Comparative physiological and transcriptome analysis provide insights into the inhibitory effect of 6-pentyl-2H-pyran-2-one on Clarireedia jacksonii.

Clarireedia spp. is a destructive phytopathogenic fungus that causes turf dollar spot of bent-grass, leading to widespread lawn death. In this study, we explored the antifungal capability of 6-pentyl-2H-pyran-2-one (6PP), a natural metabolite volatilized by microorganisms, which plays an important role in the biological control of turfgrass dollar spot. However, the mechanisms by which 6PP inhibits Clarireedia jacksonii remain unknown. In the present study, C. jacksonii mycelial growth was inhibited by the 6PP treatment and the 6PP treatment damaged cell membrane integrity, causing an increase in relative conduc-tivity. Furthermore, physiological and biochemistry assay showed that 6PP treatment can enhance reactive oxygen species (ROS) levels, malondialdehyde (MDA) content obviously increased with 6PP exposure, increased alchohol dehydrogenase (ADH) and depleted acetalde-hyde dehydrogenase (ALDH), and activated the activities of many antioxidant enzymes in C. jacksonii. Gen Ontology and Kyoto Encyclopedia of Genes and Genomes analysis revealed that some genes in C. jacksonii after 6PP treatment related to integrity of the cell wall and membrane, and oxidative stress were significantly downregulated. It is worth mentioning that the fatty acid degradation pathway is significantly upregulated, with an increase in ATP content and ATP synthase activity, which may promote fungal cell apoptosis. Moreover, we found that the expression of ABC transporters, and glutathione metabolism encoding genes were increased to respond to external stimuli. Taken together, these findings revealed the potential antifungal mechanism of 6PP against Clarireedia spp., which also provides a theoretical basis for the commercial utilization of 6PP as a green pesticide in the future.

Trichoderma atroviride LZ42 releases volatile organic compounds promoting plant growth and suppressing Fusarium wilt disease in tomato seedlings.

BACKGROUND: The promotion of plant growth and suppression of plant disease using beneficial microorganisms is considered an alternative to the application of chemical fertilizers or pesticides in the field. RESULTS: A coconut-scented antagonistic Trichoderma strain LZ42, previously isolated from Ganoderma lucidum-cultivated soil, was investigated for biostimulatory and biocontrol functions in tomato seedlings. Morphological and phylogenetic analyses suggested that strain LZ42 is closely related to T. atroviride. Tomato seedlings showed increased aerial and root dry weights in greenhouse trials after treatment with T. atroviride LZ42 formulated in talc, indicating the biostimulatory function of this fungus. T. atroviride LZ42 effectively suppressed Fusarium wilt disease in tomato seedlings, with an 82.69% control efficiency, which is similar to that of the carbendazim treatment. The volatile organic compounds (VOCs) emitted by T. atroviride LZ42 were found to affect the primary root growth direction and promote the root growth of tomato seedlings in root Y-tube olfactometer assays. The fungal VOCs from T. atroviride LZ42 were observed to significantly inhibit F. oxysporum in a sandwiched Petri dish assay. SPME-GC-MS analysis revealed several VOCs emitted by T. atroviride LZ42; the dominant compound was tentatively identified as 6-pentyl-2H-pyran-2-one (6-PP). The VOC 6-PP exhibited a stronger ability to influence the direction of the primary roots of tomato seedlings but not the length of the primary roots. The inhibitory effect of 6-PP on F. oxysporum was the highest among the tested pure VOCs, showing a 50% effective concentration (EC50) of 5.76 µL mL-1 headspace. CONCLUSIONS: Trichoderma atroviride LZ42, which emits VOCs with multiple functions, is a promising agent for the biostimulation of vegetable plants and integrated management of Fusarium wilt disease.

Inhibitory mechanism of 6-Pentyl-2H-pyran-2-one secreted by Trichoderma atroviride T2 against Cylindrocarpon destructans.

Root rot caused by Cylindrocarpon destructans is one of the most devastating diseases of Panax notoginseng, and Trichoderma species are potential agents for the biocontrol of fungal diseases. Thus, we screened a total of 10 Trichoderma isolates against C. destructans and selected Trichoderma atroviride T2 as an antagonistic strain for further research. 6-Pentyl-2H-pyran-2-one (6PP) was identified as an important active metabolite in the fermentation broth of the strain and exhibited antifungal activity against C. destructans. Transcriptome and metabolome analyses showed that 6PP significantly disturbed the metabolic homeostasis of C. destructans, particularly the metabolism of amino acids. By constructing a gene coexpression network, ECHS1 was identified as the hub gene correlated with 6PP stress. 6PP significantly downregulated the expression of ECHS1 at the transcriptional level and combined with the ECHS1 protein. Autophagy occurred in C. destructans cells under 6PP stress. In conclusion, 6PP may induce autophagy in C. destructans by downregulating ECHS1 at the transcriptional level and inhibiting ECHS1 protein activity. 6PP is a potential candidate for the development of new fungicides against C. destructans.

The volatile 6-pentyl-2H-pyran-2-one from Trichoderma atroviride regulates Arabidopsis thaliana root morphogenesis via auxin signaling and ETHYLENE INSENSITIVE 2 functioning.

Plants interact with root microbes via chemical signaling, which modulates competence or symbiosis. Although several volatile organic compounds (VOCs) from fungi may affect plant growth and development, the signal transduction pathways mediating VOC sensing are not fully understood. 6-pentyl-2H-pyran-2-one (6-PP) is a major VOC biosynthesized by Trichoderma spp. which is probably involved in plant-fungus cross-kingdom signaling. Using microscopy and confocal imaging, the effects of 6-PP on root morphogenesis were found to be correlated with DR5:GFP, DR5:VENUS, H2B::GFP, PIN1::PIN1::GFP, PIN2::PIN2::GFP, PIN3::PIN3::GFP and PIN7::PIN7::GFP gene expression. A genetic screen for primary root growth resistance to 6-PP in wild-type seedlings and auxin- and ethylene-related mutants allowed identification of genes controlling root architectural responses to this metabolite. Trichoderma atroviride produced 6-PP, which promoted plant growth and regulated root architecture, inhibiting primary root growth and inducing lateral root formation. 6-PP modulated expression of PIN auxin-transport proteins in a specific and dose-dependent manner in primary roots. TIR1, AFB2 and AFB3 auxin receptors and ARF7 and ARF19 transcription factors influenced the lateral root response to 6-PP, whereas EIN2 modulated 6-PP sensing in primary roots. These results indicate that root responses to 6-PP involve components of auxin transport and signaling and the ethylene-response modulator EIN2.

6-Pentyl-2H-pyran-2-one from Trichoderma erinaceum Is Fungicidal against Litchi Downy Blight Pathogen Peronophythora litchii and Preservation of Litchi.

The postharvest losses of litchi caused by litchi downy blight are considerably high. We identified a natural antifungal volatile pyrone, 6-pentyl-2H-pyran-2-one (6PP), synthesized by Trichoderma erinaceum LS019-2 and investigated as biocontrol for litchi downy blight and preservation. 6PP significantly inhibited the growth and sporangial germination of Peronophythora litchii, the causal agent of litchi downy blight, and caused severe cellular and intracellular destructions, as evidenced by electron microscopic analysis. Furthermore, in the treatment, the fruit kept better color, higher weight, and antioxidant activity, so it can maintain freshness and prolong shelf life. Metabolome analysis confirmed the decline of lipids and the accumulation of organic acids in litchi fruits in response to 6PP treatment. These effects from 6PP could alleviate disease effects and prolong the shelf life of litchi fruits. These findings suggested that 6PP could be a useful natural product to control downy blight disease and a new preservative of litchi fruits.

A New Insight into 6-Pentyl-2H-pyran-2-one against Peronophythora litchii via TOR Pathway.

The litchi downy blight disease of litchi caused by Peronophythora litchii accounts for severe losses in the field and during storage. While ample quantitative studies have shown that 6-pentyl-2H-pyran-2-one (6PP) possesses antifungal activities against multiple plant pathogenic fungi, the regulatory mechanisms of 6PP-mediated inhibition of fungal pathogenesis and growth are still unknown. Here, we investigated the potential molecular targets of 6PP in the phytopathogenic oomycetes P. litchii through integrated deployment of RNA-sequencing, functional genetics, and biochemical techniques to investigate the regulatory effects of 6PP against P. litchii. Previously we demonstrated that 6PP exerted significant oomyticidal activities. Also, comparative transcriptomic evaluation of P. litchii strains treated with 6PP Revealed significant up-regulations in the expression profile of TOR pathway-related genes, including PlCytochrome C and the transcription factors PlYY1. We also noticed that 6PP treatment down-regulated putative negative regulatory genes of the TOR pathway, including PlSpm1 and PlrhoH12 in P. litchii. Protein-ligand binding analyses revealed stable affinities between PlYY1, PlCytochrome C, PlSpm1, PlrhoH12 proteins, and the 6PP ligand. Phenotypic characterization of PlYY1 targeted gene deletion strains generated in this study using CRISPR/Cas9 and homologous recombination strategies significantly reduced the vegetative growth, sporangium, encystment, zoospore release, and pathogenicity of P. litchii. These findings suggest that 6PP-mediated activation of PlYY1 expression positively regulates TOR-related responses and significantly influences vegetative growth and the virulence of P. litchii. The current investigations revealed novel targets for 6PP and underscored the potential of deploying 6PP in developing management strategies for controlling the litchi downy blight pathogen.

Kairomone and Camera Trapping New Zealand Flower Thrips, Thrips obscuratus.

This project investigated how kairomone lures, camera traps, and counting software could together contribute to pest management. Images of cumulative daily catch of New Zealand Flower Thrips (NZFT) attracted to a ripe peach lactone (6-pentyl-2H-pyran-2-one; 6-PAP) were automatically loaded to the internet and compared with scanned bases checked weekly using in-house software and manual counting. Camera traps were able to provide thrips counts equivalent to delta traps, but daily and remotely. An 11-fold greater NZFT count occurred within 24 h in passive traps after polyethylene sachets loaded with 250 mg of 6-PAP were placed in trees. Intensive trapping, by placing 1, 2, 4, and 8 traps per tree (500 mg/trap), resulted in a maximum 32-fold increase in thrips per tree. While 6-PAP has proved to be a useful tool for monitoring NZFT numbers, our results suggest that it is not likely to be suitable for mass trapping. Future research should investigate NZFT behavior to better understand population movement on an area-wide basis. Camera traps can be a valuable tool for recording insect flight activity remotely, but the number of traps required for statistically reliable estimates may be prohibitive.

Development of an efficient trapping system for New Zealand flower thrips, Thrips obscuratus.

BACKGROUND: New Zealand flower thrips (NZFT), Thrips obscuratus (Crawford), is an economic pest of various horticultural crops in New Zealand and is recognised as a quarantine pest globally. Two chemical attractants (ethyl nicotinate and 6-pentyl-2H-pyran-2-one), three dispensers, three trap designs and four trap heights were investigated to determine the most effective method for monitoring NZFT. Phenology of NZFT at two locations was compared. RESULTS: 6-Pentyl-2H-pyran-2-one in a polyethylene bag dispenser was the most attractive lure formulation and exhibited high stability in release rate trials. There was no difference in NZFT catch between vertical-panel and cross-panel traps, but both caught significantly more than delta traps. However, both types of panel trap had unacceptably high by-catch of native insects. Catch of thrips increased with height from 0 to 3 m. Phenology of NZFT showed similar population trends at both locations, but with a timing difference of around 50 days. CONCLUSIONS: Delta traps containing 6-pentyl-2H-pyran-2-one in a polyethylene bag at 2 m above the ground is the recommended method for monitoring NZFT, significantly improving the sensitivity, accuracy and labour input compared with prior methods. Long-term monitoring of NZFT could lead to more accurate economic damage thresholds and timing for when to apply insecticides. © 2014 Society of Chemical Industry.