, M. M. Ekperigin, F. E. Nwilene, Y. Sere, and R. O. Onasanya, “Two pathotypes of Xanthomonas oryzae pv. oryzae virulence identified in West Africa,” Current Research in Bacteriology, vol. 2, no. 2, pp. 22–35, 2009.
[5]
K. M. K. Dewa, A. Banito, A. Onasanya, K. E. Kpemoua, and Y. Sere, “Rice bacterial blight in Togo: importance of the disease and virulence of the pathogen,” Current Research in Bacteriology, vol. 4, pp. 94–100, 2011.
[6]
S. Savary, P. S. Teng, L. Willocquet, and F. W. Nutter Jr., “Quantification and modeling of crop losses: a review of purposes,” Annual Review of Phytopathology, vol. 44, pp. 89–112, 2006.
[7]
S. U. Chaudhary, I. Javed, and H. Muzzammil, “Effectiveness of different fungicides and antibiotics against bacterial leaf blight in rice,” Journal of Agricultural Research, vol. 50, no. 1, pp. 109–117, 2012.
[8]
H. Chen, S. Wang, and Q. Zhang, “New gene for bacterial blight resistance in rice located on chromosome 12 identified from Minghui 63, an elite restorer line,” Phytopathology, vol. 92, no. 7, pp. 750–754, 2002.
[9]
S. S. Gnanamanickam, “An overview of progress in biological control,” Biomedical and Life Sciences, vol. 8, pp. 43–51, 2009.
[10]
A. Onasanya, M. M. Ekperigin, Y. Sere, F. E. Nwilene, and J. O. Ajele, “Enzyme polymorphism and genetic diversity in Xanthomonas oryzae pv. oryzae isolates causing rice bacterial leaf blight disease in West Africa,” International Journal of Agricultural Research, vol. 3, no. 3, pp. 227–236, 2008.
[11]
A. Onasanya, M. M. Ekperigin, Y. Sere, F. E. Nwilene, J. O. Ajele, and G. Oboh, “Isozyme fingerprinting and genetic differentiation of Xanthomonas oryzae pv. oryzae isolates as revealed by glucose 6-phosphate dehydrogenase (G6PH) analysis,” Biotechnology, vol. 6, no. 3, pp. 257–263, 2007.
[12]
B. O. Akanji, J. O. Ajele, A. Onasanya, and O. Oyelakm, “Genetic fingerprinting of Pseudomonas aeruginosa involved in nosocomial infection as revealed by RAPD-PCR markers,” Biotechnology, vol. 10, no. 1, pp. 70–77, 2011.
[13]
Y. Anzai, H. Kim, J. Y. Park, H. Wakabayashi, and H. Oyaizu, “Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence,” International Journal of Systematic and Evolutionary Microbiology, vol. 50, no. 4, pp. 1563–1589, 2000.
[14]
C. Srinivasa, U. Sharanaiah, and C. Shivamallu, “Molecular detection of plant pathogenic bacteria using polymerase chain reaction single-strand conformation polymorphism,” Acta Biochimica et Biophysica Sinica, vol. 44, no. 3, pp. 217–223, 2012.
[15]
W.-J. Zhao, S.-F. Zhu, X.-L. Liao, H.-Y. Chen, and T.-W. Tan, “Detection of Xanthomonas oryzae pv. oryzae in seeds using a specific TaqMan probe,” Molecular Biotechnology, vol. 35, no. 2, pp. 119–127, 2007.
[16]
A. Onasanya, P. Kiepe, A. Basso et al., “Molecular characterization and DNA fingerprinting of Xanthomonas oryzae pv oryzae isolates from climate change prone areas in East Africa,” Research Journal of Environmental Sciences, vol. 5, no. 9, pp. 772–778, 2011.
[17]
C. Gonzalez, B. Szurek, C. Manceau, T. Mathieu, Y. Sere, and V. Verdier, “Molecular and pathotypic characterization of new Xanthomonas oryzae strains from West Africa,” Molecular Plant-Microbe Interactions, vol. 20, no. 5, pp. 534–546, 2007.
[18]
A. Onasanya, A. Basso, E. Somado et al., “Development of a combined molecular diagnostic and DNA fingerprinting technique for rice bacteria pathogens in Africa,” Biotechnology, vol. 9, no. 2, pp. 89–105, 2010.
[19]
A. Basso, A. Onasanya, S. Issaka et al., “Bacterial leaf blight of rice in Niger: pathological diversity of isolates collected on irrigated lands,” Journal of Applied BioSciences, vol. 38, pp. 2551–2563, 2011.
[20]
A. Onasanya, H. D. Mignouna, and G. Thottappilly, “Genetic fingerprinting and phylogenetic diversity of Staphylococcus aureus isolates from Nigeria,” African Journal of Biotechnology, vol. 2, no. 8, pp. 246–250, 2003.
[21]
F. J. Rohlf, NTSys Pc, Version 2. 02j, Exeter Software, New York, NY, USA, 2000.
[22]
G. I. Ivchenko and S. A. Honov, “On the Jaccard similarity test,” Journal of Mathematical Sciences, vol. 88, no. 6, pp. 789–794, 1998.
[23]
é. Jakó, E. Ari, P. Ittzés, A. Horváth, and J. Podani, “BOOL-AN: a method for comparative sequence analysis and phylogenetic reconstruction,” Molecular Phylogenetics and Evolution, vol. 52, no. 3, pp. 887–897, 2009.
[24]
J. S. Ebdon and H. G. Gauch, “AMMI analysis of national turfgrass performance trials,” Crop Science, vol. 42, pp. 489–496, 2002.
[25]
T. B. Adhikari, R. C. Basnyat, and T. W. Mew, “Virulence of Xanthomonas oryzae pv. oryzae on rice lines containing single resistance genes and gene combinations,” Plant Disease, vol. 83, no. 1, pp. 46–50, 1999.
[26]
A. Onasanya, P. Kiepe, A. Basso et al., “Molecular characterization and DNA fingerprinting of Xanthomonas oryzae pv oryzae isolates from climate change prone areas in East Africa,” Research Journal of Environmental Sciences, vol. 5, pp. 772–778, 2011.
[27]
S. M. Lalitha, K. V. C. Mohan, P. Premalatha, D. G. Lalitha, Z. Usha, and F. Wayne, “Understanding the bacterial blight pathogen-combining pathotyping and molecular marker studies,” International Journal of Plant Pathology, vol. 1, no. 2, pp. 58–68, 2010.
[28]
J. Hu, Y. Zhang, W. Qian, and C. He, “Avirulence gene and insertion element-based RFLP as well as RAPD markers reveal high levels of genomic polymorphism in the rice pathogen Xanthomonas oryzae pv. oryzae,” Systematic and Applied Microbiology, vol. 30, no. 8, pp. 587–600, 2007.
[29]
D. Innes, I. R. Beacham, C. A. Beven et al., “The cryptic ushA gene (ushAc) in natural isolates of Salmonella enterica (serotype Typhimurium) has been inactivated by a single missense mutation,” Microbiology, vol. 147, no. 7, pp. 1887–1896, 2001.
[30]
S. Mongkolsuk, W. Whangsuk, M. Fuangthong, and S. Loprasert, “Mutations in oxyR resulting in peroxide resistance in Xanthomonas campestris,” Journal of Bacteriology, vol. 182, no. 13, pp. 3846–3849, 2000.
[31]
S. T. Chisholm, G. Coaker, B. Day, and B. J. Staskawicz, “Host-microbe interactions: shaping the evolution of the plant immune response,” Cell, vol. 124, no. 4, pp. 803–814, 2006.