Characterization of Erwinia amylovora causing Twig Blight of Loquat Tree in Loquat Growing Areas of Punjab and KPK, Pakistan

Alveen Mumtaz, Farah Naz, Gulshan Irshad, Riffat Hayat

Abstract


Twig blight of loquat (Eriobotrya japonica Lindl.), caused by the gram-negative enterobacterium Erwinia amylovora, is a major constraint to loquat production worldwide. Recent surveys in loquat-growing areas of Punjab and KPK, Pakistan—specifically in Chakwal (Kalar Kahar, Choa Saidan Shah), Rawalpindi (Taxila, Wah), Murree (Tret and Chattar), and Haripur (Haripur, Khanpur, and Sara i Saleh)—revealed that this disease is highly prevalent. Twig blight compromises the health and vigor of loquat trees, often leading to their death, which discourages farmers from cultivating loquat commercially. Consequently, the area under loquat cultivation and its production have both been declining. Despite the severity of the issue, no systematic studies had been conducted to address this problem until now. A survey was carried out over two consecutive years (2016 and 2017) in 10 loquat-growing locations in Punjab and KPK (three orchards per location). The incidence of twig blight was higher in 2017 compared to 2016. The highest incidence was recorded in Taxila, with 69.74% in 2016 and 78.39% in 2017, while the lowest was in Kalar Kahar (20.86%). The incidence in Kalar Kahar showed a slight increase of less than 1% from the previous year, likely due to effective control measures implemented to limit the spread of the disease. The pathogen E. amylovora was isolated from diseased samples. Out of 102 bacterial isolates, 92 strains were gram-negative, and 72 tested positive in various biochemical assays (Levan, lipase, KOVAC oxidase, glucose fermentation, catalase oxidase, growth at 39°C, production of fluorescent pigment, and pectolytic activity tests). Three highly virulent isolates were identified through pathogenicity tests and confirmed by biochemical and molecular indicators. Molecular tagging of two chromosomal genes (amsB [MN902191] and Pst-glms [MN915084]) and one plasmid gene (pAE29 [MN902192]) was performed, confirming the identity of the isolates as E. amylovora. This study represents the first report of E. amylovora infecting loquat in Pakistan.

Keywords


Bacteria; Erwinia amylovora; Loquat; Molecular tagging; Twig Blight

References


The authors declare that there are no conflicts of interest.

REFERENCES

Agrios, G. 2005. Plant Pathology. Elsevier Academic Press: San Diego, USA.

Alfano, J. R. and A. Collmer. 1997. The type III (Hrp) secretion pathway of plant pathogenic bacteria: trafficking harpins, Avr proteins, and death. Journal of bacteriology, 179: 5655-62.

Ashmawy, N. A., T. I. Zaghloul and M. A. El-Sabagh. 2015. Isolation and molecular characterization of the fire blight pathogen, Erwinia amylovora, isolated from apple and pear orchards in Egypt. Plant Pathology Journal 14: 142-47.

Bell, A. C., T. G. Ranney, T. A. Eaker and T. B. Sutton. 2005. Resistance to fire blight among flowering pears and quince. Horticultueal Science 40: 413-15.

Chiang, K.-S., H. Liu and C. Bock. 2017. A discussion on disease severity index values. Part I: Warning on inherent errors and suggestions to maximise accuracy. Annals of Applied Biology, 171: 139-54.

Dewey, F., Y. L. Wong, R. Seery, T. Hollins and S. Gurr. 1999. Bacteria associated with Stagonospora (Septoria) nodorum increase pathogenicity of the fungus. The New Phytologist, 144: 489-97.

Espinosa, I., M. Báez, M. I. Percedo and S. Martínez. 2013. Evaluation of simplified DNA extraction methods for Streptococcus suis typing. Journal Salud Animal, 35: 59-63.

Hussain, A., N. Abbasi and A. Akhtar. 2006. Fruit characteristics of different loquat genotypes cultivated in Pakistan. II International Symposium on Loquat.

Islam, M. S., R. Sultana, M. Khalekuzzaman, B. Sikdar, U. K. Acharjee, M. F. Hasan and M. A. Islam. 2017. Isolation and characterization of bacterial spot disease of citrus through biochemical approaches and its control measures. Journal of Pharmacognosy and Phytochemistry, 6: 2418-22.

Jakovljevic, V., S. Jock, Z. Du and K. Geider. 2008. Hypersensitive response and acyl‐homoserine lactone production of the fire blight antagonists Erwinia tasmaniensis and Erwinia billingiae. Microbial Biotechnology, 1: 416-24.

Johnson, K. B. 2000. Fire blight of apple and pear. The Plant Health Instructor, 2015: 43-69.

Jones, A. L. and E. L. Schnabel. 2000. The development of streptomycin-resistant strains of Erwinia amylovora. In, Fire blight: the disease and its causative agent, Erwinia amylovora.

Kabeil, S., M. Fayed, M. Amar and I. Al-Far. 2010. Antagonistic activity of some bacterial isolates against Erwinia amylovora. American-Eurasian Journal of Agricultural and Environmental Science, 8: 588-96.

Karadeniz, T. and M. Şenyurt. 2006. Pomological characterization of loquat selections of the Black Sea region of Turkey. II International Symposium on Loquat.

Khan, I. 2003. The history of loquat growing and future prospects of its commercial cultivation and marketing in Pakistan. First International Loquat Symposium Options Mediterraneennes.

Mitrev, S. and E. Kostadinovska. 2016. Isolation and molecular determination of the fire blight pathogen, Erwinia amylovora, isolated from apple trees in the Republic of Macedonia. Journal of Plant Pathology, 98: 267-82.

Nishioka, Y., S. Yoshioka, M. Kusunose, T. Cui, A. Hamada, M. Ono, M. Miyamura and S. Kyotani. 2002. Effects of extract derived from Eriobotrya japonica on liver function improvement in rats. Biological and Pharmaceutical Bulletin, 25: 1053-57.

Noviello, S., R. Gallo, M. Kelly, R. J. Limberger, K. DeAngelis, L. Cain, B. Wallace and N. Dumas. 2004. Laboratory-acquired brucellosis. Emerging infectious diseases, 10: 1848.

Piqué, N., D. Miñana-Galbis, S. Merino and J. M. Tomás. 2015. Virulence factors of Erwinia amylovora: A review. International journal of molecular sciences, 16: 12836-54.

Puławska, J. and P. Sobiczewski. 2012. Phenotypic and genetic diversity of Erwinia amylovora: The causal agent of fire blight. Trees, 26: 3-12.

Schaad, N. 1980. Laboratory guide for identification of plant pathogenic. In, Bacteria. American Phytopathol Society. Minnual, USA.

Schoofs, H., S. Delalieux, T. Deckers and D. Bylemans. 2020. Fire blight monitoring in pear orchards by unmanned airborne vehicles (UAV) systems carrying spectral sensors. Agronomy, 10: 615-24.

Shemshura, O., M. Alimzhanova, E. Ismailova, A. Molzhigitova, S. Daugaliyeva and A. Sadanov. 2020. Antagonistic activity and mechanism of a novel Bacillus amyloliquefaciens MB40 strain against fire blight. Journal of Plant Pathology, 102: 825-33.

Shoeib, A., N. Ashmawy, S. Hammad and E. Hafez. 2016. Molecular and biological identification of Erwinia amylovora Egyptian isolates compared with other German strains. Journal of Plant Physiology and Pathology 5: 1-16.

Slack, S. M., Q. Zeng, C. A. Outwater and G. W. Sundin. 2017. Microbiological examination of Erwinia amylovora exopolysaccharide ooze. Phytopathology, 107: 403-11.

Szwengiel, A., M. Czarnecka and Z. Czarnecki. 2007. Levan synthesis during associated action of levansucrase and Candida cacaoi DSM 2226 yeast. Polish Journal of Food and nutrition sciences, 57: 433-40.

Tancos, K., E. Borejsza-Wysocka, S. Kuehne, D. Breth and K. D. Cox. 2017. Fire blight symptomatic shoots and the presence of Erwinia amylovora in asymptomatic apple budwood. Plant disease, 101: 186-91.

Umesha, S., P. Richardson, P. Kong and C. Hong. 2008. A novel indicator plant to test the hypersensitivity of phytopathogenic bacteria. Journal of microbiological methods, 72: 95-97.

Vanneste, J. L. 2000. Fire blight: The disease and its causative agent, Erwinia amylovora. CAB International.

Vrancken, K., M. Holtappels, H. Schoofs, T. Deckers and R. Valcke. 2013. Pathogenicity and infection strategies of the fire blight pathogen Erwinia amylovora in Rosaceae: State of the art. Microbiology, 159: 823-32.

Wee, Y. C. and H. Keng. 1992. An illustrated dictionary of Chinese medicinal herbs. CAB International.

Yang, H.-w., M. Yu, J. H. Lee, T. Chatnaparat and Y. Zhao. 2020. The stringent response regulator (p) ppGpp mediates virulence gene expression and survival in Erwinia amylovora. Genomics, 21: 1-19.

Zhao, Y.-q., Y.-l. TIAN, L.-m. Wang, G.-m. Geng, W.-j. Zhao, B.-s. Hu and Y.-f. Zhao. 2019. Fire blight disease, a fast-approaching threat to apple and pear production in China. Journal of Integrative Agriculture, 18: 815-20.


Full Text: PDF

DOI: 10.33687/phytopath.013.01.4605

Refbacks

  • There are currently no refbacks.




Copyright (c) 2024 Alveen Mumtaz, Farah Naz, Gulshan Irshad, Riffat Hayat

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.