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Redalyc.Antifungal Potential of Crude Plant Extracts on Conidial Revista Mexicana de Fitopatología ISSN: 0185-3309 [email protected] Sociedad Mexicana de Fitopatología, A.C. México Hernández-Albíter, Rocío Citlali; Barrera-Necha, Laura Leticia; Bautista-Baños, Silvia; Bravo-Luna, Leticia Antifungal Potential of Crude Plant Extracts on Conidial Germination of Two Isolates of Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. Revista Mexicana de Fitopatología, vol. 25, núm. 2, julio-diciembre, 2007, pp. 180-185 Sociedad Mexicana de Fitopatología, A.C. Texcoco, México Disponible en: http://www.redalyc.org/articulo.oa?id=61225213 Cómo citar el artículo Número completo Sistema de Información Científica Más información del artículo Red de Revistas Científicas de América Latina, el Caribe, España y Portugal Página de la revista en redalyc.org Proyecto académico sin fines de lucro, desarrollado bajo la iniciativa de acceso abierto 180 / Volumen 25, Número 2, 2007 Antifungal Potential of Crude Plant Extracts on Conidial Germination of Two Isolates of Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. Rocío Citlali Hernández-Albíter, Laura Leticia Barrera-Necha, Silvia Bautista- Baños, and Leticia Bravo-Luna, Instituto Politécnico Nacional, Centro de Desarrollo de Productos Bióticos, Apdo. Postal 24, km 8.5 Carr. Yautepec-Jojutla, San Isidro Yautepec, Morelos, México CP 62731. Correspondencia: [email protected] (Recibido: Enero 31, 2006 Aceptado: Abril 18, 2006) Hernández-Albíter, R.C., Barrera-Necha, L.L., Bautista-Baños, espectrofotometría (ME). Los conidios de ambos aislamientos S., and Bravo-Luna, L. 2007. Antifungal potential of crude incubados en los extractos crudos de Origanum majorana, plant extracts on conidial germination of two isolates of Carica papaya, Citrus aurantium, Citrus aurantifolia, Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. Bougainvillea spectabilis, Justicia spicigera, Petroselinum Revista Mexicana de Fitopatología 25:180-185. sativum, Parthenium hysterophorus, Schinus molle y Ricinus Abstract. Crude plant extracts of leaves and stems of 40 communis mostraron porcentajes de germinación menores different plant species from the state of Morelos, Mexico, de 61% por MMO y menores de 56% por ME. Los 28 extractos were used to compare conidia germination of two isolates of crudos restantes no mostraron algún efecto antifúngico. La Colletotrichum gloeosporioides obtained from papaya fruit germinación de conidios varió de acuerdo al extracto vegetal, with anthracnose symptoms, collected from the states of aislamiento, tiempo de incubación y método de evaluación. Guerrero and Veracruz, Mexico. In general, better antifungal effect was observed with crude plant extract of night-blooming Palabras clave adicionales: Compuestos naturales, jessamine (Cestrum nocturnum) and cherimoya (Annona espectrofotometría, antracnosis. cherimola) for both isolates, after 14 or 18 h incubation, determined by optical microscopy method (OMM) and Colletotrichum gloeosporioides (Penz.) Penz. y Sacc. is the espectrophotometry method (EM). Conidia of both fungal casual agent of anthracnose, an important disease of papaya isolates incubated in crude extracts of Origanum majorana, fruit (Carica papaya L.) (Alvarez and Nishijima, 1987; Carica papaya, Citrus aurantium, Citrus aurantifolia, Snowdon, 1990). Bolkan et al. (1976) reported that rots Bougainvillea spectabilis, Justicia spicigera, Petroselinum originated by C. gloeosporioides isolated from immature and sativum, Parthenium hysterophorus, Schinus molle, and mature fruit accounted for 40 and 89% of the infection, Ricinus communis, showed less than 61% germination by respectively. Chemical control which is used to reduce OMM and less than 56% by EM. The remaining 28 crude incidence of postharvest diseases in papaya is causing the extracts did not show antifungal effect. Conidial germination development of fungal resistance to chemical products (Brent varied according to extract, isolate, time of incubation, and and Hollomon, 1998). Application of higher concentrations methodology evaluated. of chemicals in an attempt to overcome anthracnose disease, increases the risk of high levels of toxic residues, which is Additional keywords: Natural compounds, particularly serious since papaya fruit is consumed in a spectrophotometry, anthracnose. relatively short time after harvest. The exploitation of natural products such as plant extracts is believed to be safer to Resumen. Extractos crudos de hojas y tallos de 40 especies consumers and the environment. In the state of Morelos, diferentes de plantas del estado de Morelos, México, se Mexico, the antimicrobial properties of plant extracts from utilizaron para comparar la germinación conidial de dos various species have been proven to control fungal aislamientos de Colletotrichum gloeosporioides, los cuales development either in vitro or in vivo. In vitro studies carried se obtuvieron de frutos de papaya con síntomas de out by Bautista-Baños et al. (2000a) reported that among 19 antracnosis, provenientes del estado de Guerrero y Veracruz, different botanical species tested, the aqueous extract of México. En general, el mejor efecto antifúngico se observó leaves of custard apple (Annona reticulata L.) and papaya con el extracto crudo de huele de noche (Cestrum nocturnum) among various others, inhibited spore formation and y chirimoya (Annona cherimola) para ambos aislamientos germination of Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. Conidial después de 14 ó 18 h de incubación y determinado por el inhibition was also observed when C. gloeosporioides was método de microscopía óptica (MMO) y el método de grown on these two extracts (Bautista-Baños et al., 2002). Revista Mexicana de FITOPATOLOGIA/ 181 Current methods used to evaluate the efficiency of plant with a conidial suspension from Veracruz or Guerrero which extracts for control of fungi include the use of high quantity presented high percent spore germination in water. Five of materials such as nutrient media and Petri plates, which in extracts with six replicates were able to be analysed for each addition, are time consuming and expensive. Wilson et al. isolate. After 14 and 18 h incubation of the plates at 28°C (1997) tested the microtiter method to evaluate the antifungal (before 14 h incubation there was not significant differences activity of crude plant extracts against Botrytis cinerea in spore germination), the absorbancy of fungal growth in Pers.:Fr., highlighting the efficacy of this method. Then, the wells was measured with a Labsystems Multiscan EX spectrophotometry´s method was compared at the same time (Model 355) microplate reader at wavelength 492 nm which with the microscopy´s method to evaluate spore germination was used by Wilson et al. (1997). This method scans a first (counting under light microscopy). The objective of this plate of plant extracted material with the inactive conidial research was to evaluate the effect 40 crude plant extracts on suspension. The second plate reading contains the plant conidial germination of C. gloeosporioides by optical extract with the active conidial one. The percent spore microscopy and espectrophotometry. germination by the espectrophotometry method (EM) was Isolates. Colletotrichum gloeosporioides was isolated from determined with the following equations: % GIAC = (AIC) diseased papaya fruit showing anthracnose symptoms, (100% GIIC)/AAC and % GAC = 100-% GIAC, where: % GIAC harvested in the states of Veracruz and Guerrero, Mexico. = percent germination inhibition of active conidia, % GIIC = Pure monosporic cultures were maintained on potato- percent germination inhibition of inactive conidia, % GAC = dextrose-agar (PDA). To maintain pathogenicity of the percent germination of active conidia, AIC = absorbancy of fungus, periodic inoculations and reisolations from infected inactive conidia, AAC = absorbancy of active conidia. A third papaya were carried out. The parameters with significantly multi-well plate (plant extracts with an active conidia differences between Veracruz and Guerrero isolates were: suspension) was prepared and 50 µl aliquot of this suspension mycelial growth, growth rate, and germination in different was placed on a 20-mm diameter agar disk and after 14 and 18 growing media. Experiments were carried out using 10-15 days h at 28ºC stained with lactophenol acid fuchsin. The percent old cultures. conidial germination was determined at 40X (optical Plant material. Plant selection was according to the fungicidal microscopy method = OMM). Plant extracts were tested or bactericidal background previously reported by Flores- separately in lots of five and a control. The entire study Moctezuma et al. (2002). Forty different plant species consisted of eight experiments. corresponding to various plant families including medicinal, Statistical analysis. Percentage conidial germination for EM forest, bush, fruit, and vegetable plants were field collected and OMM were analysed using the statistical program Sigma or bought at local markets from different regions of the state Stat 2.0 using test of Kruskal-Wallis to obtain mean separation of Morelos (Table 1). Plant species were collected throughout (p = 0.05). the year. In the isolate from Veracruz, germination measured by EM Plant material preparation. 100 g of each of the collected was significantly lower (p = 0.05), (13-45%) than control (1- plant material were disinfested in 2% sodium hypochlorite 8%) after 14 and 18 h respectively (Table 2, Lot 1). In general, for 15 min, rinsed twice in distilled water for 15 min and
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