Bioprospecting foliar endophytic fungi of Vitis labrusca Linnaeus, Bordô and Concord cv.
- 319 Downloads
Endophytic fungi colonize the interior of plant tissues and organs, establishing an intimate mutualistic association with no visible symptoms. The fungi may help protect the plant against herbivores and pathogens, making them potentially useful endophytes in the biological control of diseases and agricultural pests. The biotechnological interest in these organisms has stimulated research related to the bioprospecting of endophytic fungi. Grapevine is among the oldest of plants cultivated by man, with the grape being one of the most highly consumed fruits in the world. Diseases cause significant damage to grape cultures, making their integrated control important to reduce the use of pesticides and, consequently, environmental and human contamination. The rustic species Vitis labrusca L. (Vitaceae), used in the preparation of juices and wines, is highly resistant to fungal diseases. We isolated leaf endophytic fungi of the Bordô and Concord cultivars (V. labrusca L.), which were ordered into 68 and 62 morpho-groups of the Bordô and Concord cultivars, respectively. We used scanning electron microscopy to confirm the presence of endophytes in the leaves. Endophytic diversity was analyzed based on sequencing the ITS1-5.8S-ITS2 region of rDNA, allowing the identification of fungi belonging to genera including Cochliobolus, Bipolaris, Fusarium, Alternaria, Diaporthe, Phoma and Phomopsis. Phylogenetic analysis confirmed the identity of the endophytes. The biotechnological potential of endophytes was tested in vitro for the control of pathogenic fungi of grapevines including Alternaria sp., Sphaceloma sp. and Glomerella sp. Inhibition percentages above 50 % as demonstrated by some isolates demonstrate their potential for biological control.
KeywordsEndophytes Grapevine Biological control Sequencing of rDNA Phylogenetic analysis
We thank the Complexo de Centrais de Apoio à Pesquisa (COMCAP/UEM) for sequencing the ITS1-5.8S-ITS2 regions and for the production of electronic scanning micrographs. We thank the EMBRAPA Uva e Vinho for the phytopathogen strains. We thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the Master’s scholarship for the first author and CAPES/PNPD-UEM postdoctoral scholarship. We thank CNPq (311534/2014-7; 447265/2014-8) and Fundação Araucária – FA (276/2014) for financial support.
- Arnold AE (2008) Endophytic fungi: hidden components of tropical community ecology. In: Carson WP, Schnitzer SA (eds) Tropical Forest Community Ecology. Wiley-Blackwell, HobokenGoogle Scholar
- Badalyan SM, Innocenti G, Garibyan NG (2002) Antagonistic activity of xylotrophic mushrooms against pathogenic fungi of cereals in dual culture. Phytopathol Mediterr 41:200–225Google Scholar
- Bernardi-Wenzel J, Garcia A, Azevedo JL, Pamphile JA (2013) Molecular characterization by amplified ribosomal DNA restriction analysis and antimicrobial potential of endophytic fungi isolated from Luehea divaricata (Malvaceae) against plant pathogenic fungi and pathogenic bacteria. Genet Mol Res 12(4):5072–5084CrossRefPubMedGoogle Scholar
- Burruano S, Alfonzo A, Lo Piccolo S, Conigliaro G, Mondello V, Torta L, Moretti M, Assante G (2008) Interaction between Acremonium byssoides and Plasmopara viticola in Vitis vinifera. Phytopathol Mediterr 47:122–131Google Scholar
- Casieri L, Hofstetter V, Viret O, Gindro K (2009) Fungal communities living in the wood of different cultivars of young Vitis vinifera plants. Phytopathol Mediterr 48:73–83Google Scholar
- Faostat—Food and Agriculture Organization of the United Nations (2012) Available from: http://faostat.fao.org/site/339/default.aspx. Accessed: 20 july 2014
- Ferreira DF (2008) SISVAR: um programa para análise e ensino de estatística. Rev Científica Symp 6:36–41Google Scholar
- Lima TEF (2010) Micobiota endofitica de Vitis labrusca L. CV Isabel no vale do Siriji, Pernambuco, Brasil. Thesis, Universidade Federal de PernambucoGoogle Scholar
- Mostert L, Crous PW, Petrini O (2000) Endophytic fungi associated with shoots and leaves of Vitis vinifera, with specific reference to the Phomopsis viticola coplex. Sydowia 52:46–48Google Scholar
- Musetti R, Vecchione A, Stringher L, Borselli S, Zulini L, Marzani C, Dambrosio M, Sanità Di Toppi L, Pertot I (2006) Inhibition of sporulation and ultrastructural alterations of grapevine downy mildew by the endophytic fungus Alternaria alternata. Phytopathology 96:689–698CrossRefPubMedGoogle Scholar
- Pamphile JA, Gai CS, Pileggi M, Rocha CLMSC, Pileggi SAV (2008a) Plant-microbe interactions between host and endophytes observed by scanning electron microscopy (SEM). In: Sorvari S, Pirttilä AM (eds) Prospects and Applications for Plant-Associated Microbes. A Laboratory Manual, part A: Bacteria. BBI (BioBien Innovations), Finland, pp 184–189Google Scholar
- Pamphile JA, Pileggi M, Gai CS, Rocha CLMSC, Pileggi SAV (2008b) Scanning electron microscopy (SEM). In: Sorvari S, Pirttilä AM (eds) Prospects and Applications for Plant-Associated Microbes. A Laboratory Manual, part A: Bacteria. BBI (BioBien Innovations), Finland, pp 9–13Google Scholar
- Rehman S, Mir T, Kour A, Qazi PH, Sultan P, Shawl AS (2011) In vitro antimicrobial studies of Nodulisporium specie: an endophytic fungus. J Yeast Fungal Res 2:53–58Google Scholar
- Reyes Chilpa R, Quiroz Vásquez RI, Jiménez Estrada M, Navarro-Ocaña A, Cassini Hernández J (1997) Antifungal activity of selected plant secondary metabolites against Coriolus versicolor. J Trop Forest Prod 3:110–113Google Scholar
- Sandhu SS, Aharwal RP, Kumar S (2014) Isolation and antibacterial property of endophytic fungi isolated from Indian medicinal plant Calotropis procera (Linn.). World J Pharm Pharm Sci 3:678–691Google Scholar
- Smith D, Onions AHS (1983) The preservation and maintenance of living fungi. Page, NorwickGoogle Scholar
- Sousa JSI (1996) Uvas para o Brasil, 2nd edn. FEALQ, PiracicabaGoogle Scholar
- Stone JK, Bacon CW, White JF (2000) An overview of endophytic microbes: endophytism defined. In: Bacon CW, White JF (eds) Microbial Endophytes. Dekker, New York, pp 3–30Google Scholar
- White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfland DH, Sninsky JJ, White TJ (eds) PCR Protocols. A Guide to Methods and Applications. Academic, New York, pp 315–322Google Scholar