The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Assessment of Resistance Induction in Mungbean against Alternaria alternata through RNA Interference

  • Hira Abbas (Department of Bioinformatics and Biotechnology, Government College University Faisalabad) ;
  • Nazia Nahid (Department of Bioinformatics and Biotechnology, Government College University Faisalabad) ;
  • Muhammad Shah Nawaz ul Rehman (Centre of Agricultural Biochemistry and Biotechnology, University of Agriculture Faisalabad) ;
  • Tayyaba Shaheen (Department of Bioinformatics and Biotechnology, Government College University Faisalabad) ;
  • Sadia Liaquat (Department of Bioinformatics and Biotechnology, Government College University Faisalabad)
  • Received : 2023.03.28
  • Accepted : 2024.01.02
  • Published : 2024.02.01
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Abstract

A comprehensive survey of mungbean-growing areas was conducted to observe leaf spot disease caused by Alternaria alternata. Alternaria leaf spot symptoms were observed on the leaves. Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). To the best of our knowledge, this study has been reported for the first time.

Keywords

Acknowledgement

We the authors acknowledge the University of Agriculture, Faisalabad.

References

  1. Adhikari, T. B., Mew, T. W. and Leach, J. E. 1999. Genotypic and pathotypic diversity in Xanthomonas oryzae pv.oryzae in Nepal. Phytopathology 89:687-694. https://doi.org/10.1094/PHYTO.1999.89.8.687
  2. Adhikari, T. B., Mew, T. W. and Teng, P. S. 1994. Phenotypic diversity of Xanthomonas oryzae pv. oryzae in Nepal. Plant Dis. 78:68-72.
  3. Ali, M. A., Wieczorek, K., Kreil, D. P. and Bohlmann, H. 2014. The beet cyst nematode Heterodera schachtii modulates the expression of WRKY transcription factors in syncytia to favour its development in Arabidopsis roots. PLoS ONE 9:e102360.
  4. Aloi, F., Riolo, M., Sanzani, S. M., Mincuzzi, A., Ippolito, A., Siciliano, I., Pane, A., Gullino, M. L. and Cacciola, S. O. 2021. Characterization of Alternaria species associated with heart rot of pomegranate fruit. J. Fungi 7:172.
  5. Aung, S. L. L., Liu, H. F., Pei, D. F., Lu, B. B., Oo, M. M. and Deng, J. X. 2020. Morphology and molecular characterization of a fungus from the Alternaria alternata species complex causing black spots on Pyrus sinkiangensis (Koerle pear). Mycobiology 48:233-239. https://doi.org/10.1080/12298093.2020.1745476
  6. Bankole, S. A. and Adebanjo, A. 2003. Mycotoxins in food in West Africa: current situation and possibilities of controlling it. Afr. J. Biotechnol. 2:254-263. https://doi.org/10.5897/AJB2003.000-1053
  7. Basim, E., Basim, H., Abdulai, M., Baki, D. and Ozturk, N. 2017. Identification and characterization of Alternaria alternata causing leaf spot of olive tree (Olea europaea) in Turkey. Crop Prot. 92:79-88. https://doi.org/10.1016/j.cropro.2016.10.013
  8. Batzer, J. C., Singh, A., Rairdin, A., Chiteri, K. and Mueller, D. S. 2022. Mungbean: a preview of disease management challenges for an alternative U.S. cash crop. J. Integr. Pest Manag. 13:4.
  9. Brar, A. S., Kaur, K., Sindhu, V. K., Tsolakis, N. and Srai, J. S. 2022. Sustainable water use through multiple cropping systems and precision irrigation. J. Cleaner Prod. 333:130117.
  10. Carrascal-Hernandez, D. C., Florez-Lopez, E., Peralta-Ruiz, Y., Chaves-Lopez, C. and Grande-Tovar, C. D. 2022. Eco-friendly biocontrol strategies of Alternaria phytopathogen fungus: a focus on gene-editing techniques. Agriculture 12:1722.
  11. Cenis, J. L. 1992. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Res. 20:2380.
  12. Chen, Y. C., Eisner, J. D., Kattar, M. M., Rassoulian-Barrett, S. L., LaFe, K., Yarfitz, S. L., Limaye, A. P. and Cookson, B. T. 2000. Identification of medically important yeasts using PCR-based detection of DNA sequence polymorphisms in the internal transcribed spacer 2 region of the rRNA genes. J. Clin. Microbiol. 38:2302-2310. https://doi.org/10.1128/JCM.38.6.2302-2310.2000
  13. Deards, M. J. and Montague, A. E. 1991. Purification and characterisation of a major allergen of Alternaria alternata. Mol. Immunol. 28:409-415. https://doi.org/10.1016/0161-5890(91)90154-C
  14. Deresa, E. M. and Diriba, T. F. 2023. Phytochemicals as alternative fungicides for controlling plant diseases: a comprehensive review of their efficacy, commercial representatives, advantages, challenges for adoption, and possible solutions. Heliyon 9:e13810.
  15. Devappa, V. and Thejakumar, M. B. 2016. Morphological and physiological studies of Alternaria alternata causing leaf spot disease of Chilli (Capsicum annuum L.). Int. J. Appl. Pure Sci. Agric. 2:762-773.
  16. Fisher, M. C., Henk, D. A., Briggs, C. J., Brownstein, J. S., Madoff, L. C., McCraw, S. L. and Gurr, S. J. 2012. Emerging fungal threats to animal, plant and ecosystem health. Nature 484:186-194. https://doi.org/10.1038/nature10947
  17. Gabriel, K. R. 1971. The biplot graphic display of matrices with application to principal component analysis. Biometrika 58:453-467. https://doi.org/10.1093/biomet/58.3.453
  18. Gabriel, M. F., Uriel, N., Teifoori, F., Postigo, I., Sunen, E. and Martinez, J. 2017. The major Alternaria alternata allergen, Alt a 1: a reliable and specific marker of fungal contamination in citrus fruits. Int. J. Food Microbiol. 257:26-30. https://doi.org/10.1016/j.ijfoodmicro.2017.06.006
  19. Habibullah, Abbas, M. and Shah, H. U. 2007. Proximate and mineral composition of mung bean. Sarhad J. Agric. 23:463-466.
  20. Hong, S. G., Cramer, R. A., Lawrence, C. B. and Pryor, B. M. 2005. Alt a 1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genet. Biol. 42:119-129. https://doi.org/10.1016/j.fgb.2004.10.009
  21. Jin, Y., Xiong, Y., Xu, C., Ren, J., Guo, Y., Zuo, Y., Zhang, Y. and Geng, X. 2022. First report of Alternaria alternata causing leaf spot on kidney bean in China. Plant Dis. 106:1531.
  22. Jung, J. H., Ha, J., Kim, M. Y., Kim, J.-E., Son, H. and Lee, S.- H. 2019. First report of Alternaria alternata causing leaf spot on mungbean (Vigna radiata) in the Republic of Korea. Plant Dis. 103:1028.
  23. Kasettranan, W., Somta, P. and Srinives, P. 2010. Mapping of quantitative trait loci controlling powdery mildew resistance in mungbean (Vigna radiata (L.) Wilczek). J. Crop Sci. Biotechnol. 13:155-161. https://doi.org/10.1007/s12892-010-0052-z
  24. Kiani, A. K., Dhuli, K., Donato, K., Aquilanti, B., Velluti, V., Matera, G., Iaconelli, A., Connelly, S. T., Bellinato, F., Gisondi, P. and Bertelli, M. 2022. Main nutritional deficiencies. J. Prev. Med. Hyg. 63(2 Suppl. 3):E93-E101.
  25. Koch, A., Biedenkopf, D., Furch, A., Weber, L., Rossbach, O., Abdellatef, E., Linicus, L., Johannsmeier, J., Jelonek, L., Goesmann, A., Cardoza, V., McMillan, J., Mentzel, T. and Kogel, K.-H. 2016. An RNAi-based control of Fusarium graminearum infections through spraying of long dsRNAs involves a plant passage and is controlled by the fungal silencing machinery. PLoS Pathog. 12:e1005901.
  26. Koch, A., Kumar, N., Weber, L., Keller, H., Imani, J. and Kogel, K.-H. 2013. Host-induced gene silencing of cytochrome P450 lanosterol C14α-demethylase-encoding genes confers strong resistance to Fusarium species. Proc. Natl. Acad. Sci. U. S. A. 110:19324-19329. https://doi.org/10.1073/pnas.1306373110
  27. Li, L., Ma, H., Zheng, F., Chen, Y., Wang, M., Jiao, C., Li, H. and Gai, Y. 2021. The transcription regulator ACTR controls ACT-toxin biosynthesis and pathogenicity in the tangerine pathotype of Alternaria alternata. Microbiol. Res. 248:126747.
  28. Lonergan, E., Pasche, J., Skoglund, L. and Burrows, M. 2015. Sensitivity of Ascochyta species infecting pea, lentil, and chickpea to boscalid, fluxapyroxad, and prothioconazole. Plant Dis. 99:1254-1260. https://doi.org/10.1094/PDIS-06-14-0620-RE
  29. Masoodi, K. Z., Lone, S. M. and Rasool, R. S. 2021. Genomic DNA extraction from Fungal Mycelium using the modified Cenis, 1992 method. In: Advanced methods in molecular biology and biotechnology, eds. by K. Z. Masoodi, S. M. Lone and R. S. Rasool, pp. 51-56. Academic Press, Cambridge, MA, USA.
  30. Matic, S., Tabone, G., Garibaldi, A. and Gullino, M. L. 2020. Alternaria leaf spot caused by Alternaria species: an emerging problem on ornamental plants in Italy. Plant Dis. 104:2275-2287. https://doi.org/10.1094/PDIS-02-20-0399-RE
  31. Mouyna, I., Henry, C., Doering, T. L. and Latge, J.-P. 2004. Gene silencing with RNA interference in the human pathogenic fungus Aspergillus fumigatus. FEMS Microbiol. Lett. 237:317-324. https://doi.org/10.1111/j.1574-6968.2004.tb09713.x
  32. Mukherjee, A. K., Rao, A. V. S., De, R. N. and Nayak, P. 1999. Genetic diversity among slow-blasting rice genotypes. Oryza 36:70-73.
  33. Nagrale, D. T., Gaikwad, A. P. and Sharma, L. 2013. Morphological and cultural characterization of Alternaria alternata (Fr.) Keissler blight of gerbera (Gerbera jamesonii H. Bolus ex J.D. Hook). J. Appl. Nat. Sci. 5:171-178.
  34. Nauanova, A. 2020. Resistance of barley varieties to the pathogens of leaf spotting on the artificial infectious background. Syst. Rev. Pharm. 11:1589-1592.
  35. Nayyar, B. G., Woodward, S., Mur, L. A. J., Akram, A., Arshad, M., Naqvi, S. M. S. and Akhund, S. 2017. The incidence of Alternaria species associated with infected Sesamum indicum L. seeds from fields of the Punjab, Pakistan. Plant Pathol. J. 33:543-553. https://doi.org/10.5423/PPJ.OA.04.2017.0081
  36. Onasanya, A., Sere, Y., Nwilene, F., Abo, M. E. and Akator, K. 2004. Reactions and resistance status of differential rice genotypes to rice yellow mottle virus, genus Sobemovirus in Cote d' Ivoire. Asian J. Plant Sci. 3:718-723. https://doi.org/10.3923/ajps.2004.718.723
  37. Price, P. P. 3rd, Purvis, M. A., Cai, G., Padgett, G. B., Robertson, C. L., Schneider, R. W. and Albu, S. 2015. Fungicide resistance in Cercospora kikuchii, a soybean pathogen. Plant Dis. 99:1596-1603. https://doi.org/10.1094/PDIS-07-14-0782-RE
  38. Rasul, F., Cheema, M. A., Sattar, A., Saleem, M. F. and Wahid, M. A. 2012. Evaluating the performance of three mungbean varieties grown under varying inter-row spacing. J. Anim. Plant Sci. 22:1030-1035.
  39. Saleem, A. and El-Shahir, A. A. 2022. Morphological and molecular characterization of some Alternaria species isolated from tomato fruits concerning mycotoxin production and polyketide synthase genes. Plants 11:1168.
  40. Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Preparation and transformation of competent E. coli. In: Molecular cloning: a laboratory manual, eds. by J. Sambrook, E. F. Fritsch and T. Maniatis, Vol. 1, pp. 1-82. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
  41. Sang, H. and Kim, J.-I. 2020. Advanced strategies to control plant pathogenic fungi by host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS). Plant Biotechnol. Rep. 14:1-8. https://doi.org/10.1007/s11816-019-00588-3
  42. Schreinemachers, P., Balasubramaniam, S., Boopathi, N. M., Ha, C. V., Kenyon, L., Praneetvatakul, S., Sirijinda, A., Le, N. T., Srinivasan, R. and Wu, M.-H. 2015. Farmers' perceptions and management of plant viruses in vegetables and legumes in tropical and subtropical Asia. Crop Prot. 75:115-123. https://doi.org/10.1016/j.cropro.2015.05.012
  43. Secic, E. and Kogel, K.-H. 2021. Requirements for fungal uptake of dsRNA and gene silencing in RNAi-based crop protection strategies. Curr. Opin. Biotechnol. 70:136-142. https://doi.org/10.1016/j.copbio.2021.04.001
  44. Singh, S. N. 1987. Response of chilli cultivars to Alternaria alternata and losses under field conditions. Farm Sci. J. 2:96-97.
  45. Skora, J., Otlewska, A., Gutarowska, B., Leszczynska, J., Majak, I. and Stepien, L. 2015. Production of the allergenic protein Alt a 1 by Alternaria isolates from working environments. Int. J. Environ. Res. Public Health. 12:2164-2183. https://doi.org/10.3390/ijerph120202164
  46. Sneath, P. H. A. and Sokal, R. R. 1973. Numerical taxonomy: the principles and practice of numerical classification. W.F. Freeman & Co., San Francisco, CA, USA. 573 pp.
  47. Tak, Y. and Kumar, M. 2020. Phenolics: a key defence secondary metabolite to counter biotic stress. In: Plant phenolics in sustainable agriculture, eds. by R. Lone, R. Shuab and A. N. Kamili, pp. 309-329. Springer, Singapore.
  48. Tarini, N. M. A., Wahid, M. H., Ibrahim, F., Yasmon, A. and Djauzi, S. 2010. Development of multiplex-PCR assay for rapid detection of Candida spp. Med. J. Indones. 19:83-87. https://doi.org/10.13181/mji.v19i2.387
  49. Toledo, A. V., Simurro, M. E. and Balatti, P. A. 2013. Morphological and molecular characterization of a fungus, Hirsutella sp., isolated from planthoppers and psocids in Argentina. J. Insect Sci. 13:18.
  50. Watanabe, M., Lee, K., Goto, K., Kumagai, S., Sugita-Konishi, Y. and Hara-Kudo, Y. 2010. Rapid and effective DNA extraction method with bead grinding for a large amount of fungal DNA. J. Food Prot. 73:1077-1084. https://doi.org/10.4315/0362-028X-73.6.1077
  51. White, T. J., Bruns, T., Lee, S. and Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: a guide to methods and applications, eds. by M. A. Innis, D. H. Gelfand, J. J. Sninsky and T. J. White, pp. 315-322. Academic Press, San Diego, CA, USA.
  52. Wise, A. A., Liu, Z. and Binns, A. N. 2006. Three methods for the introduction of foreign DNA into Agrobacterium. Methods Mol. Biol. 343:43-53. https://doi.org/10.1385/1-59745-130-4:43
  53. Woudenberg, J. H. C., Seidl, M. F., Groenewald, J. Z., de Vries, M., Stielow, J. B., Thomma, B. P. H. J. and Crous, P. W. 2015. Alternaria section Alternaria: species, formae speciales or pathotypes? Stud. Mycol. 82:1-21. https://doi.org/10.1016/j.simyco.2015.07.001
  54. Yadav, D. L., Jaisani, P. and Pandey, R. N. 2014. Identification of sources of resistance in mungbean genotypes and influence of fungicidal application to powdery mildew epidemics. Int. J. Curr. Microbiol. Appl. Sci. 3:513-519.
  55. Yang, Y., Li, R. and Qi, M. 2000. In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. Plant J. 22:543-551. https://doi.org/10.1046/j.1365-313x.2000.00760.x
  56. Zhang, Y. J., Zhang, S., Liu, X. Z., Wen, H. A. and Wang, M. 2010. A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains. Lett. Appl. Microbiol. 51:114-118. https://doi.org/10.1111/j.1472-765X.2010.02867.x
  57. Zhou, X.-F., Sun, J.-D., Zhao, Z., Lv, J., Wei, X.-W. and Cai, R. 2012. The feasibility analysis of PVX and TRV vectors as the VIGS tool for studying the gene function. Phys. Proc. 33:46-54. https://doi.org/10.1016/j.phpro.2012.05.029