Maestría en Nutrición vegetal

Permanent URI for this collectionhttp://repositorio.uta.edu.ec/handle/123456789/31989

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    Aislamiento, caracterización cultural, morfológica, patogénica e identificación de genes AVR en Cladosporium fulvum Cooke.
    (2023-12) Sánchez Ortiz, Aldo Martin; Leiva Mora, Michel
    Gray mold caused by Cladosporium fulvum was described by Cooke in 1883, which develops in nightshades, specifically in tomato crops, causing damage from the youngest leaves to the top of the plant. The objective of this research was to "Isolate and characterize culturally, morphologically and pathogenically monosporic isolates of Cladosporium fulvum" obtained from signs of gray mold on Solanum lycopersicum leaves to identify the presence or absence of Avr genes in the cantons of the province of Tungurahua (Ambato, Baños, Cevallos, Mocha, Patate, Pelileo, Píllaro, Tisaleo). For isolation, the leaf printing method was applied in potato dextrose (PDA) culture medium with gentamicin sulfate, to describe the cultural and morphological characteristics, the microculture method was used accompanied by the observation of fungal structures under an optical microscope. of transmitted light. In the pathogenic characterization, the isolates of C. fulvum were activated The conidia were counted in the Neubauer chamber of the differential cultivars previously sown on substrates and the inoculum was applied with a manual sprayer. For amplification, DNA extraction and lyophilization of each of the isolates was carried out prior to development. of conventional PCR with two denaturations at 94°C with different cycles and an annealing at 55°C and 34 cycles with a final extension of 72°C. Obtaining that in the 8 cantons of the province of Tungurahua they all showed olive green colors on the obverse, while on the back it was black, with a superficial elevation, plush texture and irregular shape, lobed edges and in some isolates there was pigmentation and perspiration liquid, the mycelial growth in the colonies was obtained with a length of 5.93 µm, width 2.8 µm, hyphae 5.37 µm and conidiophores 32.97 µm average values. In the pathogenic characterization it was evident that the degree of affectation 1 represented between 1 to 5% of the surface of leaves with signs of C. fulvum, while 2 varied from 6 to 20% in the differential cultivars (Cf0-Cf2- Cf4- Cf5-Cf6), finally in the detection of the presence or absence of genes, the presence of the genes Avr2, Avr4e, Ecp2, Ecp4 and Ecp5 and total absence of the genes Avr4, Avr9 and Ecp1 were observed. Based on the results and the tests carried out, it was possible to create bases for the selection of resistance genes for the production of S. lycopersicum hybrids in the province of Tungurahua.
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    Aislamiento, caracterización e identificación de bacterias con capacidad fijadora de nitrógeno atmosférico asociadas con la rizosfera de Solanum tuberosum L. Var. Superchola
    (2024-02) Muñoz Muñoz, Kevin Alexis; Leiva Mora, Michel
    The present research was developed with the purpose of isolating, characterizing and identifying in vitro atmospheric nitrogen-fixing bacteria associated with the rhizosphere of S. tuberosum var. Superchola; as well as to determine their effect on the growth of potato vitroplants in greenhouses. Isolates were obtained from four localities in the province of Tungurahua, which were culturally and morphologically characterized. The in vitro atmospheric nitrogen fixation capacity was determined using modified Jensen culture medium, and molecular identification was performed by PCR amplification of 16S rRNA genes. Finally, the effect of the bacterial isolates on the growth of potato vitroplants in greenhouses was determined. The results showed that in the canton of Píllaro there was the highest quantity of CFU. g-1 of soil (9.08 x 109), in the cultural characterization 87% had smooth texture, 42% flat surface, 82% smooth consistency, 42% circular shape, 84% entire margin and 95% showed brightness. In the morphological characterization of the isolates, 84% were Gram (-) bacteria and 95% were bacilli. The bacterial isolate with the highest atmospheric nitrogen-fixing potential was CC-FCAGP-BFN1 and the one with the lowest potential was CC-FCAGP-BFN15. PCR amplification of 16S rRNA genes identified the strains Pseudomonas sp. (CC-FCAGP-BFN1, CC-FCAGP-BFN16), CC-FCAGP-BFN14) and Paraburkholderia dipogonis (CC-FCAGP-BFN24), which showed a high capacity for atmospheric nitrogen fixation. Likewise, the bacterial strain CC-FCAGP-BFN1 (Pseudomonas sp.) was the only one that increased leaf area, fresh and dry mass and leaf area index of S. tuberosum vitroplants. Based on the above results, four bacterial strains were identified from the rhizosphere of S. tuberosum, one of which was the most promising for favoring vegetative growth due to its high capacity to fix atmospheric nitrogen.
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