Morphological and molecular identification of Neopestalotiopsis clavispora causing flower blight on Anthurium andraeanum in Thailand

Flower blight on anthurium (Anthurium andraeanum) was observed during August 2018 on an anthurium cultivation farm in the Songkhla Province of southern Thailand. The fungal isolate was identified as Neopestalotiopsis clavispora based on the morphology and DNA sequence of the internal transcribed spacer (ITS), translation elongation factor 1-α (tef1-α), and β-tubulin (tub) genes. The phylogenetic tree, based on the combined sequences of ITS, tef1-α, and tub, confirmed this pathogen as N. clavispora. Pathogenicity of the species was confirmed according to Koch's postulate: N. clavispora could infect anthurium. To the best of our knowledge, this is the first report of N. clavispora as a pathogen of anthurium

<i>Trichoderma asperelloides</i> PSU-P1 Induced Expression of Pathogenesis-Related Protein Genes against Gummy Stem Blight of Muskmelon (<i>Cucumis melo</i>) in Field Evaluation

Gummy stem blight caused by Stagonosporopsis cucurbitacearum is the most destructive disease of muskmelon cultivation. This study aimed to induce disease resistance against gummy stem blight in muskmelon by Trichoderma asperelloides PSU-P1. This study was arranged into two crops. Spore suspension at a concentration of 1 × 106 spores/mL of T. asperelloides PSU-P1 was applied to muskmelon to investigate gene expression. The expression of PR genes including chitinase (chi) and β-1,3-glucanase (glu) were determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and enzyme activity was assayed by the DNS method. The effects of T. asperelloides PSU-P1 on growth, yield, and postharvest quality of muskmelon fruit were measured. A spore suspension at a concentration of 1 × 106 spore/mL of T. asperelloides PSU-P1 and S. cucurbitacearum was applied to muskmelons to determine the reduction in disease severity. The results showed that the expression of chi and glu genes in T. asperelloides PSU-P1-treated muskmelon plants was 7–10-fold higher than that of the control. The enzyme activities of chitinase and β-1,3-glucanase were 0.15–0.284 and 0.343–0.681 U/mL, respectively, which were higher than those of the control (pathogen alone). Scanning electron microscopy revealed crude metabolites extracted from T. asperelloides PSU-P1-treated muskmelon plants caused wilting and lysis of S. cucurbitacearum hyphae, confirming the activity of cell-wall-degrading enzymes (CWDEs). Application of T. asperelloides PSU-P1 increased fruit weight and fruit width; sweetness and fruit texture were not significantly different among treated muskmelons. Application of T. asperelloides PSU-P1 reduced the disease severity scale of gummy stem blight to 1.10 in both crops, which was significantly lower than that of the control (2.90 and 3.40, respectively). These results revealed that application of T. asperelloides PSU-P1 reduced disease severity against gummy stem blight by overexpressed PR genes and elevated enzyme activity in muskmelon plants

UTILIZATION OF BASALT FIBERS AS A RAW MATERIAL FOR CLAY CERAMIC PRODUCTION

This research aimed to investigate the possibility of utilization basalt fibers as a raw material for ceramic production. Both quartz and feldspar were replaced partially or entirely by basalt fiber in the range of 10-25 wt%. The mixture of ceramic powders and basalt fibers were uniaxially pressed and sintered at temperatures between 1000 and 1200°C for 1 h. The substitution of basalt fibers in ceramic compositions demonstrated the positive effect on the physical and mechanical properties. The addition of basalt fibers in an appropriate amount enhance the densification and reduce sintering temperature of clay-based ceramics (CB-0) from 1200 to 1150°C. The highest density and strength were 2.40 g/cm³ and 116 MPa, respectively, when replacing feldspar and quartz with basalt up to 20 wt% (CB-20) and sintering at 1150°C

Morphological, Molecular Identification and Pathogenicity of Neoscytalidium dimidiatum Causing Stem Canker of Hylocereus polyrhizus in Southern Thailand

Red-fleshed dragon fruit (Hylocereus polyrhizus) is commonly cultivated in Thailand, especially in southern Thailand, where the weather favors plant growth and development. In 2021, stem canker of H. polyrhizus was observed in a dragon fruit plantation field in Phatthalung Province, southern Thailand. Small, orange circular spots developed on the stem of H. polyrhizus, which later became gray, and the lesion expanded with a mass of conidia. Scytalidium-like fungus was isolated from infected tissues. Based on morphology and phylogenetic analyses of internal transcribed spacer (ITS), nuclear large subunit (LSU) and &beta;-tubulin (tub) sequences of fungal isolates, the fungus was identified as Neoscytalidium dimidiatum. Pathogenicity tests revealed that this isolate caused stem canker on the stem of H. polyrhizus, similar to that observed in the field. Knowledge of the diagnosis of plant diseases is an important step for managing plant diseases and therefore, this finding provides basic information for the development of appropriate strategies to manage stem canker disease on H. polyrhizus plants

Development of a Novel Emulsion Formulation of <i>Trichoderma asperelloides</i> PSU-P1 Conidia against Stem Canker on Dragon Fruit Caused by <i>Neoscytalidium dimidiatum</i>

Stem canker on dragon fruit caused by Neoscytalidium dimidiatum causes severe losses in production of this fruit worldwide. Biological control by Trichoderma species is widely used to control several plant diseases. However, environmental conditions affect the use of biocontrol agents in the field. The development of a new formulation may offer an alternative way to address the problem of stem canker on dragon fruit caused by N. dimidiatum. In this study, we sought to develop a Trichoderma asperelloides PSU-P1 formulation that would be effective against N. dimidiatum. Three vegetable oils, two emulsifier-dispersing agents (Tween 20 and Tween 80), and one source of carbon (dextrose) were tested for carrier additives. We assessed the viability and antifungal ability of formulations incubated at ambient temperature and at 10 °C during a storage period of 1–6 months. The formulation composed of coconut oil, DW, and tween 20 in a ratio of 30:60:10 required a mixing time of 1.14 min; this was significantly faster than the mixing times of other formulations. Application of this formulation suppressed canker development; a canker area of 0.53 cm2 was recorded, compared with a control (pathogen only) area of 1.65 cm2. In terms of viability, this formulation stored at ambient temperature showed a surface area percentage of T. asperelloides PSU-P1 ranging from 64.43 to 75.7%; the corresponding range for the formulation stored at cool temperature was 70.59–75.6%. For both formulations, percentage inhibition gradually decreased from 1 to 6 months, with ranges of 59.21–77% and 60.65–76.19% for formulations incubated at ambient and cool temperatures, respectively. Our findings suggest that the formulation developed in this study prolongs the viability of T. asperelloides PSU-P1 conidia by up to 6 months, effectively inhibits N. dimidiatum in vitro, and reduces stem canker in vivo

Plant Growth Promotion and Biological Control against <i>Rhizoctonia solani</i> in Thai Local Rice Variety “Chor Khing” Using <i>Trichoderma breve</i> Z2-03

Several strains of Trichoderma are applied in the field to control plant diseases due to their capacity to suppress fungal pathogens and control plant diseases. Some Trichoderma strains also are able to promote plant growth through the production of indole-3-acetic acid (IAA). In southern Thailand, the local rice variety “Chor Khing” is mainly cultivated in the Songkhla province; it is characterized by slow growth and is susceptible to sheath blight caused by Rhizoctonia solani. Therefore, this research aimed to screen Trichoderma species with the ability to promote plant growth in this rice variety and enact biological control against R. solani. A total of 21 Trichoderma isolates were screened for indole compound production using the Salkowski reagent. The Z2-03 isolate reacted positively to the Salkowski reagent, indicating the production of the indole compound. High-performance liquid chromatography (HPCL) confirmed that Z2-03 produced IAA at 35.58 ± 7.60 μg/mL. The cell-free culture filtrate of the potato dextrose broth (CF) of Z2-03 induced rice germination in rice seeds, yielding root and shoot lengths in cell-free CF-treated rice that were significantly higher than those of the control (distilled water and culture broth alone). Furthermore, inoculation with Trichoderma conidia promoted rice growth and induced a defense response against R. solani during the seedling stage. Trichoderma Z2-03 displayed an antifungal capacity against R. solani, achieving 74.17% inhibition (as measured through dual culture assay) and the production of siderophores on the CAS medium. The pot experiment revealed that inoculation with the Trichoderma sp. Z2-03 conidial suspension increased the number of tillers and the plant height in the “Chor Khing” rice variety, and suppressed the percentage of disease incidence (PDI). The Trichoderma isolate Z2-03 was identified, based on the morphology and molecular properties of ITS, translation elongation factor 1-alpha (tef1-α), and RNA polymerase 2 (rpb2), as Trichoderma breve Z2-03. Our results reveal the ability of T. breve Z2-03 to act as a plant growth promoter, enhancing growth and development in the “Chor Khing” rice variety, as well as a biological control agent through its competition and defense induction mechanism in this rice variety

Volatile Organic Compound from Trichoderma asperelloides TSU1: Impact on Plant Pathogenic Fungi

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