RESUMEN
AIMS: This study aimed to use palm oil mill effluent (POME) as a renewable resource for the production of antifungal compounds by Streptomyces philanthi RM-1-138 against Ganoderma boninense, Ceratocystis paradoxa and Curvularia oryzae. METHODS AND RESULTS: The efficacy of antifungal compounds RM-1-138 against the three strains of fungal oil palm pathogen was evaluated both in vitro and on oil palm leaf segments. In vitro studies using confrontation tests on glucose yeast-malt extract (GYM) agar plates indicated that the strain RM-1-138 inhibited the growth of all three fungal pathogenic strains. The antifungal compounds produced in the GYM medium exhibited significantly higher inhibition (79%-100%) against the three fungal pathogens than using the diluted POME (50%) medium (80%-83% inhibition). The optimum condition for the production of antifungal compounds from the strain RM-1-138 was as following: POME of 47,966 mg L-1 chemical oxygen demand (COD), the initial pH at 7.0 and supplemented with yeast extract (0.4%). Meanwhile, severe morphological and internal abnormalities in C. oryzae hyphae were observed under a scanning electron microscope and transmission electron microscope. In vivo experiment on oil palm leaf segments indicated that the efficacy of the antifungal compounds RM-1-138 (DSI = 1.3) were not significantly difference in the suppression of Curvularia leaf spot compared with the two commercial chemical fungicides of mancozeb® (DSI = 1.0) and tetraconazole® (DSI = 1.3). CONCLUSIONS: Antifungal compounds produced by S. philanthi RM-1-138 grown in POME have the potential to inhibit fungal pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The POME (about 47 mg L-1 COD) with the initial pH of 7.0 and supplementation of 0.4% nitrogen could be used as a culture medium for the growth and production of antifungal compounds of S. philanthi RL-1-138. In addition, the antifungal compound RM-1-138 could suppress the three strains of oil palm fungal pathogen tested on oil palm leaf segment.
Asunto(s)
Fungicidas Industriales , Streptomyces , Antifúngicos/farmacología , Análisis de la Demanda Biológica de Oxígeno , Fungicidas Industriales/farmacología , Residuos Industriales/análisis , Aceite de Palma , Aceites de Plantas/farmacología , Eliminación de Residuos Líquidos/métodosRESUMEN
This is the first report to present a full-length cDNA (designated HbPR-1) encoding a putative basic HbPR-1 protein from rubber tree (Hevea brasiliensis) treated with salicylic acid. It was characterized and also expressed in Nicotiana benthamiana using Agrobacterium-mediated transient gene expression system in order to investigate the role of HbPR-1 gene in rubber tree against its oomycete pathogen Phytopthora palmivora and to produce recombinant HbPR-1 protein for microbial inhibition test. The HbPR-1 cDNA was 647 bp long and contained an open reading frame of 492 nucleotides encoding 163 amino acid residues with a predicted molecular mass of 17,681 Da and an isoelectric point (pI) of 8.56, demonstrating that HbPR-1 protein belongs to the basic PR-1 type. The predicted 3D structure of HbPR-1 was composed of four α-helices, three ß-sheets, seven strands, and one junction loop. Expression and purification of recombinant HbPR-1 protein were successful using Agrobacterium-mediated transient expression and one-step of affinity chromatography. Heterologous expression of HbPR-1 in N. benthamiana reduced necrosis areas which were inoculated with P. palmivora zoospores, indicating that the expressed HbPR-1 protein played an important role in plant resistance to pathogens. The purified recombinant HbPR-1 protein was found to inhibit 64% of P. palmivora zoospore germination on a water agar plate compared with control, suggesting that it was an antimicrobial protein against P. palmivora.