Gray mold caused by Botrytis cinerea is one of the most destructive postharvest decay of strawberry fruit. The present study aims to identify essential oils with antifungal activity against B. cinerea and the underlying mechanisms and their potential application in controlling postharvest decay. In the screening test, essential oils from Cymbopogon citratus (Cc), Thymus vulgraris (Tv), and Origanum heracleoticum (Oh) exhibited maximum inhibition of B. cinerea mycelial growth. The three essential oils altered the hyphal morphology and ultrastructure and resulted in many blebs around the hyphae. The essential oils damaged the plasma membrane of B. cinerea cells and resulted in the leakage of intercellular nucleic acids, proteins and soluble sugars. The exposure of strawberries to the vapors of these three essential oils in commercial package reduced gray mold, with Tv and Oh exhibiting strong efficiency and disease index reduction by 53.85% and 57.69%, respectively. Oh also inhibited postharvest decay and maintained fruit quality, preventing weight loss and soluble solid degradation. The study proposes using plant essential oils as an alternative to chemical fungicides in controlling the gray mold of strawberries.

Fruit development and ripening are accompanied by a large increase in cellular soluble solid contents, which results in a significant decrease in osmotic potential (DOP). Here, we report that this development-associated DOP contributes to the initiation of ripening in strawberry (Fragaria ananassa Duch., Benihoppe) fruit. We show that fruit water potential significantly decreases at the onset of ripening as a result of the DOP. Further analysis using nuclear magnetic resonance spectroscopy (NMR) indicated that the change in fruit water potential was likely caused by catabolism of large molecules in receptacle cells, and bioinformatic analysis identified a family of osmotin-like proteins (OLP) that have a potential role in osmolyte accommodation. The gene expression of more than half of the OLP members increased substantially at the onset of fruit ripening, and specifically responded to DOP treatment, consistent with a close relationship between DOP and fruit ripening. We report that the DOP induced either by mannitol or water loss, triggered fruit ripening, as indicated by the elevated expression of multiple ripening genes and diverse ripening-associated physiological parameters. Collectively, these results suggest that the DOP contributes to strawberry fruit ripening initiation.

The interactive effects of six maturity stages and refrigerated storage (chilling)/blanching (heating) treatments on the volatile profiles of ripe tomatoes were studied. A total of 42 volatiles were identified, of which 19 compounds had odor activity values equal to or greater than 1. Of those, “green” and “leafy” aroma volatiles were most abundant. Chilling and heating treatments both suppressed overall volatile production, with chilling having the greater impact, regardless of harvest maturity. However, fruit harvested at the turning stage had the least volatile suppression by chilling and heating treatments in comparison with fruit harvested earlier or later, mostly in the fatty acid- and phenylalanine-derived volatiles. Volatiles derived from amino acids were promoted by heat treatment for fruit harvested at all maturities, and those derived from carotenoid and phenylalanine pathways and harvested at advanced harvest maturities were stimulated by chilling treatment. Volatile production is generally believed to be improved by delayed harvest, with vine-ripe being optimum. However, opposite results were observed possibly because the later-harvested fruit had longer exposure to open-field weather stress. The best harvest maturity recommendation is the turning stage where fruit developed abundant volatiles and were least impacted by chilling and heating treatments.

Gray mold caused by Botrytis cinerea is a major threat to the production of strawberry. An increasing number of studies have reported that wall-associated kinase/wall-associated kinase-like (WAK/WAKL) played an important role in the recognition of oligogalacturonic acids (OGs) and the induction of plant defense, but there have been no systematic studies of FaWAK/FaWAKL in strawberry. In this study, we identified 167 FaWAK/FaWAKL gene family members within the strawberry (Fragaria×ananassa) genome. The phylogenetic analysis showed the FaWAK/FaWAKL gene family has been divided into five groups, and they were unevenly distributed on 46 chromosomes. An analysis of the cis-regulatory elements suggested the FaWAK/FaWAKL gene family was more sensitive to abscisic acid and methyl jasmonate. A total of 36 FaWAK/FaWAKL genes were activated by B. cinerea according to an RNA-seq analysis, and 8 of them strongly responded to B. cinerea and exogenous treatment with OGs, particularly FaWAK35. Transient overexpression of FaWAK35 increased the strawberry resistance to B. cinerea. This study conducted a comprehensive analysis of FaWAK/FaWAKL and provides foundational insights for further exploration of FaWAK/FaWAKL genes in strawberry resistance to B. cinerea.

Postharvest decay of strawberry (Fragaria × ananassa Duch.) is a major factor causing fruit losses. Strawberries were obtained from various harvests at cooling facilities located in Dover and Plant City, FL during the 2018-19 and 2019-20 seasons. After the fruits were incubated at 22ºC for up to 5 days (d) to promote disease development, Lasiodiplodia decay was observed at up to 3% from some harvests, exhibiting gray mycelia on small lesions that gradually covered the whole fruit. The fungus was isolated onto potato dextrose agar (PDA). Five isolates (SBD18-14, SBD18-277, SBD18-279, SBD19-02 and SBD19-57) were characterized. Fungal mycelia were initially grayish white and then gradually changed to gray to dark gray on PDA at 25oC, and later produced black pigments (Fig. S1). Pycnidia were observed from inoculated strawberries at 14 d. Isolates shared similar conidia morphology: aseptate, hyaline, ellipsoid to ovoid, measuring L × W: 24.0-34.0 (28.3) × 13.0-16.0 (14.3) μm (n =100). Mature conidia were brown, one septate, measuring L × W: 25.0-33.0 (28.8) × 13.0-16.0 (14.5) μm (n =100). The isolates were identified as Lasiodiplodia spp. morphologically (Alves et al. 2008). DNA was extracted from fungal mycelia using an OmniPrep DNA extraction kit, and PCR amplification of ITS and EF1-α genes was performed following the conditions described by White et al. (1990) with some modifications using primers ITS1F-F/ITS4-R (Gardes and Bruns, 1993; White et al., 1990) and EF1-668-F/EF1-1251-R (Alves et al., 2008), respectively. The BLASTn in GenBank showed that the sequences obtained had 99.61 to 100% homology with those of ITS (EF622077) and EF1-α (EF622057) from L. pseudotheobromae CBS116459 (an ex-type strain) (Alves et al., 2008). Sequences of the isolates have been deposited in GenBank with accessions OP326017 to OP326021 for ITS, and OP356202 to OP356206 for EF1-α. Phylogenetic analysis showed that these isolates clustered in the same clade (bootstrap value at 64) with L. pseudotheobromae (Fig. S2). Two fungal inoculum types (mycelia and conidia), two fruit inoculation methods (injury and non-injury) and five fungal isolates were used for pathogenicity tests. Fungal mycelia (2-day-old) on PDA plug (5 mm) or 10 µL of conidial suspension (106 spores/mL) was placed onto each injury (1 x 1 mm in size) or a non-injury area on the surfaces of five strawberry fruits (cv. Florida Brilliance). PDA plug alone or water drops placed on injury or non-injury areas on fruits served as respective controls. Inoculated and control fruits were incubated in a covered plastic container with 100% RH at 22ºC. The experiment was repeated twice. Decay initially appeared as soft and lightly discolored tissue at inoculation areas 2 d post-inoculation (dpi) that extended quickly thereafter. Brown to dark lesions on both injury- and non-injury fruits inoculated with conidia or mycelia were observed at 3 dpi. Decay and gray mycelia gradually developed over the whole fruit at 6 dpi, and pycnidia were observed after 14 dpi (Fig. S1). Disease incidence of 100% was observed on all tests. Control fruits did not develop decay. The results indicate that these isolates are pathogenic to strawberries and infect fruit via both non-injured and injured fruit surfaces. The inoculated fungal isolates were re-isolated, thus, fulfilling Koch's postulates. L. theobromae, Neofusicoccum parvum/N. ribis species complex causing strawberry fruit rot in Florida fields was reported (Oliveira et al., 2019), but not L. pseudotheobromae. To our knowledge, this is the first report of postharvest decay caused by L. pseudotheobromae A.J.L. Phillips, A. Alves & Crous on strawberries in Florida and in the USA, and it should be considered in strawberry disease management.

Fumonisins are polyketide-derived mycotoxins posing significant health threats to humans and animals. Among these, fumonisin B₁ (FB₁) is the most prevalent mycotoxin, primarily produced by Fusarium verticillioides, especially in maize and its derived products. Tangeretin, a polymethoxyflavonoid, has been identified as having potential medicinal properties, particularly as an antioxidant. To evaluate the antifungal and anti-mycotoxigenic properties of tangeretin and to elucidate the mechanisms underlying its inhibitory effects, assessments of fungal growth, FB₁ production, conidial germination, and cellulase activity, antioxidant capacity and enzyme activities, transcriptomic analysis and gene deletion experiments were conducted. Consequently, tangeretin significantly curtailed fungal growth and FB₁ production and provided protection against pathogenic infection on corn. It affected genes associated with fungal growth, conidial development, and antioxidant response. Furthermore, tangeretin interfered with the supply of biosynthetic substrate necessary for fumonisin production, particularly impacting pathways involved in alanine metabolism, pyruvate metabolism, fatty acid degradation, and sphingolipid metabolism. Notably, tangeretin downregulated four biosynthetic genes (Fum2, Fum3, Fum10 and Fum11) that are involved in the final steps of fumonisin formation. It likely disrupted the MAPK signaling pathway and targeted a putative NmrA- and HSCARG-like protein Fv_Tan1, which was identified as having positive effects on fungal growth and mycotoxin biosynthesis. This study presents a promising approach for controlling fumonisin contamination in agricultural settings.