Effect of light quality and light-dark cycle on sporulation patterns of the mite pathogenic fungus Neozygites floridana (Neozygitales: Entomophthoromycota), a natural enemy of Tetranychus urticae.

A controlled climatic chamber microcosm experiment was conducted to examine how light affects the hourly sporulation pattern of the beneficial mite pathogenic fungus Neozygites floridana during a 24h cyclus over a period of eight consecutive days. This was done by inoculating two-spotted spider mites (Tetranychus urticae) with N. floridana and placing them on strawberry plants for death and sporulation. Spore (primary conidia) discharge was observed by using a spore trap. Two light regimes were tested: Plant growth light of 150µmolm(-2)s(-1) for 12h supplied by high pressure sodium lamps (HPS), followed by either; (i) 4h of 50µmolm(-2)s(-1) light with similar HPS lamps followed by 8h darkness (full HPS light+reduced HPS light+darkness) or (ii) 4h of 50µmolm(-2)s(-1) red light followed by 8h darkness (full HPS light+red light+darkness). A clear difference in hourly primary conidia discharge pattern between the two different light treatments was seen and a significant interaction effect between light treatment and hour in day during the 24h cycle was observed. The primary conidia discharge peak for treatment (ii) that included red light was mainly reached within the red light hours (19:00-23:00) and the dark hours (23:00-07:00). The primary conidia discharge peak for treatment (i) with HPS light only was mainly reached within the dark hours (23:00-07:00).

Shedding the Light on Powdery Mildew: The Use of Optical Irradiation in Management of the Disease.

Ultraviolet (UV) irradiation below 300 nm may control powdery mildew in numerous crops. Depending on disease pressure, wavelength, and crop growth stage, one to three applications of 100-200 J/m2 per week at night are as effective or better than the best fungicides. Higher doses may harm the plants and reduce yields. Although red light alone or in combination with UV has a suppressive effect on powdery mildew, concomitant or subsequent exposure to blue light or UV-A strongly reduces the efficacy of UV treatments. To be effective, direct exposure of the pathogen/infection sites to UV/red light is important, but there are clear indications for the involvement of induced resistance in the host. Other pathogens and pests are susceptible to UV, but the effective dose may be phytotoxic. Although there are certain limitations, this technology is gradually becoming more used in both protected and open-field commercial production systems.

Propionate poses antivirulence activity against Botrytis cinerea via regulating its metabolism, infection cushion development and overall pathogenic factors.

Botrytis cinerea is a devastating pathogen causing gray mold in fruits and vegetables if not properly managed. Although the mechanisms remain unclear, we previously revealed that the safe food additive calcium propionate (CP) could suppress gray mold development on grapes. The present study reports that sub-lethal dose of CP (0.2 % w/v) could allow growth with substantial reprograming the genome-wide transcripts of B. cinerea. Upon CP treatment, the genes related to fungal methylcitrate cycle (responsible for catabolizing propionate) were upregulated. Meanwhile, CP treatment broadly downregulated the transcript levels of the virulence factors. Further comparative analysis of multiple transcriptomes confirmed that the CP treatment largely suppressed the expression of genes related to development and function of infection cushion. Collectively, these findings indicate that CP can not only reduce fungal growth, but also abrogate fungal virulence factors. Thus, CP has significant potential for the control of gray mold in fruit crops.

Functional Characterization of Pseudoidium neolycopersici Photolyase Reveals Mechanisms Behind the Efficacy of Nighttime UV on Powdery Mildew Suppression.

Powdery mildews can be controlled by brief exposure to ultraviolet (UV) radiation with devastating effect on their developmental stages including conidia germination. The treatment effect can be impaired by subsequent exposure to UV-A/blue light. UV-A/blue light-activated photolyase may be responsible for this and therefore we tested the function of three cryptochrome/photolyase family (CPF)-like genes (OINE01015670_T110144, OINE01000912_T103440, and OINE01005061_T102555) identified in the obligate biotrophic fungus Pseudoidium neolycopersici, the cause of tomato powdery mildew. A photolyase-deficient mutant of Escherichia coli transformed with coding sequence of OINE01000912_T103440 and exposed to brief (UV)-C treatment (peak emission at 254 nm) showed photoreactivation and cell survival when exposed to subsequent blue light, indicating complementation of photolyase activity. In contrast, the same photolyase-deficient E. coli transformed with the coding sequences of other two CPF-like genes did not survive this treatment, even though their expression were confirmed at protein level. This confirmed that OINE01000912_T103440 is a gene encoding photolyase, here named PnPHR1, with functionality similar to the native photolyase in E. coli, and classified as a class I cyclobutane pyrimidine dimer (CPD) photolyase. Modeling of the 634-amino acid sequence of PnPHR1 suggested that it is capable of binding flavin adenine dinucleotide (FAD) and methenyltetrahydrofolate (MTHF). However, spectroscopic data of the protein produced in an E. coli expression system could only reveal the presence of a reduced form of FAD, i.e., FADH- as an intrinsic chromophore. Within the tested wavelength range of 365-525 nm, the survival of photolyase-deficient mutant E. coli transformed with PnPHR1 showed a broad action spectrum from 365 to 454 nm. This was very similar to the previously characterized action spectrum for survival of P. neolycopersici conidia that had been treated with UV-C. Quantitative RT-PCR revealed that the expression of PnPHR1 in P. neolycopersici conidia was induced by UV-C, and peak expression occurred 4 h after brief UV-C treatment. The expression of PnPHR1 was repressed when incubated in red light after the UV-C treatment, but not when incubated in UV-A/blue light. The results may explain why the disease-reducing effect of short wavelength UV is impaired by exposure to UV-A and blue light.

Wavelength dependent recovery of UV-mediated damage: Tying up the loose ends of optical based powdery mildew management.

Controlled environment chamber experiments at Petri dish level were conducted to examine the wavelength and dose dependent efficacy of ultraviolet (UV) radiation, the recovery action potential of optical radiation applied concomitantly/subsequently to effective UV treatment, and the lapse time between UV treatment and subsequent exposure to recovery wavelength on germination efficiency of Oidium neolycopersici conidia. Conidia of eight- to nine-day-old colonies were dusted on water agar surface in Petri dishes and exposed to UV treatments (without lid). Immediately after UV treatments, Petri dishes were sealed and incubated in darkness or differing optical environments generated using seven different radiation sources (range 290nm to 780nm). Twenty-four hours after UV treatment, fifty conidia from each sample were assessed for germination. Compared to non-UV controls, <10% of the conidia germinated after 30s of exposure to 254nm or 283nm UV and subsequent dark incubation. Conidia germination was almost negligible if the exposure duration increased to 4min. Germination was about 60% with broad spectrum UV after 1min of exposure, and about 35% after 2 to 4min of exposure. There was no reduction of conidia germination with the exposure of ≤4min with 310nm. With the tested wavelength and dose ranges, germination recovery was effective in the 350nm to 500nm range. Germination efficiency of conidia treated with effective UV was significantly higher (>73%) if incubated subsequently in the 350nm to 500nm range (germination recovery). Furthermore, germination recovery depends on the characteristics of UV treatment (wavelength, and duration of exposure) and the lapse time between UV treatment and subsequent exposure to optical radiation in the recovery range. The findings of this study provide key criteria for wavelength selection, combination and application time in the optical radiation range, enabling improved design of optical based management strategies against powdery mildews.

Daily light integral and day light quality: Potentials and pitfalls of nighttime UV treatments on cucumber powdery mildew.

Nighttime ultraviolet (UV) radiation, if applied properly, has a significant potential for management of powdery mildews in many crop species. In this study, the role of growth light duration, irradiance, a combination of both (daily light integral) and light spectral quality (blue or red) on the efficacy of UV treatments against powdery mildew caused by Podosphaera xanthii and the growth performance of cucumber plants was studied in growth chambers. Increasing daily light integral provided by high-pressure sodium lamps (HPS) decreased efficacy of nighttime UV treatments against P. xanthii, but it increased plant growth. Furthermore, the efficacy of nighttime UV decreased when day length was increased from 16 to 20h at a constant daily light integral. The efficacy of nighttime UV increased if red light was applied after UV treatment, showing the possibility of day length extension without reducing the effect of UV. Increasing the dose of blue light during daytime reduced the efficacy of nighttime UV in controlling the disease, whereas blue deficient growth light (<6% of blue) caused UV mediated curling of young leaves. Furthermore, application of blue light after nighttime UV reduced its disease control efficacy. This showed the importance of maintaining a minimum of blue light in the growth light before nighttime UV treatment. Findings from this study showed that optimization of nighttime UV for management of powdery mildew is dependent on the spectral composition of the photosynthetically active radiation.

Determination of UV action spectra affecting the infection process of Oidium neolycopersici, the cause of tomato powdery mildew.

Oidium neolycopersici, the cause of powdery mildew in tomato, was exposed to UV radiation from 250 to 400 nm for 1, 12, or 24 min. Radiation ≤ 280 nm strongly reduced conidial germination, hyphal expansion, penetration attempt and infection of O. neolycopersici. From 290 to 310 nm the effect depended on duration of exposure, while there was no effect ≥ 310 nm. There were no significant differences within the effective UV range (250-280 nm). Conidial germination on a water agar surface was <20% or around 40%, respectively, if samples were exposed for 1 min within the effective UV range followed by 24h or 48 h incubation. Twelve or 24 min exposure reduced germination to close to nil. A similar trend occurred for germination of conidia on leaf disks on water agar in Petri dishes. The effective UV range significantly reduced all subsequent developmental stages of O. neolycopersici. There was no cytoplasmic mitochondrial streaming in conidia exposed to the effective UV range, indicating that there may be a direct effect via cell cycle arrest. There was no indication of reactive oxygen species involvement in UV mediated inhibition of O. neolycopersici. Optical properties of O. neolycopersici indicated that the relative absorption of UV was high within the range of 250 to 320 nm, and very low within the range of 340 to 400 nm. Identification of UV wavelengths effective against O. neolycopersici provides a future basis for precise disease control.