Application of Thymol Vapors to Control Postharvest Decay Caused by Penicillium digitatum and Lasiodiplodia theobromae in Grapefruit.

Two of the major postharvest diseases impacting grapefruit shelf life and marketability in the state of Florida (USA) are stem-end rot (SER) caused by Lasiodiplodia theobromae and green mold (GM) caused by Penicillium digitatum. Here, we investigated the in vitro and in vivo efficacy of vapors of thymol, a natural compound found in the essential oil of various plants and the primary constituent of thyme (Thymus vulgaris) oil, as a potential solution for the management of GM and SER. Thymol vapors at concentrations lower than 10 mg L-1 significantly inhibited the mycelial growth of both pathogens, causing severe ultrastructural damage to P. digitatum conidia. In in vivo trials, the incidence and lesion area of GM and SER on inoculated grapefruit were significantly reduced after a 5 d exposure to 50 mg L-1 thymol vapors. In addition, the in vitro and in vivo sporulation of P. digitatum was suppressed by thymol. When applied in its vapor phase, thymol had no negative effect on the fruit, neither introducing perceivable off-flavor nor causing additional weight loss. Our findings support the pursuit of further studies on the use of thymol, recognized as safe for human health and the environment, as a promising strategy for grapefruit postharvest disease management.

Overcoming lemon postharvest molds caused by Penicillium spp. multiresistant isolates by the application of potassium sorbate in aqueous and wax treatments.

Penicillium digitatum and Penicillium italicum are the main causal agents of postharvest diseases in lemon. Over the last decades, the appearance of isolates resistant to the main commercial fungicides has been considered one of the most serious problems for the citrus industry. In this work, potassium sorbate (KS) was evaluated as an alternative to chemical fungicides to control postharvest diseases caused by Penicillium isolates resistant to imazalil, thiabendazol, and pyrimethanil. In vitro assays showed that 1% KS inhibited conidia germination and mycelial growth of sensitive and resistant P. digitatum and P. italicum isolates, being this effect stronger at pH 5 than at pH 9. In curative treatments, the immersion of inoculated lemons in 1% KS aqueous solution for 30 s reduced green and blue molds incidences by around 80%. No wound protection effect was observed when wounded lemons were immersed in 3% salt solution before inoculation. Noteworthy, the inclusion of KS in a commercial wax coating effectively controlled green and blue molds, even in decays caused by fungicide resistance isolates. Together, results encourage the use of KS in lemon postharvest treatments to contribute to the management of resistant strains, which represent a major challenge in packinghouses worldwide. PRACTICAL APPLICATION: The use of KS in citrus postharvest treatments would help producers to reduce spoilage caused by Penicillium fungicide-resistant strains. The inclusion of this generally recognized as safe compound in wax coatings improves its persistence on the fruit surface, keeping product quality during long-term overseas transport. In sum, KS constitutes an affordable and eco-friendly option for controlling postharvest molds in lemon fruit.

Inhibition of the lemon brown rot causal agent Phytophthora citrophthora by low-toxicity compounds.

BACKGROUND: Phytophthora spp., soil-borne oomycetes, cause brown rot (BR) on postharvest lemons. The management of this disease is based on cultural practices and chemical control using inorganic salts of limited efficacy. In the search for new alternatives, the aim of this work was to evaluate the effect of low-toxicity compounds to inhibit the growth of P. citrophthora and to control BR disease on lemons. Sodium bicarbonate, potassium sorbate, polyhexamethylene guanidine, Ascophyllum nodosum extract and a formulation containing phosphite salts plus A. nodosum (P+An) were evaluated. RESULTS: All tested products inhibited mycelial growth, sporangia formation and zoospore germination of P. citrophthora in vitro. In postharvest applications on artificially inoculated lemons, only P+An exhibited a BR curative effect, with incidence reduction of around 60%. When this formulation was applied in field treatments, BR incidence was reduced by 40% on lemons harvested and inoculated up to 30 days post application. CONCLUSION: Our results demonstrate the in vitro direct anti-oomycete effect of low-toxicity compounds and the in vivo efficacy of P+An formulation to control BR, encouraging the incorporation of the latter in the management of citrus BR. © 2020 Society of Chemical Industry.

Polyhexamethylene guanidine as a fungicide, disinfectant and wound protector in lemons challenged with Penicillium digitatum.

Citrus green mold, a postharvest disease caused by Penicillium digitatum, provokes important economic losses on lemon production. Here, the effectiveness of polyhexamethylene guanidine (PHMG) to inhibit P. digitatum growth and to control green mold on artificially infected lemons was evaluated. At sublethal concentrations, PHMG inhibited conidia germination and infectivity (5 mg L-1), and mycelial growth (50 mg L-1). Viability of conidia was completely suppressed by treatment with 500 mg L-1 PHMG. In this condition, membrane integrity loss, cell wall disruption and ultrastructural alterations were detected, as well as conidia distortion, deformation and collapse. In artificially inoculated lemons, a 30 s-immersion in 500 mg L-1 PHMG completely inhibited green mold. PHMG also exhibited a high disinfectant activity, even in the presence of 1% organic matter, with a better performance than the standard NaClO disinfectant. In addition, 500 mg L-1 PHMG protected wounds against infection. Taken together, our results indicate that PHMG is a promising fungicide for the postharvest control of green mold in lemon packinghouses.

UVA Photoactivation of Harmol Enhances Its Antifungal Activity against the Phytopathogens Penicillium digitatum and Botrytis cinerea.

Phytopathogenic fungi responsible for post-harvest diseases on fruit and vegetables cause important economic losses. We have previously reported that harmol (1-methyl-9H-pyrido[3,4-b]indol-7-ol) is active against the causal agents of green and gray molds Penicillium digitatum and Botrytis cinerea, respectively. Here, antifungal activity of harmol was characterized in terms of pH dependency and conidial targets; also photodynamic effects of UVA irradiation on the antimicrobial action were evaluated. Harmol was able to inhibit the growth of both post-harvest fungal disease agents only in acidic conditions (pH 5), when it was found in its protonated form. Conidia treated with harmol exhibited membrane integrity loss, cell wall disruption, and cytoplasm disorganization. All these deleterious effects were more evident for B. cinerea in comparison to P. digitatum. When conidial suspensions were irradiated with UVA in the presence of harmol, antimicrobial activity against both pathogens was enhanced, compared to non-irradiated conditions. B. cinerea exhibited a high intracellular production of reactive oxygen species (ROS) when was incubated with harmol in irradiated and non-irradiated treatments. P. digitatum showed a significant increase in ROS accumulation only when treated with photoexcited harmol. The present work contributes to unravel the antifungal activity of harmol and its photoexcited counterpart against phytopathogenic conidia, focusing on ROS accumulation which could account for damage on different cellular targets.

Antifungal activity of ß-carbolines on Penicillium digitatum and Botrytis cinerea.

ß-carbolines (ßCs) are alkaloids widely distributed in nature that have demonstrated antimicrobial properties. Here, we tested in vitro six ßCs against Penicillium digitatum and Botrytis cinerea, causal agents of postharvest diseases on fruit and vegetables. Full aromatic ßCs (harmine, harmol, norharmane and harmane) exhibited a marked inhibitory effect on conidia germination at concentrations between 0.5 and 1 mM, while dihydro-ßCs (harmalina and harmalol) only caused germination delay. Harmol showed the highest inhibitory effect on both fungal pathogens. After 24 h of exposure to 1 mM harmol, conidia revealed a severe cellular damage, exhibiting disorganized cytoplasm and thickened cell wall. Harmol antimicrobial effect was fungicidal on B. cinerea, while it was fungistatic on P. digitatum. Conidia membrane permeabilization was detected in treatments with harmol at sub-inhibitory and inhibitory concentrations, for both pathogens. In addition, residual infectivity of P. digitatum on lemons and B. cinerea on blueberries was significantly reduced after exposure to this alkaloid. It also inhibited mycelial growth, preventing sporulation at the highest concentration tested. These results indicate that harmol might be a promising candidate as a new antifungal molecule to control causal agents of fruit diseases.