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Diagnosis and Integrated Management of Fruit Rot in Cucurbita argyrosperma, Caused by Sclerotium rolfsii.

ABSTRACT: Fruit rot is the principal phytopathological problem of pipiana pumpkin (Cucurbita argyrosperma Huber) in the state of Guerrero. The aims of this research were to 1) identify the causal agent of southern blight on pumpkin fruits by morphological, pathogenic, and molecular analysis (ITS1, 5.8S, ITS2); 2) evaluate in vitro Trichoderma spp. strains and chemical fungicides; and 3) evaluate under rainfed field conditions, the strains that obtained the best results in vitro, combined with fungicides during two crop cycles. Number of commercial and non-commercial fruits at harvest, and seed yield (kg ha^-1) were registered. Morphological, pathogenic and molecular characterization identified Sclerotium rolfsii as the causal agent of rot in pipiana pumpkin fruits. Now, in vitro conditions, the highest inhibition of S. rolfsii were obtained by Trichoderma virens strain G-41 (70.72%), T. asperellum strain CSAEGro-1 (69%), and the fungicides metalaxyl (100%), pyraclostrobin (100%), quintozene (100%), cyprodinil + fludioxonil (100%), and prochloraz (100%). Thiophanate-methyl only delayed growth (4.17%). In field conditions, during the spring-summer 2015 cycle, T. asperellum strain CSAEGro-1 + metalaxyl, and T. asperellum + cyprodinil + fludioxonil, favored the highest number of fruits and seed yield in the crop.

SUBMITTER: Najera JFD

PROVIDER: S-EPMC5985643 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Peanut stem rot, caused by Sclerotium rolfsii, considerably affects crop productivity. Application of chemical fungicides harms the environment and induces drug resistance. Biological agents are valid and eco-friendly alternatives to chemical fungicides. Bacillus spp. are important biocontrol agents that are now widely used against several plant diseases. This study aimed to evaluate the efficacy and mechanism of a potential biocontrol agent Bacillus sp. for controlling peanut stem rot caused by S. rolfsii. Here, we isolated a strain of Bacillus from pig biogas slurry that considerably inhibits the radial growth of S. rolfsii. The strain CB13 was identified as Bacillus velezensis on the basis of morphological, physiological, biochemical characteristics and phylogenetic trees based on the 16S rDNA and gyrA, gyrB, and rpoB gene sequences. The biocontrol efficacy of CB13 was evaluated on the basis of colonization ability, induction of defense enzyme activity, and soil microbial diversity. The control efficiencies of B. velezensis CB13-impregnated seeds in four pot experiments were 65.44, 73.33, 85.13, and 94.92%. Root colonization was confirmed through green fluorescent protein (GFP)-tagging experiments. The CB13-GFP strain was detected in peanut root and rhizosphere soil, at 104 and 108 CFU/g, respectively, after 50 days. Furthermore, B. velezensis CB13 enhanced the defense response against S. rolfsii infection by inducing defense enzyme activity. MiSeq sequencing revealed a shift in the rhizosphere bacterial and fungal communities in peanuts treated with B. velezensis CB13. Specifically, the treatment enhanced disease resistance by increasing the diversity of soil bacterial communities in peanut roots, increasing the abundance of beneficial communities, and promoting soil fertility. Additionally, real-time quantitative polymerase chain reaction results showed that B. velezensis CB13 stably colonized or increased the content of Bacillus spp. in the soil and effectively inhibited S. rolfsii proliferation in soil. These findings indicate that B. velezensis CB13 is a promising agent for the biocontrol of peanut stem rot.

Project description:Peanut stem white rot caused by Sclerotium rolfsii Sacc. is a soil-borne disease that is widely prevailing across peanut farms, leading to serious economic losses. Screening for biocontrol agents against this pathogen is urgent. In this research, 166 fungal isolates including 136 isolates of S. rolfsii and 30 isolates of antagonistic endophytic fungi were obtained from a total of 220 samples collected from peanut farms in Guilan province, Iran. After morphological and molecular identification, six superior endophytic isolates were finally selected for the in vitro and greenhouse trials, including four isolates from Trichoderma viride, Trichoderma virens, Penicillium decaturense, and Aspergillus flavus and two isolates from Penicillium rubens. Four methods of biocontrol were used during the in vitro phase, i.e., dual culture, volatile metabolites assay, non-volatile metabolites assay (culture extract), and slide culture. It was found that T. virens had the highest capability of suppressing the mycelial growth of S. rolfsii in the dual culture method (90.98%). As for the volatile metabolites assay, the most effective isolates in inhibiting the pathogen's mycelial growth were P. rubens (MN395854.1) and A. flavus (84.30% and 73.50% inhibition, respectively). In the non-volatile metabolites method, the isolates that performed the best in suppressing the mycelial growth of S. rolfsii were T. viride and P. rubens (MN395854.1) with 91.80% and 90.20% inhibitory effects, respectively. On the other hand, in the slide culture method, all isolates, except for T. virens and T. viride, successfully controlled the development of S. rolfsii hyphae. The greenhouse trials also supported the effectiveness of endophytic fungi in controlling the pathogen on the host plants. According to the results, T. viride, A. flavus, and P. rubens (MN395854.1) were 44%, 42%, and 38% effective in alleviating the disease incidence and severity. Moreover, the application of these antagonistic fungi in the greenhouse conditions increased the height, fresh weight, and dry weight of the Arachis hypogaea plants infected with the disease causal agent compared to the plants treated only with the pathogen. The results of the in vitro and greenhouse experiments revealed that the endophytic fungi occurring in the natural microbiota of peanut are capable of bio-controlling S. rolfsii, the causal agent of peanut stem white rot disease. These findings shed new insights into the possible resistance induction in A. hypogaea plants through biological protection.

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Diagnosis and Integrated Management of Fruit Rot in Cucurbita argyrosperma, Caused by Sclerotium rolfsii.

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