Molecular and Genotoxicity Effect of Certain aqueous Plant Extracts in Helminthosporium Rostratum As a Phytopathogenic Fungi
Author(s)
Fatimah Alotibi , Eman Ashour , Rafa S. Almeer , Ekram A. Mohamed ,
Download Full PDF Pages: 47-65 | Views: 760 | Downloads: 207 | DOI: 10.5281/zenodo.3986607
Abstract
The development of natural antifungal treatments by plant extracts is of great importance because of their safe effects on both the human and animal health, besides their antifungal effects against different Phytopathogenic Fungi. The current study aimed to explore the molecular and genotoxicity effect of two aqueous extracts from wild plants Rumex vesicarius L. and Ziziphus spina-christi aqueous extracts on the leaf spots fungi, Helminthosporium rostratum. The fungal strain was treated with 25 mg/ml of each extract then the alteration in both of the amino acids and DNA of the fungus was assessed by different experiments including, Proteomic analysis by SDS-Page, the total amino acids analysis by the high-performance liquid chromatography (HPLC), DNA profiles analysis by Restriction fragment length polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD), and COMET assay. The results showed that 25 unique bands were produced by the plant's extracts, despite, Ziziphus extract have more effective than Rumex. Besides, the treatments decreased the total amino acid contents dried biomass of H. rostratum. Plant Extracts affects the DNA composition wherein RFLP, Cla I restriction enzyme could produce 100% polymorphic bands post-treatment, in RAPD, seven polymorphic unique bands were obtained with P-12 primer, and finally, COMET assay showed an observed tailed DNA in the treated fungal nuclei. We concluded that both of the two plants aqueous extracts could induce successful changes in the amino acid composition besides a remarkable genotoxic effect on H. Rostratum which highlights their antifungal effect
Keywords
Rumex vesicarius L; Ziziphus spina-christi; Helminthosporium Rostratum, Essential and non-essential amino acids; RFLP; RAPD; COMET assay.
References
i. Aamir, S., Sutar, S., Singh, S.K., Baghela, A., 2015. A rapid and efficient method of fungal genomic DNA extraction, suitable for PCR based molecular methods. Plant Pathology & Quarantine, 5, 74–81.
ii. Abd-Elaah, A.G., Ahmed, S.S., 2005. Effect of fluconazole on mycelial growth and protein profiles of some fungal species isolated from molasses. Assiut University Journal of Botany, 34,131–145.
iii. Abd-Elaah, G.A., Abo-Amer, A.E., Soliman, S.A., 2006. Protein patterns and mycelial growth of dermatophytic fungi affected by desert plant extracts. International Journal of Agriculture and Biology, 8, 434-439.
iv. Abu-Taleb, A.M., El-Deeb, K., El-Otibi, F.O. 2011. Assessment of antifungal activity of Rumex vesicarius L. and Ziziphus spina-christi(L) wild extract against two phytopathogenic fungi. African Journal of Microbiology Research, 5,1001-1011.
v. Ahmad, R., Ahmad, M., Mehjabeen, Jahan, N. Phytochemical screening and anti-oxidant activity of the two plants Ziziphus oxyphylla Edgew and Cedrela serrata Royle. Pakistan Journal of Pharmaceutical Sciences, 27,1477-1482.
vi. Al-Ali, S., Al-Judaibi, A., 2019. Effects of Natural Products and Essential Oils on Pathogenic Fungi. Acta Scientific Microbiology, 2,149-157.
vii. Belmokhtar, Z., Yassine, M., Abdelkader, H., Zohra, F., Samira, G., Lamia, M., Mezmez Ryad, M., Meriem, K.H., 2020. Evaluation of antifungal, antioxidant and phytochemical screening of Ziziphus jujuba Miller leaves from the northwestern region of Algeria. International Journal of Biological and Chemical Sciences, 17,1-9.
viii. Cohen, S.A., Meys, M., Travin, T.L.,1989. The Pico Tag Method: A Manual of Advanced Techniques for Amino Acid Analysis. Waters Chromatography Division, Millipore Corporation, USA.
ix. Coumans, J.V.F., Harvey, J., Backhouse, D., Poljak, A., Raftery, M.J., Nehl, D., Katz, M. E. Pereg, L., 2011. Proteomic assessment of hostassociated microevolution in the fungus Thielaviopsis basicola. Environmental Microbiology, 13,576–588.
x. Cunha, M.G., Rizzo, D.M. 2003. Development of Fungicide Cross Resistance in Helminthosporium solani Populations from California. Plant Disease, 87,798-803.
xi. da Silva, A.F., da Rocha, C.Q., da Silva, L.C.N., Carvalho Júnior, A.R., Mendes, I.N.F.V., de Araruna, A.B., Motta, E.P., Silva, R.S., Campos, C.D.L., Farias, J.R., Oliveira, A.D.S., Silva, D.H.D.S., Nascimento, F.R.F., Guerra, R.N.M., Monteiro, C.A., 2020. Antifungal and Antivirulence Activities of Hydroalcoholic Extract and Fractions of Platonia insignis Leaves against Vaginal Isolates of Candida Species. Pathogens, 9, E84.
xii. da Silva, A. P., Nascimento da Silva, L. C., Martins da Fonseca, C. S., de Araújo, J. M., Correia, M. T., Cavalcanti, M., Lima, V. L., 2016. Antimicrobial Activity and Phytochemical Analysis of Organic Extracts from Cleome spinosa Jaqc. Frontiers in microbiology, 7, 963.
xiii. Daughaday, W.H., Lowry, O.H., Rosebrough, N.J., Fields, W.S., 1952. Determination of cerebrosepiral fluid protein with the folin phenol reagent. Journal of Laboratory and Clinical Medicine, 39,663.
xiv. Davies, M.J., 2016. Protein oxidation and peroxidation. Biochemical Journal, 473,805–825.
xv. Desikan, S., Narayanan, S., 2015. Genetic markers, genotyping methods & next generation sequencing in Mycobacterium tuberculosis. Indian Journal of Medical Research, 141,761–774.
xvi. Diba, K., Mirhendi, H., Kordbacheh, P., Rezaie, S., 2014. Development of RFLP-PCR method for the identification of medically important Aspergillus species using single restriction enzyme MwoI. Brazilian Journal of Microbiology, 45,503–507.
xvii. Ezeonu, C.S., Imo, C., Agwaranze, D.I., Iruka, A., Joseph, A., 2018. Antifungal effect of aqueous and ethanolic extracts of neem leaves, stem bark and seeds on fungal rot diseases of yam and cocoyam. Chemical and Biological Technologies in Agriculture, 5,18.
xviii. Fiorini, A., Rosado, F. R., Bettega, E. M., Melo, K. C., Kukolj, C., Bonfim-Mendonça, P., Shinobu-Mesquita, C. S., Ghiraldi, L. D., Campanerut, P. A., Capoci, I. R., Godoy, J. S., Ferreira, I. C., Svidzinski, T. I., 2016. Candida albicans PROTEIN PROFILE CHANGES IN RESPONSE TO THE BUTANOLIC EXTRACT OF Sapindus saponariaL. Revista do Instituto de Medicina Tropical de Sao Paulo, 58, 25.
xix. Forni, D., Martin, D., Abujaber, R., Sharp, A. J., Sironi, M., Hollox, E. J., 2015. Determining multiallelic complex copy number and sequence variation from high coverage exome sequencing data. BMC genomics, 16, 891.
xx. Garbe, E., Vylkova, S., 2019. Role of Amino Acid Metabolism in the Virulence of Human Pathogenic Fungi. Current Clinical Microbiology Reports, 6,108–119.
xxi. Gatto, M.A., Ippolito, A., Linsalata, V., Cascarano, N.A., Nigro, F., Vanadiaa, S. Di Venerea, D., 2011. Activity of extracts from wild edible herbs against postharvest fungal diseases of fruit and vegetables. Postharvest Biology and Technology, 61,72–82.
xxii. Guan, F., Jin, Y.T., Zhao, J., Xu, A.C., Luo Y.Y., 2018. A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification. Journal of Analytical Methods in Chemistry, 2018,5890140.
xxiii. Idris, O. A., Wintola, O. A., Afolayan, A. J., 2019. Evaluation of the Bioactivities of Rumex crispus L. Leaves and Root Extracts Using Toxicity, Antimicrobial, and Antiparasitic Assays. Evidence-based complementary and alternative medicine : eCAM, 2019, 6825297.
xxiv. Irfan, M., Yang, S., Yuxin, L., Sun, J. X., 2017. Genetic diversity analysis of Morchella sp.by RAPD. Molecular biology research communications, 6(1), 27–31.
xxv. Kawaguchi, S., Nakamura, T., Yamamoto, A., Honda, G., Sasaki, Y. F., 2010. Is the comet assay a sensitive procedure for detecting genotoxicity?. Journal of nucleic acids, 2010, 541050.
xxvi. Khaleel, S.M.J., Jaran, A.S., Haddadin, M.S.Y., 2016. Evaluation of Total Phenolic Content and Antioxidant Activity of Three Leaf Extracts of Ziziphus spina-christi (Sedr) Grown in Jordan. Journal of Advances in Medicine and Medical Research, 14,1-8.
xxvii. Krisko, A., & Radman, M., 2019. Protein damage, ageing and age-related diseases. Open biology, 9(3), 180249.
xxviii. Kumar, V., Sharma, A., Kaur, R., Thukral, A.K., Bhardwaj, R., Ahmad, P., 2017. Differential distribution of amino acids in plants. Amino Acids, 49,821-869.
xxix. Kusai, N.A., Azmi, M.M., Zainudin, N.A., Yusof, M.T., Razak, A.A., 2016. Morphological and molecular characterization, sexual reproduction, and pathogenicity of Setosphaeria rostrata isolates from rice leaf spot. Mycologia, 108,905-914.
xxx. Kuzdraliński, A., Kot, A., Szczerba, H., Nowak, M., Muszyńska, M., 2017. A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat. Journal of Molecular Microbiology and Biotechnology, 27.175-189.
xxxi. Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 277,680-685.
xxxii. Langie, S. A., Azqueta, A., Collins, A. R., 2015. The comet assay: past, present, and future. Frontiers in genetics, 6, 266.
xxxiii. Lehmann, P. F., Lin, D., Lasker, B. A., 1992. Genotypic identification and characterization of species and strains within the genus Candida by using random amplified polymorphic DNA. Journal of clinical microbiology, 30(12), 3249–3254.
xxxiv. Mahboob, S., Al-Ghanim, K. A., Al-Misned, F., Al-Balawi, H., Ashraf, A., Al-Mulhim, N., 2019. Genetic diversity in tilapia populations in a freshwater reservoir assayed by randomly amplified polymorphic DNA markers. Saudi journal of biological sciences, 26(2), 363–367.
xxxv. Mahlo, S. M., Chauke, H. R., McGaw, L., Eloff, J., 2016. ANTIOXIDANT AND ANTIFUNGAL ACTIVITY OF SELECTED MEDICINAL PLANT EXTRACTS AGAINST PHYTOPATHOGENIC FUNGI. African journal of traditional, complementary, and alternative medicines: AJTCAM, 13(4), 216–222.
xxxvi. Mouradov, A., Spangenberg, G., 2014. Flavonoids: a metabolic network mediating plants adaptation to their real estate. Frontiers in plant science, 5, 620.
xxxvii. Nybom, H., Weising, K., Rotter, B., 2014. DNA fingerprinting in botany: past, present, future. Investigative genetics, 5, 1.
xxxviii. Olive, P.L., Banáth, J.P., 2006. The comet assay: a method to measure DNA damage in individual cells. Nature Protocols, 1,23-29.
xxxix. Pontes, F.C., Abdalla, V.C.P., Imatomi, M., Fuentes, L.F.G., Gualtieri, S.C.J., 2019. Antifungal and antioxidant activities of mature leaves of Myrcia splendens (Sw.) DC. Brazilian Journal of Biology. 79,127-132.
xl. Prasad, K., 2017. HPLC Analysis of Amino Acid and Antioxidant Composition of Three Medicinal Plants of (Pithoragarh) Uttarakhand Himalayas. Journal of Analytical & Pharmaceutical Research, 6,00186.
xli. Reinoso, E.B., Bettera, S.G., 2016. Random amplified polymorphic DNA PCR in the teaching of molecular epidemiology. Biochemistry and Molecular Biology Education, 44,391-366.
xlii. Rosenzweig, N., Hanson, L.E., Clark, G., Franc, G.D., Stump, W.L., Jiang, Q.W., Stewart, J., Kirk, W.W., 2015. Use of PCR-RFLP Analysis to Monitor Fungicide Resistance in Cercospora beticola Populations from Sugarbeet (Beta vulgaris) in Michigan, United States. Plant Disease, 99,355-362.
xliii. Saarela, J. M., Burke, S. V., Wysocki, W. P., Barrett, M. D., Clark, L. G., Craine, J. M., Peterson, P. M., Soreng, R. J., Vorontsova, M. S., Duvall, M. R., 2018. A 250 plastome phylogeny of the grass family (Poaceae): topological support under different data partitions. PeerJ, 6, e4299.
xliv. Sahu, A.K., Katageri, I.S., Jadhav, M.P., Vamadevaiah, H.M., 2017. A Simple, Rapid and Effective Protocol for Extraction of Total Plant Proteins from Cotton Leaf. International Journal of Current Microbiology and Applied Sciences, 6, 2968-2975.
xlv. Salem, M.S.Z., 2016. Pathogenetics. An introductory review. Egyptian Journal of Medical Human Genetics, 17,1-23.
xlvi. Sharma, V.K., Graham, N.J.D., 2010. Oxidation of Amino Acids, Peptides and Proteins by Ozone: A Review. The Journal of the International Ozone Association, 32,81-90.
xlvii. Shomeet, A.M.M., El-Samadisy, A.M.I., El-Kholy, R.M.A., Ibrahim, I.S., 2018. Effect of fungicides, antioxidants and their mixtures on mycelial growth of some fungi. Middle East Journal of Applied Sciences, 8,647-655.
xlviii. Stevenson, P.C., Isman, M.P., Belmaina, S.R., 2017. Natural products derived from higher plants have contributed to pest control in their own right either as crude extracts or in purified form and as starting materials for semi-synthetic derivatives. Industrial Crops and Products, 110,2-9.
xlix. Swapna, P.K., Lalchand, P.D., 2016. Fungal Biodiversity of a Library and Cellulolytic Activity of Some Fungi. Indian Journal of Pharmaceutical Sciences, 78,849-854
l. Tembo, Y., Mkindi, A. G., Mkenda, P. A., Mpumi, N., Mwanauta, R., Stevenson, P. C., Ndakidemi, P. A., Belmain, S. R., 2018. Pesticidal Plant Extracts Improve Yield and Reduce Insect Pests on Legume Crops Without Harming Beneficial Arthropods. Frontiers in plant science, 9, 1425.
li. Voglmayr, H., & Jaklitsch, W. M., 2017. Corynespora, Exosporium and Helminthosporium revisited - New species and generic reclassification. Studies in mycology, 87, 43–76.
Cite this Article: