The tangerine pathotype of Alternaria alternata is the aetiological agent of Alternaria brown spot on tangerines. In the state of Paraíba, Brazil, its occurrence on “Dancy ” tangerine trees is associated with genetic aspects as well as the influence of environmental conditions on reproduction and dissemination within and between populations. The aim of the present study was to evaluate the diversity of isolates of this pathogen using morphophysiological and molecular markers. For the analysis of mycelial growth and sporulation, 3 0 isolates from different locations were examined at 24-hour intervals until the seventh day, when the spores were quantified. The 3 0 isolates were characterised based on molecular markers (ISSR) and genetic similarity (Jaccard index). A factor arrangement was used: 30 isolates, four media (ODA, PDA, LEA and V8), three light regimes (continuous dark, alternating light and continuous light) and three temperatures (15°C, 25°C and 35°C), with 12 repetitions. Groups 1, 2 and 3 presented low genetic variability. Group 4 showed high genetic variability of the isolates obtained from the Massaranduba (state of Paraíba-Brazil) producing region and higher mycelial growth and sporulation of A. alternata . The continuous light regime and the temperature 25°C in PDA and V8 media were the ideal conditions for the mycelial growth and sporulation, respectively, of the isolates of A. alternata .
Keywords<i>Citrus tangerine</i>Alternaria Brown SpotFungal VariabilityEnvironment FactorsISSR Markers
Whiteside, J.O. (1976) A Newly Recorded Alternaria Induced Brown Spot Disease on Dancy Tangerines in Florida. Plant Disease Report, 3, 326-329.
Canihos, Y., Erkilic, A. and Timmer, L.W. (1997) First Report of Alternaria Brown Spot of Minneola tangelo in Turkey. Plant Disease, 81, 1214. https://doi.org/10.1094/PDIS.1997.81.10.1214B
Vicent, A., Armengol, J., Sales, R., GarciaJimenez, J. and Alfaro-Lassala, F. (2000) First Report of Alternaria Brown Spot of Citrus in Spain. Plant Disease, 84, 1044. https://doi.org/10.1094/PDIS.2000.84.9.1044B
Peres, N.A. and Timmer, L.W. (2006) Evaluation of the Alter-Rater Model for Spray Timing for Control of Alternaria Brown Spot on Murcott tangor in Brazil. Crop Protection, 25, 454-460. https://doi.org/10.1016/j.cropro.2005.07.010
Wang, X.F., Li, Z.A., Tang, K.Z., Zhou, C.Y. and Yi, L. (2010) First Report of Alternaria Brown Spot of Citrus Caused by Alternaria alternata in Yunnan Province, China. Plant Disease, 94, 375. https://doi.org/10.1094/PDIS-94-3-0375C
Ghasemloee, M. and Niazmand, A.R. (2015) Study of A. alternata Associated with Citrus spp. Journal of Pure and Applied Microbiology, 9, 473-482.
Akimitsu, K., Peever, T.L. and Timmer, L.W. (2003) Molecular, Ecological and Evolutionary Approaches to Understanding Alternaria Diseases of Citrus. Molecular Plant Pathology, 1, 435-446. https://doi.org/10.1046/j.1364-3703.2003.00189.x
Azevedo, F.A., Martelli, I.B., Polydoro, D.A., Pacheco, C.A., Schinor, E.H. and Batianel, M. (2015) Positive Relationship between Citrus Leaf Miner and Alternaria Brown Spot. Ciência Rural, 45, 1160-1163. https://doi.org/10.1590/0103-8478cr20141248
Bassimba, D.D.M., Mira, J.L. and Vicent, A. (2014) Inoculum Sources, Infection Periods, and Effects of Environmental Factors on Alternaria Brown Spot of Mandarin in Mediterranean Climate Conditions. Plant Disease, 98, 409-417. https://doi.org/10.1094/PDIS-09-13-0956-RE
Dewdney, M.M. (2016) Alternaria Brown Spot, Alternaria Diseases in Citrus, SP 043 Florida Citrus Pest Management Guide. http://edis.ifas.ufl.edu/cg021
Woudenberg, J.H.C., Seidl, M.F., Groenewald, J.Z., Vries, M., Stielow, M.B., Thomma, B.P.H.J. and Crous, P.W. (2015) Alternaria Section Alternaria: Species, Formae Speciales or Pathotypes? Studies in Mycology, 1, 1-21. https://doi.org/10.1016/j.simyco.2015.07.001
Silva, J.L. and Teixeira, R.N.V. (2012) Sporulation and Mycelial Growth of Fusarium solani in Different Culture Media and Steady Bright. Revista Agroambiente, 6, 47-52.
Dini-Andreote, F., Pietrobon, V.C., Dini-Andreote, F., Romao, A.S., Spósito, M.B. and Araújo, W.L. (2009) Genetic Variability of Brazilian Isolates of Alternaria alternata Detected by AFLP and RAPD Techniques. Brazilian Journal of Microbiology, 40, 670-677. https://doi.org/10.1590/S1517-83822009000300032
Souza, D.C.L. (2015) Molecular Techniques for Characterization and Conservation of Medicinal and Aromatic Plants: A Review. Revista Brasileira de Plantas Medicinais, 17, 495-503. https://doi.org/10.1590/1983-084X/13_071
Canihos, Y., Peever, T.L. and Timmer, L.W. (1999) Temperature, Leaf Wetness, and Isolate Effects on Infection of Minneola Tangelo Leaves by Alternaria sp. Plant Disease, 83, 429-433. https://doi.org/10.1094/PDIS.1999.83.5.429
Pulz, P. and Massola Junior, N.S. (2009) Effect of Culture Media and Physical Factors on Growth and Sporulation of Alternaria dauci and A. solani. Summa Phytopathologica, 35, 121-126. https://doi.org/10.1590/S0100-54052009000200007
Salgado, A.P.S.P., Cardoso, M.G., Souza, P.E., Souza, J.A., Abreu, C.M.P. and Pinto, J.E.B.P. (2003) Fungitoxic Activity Evaluation of Essencial Leaf Oils of Eucalyptus on Fusarium oxysporum, Botrytis cinerea and Bipolaris sorokiniana. Ciência e Agrotecnologia, 27, 249-254. https://doi.org/10.1590/S1413-70542003000200001
Gomes-Oliveira, I.V. and Matsumura, A.T.S. (2001) Technique to Obtain of Alternaria solani Culture Filtrate. Tropical Plant Pathology, 26, 1.
Waals, J.E.V., Korsten, L. and Slippers, B. (2004) Genetic Diversity among Alternaria solani Isolates from Potatoes in South Africa. Plant Disease, 88, 959-964. https://doi.org/10.1094/PDIS.2004.88.9.959
Raeder, V. and Broda, P. (1987) Rapid Preparation of DNA from Filamentous Fungi. Letters of Applied Microbiology, 1, 17-20. https://doi.org/10.1111/j.1472-765X.1985.tb01479.x
Jaccard, P. (1901) étude Comparative de la distribuition florale dans une portion des Alpes et des Jura. Bulletin de la Société Vaudoise des Sciences Naturelles, 7, 547-579.
Nei, M. (1973) Analysis of Gene Diversity in Subdivided Populations. Proceedings of the National Academy of Sciences, 70, 3321-3323. https://doi.org/10.1073/pnas.70.12.3321
Nei, M. (1977) F-Statistics and Analysis of Gene Diversity in Subdivided Populations. Annals of Human Genetics, 41, 225-233. https://doi.org/10.1111/j.1469-1809.1977.tb01918.x
Nei, M. (1987) Molecular Evolutionary Genetics. Columbia University Press, New York.
SAS Institute (2011) SAS/STAT Versao 9.3: User’s Guide. SAS Institute Inc., 8621.
Lukens, R.L. (1963) Photo-Inhibition of Sporulation in Alternaria solani. American Journal of Botany, 50, 721-724. https://doi.org/10.2307/2440051
Aragaki, M. (1962) Quality of Radiation Inhibitory to Sporulation of Alternaria tomato. Phytopathology, 52, 1227-1228.
Trione, E.J. and Leach, C.M. (1969) Ligth-Induced Sporulation and Sporogenic Substances in Fungi. Phytopathology, 59, 1077-1083.
Hanada, R.E., Gasparotto, L. and Pereira, J.C.R. (2002) Sporulation of Mycosphaerella fijiensis in Different Culture Media. Tropical Plant Pathology, 27, 170-173.
Brunelli, K.R., Fazza, A.C., Athayde Sobrinho, C. and Camargo, L.E.A. (2006) Effect of Culture Media and Light Exposure on the Sporulation of Cercospora zeae-maydis. Summa Phytopathologica, 32, 92-94. https://doi.org/10.1590/S0100-54052006000100016
Meng, J.W., Zhu, W., He, M.H., Wu, E.J. Yang, L.N., Shang, L.P. and Zhan, J. (2015) High Genotype Diversity and Lack of Isolation by Distance in the Alternaria solani Populations from China. Plant Pathology, 64, 434-441. https://doi.org/10.1111/ppa.12275
Satyanarayana, K. and Sadasiva Reddy, C. (1986) A New and Cheap Medium Supporting the Sporulation of Pyricularia oryzae. Indian Journal Mycological Plant Pathology, 163, 329-330.
Leao, E.U., Santos, G.R., Sarmento, Reis, M.R. and Chagas Júnior, A.F. (2012) Mycelial Growth and Conidial Production of Ascochyta cucumis in Different Culture Media and Light Regimen. Bioscience Journal, 28, 325-331.
Choudhary, S., Ghasolia, R.P., Yadav, A.L., Kakraliya, G.L. and Nasanwa, R. (2017) Effect of Physical Factors on Growth of Alternaria alternata Inciting Leaf Blight of Isabgol. Chemical Science Review and Letters, 22, 857-863.
Queiroz, F.M., Batista, U.G. and Brommonschenkel, S.H. (2004) Evaluation of Culture Media on the Mycelial Growth and Sporulation of Alternaria brasiliensis. Tropical Plant Pathology, 29, 541-543.
Carvalho, D.D.C., Alves, E., Camargo, R.B., Oliveira, D.F., Scolforo, J.R.S., Carvalho, D.A. and Batista, T.R.S. (2011) Plant Extracts to Control Alternaria alternata in Murcott Tangor Fruits. Revista Iberoamericana de Micología, 28, 173-178. https://doi.org/10.1016/j.riam.2011.05.001
Ghasemloee, M. and Niazmand, A.R. (2014) Genetic Diversity of Alternaria alternate Associated with Citrus spp. in Southern Iran Based on RAPD-PCR. Journal of Pure and Applied Microbiology, 9, 10-12.
Souza, B.O., Casela, C.R., Nunes, J.A. and Castro, H.A. (2013) Management of Sorghum Anthracnose through Diversification of Genetic Resistance in Host Population. Tropical Plant Pathology, 38, 20-27. https://doi.org/10.1590/S1982-56762013000100003
Adachi, Y., Watanabe, H., Tababe, K., Doke, N., Nishimura, S. and Tsuge, T. (1993) Nuclear Ribosomal DNA as a Probe for Genetic Variability in the Japanese Pear Pathotype of Alternaria alternate. Applied and Environmental Microbiology, 59, 3197-3205.
Nodari, R.O. and Guerra, M.P. (2015) Agroecology: Research Strategies and Values. Estudos Avancados, 29, 183-207. https://doi.org/10.1590/S0103-40142015000100010
Michereff, S.J., Noronha, M.A., Rocha Jr., O.M., Silva, J.A. and Mizubuti, E.S.G. (2003) Variability of Alternaria brassicicola Strains in the State of Pernambuco, Brazil. Tropical Plant Pathology, 28, 656-663.