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.

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.

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.

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.

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.

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.

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.

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.
CONCLUSIONS: 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.

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.

For sale in the domestic market, Hass avocados are kept in non-biodegradable plastic nets and stored at an ambient temperature. The fungus Colletotrichum gloeosporioides can cause important losses at this stage. Consequently, formulations based on polylactic acid (PLA)/poly (butylene adipate-co-terephthalate) (PBAT) 60/40 biodegradable blends added with pine essential oil (PEO) at 10, 12, 14, and 20% and coated with 1% chitosan (CH) were used for the elaboration of nets, which were evaluated on C. gloeosporioides and Rhizopus stolonifer on nutrient media and fruit and on the ripening behavior of Hass avocados at ambient temperature. The spore germination stage of C. gloeosporioides was the most extensively damaged (78% inhibition). The incidence of anthracnose was notably reduced by almost 80% in the avocados stored in the 60/40 PLA/PBAT nets coated with 1% CH. The overall values regarding weight loss, °Brix, and dry matter were c.a. 23.5%, 7.5, and 24.5%, respectively. The changes in firmness, color, and CO2 production were particularly associated with the initial harvest index and storage temperature rather than with the nets. In future research, essays on nets should include evaluations at the commercial levels.