Characterization of Alternaria brassicae causing black leaf spot disease of cabbage (Brassica oleracea var. capitata) in the southern part of Kazakhstan
Nurdana Salybekova Nurtaevna
Department of Biology, Akhmet Yasawi International Kazakh-Turkish University, Turkistan, KazakhstanEsin Basim
Organic Agriculture Department, Technical Science Vocational School, Akdeniz University, Antalya, TurkeyHüseyin Basim
Department of Plant Protection, Faculty of Agriculture, Akdeniz University, Antalya, TurkeyGulmira Turmetova Zhusupovna
Department of Biology, Akhmet Yasawi International Kazakh-Turkish University, Turkistan, KazakhstanAbstract
Cabbage plants showing symptoms of leaf spot were detected from various fields in the Almaty region of Kazakhstan in the winter seasons of 2015 and 2016. The disease incidences of approximately 50% were recorded in various fields visited in the Almaty region. The pathogen was aseptically isolated from the symptomatic leaves and maintained in an in vitro culture media. Morphological characteristics and sporulation of the fungus was determined under both light and electron microscopy. The extracted genomic DNA of the fungi was subjected to Polymerase Chain Reaction (PCR) using ABCsens/ABCrev and ITS1/ITS4 primers amplifying ABC transporter (Atr1) gene and the internal transcribed spacer regions, respectively. The amplified products of PCR were sequenced, aligned, blasted and compared for similarity with other species in the NCBI GenBank. The cluster analysis result showed 99% homology with related fungi retrieved from the NCBI GenBank for the ITS region. The fungal isolate was pathogenic towards twenty-two-day-old plants, namely, Brassica oleracea, Lycopersicon esculentum, Solanum melongena, and was established as the causal agent of leaf spot on these plants. This is the first record implicating A. brassicae for black leaf spot disease of cabbage in Kazakhstan.
Keywords:
cabbage, Alternaria brassicae, ITS, PCRReferences
Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J. (1990). Basic local alignment search tool. J. Mol. Biol., 215, 403−410.
Bansal, V.K., Kharbanda, P.D., Stringam, G.R., Thiagarajah, M.R., Tewari, J.P. (1994). A comparison of greenhouse and field screening methods for blackleg resistance in doubled haploid lines of Brassica napus. Plant Dis., 78, 276–281.
Chillali, M., Wipf, D., Guillaumin, J.J., Mohammed, C., Botton, B. (1998). Delineation of the European Armillaria species based on the sequences of the internal transcribed spacer (ITS) of ribosomal DNA. New Phytol., 138, 553–561.
Cooke, D.Е.L., Forster, J.W., Jenkins, P.D., Jones, D.G., Lewis, D.M. (1998). Analysis of intraspecific and interspecific variation in the genus Alternaria by the use of RAPD-PCR. Ann. Appl. Biol., 132(2), 197–209.
Conn, K.L., Tewari, J.P., Awasthi, R.P. (1990). A disease assessment key for Alternaria blackspot in rapeseed and mustard. Can. Plant Dis. Sur., 70(1), 19–22.
D’Antuono, L.F., Elementi, S., Neri, R. (2007). Sensory attributes, health promoting aspects and new uses of edible Brassicaceae. Acta Hortic., 741, 65–72.
Guillemette, T., Iacomi-Vasilescu, B., Simoneau, P. (2004). Conventional and real-time PCR based assay for detecting pathogenic Alternaria brassicae in cruciferous seed. Plant Dis., 88(5), 490–496.
Hall, T.A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98NT. Nucl. Acids Sym. Ser., 41, 95–98.
Hong, S.G., Cramer, R.A., Lawrence, C.B., Pryor, B.M. (2005). Alt a1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genet. Biol., 42(2), 119–129.
Jasalavich, C.A., Morales, V.M., Pelcher, L.E., Seguin-Swartz, G. (1995). Comparison of nuclear ribosomal DNA sequences from Alternaria species pathogenic to crucifers, Mycol. Res., 99(5), 604–614.
Jukes, T.H., Cantor, C.R. (1969). Evolution of protein molecules. In: Mammalian protein metabolism, Munro, H.N. (ed.). Vol. III. Academic Press, New York, 21–132.
Jung, D.S., Na, Y.J., Ryu, K.H. (2002). Phylogenic analysis of Alternaria brassicicola producing bioactive metabolites. J. Microbiol., 40, 289–294.
Kordalewska, M., Brillowska-Dąbrowska, A., Jagielski, T., Dworecka-Kaszak, B. (2015). PCR and real-time PCR assays to detect fungi of Alternaria alternata species. Acta Biochim. Pol., 62(4), 707–712.
Kumar, V., Haldar, S., Pandey, K.K., Singh, R.P., Singh, A.K., Singh, P.C. (2008). Cultural, morphological, pathogenic and molecular variability amongst tomato isolates of Alternaria solani in India. World J. Microb. Biot., 24(7), 1003–1009.
Kusaba, M., Tsuge, T. (1995). Phologeny of Alternaria fungi known to produce host-specific toxins on the basis of variation in internal transcribed spacers of ribosomal DNA. Curr. Genet., 28, 491–498.
Maringoni, A.C. (2005). Doenças das crucíferas (brócolis, couve, couve-chinesa, couve-flor, rabanete, repolho e rúcula). In: Manual de fitopatologia: doenças das plantas cultivadas, Kimati, H., Amorin, L., Bergamin, F.A., Camarg, L., Rezende, J. (eds.). 3rd ed. Sao Paulo, 285–287.
Meena, P., Chattopadhyay, C., Kumar, V.R., Meena, R.L., Rana, U.S. (2005). Spore behaviour in atmosphere and trends in variability of Alternaria brassicae population in India. J. Mycol. Plant Pathol., 35, 511.
Meena, P.D., Rani, A., Meena, R., Sharma, P., Gupta, R., Chowdappa, P. (2012). Aggressiveness, diversity and distribution of Alternaria brassicae isolates infecting oilseed brassica in India. Afr. J. Microbiol. Res., 6(24), 5249–5558.
Pandey, K.K., Pande, P.K., Satpathy, S. (2002). Integrated management of disease and insects of tomato, hot pepper and cole crops. IIVR Varanasi. Tech. Bull., 9, 22 pp.
Patni, C.S., Kolte, S.J., Awasthi, R.P. (2005). Cultural variability of Alternaria brassicae, causing Alternaria blight of mustard. Ann. Plant Physiol., 19, 231–242.
Polizotto, R., Andersen, B., Martini, M., Grisan, S., Assante, G., Musetti, R. (2012). A polyphasic approach for the characterization of endophytic Alternaria strains isolated from grapevines. J. Microbiol. Methods, 88(1), 162–171.
Sami, J.M., Marissônia, A.N., Maria, S.X.F., Marcos, P.S. Câmara, Ailton, R. (2012). Survey and prevalence of species causing Alternaria leaf spots on brassica species in Pernambuco. Hortic. Bras., 30(2), http://dx.doi.org/10.1590/S0102-05362012000200027
Sharma, M., Deep, S., Bhati, D.S., Chowdappa, P., Selvamani, R., Sharma, P. (2013). Morphological, cultural, pathogenic and molecular studies of Alternaria brassicae infecting cauliflower and mustard in India. Afr. J. Microbiol. Res., 7(26), 3351–3363.
Simmons, E.G. (1992). Alternaria taxonomy: current status, viewpoint, challenge. In: Alternaria: biology, plant diseases and metabolites, Chelkowski, J., Visconti, A. (eds.). Elsevier, Amsterdam, 1–35.
Simmons, E.G., Roberts, R.G. (1993). Alternaria themes and variations (73). Mycotaxon, 48, 109–140.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S. (2011). MEGA5: molecular evolutionary genetic analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol., 28(10), 2731–2739.
Thomma, B.P.H.J. (2003). Alternaria spp.: from general saprophyte to specific parasite. Mol. Plant Pathol., 4(4), 225–236.
Weising, K., Nybom, H., Wolff, K., Meyer, W. (1995). DNA Fingerprinting in plants and fungi. CRC Press, Boca Raton, 336 pp.
White, T.J., Bruns, T., Lee, S., Taylor, J.W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols.
A guide to methods and applications. Academic Press, New York, 315–322.
Yu, S.H., Cho, H.S., Kim, B.R., Park, M.S. (2003). Morphological and molecular characterization of Alternaria isolates from Solanaceous crops. Korean
J. Mycol., 31, 103–113.
Department of Biology, Akhmet Yasawi International Kazakh-Turkish University, Turkistan, Kazakhstan
Organic Agriculture Department, Technical Science Vocational School, Akdeniz University, Antalya, Turkey
Department of Plant Protection, Faculty of Agriculture, Akdeniz University, Antalya, Turkey
Department of Biology, Akhmet Yasawi International Kazakh-Turkish University, Turkistan, Kazakhstan
License
Articles are made available under the conditions CC BY 4.0 (until 2020 under the conditions CC BY-NC-ND 4.0).
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.
The author signs a statement of the originality of the work, the contribution of individuals, and source of funding.
