PCR-RFLP ANALYSIS OF THE 16S RRNA AND ITS REGIONS IN BACTERIAL BLIGHT (XANTHOMONAS AXONOPODIS PV. PUNICAE) ACROSS POMEGRANATE FARMS IN KURDISTAN REGIONS-IRAQ.
DOI:
https://doi.org/10.36103/ijas.v54i2.1707Keywords:
survey , infected plants , phylogenetic genetic similarity ITS region, 16S rRNAAbstract
Bacterial blight caused by Xanthomonas axonopodis pv. punicae (Xap) is a major biotic diseases in pomegranate (Punica granatum L.). In the field survey which lasted 2 year in six different geographical locations, Xap strains from the severely infected plant material collected including DuhokCcenter, around Erbil, Zaxo, south of Duhok, Akre, and Amedi. Xanthomonas was detected from infected plant material and its identity was confirmed by morphological, Microbial and Molecular Characterization. To study its genetic variability and phylogenetic relationship two important loci were targeted namely ITS region of 16s rDNA, 16S rRNA and then a PCR-RFLP was performed for PCR product of 16s rRNA loci using Hinf I, Hae III, and Rsa I restriction enzymes. The PCR-RFLP showed the genetic similarity coefficient ranging from 1.00 to 3.73, and the dendrogram grouped all tested strains in 4 clusters. The result revealed that this disease is in blooming stage in the country which was thought that has not been recorded before in pomegranate. To the best of our knowledge this is the first record of Xap. In Kurdistan/ Iraq therefore, further studies are needed to be performed to manage this hazardous bacterium.
References
Adhikari T.B, C.M. Vera Cruz ,Q. Zhang , R.J.Nelson , D.Z. Skinner , T.W. Mew , and T.E. Leach. 1995 Genetic diversity of Xanthomonas oiyzae pv. oryzae in Asia. Appl Environ Microbiol 61:966–971.
Aljabary, A. M. A. O., A.A.Fatih and Z.M. Ahmad. 2022. Response postharvest quality of pomegranates to thermal treatments, immersing in black seed and flaxseed oils. Iraqi Journal of Agricultural Sciences –2020:53(5):1048-1057. https://doi.org/10.36103/ijas.v53i5.1618
Anonymous 2008. Progress Report of Network Project on Mitigating The Bacterial Blight Disease Of Pomegranate in Maharashtra, Karnataka and Andhra Pradesh. National Research Centre on Pomegranate (ICAR), Solapur-413 006, India
Anonymous 2006.. Diagnosis Of Bacterial Blight Of Pomegranate And Its Management- Training Manual, National Research Centre on Pomegranate, Solapur, 2006
Bogdanove AJ, et al. 2011. Two new complete genome sequences offer insight into host and tissue specificity of plant pathogenic Xanthomonas spp. J. Bacteriol. 193:5450–5464
Chand R,and R. Kishun, 1991. Studies on bacterial blight (Xanthomonas campestris pv. punicae) of pomegranate. Indian Phytopathology. 44(3):370-372
Fawcett HS, and AE Jenkins . 1933. Records of citrus canker from herbarium specimens of the genus Citrus in England and the United States. Phytopathology 23:820–824
GAO, R., WANG, J., ZHU, T., JIA, X., & LI, X. .2018. Identification and Molecular Characterization of a Phytoplasma Associated with Pomegranate Fasciation Disease. Horticultural Plant Journal, 4(1), 30–34. doi:10.1016/j.hpj.2018.01.001
Hamdan, A. Q. and F. F. Jomaa. 2020. Response of pomegranate " cv. wonderful" transplants to mineral nutrition and gibberrelic acid. Iraqi Journal of Agricultural Sciences –2020:51(1):339-346. https://doi.org/10.36103/ijas.v51i1.933
Hauben L, L. Vauterin, J. Swings, and ERB Moore. 1997 Comparison of 16S ribosomal DNA sequences of all Xanthomonas species. Int J Syst Bacteriol 47:328–335
.Hayward, A. C. 1993. The hosts of Xanthomonas. In: Xanthomonas (Swings JG. and Civerolo EL eds). London: Chapman and Hall,; pp 1–119
Hingorani MK, and NJ.Singh. 1959. Xanthomonas punicae sp. now on Punica granatum. Indian J. Agric. Sci. 29:45–48
Jomaa, F. F., and A. A. Ali. 2016. Effect of foliar application of potassium zinc and gibberelic acid on leaves and peel content of mineral elements of pomegranate. Iraqi Journal of Agricultural Sciences –47(2): 533-542. https://doi.org/10.36103/ijas.v47i2.599
Kale, P., Chimote, V., Raghuwanshi, K., Kale, A., Jadhav, A., and S. Borkar. 2012. Microbial, biochemical, pathogenicity and molecular characterization of Xanthomonas axonopodis pv. punicae from pomegranate. Journal Pure Applied Microbiology, 6(4): 1699-1706
Lazeeza, S. O. 2021. Antioxidant activity of pomegranate, Iraqi journal of agricultural sciences, 52(1):196–203. https://doi.org/10.36103/ijas.v52i1.1251
Lee JS, KS Ko and H.S. Jung . 2000. Phylogenetic analysis of Xylaria based on nuclear ribosomal ITS1-5.8S-ITS2 sequences. FEMS Microbiology Letters 2000; 187:89-93
Li W, Q. Song , RH. Brlansky and JS. Hartung. 2007. Genetic diversity of citrus bacterial canker pathogens preserved in herbarium specimens. Proc. Natl. Acad. Sci. U. S. A. 104:18427–18432
Lievens B, M. Brouwer, ACRC.Vanachter , CA Levesque, BPA. Cammue and BPHJ. Thomma. 2006.: Quantitative assessment of phytopathogenic fungi in various substrates using a DNA macroarray. Environ Microbiol 2005,7 (11):1698–710. 10.1111/j.1462-2920. 2005.00816.x
Mondal, K. K., Rajendran, T., Phaneendra, C., Mani, C., Sharma, J., Shukla, R., and A. Kumar. 2012. The reliable and rapid polymerase chain reaction (PCR) diagnosis for Xanthomonas axonopodis pv. punicae in pomegranate. African Journal of Microbiology Research, 6(30): 5950-5956
Mondal, K. K., Verma, G., and C. Mani. 2013. Phylogenetic relatedness of Xanthomonas axonopodis pv. punicae, the causal agent of bacterial blight of pomegranate based on two loci, 16S rRNA and gyrB. Annals of Microbiology, 63(2): 801-804
Parkinson N, C. Cowie J. Heeney and D. Stead .2009. Phylogenetic structure of Xanthomonas determined by comparison of gyrB sequences. International journal of systematic and evolutionary microbiology. 2009; 59:264-274
Raghuwanshi K.S, B.A Hujare, V.P, Chimote and S.G. Borkar Characterization of Xanthomonas axonopodis pv. punicae isolates from western Maharashtra and their sensitivity to chemical treatments. The Bio scan. 2013; 8(3):845-850
Rohlf F. J. 1998 Numerical Taxonomy and Multivariate Analysis System, Version 2.0 (Exeter Software, New York), 1998
Schaad N . 1992. Laboratory Guide for Identification of Plant Pathogenic Bacteria. Second Ed. International Book Distributing Co, Lucknow, 1992, 165
Sharma, K., J. Sharma, . and V. Jadhav, 2015. In Recent Advances in Diagonosis and Management of plant Disease 119–126 (Springer 2015).
Sharma, V., S. Midha, M., Ranjan, A.K.., Pinnaka, and P.B. Patil, 2012. Genome
Sequence of Xanthomonas axonopodis pv. punicae Strain LMG 859. Journal of Bacteriology, 194(9), 2395–2395. doi:10.1128/jb.00181-12
Weigand F., Baum and S. Udupa. 1993.. DNA Molecular Marker Techniques. Technical Manual No.20. International Center for Agricultural Research in the Dry Areas (ICARDA). Aleppo, Syria.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
2.jpg)
2.jpg)
