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Vermicompost (VC) produced by African nightcrawler earthworms (Eudrilus eugeniae) is a natural fertilizer with a rich microbial community. Trichoderma asperelloides PSU-P1 is an effective antagonistic microorganism with multifaceted activity mechanisms. This research aimed to develop Trichoderma-bioenriched vermicompost (TBVC) to promote plant growth and induce the defense response in the Thai rice variety "Chor Khing". T. asperelloides PSU-P1 was tested against Rhizoctonia solani, the pathogen of sheath blight disease, using a dual-culture assay. The results showed that T. asperelloides PSU-P1 effectively inhibited R. solani in vitro growth by 70.48%. The TBVC was prepared by adding a conidial suspension (108 conidia/mL) to vermicompost. The viability of Trichoderma persisted in the vermicompost for 6 months and ranged from 1.2 to 2.8 × 107 CFU/mL. Vermicompost water extracts significantly enhanced seed germination, root length, and shoot length compared to a control group (p < 0.05). Plants that received the TBVC displayed significantly longer shoot and root lengths and higher total chlorophyll content than control plants (p < 0.05). The TBVC induced defense response by increasing the enzyme activity of peroxidase (POD) and polyphenol oxidase (PPO) in comparison with control plants. Rice grown in the TBVC had a significantly reduced incidence of sheath blight caused by R. solani in comparison with control rice (p < 0.05). Furthermore, the fungal community of rice plants was analyzed via the high-throughput next-generation sequencing of the internal transcribed spacer (ITS). The fungal community in the TBVC had greater alpha diversity than the community in the VC. Phylum Ascomycota was dominant in both samples, and a heat map showed that Trichoderma was more prevalent in the TBVC than in the VC. Our results indicate that the enrichment of VC with Trichoderma increases growth, enhances the defense response, and reduces the incidence of sheath blight disease in the Thai rice variety "Chor Khing". [ABSTRACT FROM AUTHOR]

Flower blight caused by Neopestalotiopsis clavispora is an emerging disease of flamingo flower (Anthurium andraeanum Lind.) that negatively impacts flower production. The use of rhizosphere fungi as biocontrol agents is an alternative way to control this disease instead of using synthetic fungicides. This research aimed to screen the potential of rhizosphere fungi, Trichoderma spp., with diverse antifungal abilities to control N. clavispora and to reduce flower blight in flamingo flowers. A total of ten isolates were tested against N. clavispora by dual culture assay, and T1-02 was found to be the most effective isolate against N. clavispora, with inhibition of 78.21%. Morphology and molecular phylogeny of multiple DNA sequences of the genes, the internal transcribed spacer (ITS), translation elongation factor 1-α (tef1-α), and RNA polymerase 2 (rpb2) identified isolate T1-02 as Trichoderma virens. Sealed plate method revealed T. virens T1-02 produced volatile antifungal compounds (VOCs) against N. clavispora, with inhibition of 51.28%. Solid-phase microextraction (SPME) was applied to trap volatiles, and GC/MS profiling showed VOCs emitted from T. virens T1-02 contained a sesquiterpene antifungal compound—germacrene D. The pre-colonized plate method showed that T. virens T1-02 aggressively colonized in tested plates with inhibition of 100% against N. clavispora, and microscopy revealed direct parasitism onto fungal hyphae. Furthermore, the application of T. virens T1-02 spore suspension reduced the disease severity index (DSI) of flower blight in flamingo flowers. Based on the results from this study, T. virens T1-02 displays multiple antagonistic mechanisms and has the potential ability to control flower blight of flamingo flowers caused by N. clavispora. [ABSTRACT FROM AUTHOR]

Strawberry anthracnose is a serious disease, and fungicides are currently widely used by farmers. Thus, biological control is a good alternative. This study aims to identify the species of endophytic Streptomyces CINv1 that was previously isolated from Cinnamomum verum J. Presl. and to evaluate its properties as a biocontrol agent, plant growth promoter, and plant colonizing endophyte. This strain was identified by analysis of its 16S rRNA gene sequences, and the result shows 100% similarity to Streptomyces albus CINv1. The CINv1 strain displayed high resistance (81.83%) against Colletotrichum sp. isolate CA0110, as tested by the dual culture technique. Additionally, inhibited pathogen growth on IMA-2 agar was observed under a compound microscope. The results demonstrated swelling, bulbousness, and cytoplasmic aggregation of abnormal hyphal, which were confirmed by SEM as well. Furthermore, the functional media used to evaluate plant growth-promoting properties, including nitrogen fixation, phosphate solubilization, and siderophore production, yielded positive results. Analyses of plant hormones by HPLC found their ability to produce indole-3-acetic acid (IAA). Thus, a biological control trial in greenhouse conditions was conducted by spraying a spore suspension of the strain onto strawberry seedlings once a week, which showed a significant reduction in disease severity. After the seventh spraying, the assessment of the number of leaves and canopy height of the seedling showed significant promotion. In addition, the CINv1 strain established a mutualistic interaction with the plant cells through colonization inter-and intracellularly in strawberry roots, leaves, and petioles. Moreover, using LC-MS/MS to analyze the secondary metabolites of this strain, various groups of compounds were found that could potentially benefit pharmaceutical and agricultural uses. [ABSTRACT FROM AUTHOR]

Cercospora leaf spot disease is a serious problem for lettuce cultivation worldwide. Cercospora lactucae-sativa, the causative agent of leaf spot disease on lettuce, was treated with non-thermal atmospheric pressure gliding arc (GA), tesla coil (TC) and dielectric barrier discharge (DBD) plasmas for the in vitro fungal inactivation of both mycelial growth and conidial germination. The fungus was exposed to the three plasmas individually for 5, 10, 15 and 20 min. The results showed that DBD plasma inactivated fungal growth during all exposure periods, and the highest inhibitory effect was caused by exposure to DBD plasma for 20 min, at 93.33% inhibition. The germination of fungal conidia was completely inactivated after exposure to all three non-thermal plasmas for 5 min, as observed 4 and 24 h after incubation. The pathogenesis of C. lactucae-sativa on lettuce after plasma treatments for 5 min was examined by spraying an inoculation of the treated conidia on lettuce. The results showed that all three plasmas reduced the disease incidence and severity compared to the non-treated control. Therefore, the non-thermal atmospheric pressure GA, TC and DBD plasmas have antifungal potential for the inactivation of C. lactucae-sativa, making them an interesting novel technology for plant disease control. [ABSTRACT FROM AUTHOR]

Athelia rolfsii is one of the most destructive and aggressive fungal pathogens worldwide and causes southern blight disease of lettuce. A nonthermal atmospheric-pressure dielectric barrier discharge (DBD) plasma has attracted interest as an alternative control method to chemical usage because of its antimicrobial activity. Exposure of A. rolfsii to DBD plasma for 5, 10, 15, and 20 min resulted in in vitro fungal inhibition of mycelial discs and sclerotia. The results showed that DBD plasma exposure for 10 min completely inhibited fungal growth of mycelial discs, whereas exposure for over 20 min was required to inhibit the hyphal growth of sclerotia. Scanning electron microscopy (SEM) observations of mycelia and sclerotia abnormalities revealed laceration and damage of both mycelia and sclerotia. In addition, disease incidence and severity were reduced in mycelial and sclerotia inoculation following DBD plasma exposure for 15 and 20 min, respectively, compared with the positive control. In conclusion, the DBD plasma demonstrates antifungal activity against A. rolfsii via inhibition of fungal growth and reduction in disease incidence and severity. Therefore, DBD plasma has the potential to be applied in controlling southern blight disease of lettuce. [ABSTRACT FROM AUTHOR]

Sclerotium rot causes damping-off and stem rot in seedlings and mature mungbeans, which negatively impacts cultivation. The use of a rhizobacterium to control soil-borne diseases is an alternative method to the excess use of synthetic fungicides; therefore, this study aims to screen rhizosphere actinobacteria with fungicidal activities against Sclerotium rolfsii, the pathogen that causes sclerotium rot in mungbeans. Primary screening showed that the Streptomyces sp. isolate Z1-04-02 displayed the highest effectiveness against S. rolfsii in dual culture plates, with a percentage inhibition of 74.28%. An assay containing enzymes that degrade cell walls, of the cell-free culture filtrate (CF) of Z1-04-02, showed that the activities of chitinase and β-1,3-glucanase were 0.0209 and 1.0210 U/mL, respectively, which was significantly higher than that of the control (media alone). The cell-free CF of Z1-04-02, incubated at 37 °C and 100 °C, using agar well diffusion, effectively inhibited the growth of S. rolfsii with inhibition percentages of 37.78% and 27.78%, respectively. Solid-phase microextraction (SPME) was applied to trap volatiles released from Z1-04-02 and gas chromatography–mass spectrometry (GC/MS); volatile antifungal compounds were tentatively identified as bicyclic monoterpene (1R)-(-)-myrtenal. The application of the cell-free CF, and the spore suspension of Z1-04-02, showed disease severity indexes (DSIs) of 12.5% and 8.25%, respectively, which were significantly lower than those showing inoculation by S. rolfsii alone. The identification of this strain by morphology, biochemistry tests, and 16s rDNA sequences revealed that Z1-04-02 was Streptomyces albulus. This finding revealed that S. albulus Z1-04-02 displayed diverse fungicidal activities against S. rolfsii, and it has the potential to act as a biological control agent in terms of inhibiting sclerotium rot in mungbeans. [ABSTRACT FROM AUTHOR]

We detected tobacco mosaic virus (TMV), a member of the genus Tobamovirus and one of the most significant plant-infecting viruses, for the first time in a chrysanthemum in Thailand using reverse-transcription polymerase chain reaction (RT–PCR). The TMV-infected chrysanthemum leaves exhibited mosaic symptoms. We conducted a sequence analysis of the coat protein (CP) gene and found that the TMV detected in the chrysanthemum had 98% identity with other TMV isolates in GenBank. We carried out bioassays and showed that TMV induced mosaic and stunting symptoms in inoculated chrysanthemums. We observed the rigid rod structure of TMV under a transmission electron microscope (TEM). To enhance the speed and sensitivity of detection, we developed a colorimetric RT loop-mediated isothermal amplification (LAMP) technique. We achieved LAMP detection after 30 min incubation in isothermal conditions at 65 °C, and distinguished the positive results according to the color change from pink to yellow. The sensitivity of the LAMP technique was 1000-fold greater than that of RT–PCR, and we found no cross-reactivity with other viruses or viroids. This is the first reported case of a TMV-infected chrysanthemum in Thailand, and our colorimetric RT–LAMP TMV detection method is the first of its kind. [ABSTRACT FROM AUTHOR]

Soil microorganisms are well studied for their beneficial effects on plant growth and their impact on biocontrol agents. The production of volatile antifungal compounds emitted from soil fungi is considered to be an effective ability that can be applied in biofumigants in the control of plant diseases. A soil fungus, Trichoderma asperelloides TSU1, was isolated from flamingo flower cultivated soil and identified on the basis of the morphology and molecular analysis of the internal transcribed spacer (ITS), rpb2, and tef1-α genes. To test T. asperelloides TSU1-produced volatile organic compounds (VOCs) with antifungal activity, the sealed plate method was used. The VOCs of T. asperelloides TSU1 inhibited the mycelial growth of fungal pathogens that were recently reported as emerging diseases in Thailand, namely, Corynespora cassiicola, Fusarium incarnatum, Neopestalotiopsis clavispora, N. cubana, and Sclerotium rolfsii, with a percentage inhibition range of 38.88–68.33%. Solid-phase microextraction (SPME) was applied to trap VOCs from T. asperelloides TSU1 and tentatively identify them through gas chromatography–mass spectrometry (GC/MS). A total of 17 compounds were detected in the VOCs of T. asperelloides TSU1, and the dominant compounds were identified as fluoro(trinitro)methane (18.192% peak area) and 2-phenylethanol (9.803% peak area). Interestingly, the commercial 2-phenyethanol showed antifungal activity against fungal pathogens that were similar to the VOCs of T. asperelloides TSU1 by bioassay. On the basis of our study’s results, T. asperelloides TSU1 isolated from soil displayed antifungal abilities via the production of VOCs responsible for restricting pathogen growth. [ABSTRACT FROM AUTHOR]