First report of Rhizopus stolonifer causing flower rot of yellow passion fruit (Passiflora edulis f. flavicarpa Deg.) in China.

The genus Passiflora, commonly known as passion fruit, originated in South America, is an economically important horticulture crop and widely distributed in the tropics and subtropics. Yellow passion fruit (Passiflora edulis f. flavicarpa) and purple passion fruit (Passiflora edulis f. edulis) are the two most planted species (Santos-Jiménez et al., 2022), which have been largely cultivated in southern China. The average annual production reaches 600,000 tons, of which yellow fruit accounts for more than 70% (Zhou et al., 2022). In 2022 to 2023, a disease caused flower rot severely in passion fruit plantations. The incidence rate was generally 10% in purple passion fruit, with an incidence up to 60% in yellow passion fruit 'Qinmi No. 9'. Flower rot occurs mainly in the rainy season, especially during periods of prolonged rain. Infected flowers had black patches that were water-soaked on the interior of the flower bud. The patches covered the entire flower bud, and fluffy mycelium and sporangia developed, which caused the flower bud rotten and abscised easily. Five symptomatic flowers from Wuhua, Guangdong (23°23'N, 115°18'E) and 8 symptomatic flowers from Shangsi, Guangxi (21°15'N, 107°98'E) of 'Qinmi No. 9' were collected during flowering period in 2022 and 2023. Diseased flower pieces were surface-sterilized with 70% ethanol for 2 to 3 min, rinsed with sterile distilled water 3 times, and placed on PDA medium at 25℃ in darkness. Four and 6 fungal isolates with similar morphology were isolated from the infected samples of Wuhua and Shangsi, respectively. Two isolates, PRFJ01 from Wuhua and PRGX02 from Shangsi, were randomly selected for further study. Purified fungal colonies at the age of 3 days accompany with diffuse cottony mycelia, turned white to gray later. The mycelia were hyaline and aseptate. Sporangiophores with 0.56 (0.22~1.10) mm in length and 6.1 (3.18~10.87) µm in width (n=100) were erect, light brown, and had rhizoids and stolons at their bases. Sporangia with 48.0 (23.45~92.85) µm in diameter (n=100) were dark-colored, near spherical and having dark ovoid sporangiospores with 3.56 (2.34~6.39) µm × 2.82 (1.73~4.70) µm (n=100). The morphology of the fungus were identical to Rhizopus stolonifer (Ehrenb.) Vuill (Haque et al. 2023). The two isolates were molecularly identified using genomic regions of 28S large ribosomal subunit (LSU) with NL1 and LR3 primers (Cruz-Lachica et al., 2018). The phylogenetic trees revealed the sequences of PRFJ01 (OR801560.1) and PRGX02 (OR801561.1) were 100% and 99% identical to R. stolonifer (MK705761.1 and KC412868.1), respectively. Pathogenicity tests were conducted on healthy flowers and leaves of 5-month-old grafted 'Qinmi No. 9' plants. Mycelial plugs with 5-mm diameter were placed on the flowers and leaves. Three plants were performed for each of the isolates, and the test was repeated twice. The inoculated plants were moisturized with plastic bags. Healthy flowers and leaves inoculated with sterile PDA plugs were used as control. Typical symptoms were observed on inoculated plants after 2 days. The dark grey mycelia and sporangia covered the entire flower after 4 days inoculation. The flower bud became putrid and the flower stalk split off. Lesions on leaves expanded accompany with numerous aerial mycelium. However, the controls were symptomless. R. stolonifer was reisolated from inoculated tissues. Previously, flower rot on passion fruit caused by R. stolonifer has only been recorded in Brazil (Ploetz, 2003). To our knowledge, this is the first report of R. stolonifer causing flower rot on passion fruit in China.

Complete genome sequence of a novel double-stranded RNA virus infecting the phytopathogenic fungus Rhizopus stolonifer.

In this study, we report the complete genome sequence of a novel toti-like virus, tentatively named "Rhizopus stolonifer double-stranded RNA virus 1" (RsDSV1), identified from a phytopathogenic fungal agent of apple fruit rot disease, Rhizopus stolonifer strain A2-1. RsDSV1 has a double-stranded RNA genome. The complete RsDSV1 genome is 5178 nucleotides (nt) in length and contains two open reading frames (ORFs) encoding a putative coat protein (CP) and an RNA-dependent RNA polymerase (RdRp). Phylogenetic analysis based on RdRp and CP amino acid sequences revealed that RsDSV1 is closely related to unclassified members of the family Totiviridae. In stress-inducing Vogel's minimal and sodium dodecyl sulfate-containing media, hyphal growth of A2-1 was suppressed, but the accumulation of RsDSV1 RNA increased, indicating that stresses promote RsDSV1 replication. To our knowledge, this is the first report of a mycovirus found in R. stolonifer.

Host susceptibility factors render ripe tomato fruit vulnerable to fungal disease despite active immune responses.

The increased susceptibility of ripe fruit to fungal pathogens poses a substantial threat to crop production and marketability. Here, we coupled transcriptomic analyses with mutant studies to uncover critical processes associated with defense and susceptibility in tomato (Solanum lycopersicum) fruit. Using unripe and ripe fruit inoculated with three fungal pathogens, we identified common pathogen responses reliant on chitinases, WRKY transcription factors, and reactive oxygen species detoxification. We established that the magnitude and diversity of defense responses do not significantly impact the interaction outcome, as susceptible ripe fruit mounted a strong immune response to pathogen infection. Then, to distinguish features of ripening that may be responsible for susceptibility, we utilized non-ripening tomato mutants that displayed different susceptibility patterns to fungal infection. Based on transcriptional and hormone profiling, susceptible tomato genotypes had losses in the maintenance of cellular redox homeostasis, while jasmonic acid accumulation and signaling coincided with defense activation in resistant fruit. We identified and validated a susceptibility factor, pectate lyase (PL). CRISPR-based knockouts of PL, but not polygalacturonase (PG2a), reduced susceptibility of ripe fruit by >50%. This study suggests that targeting specific genes that promote susceptibility is a viable strategy to improve the resistance of tomato fruit against fungal disease.

Antifungal activity screening for mint and thyme essential oils against Rhizopus stolonifer and their application in postharvest preservation of strawberry and peach fruits.

AIMS: With a goal to identify specific essential oils that can control postharvest Rhizopus rot on strawberry and peach fruits, we performed screening for 26 essential oils based on their antifungal activity against Rhizopus stolonifer in vitro and investigated the underlying mechanism. METHODS AND RESULTS: Mentha spicata (Ms), Mentha piperita (Mp), Thymus vulgaris CT carvacrol (Tc) and Thymus vulgaris CT thymol (Tt) exhibited strong inhibition on R. stolonifer growth in the screening. These essential oils increased plasma membrane permeability of R. stolonifer and resulted in the outflow of intercellular electrolyte, nucleic acid, protein and soluble sugar. Morphology of R. stolonifer mycelia was greatly altered by these essential oils. Hyphae treated by these essential oils exhibited high accumulation of superoxide anion and malonaldehyde. Combination of these essential oils in commercial package reduced Rhizopus rot on strawberry and peach fruits, with Mp showing the strongest efficiency. CONCLUSION: Ms, Mp, Tc and Tt essential oils inhibited R. stolonifer growth by targeting its plasma membrane and reduced Rhizopus rot on strawberry and peach fruits. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides potential applications of natural plant extract, as alternatives to tradition fungicides, to control postharvest decay on fruits and vegetables.

Iturin A Induces Resistance and Improves the Quality and Safety of Harvested Cherry Tomato.

The soft rot disease caused by Rhizopus stolonifer is an important disease in cherry tomato fruit. In this study, the effect of iturin A on soft rot of cherry tomato and its influence on the storage quality of cherry tomato fruit were investigated. The results showed that 512 µg/mL of iturin A could effectively inhibit the incidence of soft rot of cherry tomato fruit. It was found that iturin A could induce the activity of resistance-related enzymes including phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), glucanase (GLU), and chitinase (CHI), and active oxygen-related enzymes including ascorbate peroxidases (APX), superoxide dismutases (SOD), catalases (CAT), and glutathione reductase (GR) of cherry tomato fruit. In addition, iturin A treatment could slow down the weight loss of cherry tomato and soften the fruit. These results indicated that iturin A could retard the decay and improve the quality of cherry tomato fruit by both the inhibition growth of R. stolonifera and the inducing the resistance.

Effect of combined Bacillomycin D and chitosan on growth of Rhizopus stolonifer and Botrytis cinerea and cherry tomato preservation.

BACKGROUND: Synthetic fungicides are most commonly used for controlling postharvest disease of fruit, although they can cause the emergence of drug-resistant strains, environmental pollution and fruit safety issues. Bacillomycin D (BD), a novel antifungal lipopeptide, and chitosan (CTS) are applied for the preservation of cherry tomato. RESULTS: The combination of BD and CTS showed an additive inhibition on the growth of Rhizopus stolonifer and Botrytis cinerea compared to that of its individual compound. In addition, BD + CTS reduced the incidence of soft rot and gray mold in cherry tomato caused by R. stolonifer and B. cinerea, respectively. Tomato treated with BD + CTS exhibited a lower weight loss and higher firmness and higher contents of total soluble solids, titratable acidity and ascorbic acid compared to those treated with sterile water (control). The kinetics models demonstrated that the shelf life of cherry tomato treated with BD + CTS could be extended by approximately 15 days longer than the control. CONCLUSION: The utilization of BD + CTS provided a novel strategy for reducing postharvest fungal rot and maintaining the storage quality of cherry tomato. © 2020 Society of Chemical Industry.

Control of nectarine fruits postharvest fungal rots caused by Botrytis Cinerea and Rhizopus Stolonifer via some essential oils.

Nectarines (Prunus persica L. Bath) are very sensitive fruit to fungal infection. Today, the control of postharvest fruit diseases with essential oils (EOs) has been significantly noticed as a novel trend in biological preservation. In this study, volatile compounds of Cinnamon zeylanicum (CEO), Zataria multiflora (ZEO), and Satureja khuzestanica (SEO) were analyzed by Gas Chromatography-Mass spectroscopy. Also, the in vitro antifungal activities of EOs against Botrytis cinerea and Rhizopus stolonifer were evaluated at different concentrations. The in vivo antifungal activity of these EOs on artificially infected nectarine fruits was also considered. The major components were Thymol (32.68%) and Carvacrol (30.57%) for ZEO, cinnamaldehyde (80.82%) for CEO, and carvacrol (38.43%) for SEO. The application of different concentrations showed a decreasing trend in the fungus radial growth in all EOs. In the in vitro experiments, ZEO and CEO exhibited more significant mycelial inhibition results and reduction of the IC50, MIC and MFC values against Botrytis cinerea and Rhizopus stolonifer, respectively. However, in the in vitro experiments, none of the treatments were capable of completely inhibiting the growth of the fungi. According to the results of this study, ZEO and CEO could reduce the damage caused by these fungi.

Bacillomycin D inhibits growth of Rhizopus stolonifer and induces defense-related mechanism in cherry tomato.

The inhibitory effect of Bacillomycin D, a cyclic lipopeptide, on Rhizopus stolonifer colonization of cherry tomato was studied, and its possible mechanism of action was explored. Bacillomycin D showed a direct inhibitory effect on R. stolonifer spore germination and mycelial growth in vitro. It conferred both a direct inhibitory effect on R. stolonifer growth in cherry tomato in vivo and induced host resistance in cherry tomato. Moreover, Bacillomycin D treatment significantly increased the activities of plant defense-related enzymes, including chitinase (CHI), ß-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), and peroxidase (POD). Real-time PCR (RT-PCR) showed that defense-related genes involved in the salicylic acid defense signaling pathway and genes encoding pathogenesis-related proteins were up-regulated in Bacillomycin D treatment. Furthermore, Bacillomycin D-C16 resulted in direct inhibition and a remarkable induced resistance to R. stolonifer which was higher than as induced by Bacillomycin D-C14. Together, the data indicated that Bacillomycin D can control the growth of R. stolonifer through both the direct inhibition of the fungus and the activation of defense-related genes and enzymes in cherry tomato.

Carvacrol and eugenol effectively inhibit Rhizopus stolonifer and control postharvest soft rot decay in peaches.

AIMS: This study aimed to investigate the antifungal mechanism of carvacrol and eugenol to inhibit Rhizopus stolonifer and the control of postharvest soft rot decay in peaches. METHODS AND RESULTS: To investigate the antifungal mechanism, the effects of carvacrol and eugenol on the mycelium growth, leakages of cytoplasmic contents, mycelium morphology, cell membrane and membrane composition of R. stolonifer were studied. Carvacrol and eugenol both exhibited dose-dependent antifungal activity against R. stolonifer, carvacrol at a concentration of 2 µl per plant and eugenol at a concentration of 4 µl per plant inhibited fungal growth completely. The two essential oils (EOs) increased cell membrane penetrability and caused the leakage of cytoplasm, nucleic acid and protein content. The observation using scanning electron microscopy and fluorescent microscopy showed modification of the hyphal morphology and breakage of the cell plasma membrane. Decreased ergosterol contents confirmed that the two EOs could destroy the membrane of R. stolonifer. For the in vivo test, the inhibition of soft rot disease and the induction of defence-related enzymes were investigated. Carvacrol and eugenol significantly reduced the incidence and severity of soft rot decay in inoculated peaches. The best treatments for controlling soft rot decay were obtained at 0·5 µl l-1 for carvacrol and 1 µl l-1 for eugenol. The activities of defence-related enzymes in peaches were also enhanced by fumigation with two EOs. CONCLUSION: This study showed that carvacrol and eugenol could effectively inhibit the growth of R. stolonifer in vitro and successfully control the incidence of soft rot decay in honey peaches. SIGNIFICANCE AND IMPACT OF THE STUDY: The above findings may be the main antifungal mechanism of carvacrol and eugenol on R. stolonifer. Furthermore, carvacrol and eugenol are helpful for their commercial application on the preservation of fresh fruit.

Transcriptional regulator XYR1 activates the expression of cellobiose synthase to promote the production of cellulase from glucose.

OBJECTIVE: To investigate the transcriptional regulation of cellobiose synthase (CBS) in Rhizopus stolonifer. RESULTS: Transcription factor XYR1 was identified as responsible for the activation of cbs. In comparison with wild-type R. stolonifer, the deletion of XYR1 resulted in transcriptional down-regulation of cbs by approximately 40%, while XYR1 over-expression increased cbs transcription up to 175%. The highest FPA activity (1.8 IU/ml) was obtained in the XYR1-overexpressing strain OExyr1 cultivated in a 2% (m/V) glucose media, corresponding to a 96% increase compared with that of the parent strain (0.92 IU/ml). Moreover, cellulase synthesis was inhibited after cbs-inactivation mutation in OExyr1. CONCLUSION: XYR1 directly activates the transcription of cbs to promote cellulase production in R. stolonifer utilizing glucose as a substrate.

Effects of chitosan and oligochitosan on development and mitochondrial function of Rhizopus stolonifer.

The antifungal activities of chitosan and oligochitosan have been used to control postharvest decay of the fruits. The effect of chitosan and oligochitosan on mycelium growth, spore germination, and mitochondrial function of Rhizopus stolonifer was evaluated in order to establish a connection between fungus development and the main organelle in charge to provide energy to the cell. The mycelium growth of R. stolonifer was significantly reduced on minimum media amended with chitosan or oligochitosan. The highest antifungal indexes were obtained on media containing chitosan or oligochitosan at 2.0 mg ml(-1). Microscopic observation showed that chitosan and oligochitosan affected the spore germination and hyphae morphology. Both polymers increased oxygen consumption of R. stolonifer. Respiratory activity was restored with NADH in permeabilized treated and untreated cells, and was inhibited with rotenone and flavones. Complex III and IV were inhibited by antimycin A and cyanide, respectively, in treated and untreated cells. Chitosan and oligochitosan increased NADH dehydrogenase activity in isolated mitochondria. However, there were not changes in the cytochrome c oxidase and ATPase activities by effect of these polymers. These results suggest that both chitosan and oligochitosan affect the development of R. stolonifer and might be implicated in the mitochondrial dysfunction.

Rhizopus stolonifer and related control strategies in postharvest fruit: A review.

Rhizopus stolonifer is one of the main pathogens in postharvest storage logistics of more than 100 kinds of fruit, such as strawberries, tomatoes and melons. In this paper, the research on the morphology and detection, pathogenicity and infection mechanism of Rhizopus stolonifer was reviewed. The control methods of Rhizopus stolonifer in recent years was summarized from three dimensions of physics, chemistry and biology, including the nanomaterials, biological metabolites, light control bacteria, etc. Future direction of postharvest Rhizopus stolonifer infection control was analyzed from two aspects of pathogenic mechanism research and new composite technology. The information provided in this review will help researchers and technicians to deepen their understanding of the pathogenicity of Rhizopus stolonifer, and develop more effective control methods in the future.

Antifungal efficacy of chitosan extracted from shrimp shell on strawberry (Fragaria × ananassa) postharvest spoilage fungi.

The strong demand for biological materials in the food industry places chitosan at the forefront of other biopolymers. The present study aims to evaluate the antifungal properties of chitosan extracted from shrimp shell waste (Parapenaeus longirostris) against post-harvest strawberry (Fragaria × ananassa) spoilage fungi. The physicochemical characteristics (DD, Mw, and solubility) of extracted chitosan were determined. In addition, functional characteristics were studied by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The antifungal effect of chitosan on mycelial growth and spore germination of Aspergillus niger, Botrytis cinerea, Fusarium oxysporum, and Rhizopus stolonifer was evaluated. Yield, degree of deacetylation, molecular weight, and solubility were 21.86%, 83.50%, 180 kDa, and 80.10%, respectively. A degree of deacetylation of 81.27% was calculated from the FTIR spectrum and a crystallinity index of 79.83% was determined from the X-ray diffraction pattern. SEM images of extracted chitosan showed a combination of fibrous and porous structure. At 3% chitosan, mycelial growth inhibition rates of A. niger, B. cinerea, F. oxysporum, and R. stolonifer ranged from 81.37% to 92.70%. At the same chitosan concentration, the percentages of spore germination inhibition of the isolated fungi ranged from 65.47% to 71.48%. The antifungal activity was highly dose-dependent. As a natural polymer, chitosan offers a convincing alternative to synthetic antimicrobials for the post-harvest preservation of strawberries. Its potential lies in its ability to inhibit the growth of spoilage fungi.

Bacillus subtilis Edible Films for Strawberry Preservation: Antifungal Efficacy and Quality at Varied Temperatures.

Fungal infestations, particularly from Rhizopus stolonifer, pose significant post-harvest challenges for strawberries, compromising their shelf life and quality. Traditional preservation methods, including refrigeration, offer limited protection against such pathogens. This study introduces an innovative approach, utilizing edible films infused with Bacillus subtilis strains GOS 01 B-67748 and HFC 103, known for their antifungal properties. We demonstrate that these bioactive films not only inhibit fungal growth effectively but also enhance the preservation of strawberries at varying temperatures. The inclusion of Bacillus subtilis in edible films represents a significant advancement in extending the viability of strawberries, surpassing the efficacy of conventional methods. Our findings suggest a promising avenue for natural, safe food preservation techniques, aligning with current consumer preferences for additive-free products. This research contributes to the broader understanding of microbial-based food preservation strategies, offering potential applications across a range of perishable commodities.

Ultrasound-assisted fabrication of chitosan-hydroxypropyl methylcellulose nanoemulsions loaded with thymol and cinnamaldehyde: Physicochemical properties, stability, and antifungal activity.

This study investigated the influence of chitosan (CH) and hydroxypropyl methylcellulose (H), along with ultrasound power, on the physicochemical properties, antifungal activity, and stability of oil-in-water (O/W) nanoemulsions containing thymol and cinnamaldehyde in a 7:3 (v/v) ratio. Eight O/W formulations were prepared using CH, H, and a 1:1 (v/v) blend of CH and H, both with and without ultrasonication (U). Compared to untreated samples, U-treated nanoemulsions had lower droplet sizes (433-301 nm), polydispersity index (0.42-0.47), and zeta potential (-0.42-0.77 mV). The U treatment decreased L* and b* values, increased a* color attribute values, and increased apparent viscosity (0.26-2.17) at the same shear rate. After 28 days, microbiological testing of nanoemulsions treated with U showed counts below the detection limits (< 2 log CFU mL-1). The U-treated nanoemulsions exhibited stronger antifungal effects against R. stolonifer, with the NE/CH-U and NE/CH-H-U formulations demonstrating the lowest minimum inhibitory and fungicidal concentrations, measured at 0.12 and 0.24 µL/mL, respectively. On day 28, U-treated nanoemulsions demonstrated higher ionic, thermal, and physical stability than untreated samples. These findings suggest that the stability and antifungal efficacy of polysaccharide-based nanoemulsions may be improved by ultrasonic treatment. This study paves the way for innovative, highly stable nanoemulsions.

Analysis of functional ingredients of tempe-like fermented Moringa oleifera seeds (Moringa tempe) prepared with Rhizopus species.

Tempe is a fermented food prepared by fermenting soybeans with Rhizopus species. However, there have recently been concerns about the stable supply of raw soybeans due to global warming and other factors. Moringa is a plant whose cultivation area is expected to expand in the future, and its seeds contain abundant proteins and lipids, and thus could be used as an alternative to soybeans. To develop a novel functional Moringa food, we fermented dehulled Moringa seeds with Rhizopus oligosporus and Rhizopus stolonifer using the solid fermentation method of tempe and investigated changes in the functional components, such as free amino acids and polyphenols, of the respective obtained Moringa tempe Rm and Rs. After 45 h of fermentation, the total content of free amino acids, mainly including gamma-aminobutyric acid and l-glutamic acid, in Moringa tempe Rm was about three times higher, while that in Moringa tempe Rs was almost the same, compared to that in unfermented Moringa seeds. Moreover, after 70 h of fermentation, both Moringa tempe Rm and Rs had approximately four times higher polyphenol content and significantly higher antioxidant activity than did unfermented Moringa seeds. Further, the content of each residual chitin-binding protein of defatted Moringa tempe Rm and Rs was almost the same as that of unfermented Moringa seeds. Taken together, Moringa tempe was rich in free amino acids and polyphenols, exhibited better antioxidant activity, and retained the levels of its chitin-binding proteins, suggesting that Moringa seeds could be used as an alternative to soybean for tempe preparation.

Biocatalysis of triglycerides transesterification using fungal biomass: a biorefinery approach.

BACKGROUND: The use of microbial biomasses, such as fungal biomass, to catalyze the transesterification of triglycerides (TG) for biodiesel production provides a sustainable, economical alternative while still having the main advantages of expensive immobilized enzymes. RESULTS: Biomasses of Aspergillus flavus and Rhizopus stolonifera were used to catalyze the transesterification of TG in waste frying oil (WFO). Isopropanol as an acyl-acceptor reduced the catalytic capability of the biomasses, while methanol was the most potent acyl-acceptor with a final fatty acid methyl ester (FAME) concentration of 85.5 and 89.7%, w/w, for R. stolonifer and A. flavus, respectively. Different mixtures of the fungal biomasses were tested, and higher proportions of A. flavus biomass improved the mixture's catalytic capability. C. sorokiniana cultivated in synthetic wastewater was used as feedstock to cultivate A. flavus. The biomass produced had the same catalytic capability as the biomass produced in the control culture medium. Response surface methodology (RSM) was adopted using central composite design (CCD) to optimize the A. flavus biomass catalytic transesterification reaction, where temperature, methanol concentration, and biomass concentration were selected for optimization. The significance of the model was verified, and the suggested optimum reaction conditions were 25.5 °C, 250 RPM agitation with 14%, w/w, biomass, 3 mol/L methanol, and a reaction duration of 24 h. The suggested optimum conditions were tested to validate the model and a final FAME concentration of 95.53%. w/w was detected. CONCLUSION: Biomasses cocktails might be a legitimate possibility to provide a cheaper technical solution for industrial applications than immobilized enzymes. The use of fungal biomass cultivated on the microalgae recovered from wastewater treatment for the catalysis of transesterification reaction provides an additional piece of the puzzle of biorefinery. Optimizing the transesterification reaction led to a valid prediction model with a final FAME concentration of 95.53%, w/w.

Development of a novel 1-octen-3-ol-loaded agar/curdlan hydrogel for inhibiting peach fruit diseases.

Our previous study found that 1-octen-3-ol fumigation treatment could effectively induce the resistance of peach fruit diseases. However, 1-octen-3-ol is a liquid fumigant, which is not conducive to storage and application. Herein, the gel of 1 % agar compound with 1 % curdlan was used as a novel material for covering 1-octen-3-ol. The interaction of agar and curdlan was promoted by adding 1-octen-3-ol, leading to a higher thermostability compared to single-component antibacterial gels. Moreover, 1-octen-3-ol resulted in changes in the internal structure and mechanical properties of gel to form a pore-like structure, which is beneficial to the retention and release of 1-octen-3-ol. Additionally, the 2 % agar gel containing 1-octen-3-ol had the best inhibitory effect on the mycelial growth of Monilinia fructicola and Rhizopus stolonifer in vitro, and the compound hydrogel of 1 % agar and 1 % curdlan with 1-octen-3-ol could most effectively inhibit brown rot and soft rot caused by these two pathogens in vivo. Overall, the data indicated that the novel 1-octen-3-ol-loaded agar/curdlan hydrogels could effectively retain and release 1-octen-3-ol, and induce the resistance of peach fruit diseases.

Miltefosine: A Repurposing Drug against Mucorales Pathogens.

Mucorales are a group of non-septated filamentous fungi widely distributed in nature, frequently associated with human infections, and are intrinsically resistant to many antifungal drugs. For these reasons, there is an urgent need to improve the clinical management of mucormycosis. Miltefosine, which is a phospholipid analogue of alkylphosphocholine, has been considered a promising repurposing drug to be used to treat fungal infections. In the present study, miltefosine displayed antifungal activity against a variety of Mucorales species, and it was also active against biofilms formed by these fungi. Treatment with miltefosine revealed modifications of cell wall components, neutral lipids, mitochondrial membrane potential, cell morphology, and the induction of oxidative stress. Treated Mucorales cells also presented an increased susceptibility to SDS. Purified ergosterol and glucosylceramide added to the culture medium increased miltefosine MIC, suggesting its interaction with fungal lipids. These data contribute to elucidating the effect of a promising drug repurposed to act against some relevant fungal pathogens that significantly impact public health.

Chemical composition and antifungal activity of Zanthoxylum riedelianum stem bark essential oil.

The phytopathogenic fungi Sclerotinia sclerotiorum and Rhizopus stolonifer infect different crop plants. Essential oils have been used as an alternative to chemical control methods. Therefore, the objective of this study was to analyze the essential oil chemical composition of the stem bark of Zanthoxylum riedelianum (ZREO) and evaluate its antifungal potential. The ZREO obtained by hydrodistillation was analyzed using gas chromatography coupled to mass spectrometry and evaluated for in vitro antifungal activity. The major components identified in the chemical analysis were E-nerolidol (67.21%), α-selinene (14.94%), and ß-selinene (7.41%). The antifungal potential of ZREO against S. sclerotiorum and R. stolonifer was detected at all concentrations evaluated in a dose-dependent manner. The best results were against R. stolonifer, with the concentration of 150 µLmL-1 inhibiting more than 80% of mycelial growth. On the basis of the current knowledge, this study describes for the first time the chemical composition and their antifungal activity.