The Arrestin-like Protein palF Contributes to Growth, Sporulation, Spore Germination, Osmolarity, and Pathogenicity of Coniella vitis.

Coniella vitis is a dominant phytopathogen of grape white rot in China, significantly impacting grape yield and quality. Previous studies showed that the growth and pathogenicity of C. vitis were affected by the environmental pH. Arrestin-like protein PalF plays a key role in mediating the activation of an intracellular-signaling cascade in response to alkaline ambient. However, it remains unclear whether palF affects the growth, development, and virulence of C. vitis during the sensing of environmental pH changes. In this study, we identified a homologous gene of PalF/Rim8 in C. vitis and constructed CvpalF-silenced strains via RNA interference. CvpalF-silenced strains exhibited impaired fungal growth at neutral/alkaline pH, accompanied by reduced pathogenicity compared to the wild-type (WT) and empty vector control (CK) strains. The distance between the hyphal branches was significantly increased in the CvpalF-silenced strains. Additionally, CvpalF-silenced strains showed increased sensitivity to NaCl, H2O2, and Congo red, and decreased sensitive to CaSO4. RT-qPCR analysis demonstrated that the expression level of genes related to pectinase and cellulase were significantly down-regulated in CvpalF-silenced strains compared to WT and CK strains. Moreover, the expression of PacC, PalA/B/C/F/H/I was directly or indirectly affected by silencing CvpalF. Additionally, the expression of genes related to plant cell wall-degrading enzymes, which are key virulence factors for plant pathogenic fungi, was regulated by CvpalF. Our results indicate the important roles of CvpalF in growth, osmotolerance, and pathogenicity in C. vitis.

Genome Analysis of Pseudomonas viciae G166 Conferring Antifungal Activity in Grapevine.

Grapevine (Vitis vinifera) is one of the major economic fruit crops but suffers many diseases, causing damage to the quality of grapes. Strain G166 was isolated from the rhizosphere of grapevine and was found to exhibited broad-spectrum antagonistic activities against fungal pathogens on grapes in vitro, such as Coniella diplodiella, Botrytis cinerea, and Colletotrichum gloeosporioides. Whole-genome sequencing revealed that G166 contained a 6,613,582 bp circular chromosome with 5749 predicted coding DNA sequences and an average GC content of 60.57%. TYGS analysis revealed that G166 belongs to Pseudomonas viciae. Phenotype analysis indicated that P. viciae G166 remarkably reduced the severity of grape white rot disease in the grapevine. After inoculation with C. diplodiella, more H2O2 and MDA accumulated in the leaves and resulted in decreases in the Pn and chlorophyll content. Conversely, G166-treated grapevine displayed less oxidative damage with lower H2O2 levels and MDA contents under the pathogen treatments. Subsequently, G166-treated grapevine could sustain a normal Pn and chlorophyll content. Moreover, the application of P. viciae G166 inhibited the growth of mycelia on detached leaves and berries, while more disease symptoms occurred in non-bacterized leaves and berries. Therefore, P. viciae G166 served as a powerful bioagent against grape white rot disease. Using antiSMASH prediction and genome comparisons, a relationship between non-ribosomal peptide synthase clusters and antifungal activity was found in the genome of P. viciae G166. Taken together, P. viciae G166 shows promising antifungal potential to improve fruit quality and yield in ecological agriculture.

A Simulation Study to Reveal the Epidemiology and Aerosol Transmission Characteristics of Botrytis cinerea in Grape Greenhouses.

Most previously studies had considered that plant fungal disease spread widely and quickly by airborne fungi spore. However, little is known about the release dynamics, aerodynamic diameter, and pathogenicity threshold of fungi spore in air of the greenhouse environment. Grape gray mold is caused by Botrytis cinerea; the disease spreads in greenhouses by spores in the air and the spore attaches to the leaf and infects plant through the orifice. In this study, 120 µmol/L propidium monoazide (PMA) were suitable for treatment and quantitation viable spore by quantitative real-time PCR, with a limit detection of 8 spores/mL in spore suspension. In total, 93 strains of B. cinerea with high pathogenicity were isolated and identified from the air samples of grapevines greenhouses by a portable sampler. The particle size of B. cinerea aerosol ranged predominately from 0.65-3.3 µm, accounting for 71.77% of the total amount. The B. cinerea spore aerosols were infective to healthy grape plants, with the lowest concentration that could cause disease being 42 spores/m3. Botrytis cinerea spores collected form six greenhouse in Shandong Province were quantified by PMA-qPCR, with a higher concentration (1182.89 spores/m3) in May and June and a lower concentration in July and August (6.30 spores/m3). This study suggested that spore dispersal in aerosol is an important route for the epidemiology of plant fungal disease, and these data will contribute to the development of new strategies for the effective alleviation and control of plant diseases.