Comparison analysis of bioactive metabolites in soybean, pea, mung bean, and common beans: reveal the potential variations of their antioxidant property.

This study showed the significantly differences of basic nutrients and metabolite compounds in nine types of beans involved in soybean, mung bean, pea, and common beans. The metabolomics results showed that serval metabolites such as histidine, proline, 3-alanine, and myricetin which could be used to identify different beans. The random forest model showed that amino acid and fatty acid could be used as special indexes to distinguish different types of beans in practice. The different expressed metabolites among different types of beans were involved in various pathways including alanine, aspartate and glutamate metabolism, arginine and proline metabolism, and purine metabolism. The antioxidant activity was significantly different among different types of beans, and the contents of amino acid, coumarin, and polyphenol contributed the antioxidant activities of beans. Together, these results will provide a comprehensive understanding of metabolites in different types of beans and theoretical guideline for the future application of beans.

Bioactive sulforaphane from cruciferous vegetables: advances in biosynthesis, metabolism, bioavailability, delivery, health benefits, and applications.

Chronic inflammation-induced diseases (CID) are the dominant cause of death worldwide, contributing to over half of all global deaths. Sulforaphane (SFN) derived from cruciferous vegetables has been extensively studied for its multiple functional benefits in alleviating CID. This work comprehensively reviewed the biosynthesis, metabolism, bioavailability, delivery, health benefits, and applications of SFN and its potential mechanisms against CID (e.g., cancer, obesity, type 2 diabetes, et al.), and neurological disorders based on a decade of research. SFN exerts its biological functions through the hydrolysis of glucosinolates by gut microbiota, and exhibits rapid metabolism and excretion characteristics via metabolization of mercapturic acid pathway. Microencapsulation is an important way to improve the stability and targeted delivery of SFN. The health benefits of SNF against CID are attributed to the multiple regulatory mechanisms including modulating oxidative stress, inflammation, apoptosis, immune response, and intestinal homeostasis. The clinical applications of SFN and related formulations show promising potential; however, further exploration is required regarding the sources, dosages, toxicity profiles, and stability of SFN. Together, SFN is a natural product with great potential for development and application, which is crucial for the development of functional food and pharmaceutical industries.

Characterization of two SGNH family cell death-inducing proteins from the horticulturally important fungal pathogen Botrytis cinerea based on the optimized prokaryotic expression system.

Botrytis cinerea is one of the most destructive phytopathogenic fungi, causing significant losses to horticultural crops. As a necrotrophic fungus, B. cinerea obtains nutrients by killing host cells. Secreted cell death-inducing proteins (CDIPs) play a crucial role in necrotrophic infection; however, only a limited number have been reported. For high-throughput CDIP screening, we optimized the prokaryotic expression system and compared its efficiency with other commonly used protein expression systems. The optimized prokaryotic expression system showed superior effectiveness and efficiency and was selected for subsequent CDIP screening. The screening system verified fifty-five candidate proteins and identified two novel SGNH family CDIPs: BcRAE and BcFAT. BcRAE and BcFAT exhibited high expression levels throughout the infection process. Site-directed mutagenesis targeting conserved Ser residues abolished the cell death-inducing activity of both BcRAE and BcFAT. Moreover, the transient expression of BcRAE and BcFAT in plants enhanced plant resistance against B. cinerea without inducing cell death, independent of their enzymatic activities. Our results suggest a high-efficiency screening system for high-throughput CDIP screening and provide new targets for further study of B. cinerea-plant interactions.

A comparative metabolomics analysis of phytochemcials and antioxidant activity between broccoli floret and by-products (leaves and stalks).

Leaves and stalks, which account for about 45% and 25% of broccoli biomass, respectively, are usually discarded during broccoli production, leading to the waste of green resources. In this study, the phytochemical composition and antioxidant capacity of broccoli florets and their by-products (leaves and stalks) were comprehensively analyzed. The metabolomics identified several unique metabolites (e.g., scopoletin, Harpagoside, and sinalbin) in the leaves and stalks compared to florets. Notably, the leaves were found to be a rich source of flavonoids and coumarins, with superior antioxidant capacity. The random forest model and correlation analysis indicated that flavonoids, coumarin, and indole compounds were the important factors contributing to the antioxidant activity. Moreover, the stalks contained higher levels of carbohydrates and exhibited better antioxidant enzyme activity. Together, these results provided valuable data to support the comprehensive utilization of broccoli waste, the development of new products, and the expansion of the broccoli industry chain.

Transcriptional landscape of pathogen-responsive lncRNAs in tomato unveils the role of hydrolase encoding genes in response to Botrytis cinerea invasion.

LncRNAs have gained increasing attention owing to their important regulatory roles on growth and stress responses of plants. However, the mechanisms underlying the functions of lncRNAs in fruit-pathogen interaction are still largely unknown. In this study, a total of 273 lncRNAs responding to Botrytis cinerea infection were identified in tomato fruit, among which a higher percentage of antisense lncRNAs were targeted to the genes enriched in hydrolase activity. To ascertain the roles of these lncRNAs, seven hydrolase-related transcripts were transiently knocked-down by virus-induced gene silencing. Silencing of lncRNACXE20 reduced the expression level of a carboxylesterase gene, further enhancing the resistance of tomato to B. cinerea. In contrast, silencing of lncRNACHI, lncRNAMMP, lncRNASBT1.9 and lncRNAPME1.9 impaired the resistance to B. cinerea, respectively. Further RT-qPCR assay and enzymatic activity detection displayed that the attenuated resistance of lncRNAMMP and lncRNASBT1.9-silenced plants was associated with the inhibition on the expression of JA-related genes, while the decreased resistance of lncRNACHI-silenced plants resulted in reduced chitinase activity. Collectively, these results may provide references for deciphering the mechanisms underlying specific lncRNAs to interfere with B. cinerea infection by regulating the expression of defence-related genes or affecting hydrolase activity.

Set1/COMPASS regulates growth, pathogenicity, and patulin biosynthesis of Penicillium expansum via H3K4 methylation and the interaction with PeVelB.

INTRODUCTION: Penicillium expansum is a harmful plant fungal pathogen that causes blue mold disease and produces mycotoxin patulin, leading to huge economic losses and food safety hazard. Set1 associated complex Set1/COMPASS deposits the methylation at lysine 4 of histone H3, which is associated with gene expression in diverse biological processes of fungi. However, the function and underlying mechanisms of Set1/COMPASS are poorly defined in P. expansum. OBJECTIVES: The study aimed to identify Set1/COMPASS and investigate its regulation mechanisms on growth, pathogenicity, and patulin biosynthesis of P. expansum. METHODS: Analyses of phylogenetic relationship, conserved structural domain, and gene deletion were used to identify components of Set1/COMPASS. Phenotype analysis and stress tolerance test of gene deletion mutants were conducted to analyze the function of these components. Yeast two-hybrid, Co-Immunoprecipitation (Co-IP), and point mutation were performed to verify the protein interaction. Western blot was conducted for detection of H3K4 methylation levels. RESULTS: P. expansum owns six components of Set1/COMPASS besides PeSet1. Absence of each component resulted in reduction of H3K4 methylation levels and impaired growth, pathogenicity, and patulin biosynthesis, as well as altered stress responses of P. expansum. One component PeBre2p was found to interact with the conserved global regulator PeVelB (VelvetLike protein B) at Asp294 of PeBre2p. This interaction affected fungal growth and utilization of fructose, lactose, glycine, and proline in P. expansum. CONCLUSION: This study revealed the important roles of Set1/COMPASS in P. expansum and clarified for the first time the combined regulation of PeBre2p and PeVelB in fungal growth and nutrition utilization. These results will provide potential targets for the control of blue mold disease.

PeAP1-mediated oxidative stress response plays an important role in the growth and pathogenicity of Penicillium expansum.

Penicillium expansum is the causal agent of post-harvest blue mold in various fruits and serves as a model for understanding fungal pathogenicity and mycotoxin production. The relevance of oxidative stress response in the growth and virulence of P. expansum has been largely unexplored. Here, we identify the transcriptional factor PeAP1 as a regulator of oxidative stress response in P. expansum. Gene expression and protein abundance of PeAP1, as well as its nuclear localization, are specifically induced by H2O2. Deletion of PeAP1 results in increased sensitivity to H2O2, and PeAP1 mutants exhibit a variety of defects in hyphal growth and virulence. PeAP1 prevents the accumulation of both intracellular H2O2 during vegetative growth and host-derived H2O2 during biotrophic growth. Application of an antioxidant glutathione and a NADPH oxidase inhibitor, diphenylene iodonium, to the PeAP1 mutant partially restored fungal growth and virulence. RNA sequencing analysis revealed 144 H2O2-induced PeAP1 target genes, including four antioxidant-related genes, PeGST1, PePrx1, PePrx2, and PeTRX2, that were also demonstrated to be involved in oxidative stress response and/or virulence. Collectively, our results demonstrate the global regulatory role of PeAP1 in response to oxidative stress and provide insights into the critical role of the PeAP1-mediated oxidative stress response to regulate growth and virulence of P. expansum. IMPORTANCE Reactive oxygen species are the core of host plant defense and also play a vital role in the successful invasion of host plants by pathogenic fungi. Despite its importance, the relevance of oxidative stress response in fungal growth and virulence is poorly understood in P. expansum. In this study, we reveal that the transcription factor PeAP1 acts as a central regulator of oxidative stress response in P. expansum and that there is a major link between PeAP1-mediated oxidative stress response and fungal growth and virulence. To explore the underlying mechanisms, we performed comparative transcriptomic studies and identified a number of H2O2-induced PeAP1 target genes, including four novel ones, PePrx1, PePrx2, PeGST1, and PeTRX2, whose functions were linked to PeAP1 and pathogenicity. These findings provide novel insights into the regulation mechanism of PeAP1 on growth and virulence, which might offer promising targets for control of blue mold and patulin contamination.

Ena Proteins Respond to PacC-Mediated pH Signaling Pathway and Play a Crucial Role in Patulin Biosynthesis.

Penicillium expansum is a main producer of patulin that causes severe postharvest decay and food safety issues in the fruit industry. Development, pathogenicity, and patulin production of P. expansum are strongly influenced by the PacC-pH signaling pathway. Global transcription factor PacC regulates various fungal biological processes through a complicated molecular network. In the present study, three Ena family genes (PeEnas), PeEnaA, PeEnaB, and PeEnaC, as important downstream targets of PePacC, were identified in P. expansum. Deletion of PeEnaA, PeEnaB, and PeEnaC showed little effect on mycelial growth under alkaline or high salinity conditions, but double and triple deletion of these genes impaired the virulence of P. expansum on apple fruit. Notably, patulin biosynthesis of P. expansum was distinctly inhibited in the deletion mutants of PeEnas. PeEnas regulated expressions of the patulin gene cluster, AP1, CreA, Sge1, and Hog1 at the transcriptional level and played roles in maintaining membrane potential. Overexpression of PeEnaC in ΔPePacC restored the patulin production defect of ΔPePacC. Our results indicated that, as downstream targets of PePacC, the PeEna family proteins play a crucial role in patulin biosynthesis in P. expansum.

Botrytis cinerea.

Chen, Zhang et al. introduce the necrotrophic fungal plant pathogen Botrytis cinerea more commonly known as gray mold.

Solanum lycopersicum, a Model Plant for the Studies in Developmental Biology, Stress Biology and Food Science.

Fruits, vegetables and other plant-derived foods contribute important ingredients for human diets, and are thus favored by consumers worldwide. Among these horticultural crops, tomato belongs to the Solanaceae family, ranks only secondary to potato (S. tuberosum L.) in yields and is widely cultivated for fresh fruit and processed foods owing to its abundant nutritional constituents (including vitamins, dietary fibers, antioxidants and pigments). Aside from its important economic and nutritional values, tomato is also well received as a model species for the studies on many fundamental biological events, including regulations on flowering, shoot apical meristem maintenance, fruit ripening, as well as responses to abiotic and biotic stresses (such as light, salinity, temperature and various pathogens). Moreover, tomato also provides abundant health-promoting secondary metabolites (flavonoids, phenolics, alkaloids, etc.), making it an excellent source and experimental system for investigating nutrient biosynthesis and availability in food science. Here, we summarize some latest results on these aspects, which may provide some references for further investigations on developmental biology, stress signaling and food science.

A data processing pipeline for petroleomics based on liquid chromatography-high resolution mass spectrometry.

Online liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) has attracted much attention in the molecular characterization of crude oil. Neither open access nor commercially available petroleomics tools were developed specifically to process LC-HRMS data. Here, a novel data processing pipeline was specifically designed for LC-HRMS-based petroleomics data. A customizable formula database was established deriving from the detected sample, which could avoid the interference caused by a large number of redundant molecules in a conventionally theoretical molecular database. Molecular formula candidates were assigned by the formula database using a low noise threshold, and false-positive assignments were eliminated by the chromatographic retention behaviors. Multi-dimensional information was obtained, including heteroatom class, double bond equivalent (DBE), carbon number, retention time, and MS/MS spectra. The developed method was compared with a popular petroleomics software, similar relative abundance class distribution was obtained, and much more formulas of low abundant components were uniquely extracted by the developed method. Finally, it was applied to reveal variation between feed and product oils in hydrodenitrogenation. Significantly compositional and structural differences were revealed. The developed method provides a useful pipeline for molecular data mining of petroleum samples.

Molecular mechanisms underlying multi-level defense responses of horticultural crops to fungal pathogens.

The horticultural industry helps to enrich and improve the human diet while contributing to growth of the agricultural economy. However, fungal diseases of horticultural crops frequently occur during pre- and postharvest periods, reducing yields and crop quality and causing huge economic losses and wasted food. Outcomes of fungal diseases depend on both horticultural plant defense responses and fungal pathogenicity. Plant defense responses are highly sophisticated and are generally divided into preformed and induced defense responses. Preformed defense responses include both physical barriers and phytochemicals, which are the first line of protection. Induced defense responses, which include innate immunity (pattern-triggered immunity and effector-triggered immunity), local defense responses, and systemic defense signaling, are triggered to counterstrike fungal pathogens. Therefore, to develop regulatory strategies for horticultural plant resistance, a comprehensive understanding of defense responses and their underlying mechanisms is critical. Recently, integrated multi-omics analyses, CRISPR-Cas9-based gene editing, high-throughput sequencing, and data mining have greatly contributed to identification and functional determination of novel phytochemicals, regulatory factors, and signaling molecules and their signaling pathways in plant resistance. In this review, research progress on defense responses of horticultural crops to fungal pathogens and novel regulatory strategies to regulate induction of plant resistance are summarized, and then the problems, challenges, and future research directions are examined.

Organic acid, a virulence factor for pathogenic fungi, causing postharvest decay in fruits.

Decay due to fungal infection is a major cause of postharvest losses in fruits. Acidic fungi may enhance their virulence by locally reducing the pH of the host. Several devastating postharvest fungi, such as Penicillium spp., Botrytis cinerea, and Sclerotinia sclerotiorum, can secrete gluconic acid, oxalic acid, or citric acid. Emerging evidence suggests that organic acids secreted by acidic fungi are important virulence factors. In this review, we summarized the research progress on the biosynthesis of organic acids, the role of the pH signalling transcription factor PacC in regulating organic acid, and the action mechanism of the main organic acid secreted via postharvest pathogenic fungi during infection of host tissues. This paper systematically demonstrates the relationships between tissue acidification and postharvest fungal pathogenicity, which will motivate the study of host-pathogen interactions and provide a better understanding of virulence mechanisms of the pathogens so as to design new technical strategies to prevent postharvest diseases.

DNA Methyltransferases Regulate Pathogenicity of Botrytis cinerea to Horticultural Crops.

Botrytis cinerea is one of the most destructive fungal pathogens that cause gray mold rot in horticultural products, including fresh fruits, vegetables, and flowers, leading to serious economic losses. B. cinerea is difficult to control because it has strong stress resistance and complex infection modes. The pathogenic mechanisms of B. cinerea have been revealed at multiple levels, but little is known at the epigenetic level. In this study, we first revealed the important role of DNA methyltransferases in regulating the development and pathogenicity of B. cinerea. We showed that two DNA methyltransferases, BcDIM2 and BcRID2, showed a strong synergistic effect in regulating the pathogenicity of B. cinerea. The double knockout mutant △Bcdim2rid2 showed slower mycelial growth, lower spore germination, attenuated oxidative tolerance, and complete pathogenicity loss on various hosts, which is related to the reduced expression of virulence-related genes in △Bcdim2rid2 and the induced resistance of the host. Although B. cinerea has multiple DNA methyltransferases, the global methylation level is very low, and few 5mC sites can be detected by BS-seq. These results first revealed the important role and the action mode of DNA methyltransferases in B. cinerea.

PeMetR-mediated sulfur assimilation is essential for virulence and patulin biosynthesis in Penicillium expansum.

Penicillium expansum, as the causal agent of blue mould and a main producer of mycotoxin patulin, is a global concern for economic and food safety. To date, the nutritional requirements of the pathogen during infection and patulin biosynthesis are poorly understood. Here, we genetically characterized the role of the bZIP transcription factor PeMetR in sulfur metabolism, virulence and patulin biosynthesis of P. expansum. The PeMetR regulator is crucial for normal germination and growth on inorganic S-sources but dispensable for utilization of organic S-sources. Accordingly, it is involved in regulating the expression of genes in sulfur assimilation pathway rather than methionine metabolic processes. Disruption of PeMetR resulted in a complete loss of virulence on various fruits. Additionally, the mutant showed a remarkably reduced ability to produce patulin. Exogenous methionine could partially or completely rescue the impaired phenotypes of the mutant. Inactivation of the sulfur assimilation pathway genes, PesA, PesB, PesC, PesF, generated growth, virulence and patulin production defects similar to those of ΔPeMetR. Overall, our study provides evidence that PeMetR-mediated sulfur assimilation is essential for growth and infection and shows for the first time that regulation of sulfur assimilation affects biosynthesis of an important mycotoxin patulin in P. expansum.

Molecular basis of pathogenesis of postharvest pathogenic Fungi and control strategy in fruits: progress and prospect.

The disease caused by pathogenic fungi is the main cause of postharvest loss of fresh fruits. The formulation of disease control strategies greatly depends on the understanding of pathogenic mechanism of fungal pathogens and control strategy. In recent years, based on the application of various combinatorial research methods, some pathogenic genes of important postharvest fungal pathogens in fruit have been revealed, and their functions and molecular regulatory networks of virulence have been explored. These progresses not only provide a new perspective for understanding the molecular basis and regulation mechanism of pathogenicity of postharvest pathogenic fungi, but also are beneficial to giving theoretical guidance for the creation of new technologies of postharvest disease control. Here, we synthesized these recent advances and illustrated conceptual frameworks, and identified several issues on the focus of future studies.

Molecular basis for optimizing sugar metabolism and transport during fruit development.

Sugars are fundamental metabolites synthesized in leaves and further delivered to fruit in fruit crops. They not only provide "sweetness" as fruit quality traits, but also function as signaling molecules to modulate the responses of fruit to environmental stimuli. Therefore, the understanding to the molecular basis for sugar metabolism and transport is crucial for improving fruit quality and dissecting responses to abiotic/biotic factors. Here, we provide a review for molecular components involved in sugar metabolism and transport, crosstalk with hormone signaling, and the roles of sugars in responses to abiotic and biotic stresses. Moreover, we also envisage the strategies for optimizing sugar metabolism during fruit quality maintenance.

Luteolin-induced activation of the phenylpropanoid metabolic pathway contributes to quality maintenance and disease resistance of sweet cherry.

Redox imbalance and fungal infection are major causes for quality deterioration and postharvest decay of fruit. Therefore, it is crucial to activate intrinsic antioxidative capacity and disease responses for fruit quality maintenance. Although plant-derived flavonoids have been reported for health-promoting benefits, their roles in the maintenance of fruit quality remains largely unexplored. Here, we exogenously applied luteolin, a flavonoid substance, and further examined its efficacy in maintaining fruit quality and inhibiting fungal diseases in sweet cherry. The results showed that 100 or 200 mg/L luteolin maintained better organoleptic quality and decreased disease incidence during storage. Biochemical assays revealed that luteolin activated the phenylpropanoid metabolic pathway and improved antioxidative capacity, thereby elevating total anthocyanin and flavonoid contents. Notably, luteolin inhibited mycelial growth of fungal pathogens and reduced patulin yield by Penicillium expansum. Collectively, these results suggest that luteolin is a promising alternative for maintaining better fruit quality and ameliorating disease resistance.

Reactive oxygen species: A generalist in regulating development and pathogenicity of phytopathogenic fungi.

Reactive oxygen species (ROS) are small molecules with high oxidative activity, and are usually produced as byproducts of metabolic processes in organisms. ROS play an important role during the interaction between plant hosts and pathogenic fungi. Phytopathogenic fungi have evolved sophisticated ROS producing and scavenging systems to achieve redox homeostasis. Emerging evidences suggest that ROS derived from fungi are involved in various important aspects of the development and pathogenesis, including formation of conidia, sclerotia, conidial anastomosis tubes (CATs) and infectious structures. In this mini-review, we summarize the research progress on the redox homeostasis systems, the versatile functions of ROS in the development and pathogenesis of phytopathogenic fungi, and the regulation effects of exogenous factors on intercellular ROS and virulence of the fungal pathogens.

Molecular basis and regulation of pathogenicity and patulin biosynthesis in Penicillium expansum.

Penicillium expansum is a necrotrophic plant pathogen with a wide range of fruit hosts. It causes blue mold rot during fruit storage, transport, and sale, resulting in huge economic losses to the fruit industry. Moreover, this pathogen is also the main producer of patulin, a toxic secondary metabolite that contaminates fruit and fruit-derived products and impairs human health. Therefore, understanding molecular basis of the pathogenicity and patulin biosynthesis of the fungal pathogen has important scientific significance and also plays an important guiding role in the research and development of new control technologies. Here, we comprehensively summarize the recent research progress, particularly regarding the molecular aspects of pathogenicity, patulin biosynthesis, and the related regulatory mechanisms, as well as control technologies for blue mold rot in the fruit industry.