Pcr Based Detection Of Xanthomonas Citri Subsp. Citri From Acid Lime Using Sequencing Of Signature Genes

Authors

  • S S Isokar Plant Pathology Section, College of Agriculture, Nagpur.
  • S B Bramhankar Plant Pathology Section, College of Agriculture, Nagpur.
  • A K Das ICAR-CCRI, Amravati Road, Nagpur.
  • G T Dinkwar Plant Pathology Section, College of Agriculture, Nagpur.
  • A A Labhashetwar Plant Pathology Section, College of Agriculture, Nagpur.

DOI:

https://doi.org/10.48165/

Keywords:

Acid lime, Canker, PCR, Xanthomonas citri subsp. citri

Abstract

Xanthomonas citri subsp. citri causes citrus canker, which is one of the most serious diseases. The bacterium infects the twigs, petioles, fruit stalks, and fruits, causing acid lime loss in both quality and quantity. On the nutrient agar medium, Xanthomonas citri subsp. citri (Xcc) was successfully isolated, from different agroclimatic zones of Maharashtra. All of the Xcc isolates' pathogenic potential was confirmed, and it was found that isolates Xcc-6, Xcc-7, Xcc-10, Xcc-11, and Xcc-12 were highly pathogenic to commence minute canker lesions and fully developed symptoms after 17-22 days. Molecular detection by polymerase chain reaction with speciûc primers J-pth 1/Jpth-2, J Rxg/Rxc2,Xac-01/Xac-02, M+/M- and Ms+/Ms- was performed on fourteen isolates. The patterns showed by PCR analysis were identical for all the isolates tested confirming their identity as Xanthomonas citri subsp. citri. The highly virulent isolates were sent to the Agri-genome Pvt. Ltd., Hyderabad for sequencing. The result of DNA sequencing were searched against other sequences deposited in the NCBI gene bank database using NCBI-BLAST and all five isolates showed 100 per cent similarity with Xanthomonas citri subsp. citri genome. 

References

Abhang, P. B., M. V. Totawar and R. Kadam, 2015: Biochemical characterization of Xanthomonas axonopodispv.citri for identification of citrus canker disease. J. Basic Sci., 10:30-33.

Anonymous, 2015: Horticulture statistics at a glance,

NHM, Govt. of India, pp.237.

Bodnar A. M., G. Santillana, B. V. Mavrodieva, B. Z. Liu, B. M. Nakhla and D. W. Gabriela, 2017: Complete genome sequences of three Xanthomonas citri strains from Texas. American Soc. of Microbiol., 5(28):52-54.

Cubero, J., J. H. Graham and T. R. Gottwald, 2002: Quantitative PCR method for diagnosis of citrus bacterial canker. App. and Env. Microbiol., 67(7):2849- 2852.

Delcourt, S., C. Vernière, C. Boyer, O. Pruvost, B. Hostachy, and I. Robène-Soustrade, 2013: Revisiting

the specificity of PCR primers for diagnostics of Xanthomonas citripv. citri by experimental and in silico analyses. Plant Dis.,97:373-378.

Filho, H. D., M. A. Takita, A. A. de Souza, J. R. Neto, S. A. L. Deste 'fano, J. S. Hartung and M. A. Machado, 2005: Primers based on the rpf gene region provide improved detection of Xanthomonas axonopodis pv. citri in naturally and artiûcially infected citrus plants. J. Appl. Micro., 100:279–285.

Giri, G. K., R. M. Gade, A. R. Gulhane and Supriya Das, 2008: Efficacy of bio-agents, botanicals, and chemicals against citrus canker. J. Pl. Dis. Sci., 3(2): 249-250.

Hauben, L., L. Vauterin, J. Swings and E. R. B. Moore, 1997: Comparison of 16s ribosomal DNA sequences of all Xanthomonas species. Int. J. Syst. Bacteriology, 47(2):328-335.

Henson, J. and R. French, 1993: The polymerase chain reaction and plant disease dignosis. Annu. Rev. Phytopathol., 31: 81-109.

Jabeen, R., U. R. Saeed, Rahman and A. Rais, 2011: Evaluating BLB resistance / aggressiveness in rice through best inoculums concentration, inoculation and application methods. Pakistan J. Bot., 43 (5): 2635-2638.

www.appjpds.org

Katkar, M., K. S. Raghuwanshi, V. P. Chimote and S. G. Borkar, 2016: Pathological, biochemical and molecular diversity amongst the isolates of Xanthomonas axonopodispv. citri causing the citrus canker in acid lime from the different agro-climatic region of India. Int. J. Environ. Agri. Biotech., 1(2): 2456-2478.

Midha, S., M. Ranjan, V. Sharma, A. K. Pinnaka, and P. B. Patil, 2012: Genome Sequence of Xanthomonas citripv. mangiferaeindicae strain LMG 941. J. Bacteriol., 194(11):3031.

Mubin, M., H. M. I. Arshad, Y. Iftikhar, M. I. Ullah and I.

Bilqees, 2015: Bio-chemical characterization of Xanthomonas axanopodis pv. citri: A gram negative bacterium causing citrus canker. Int. J. Sci. Nature, 6(2): 151-154.

Nehal, J., A. M. Khan, A. A. Khan, M. Mubin, H. M. Tahir and J. Iqbal, 2017: Development and optimization of molecular technique for diagnosis of citrus canker in citrus cultivars. Bio. Pak., 63 (2):135-139.

Schoulties, C. L., E. L. Civerolo, J. W. Miller and R. E. Stall, 1987: Citrus canker in Florida. Plant Dis., 71:388-395.

Toth, I. K., L. J. Hyman, R. Taylor and P. R. J. Birch, 1998:

PCR-based detection of Xanthomonas campestris pv. phaseoli var. fuscans in plant material and its differentiation from X. campestris pv. phaseoli. J. App. Microb., 85:327-336

Published

2021-10-20

How to Cite

Pcr Based Detection Of Xanthomonas Citri Subsp. Citri From Acid Lime Using Sequencing Of Signature Genes . (2021). Journal of Plant Disease Sciences, 16(2), 160–165. https://doi.org/10.48165/