Secondary Metabolite Biosynthesis Pathways Research Articles

Exogenous glutathione (GSH/GSSG) promoted the synthesis of patchoulol and pogostone, main active components of Pogostemon cablin (Patchouli)

Glutathione redox pair (GSH/GSSG) is the main characterizing substance that regulates the redox balance in cells and affects several physiological processes, such as plant metabolism, antioxidants, and detoxification. Pogostemon cablin was treated with different concentrations of reduced glutathione (GSH) and oxidized glutathione (GSSG), and it was found that 2 mM GSH and 4 mM GSSG treatments promoted the synthesis of patchoulol and pogostone. A total of 6620 significant differentially expressed gene were identified by transcriptome analysis. KEGG analysis showed that DEGs were mainly enriched in metabolic pathways, secondary metabolite biosynthesis, carbon metabolism, and phytohormone signaling pathways; 388 DEGs in the 2 mM GSH (11d) group, and 254 DEGs in the 4 mM GSSG (11d) group, were related to the biosynthesis of secondary metabolites. Further analysis by Gene Set Enrichment Analysis (GSEA) revealed that there were 139 and 83 DEGs associated with terpene synthesis between the two treatment groups, respectively, and the expression of key genes in the synthesis pathway (11 HMGR, 4 DXR, 2 GPPS, and 8 PTS) was up-regulated, which may be the result of transcription factors regulating their expression. The present study preliminarily showed that glutathione redox pair induced the expression of genes related to the terpene biosynthesis pathway, thereby promoting the biosynthesis of patchoulol and pogostone, which provides a reference basis for the technique of regulating and controlling the content of medicinal constituents of patchouli.

Elucidating the downstream pathways triggered by H2S signaling in Arabidopsis thaliana under drought stress via transcriptome analysis

ABSTRACT Hydrogen sulfide (H2S) is a crucial signaling molecule in plants. Recent studies have shown that H2S plays an equally important role as nitric oxide (NO) and hydrogen peroxide (H2O2) in plant signaling. Previous studies have demonstrated the involvement of H2S in regulating drought and other stressful environmental conditions, but the exact downstream molecular mechanisms activated by the H2S signaling molecule remain unclear. In this study, we conducted a comprehensive genome-wide transcriptomic analysis of both wild type (WT) and double mutant (lcd/des1). Arabidopsis thaliana plants were exposed to 40% polyethylene glycol (PEG) to induce drought stress and 20 µM sodium hydrosulfide (NaHS). The resulting transcriptome data were analyzed for differentially significant genes and their statistical enrichments in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The results indicated significant upregulation of genes related to photosynthesis, carbon fixation, plant secondary metabolite biosynthesis, inositol and phosphatidylinositol signaling pathways, and stress-responsive pathways in mutant plants under drought stress. Mutant plants with impaired H2S signaling mechanisms displayed greater susceptibility to drought stress compared to wild-type plants. In summary, all findings highlight the pivotal role of H2S signaling in stimulating other drought-responsive signaling pathways.

Differences in flavor characteristics of rounds of Sauce-based Baijiu based on GC-IMS, GC-MS, and electronic tongue

The volatile flavor and metabolic pathway differences of rounds of Sauce-based Baijiu (RSBs, R-1∼R-7) were analyzed using HS-GC-IMS and HS-SPME-GC-MS. Firstly, 18 (R-1, R-2, R-4, R-6, and R-7), 17 (R-3), and 19 (R-5) characteristic flavor compounds were identified based on GC-IMS by the relative odor activity value (ROAV) analysis. Secondly, a total of 751 metabolites were identified using GC-MS, and the differential metabolites were screened for pathway enrichment analysis by variable importance in projection (VIP) > 1 combined with fold change (FC) ≥ 2 or ≤ 0.5. The results demonstrated that unsaturated fatty acid metabolism, fatty acid metabolism, terpene metabolism, and secondary metabolite biosynthesis pathways were the most significant metabolic pathways influencing the flavor of basic baijiu, which primarily resulted in alterations in the composition and concentration of metabolites, including fatty acids and terpenes. Furthermore, RSBs demonstrated distinctive taste attributes, the addition of R-1 was found to markedly influence the astringency of baijiu (p < 0.05), while no notable impact was observed on the acidity or aftertaste. This study elucidates the flavor characteristics of RSBs, which provides a reference for the flavor quality regulation and blending of baijiu.

Metabolomics Reveal the Chemical Characteristic of Cigar Tobacco Leaves During Air-Curing Process

Air-curing is a critical process in the production of cigar tobacco leaves. This process involves the vital degradation and transformation of macromolecular substances into micromolecular chemical compounds. In this study, three different varieties of tobacco leaves harvested from the Wuzhishan and Danzhou areas of Hainan, China were used as materials. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was employed to investigate the chemical changes before and after air-curing in the metabolites. Metabolic analysis revealed significant stepwise alterations in the tobacco leaf metabolome during the air-curing process. A total of 172 differential metabolites were identified as significant contributors in the biosynthesis pathways of flavonoids, flavone, flavonol, and other secondary metabolites, among the total analyzed 1841 metabolites obtained from the tobacco leaves. Notably, the contents of flavonoid metabolites, including kaempferol, rutin, chlorogenic acid, and quercetin, exhibited a significant decrease throughout the air-curing period. Correlation analysis indicated a strong association between the biosynthesis of flavonoids and the contents of chlorogenic acid and kaempferol, suggesting their role in regulation the formation of yellow and brown colors in tobacco leaves. These findings provide valuable insights for future investigations into the quality, commercial value, and chemical properties of cigar tobacco.

Enhanced triacylglycerol metabolism contributes to the efficient biosynthesis of spinosad in Saccharopolyspora spinosa

Triacylglycerol (TAG) is crucial for antibiotic biosynthesis derived from Streptomyces, as it serves as an important carbon source. In this study, the supplementation of exogenous TAG led to a 3.92-fold augmentation in spinosad production. The impact of exogenous TAG on the metabolic network of Saccharopolyspora spinosa were deeply analyzed through comparative proteomics. To optimize TAG metabolism and enhance spinosad biosynthesis, the lipase-encoding genes lip886 and lip385 were overexpressed or co-expressed. The results shown that the yield of spinosad was increased by 0.8-fold and 0.4-fold when lip886 and lip385 genes were overexpressed, respectively. Synergistic co-expression of these genes resulted in a 2.29-fold increase in the yield of spinosad. Remarkably, the combined overexpression of lip886 and lip385 in the presence of exogenous TAG elevated spinosad yields by 5.5-fold, led to a drastic increase in spinosad production from 0.036 g/L to 0.234 g/L. This study underscores the modification of intracellular concentrations of free fatty acids (FFAs), short-chain acyl-CoAs, ATP, and NADPH as mechanisms by which exogenous TAG modulates spinosad biosynthesis. Overall, the findings validate the enhancement of TAG catabolism as a beneficial strategy for optimizing spinosad production and provide foundational insights for engineering secondary metabolite biosynthesis pathways in another Streptomyces.

Cold-stress induced metabolomic and transcriptomic changes in leaves of three mango varieties with different cold tolerance

BackgroundMango (Mangifera indica L.) is grown in Hainan, Guangdong, Yunnan, Sichuan, and Fujian provinces and Guanxi autonomous region of China. However, trees growing in these areas suffer severe cold stress during winter, which affects the yield. To this regard, data on global metabolome and transcriptome profiles of leaves are limited. Here, we used combined metabolome and transcriptome analyses of leaves of three mango cultivars with different cold stress tolerance, i.e. Jinhuang (J)—tolerant, Tainung (T) and Guiremang No. 82 (G)—susceptible, after 24 (LF), 48 (MF) and 72 (HF) hours of cold.ResultsA total of 1,323 metabolites belonging to 12 compound classes were detected. Of these, amino acids and derivatives, nucleotides and derivatives, and lipids accumulated in higher quantities after cold stress exposure in the three cultivars. Notably, Jinhuang leaves showed increasing accumulation trends of flavonoids, terpenoids, lignans and coumarins, and alkaloids with exposure time. Among the phytohormones, jasmonic acid and abscisic acid levels decreased, while N6-isopentenyladenine increased with cold stress time. Transcriptome analysis led to the identification of 22,526 differentially expressed genes. Many genes enriched in photosynthesis, antenna proteins, flavonoid, terpenoid (di- and sesquiterpenoids) and alkaloid biosynthesis pathways were upregulated in Jihuang leaves. Moreover, expression changes related to phytohormones, MAPK (including calcium and H2O2), and the ICE-CBF-COR signalling cascade indicate involvement of these pathways in cold stress responses.ConclusionCold stress tolerance in mango leaves is associated with regulation of primary and secondary metabolite biosynthesis pathways. Jasmonic acid, abscisic acid, and cytokinins are potential regulators of cold stress responses in mango leaves.

Peribacillus frigoritolerans T7-IITJ, a potential biofertilizer, induces plant growth-promoting genes of Arabidopsis thaliana.

This study aimed to isolate plant growth and drought tolerance-promoting bacteria from the nutrient-poor rhizosphere soil of Thar desert plants and unravel their molecular mechanisms of plant growth promotion. Among our rhizobacterial isolates, Enterobacter cloacae C1P-IITJ, Kalamiella piersonii J4-IITJ, and Peribacillus frigoritolerans T7-IITJ, significantly enhanced root and shoot growth (4-5-fold) in Arabidopsis thaliana under PEG-induced drought stress. Whole genome sequencing and biochemical analyses of the non-pathogenic bacterium T7-IITJ revealed its plant growth-promoting traits, viz., solubilization of phosphate (40-73µg/ml), iron (24±0.58mm halo on chrome azurol S media), and nitrate (1.58±0.01µg/ml nitrite), along with production of exopolysaccharides (125±20µg/ml) and auxin-like compounds (42.6±0.05µg/ml). Transcriptome analysis of A. thaliana inoculated with T7-IITJ and exposure to drought revealed the induction of 445 plant genes (log2fold-change>1, FDR<0.05) for photosynthesis, auxin and jasmonate signalling, nutrient uptake, redox homeostasis, and secondary metabolite biosynthesis pathways related to beneficial bacteria-plant interaction, but repression of 503 genes (log2fold-change < -1) including many stress-responsive genes. T7-IITJ enhanced proline 2.5-fold, chlorophyll 2.5-2.8-fold, iron 2-fold, phosphate 1.6-fold, and nitrogen 4-fold, and reduced reactive oxygen species 2-4.7-fold in plant tissues under drought. T7-IITJ also improved the germination and seedling growth of Tephrosia purpurea, Triticum aestivum, and Setaria italica under drought and inhibited the growth of two plant pathogenic fungi, Fusarium oxysporum, and Rhizoctonia solani. P. frigoritolerans T7-IITJ is a potent biofertilizer that regulates plant genes to promote growth and drought tolerance.

Metabolism and transcriptional regulation in chilling injury development of nectarine fruit during postharvest cold storage

Peach fruit is susceptible to chilling injury during postharvest long-term cold storage, but the understanding of metabolic and transcriptional reaction of nectarine cultivar fruit to cold storage remains incomplete. In this study, nectarine fruit was used for a low temperature treatment, which gradually leading to the deterioration of fruit quality. Comparative metabolome and transcriptome analyses were performed to comprehensively understand the metabolic and transcriptional changes occurring during cold storage in nectarine fruit. The result showed that numerous primary and secondary metabolites exhibited increased levels following the low-temperature treatment. And there were 3400 differentially expressed genes continuously regulated by cold storage, which were mostly enriched in metabolic and secondary metabolite biosynthesis pathways. The combined pathways enrichment of differential metabolites and genes were primarily centered on secondary metabolite biosynthesis. Correspondingly, a strong correlation was observed between changes in secondary metabolites and the expression of key enzymes including 4-coumarate CoA ligase (4CL), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT), flavonol synthase (FLS), dihydroflavonol reductase (DFR), and UDP-glycose flavonoid glycosyltransferase (UFGT), which were involved in flavonoids biosynthesis. Additionally, several fruit quality characteristics (carotenoids, cell wall components, antioxidant enzymes) and stress related signaling genes were markedly affected by cold storage. These findings offer new insights into the networks governing metabolic and transcriptional regulation during nectarine fruit cold storage.

Endophytic fungi improved wheat resistance to Rhopalosiphum padi by decreasing its feeding efficiency and population fitness

The improvement of crop resistance to insect using endophytic fungi is an environmentally friendly and sustainable strategy for agricultural pest control. Clarifying the efficacy and mechanism of endophytic fungi in improving crop resistance to pest offers the opportunity for biological control. In this study, changes in the transcriptome and defense compounds of wheat inoculated with endophytic fungal strains (i.e., YC and BB) were evaluated, and the efficacy of endophytic fungi in improving wheat resistance to Rhopalosiphum padi was studied. The results showed that the numbers of upregulated differentially-expressed genes (DEGs) in wheat plants inoculated with endophytic fungal strains YC and BB were higher than those of the downregulated DEGs, irrespective of R. padi infestation. Defense-related metabolic pathways, such as plant hormone signal transduction and secondary metabolite biosynthesis pathways were significantly enriched. Endophytic fungal strains YC and BB significantly increased jasmonic acid, DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one), total flavone, and tannin contents in wheat plants (P < 0.05) but decreased salicylic acid content. Variations in the contents of defense compounds were significantly correlated with decreased feeding, development, and reproduction of R. padi fed on wheat plants inoculated with strains YC and BB (|r| = 0.68–0.91, P < 0.05). The results suggested that endophytic fungi significantly decreased the feeding efficiency and population fitness [YC: (−11.13%) – (−22.07%); BB: (−10.98%) – (−22.20%)] of R. padi by altering the phytohormone pathway and secondary metabolite biosynthesis in wheat plants. This study helps in understanding of the efficacy of endophytic fungi in improving wheat resistance to insect and will be conducive to integrated pest management.

Exploring novel insights: Methyl jasmonate treatment reveals novel lncRNA-mediated regulation of secondary metabolite biosynthesis pathways in Echinacea purpurea

RNA-seq analysis has revolutionized the discovery of long non-coding RNAs (lncRNAs) in medicinal plants like Echinacea purpurea (L.) Moench, commonly known as purple coneflower. These lncRNAs potentially play crucial roles in regulating the biosynthesis of bioactive secondary metabolites. Our study aimed to identify regulatory lncRNAs involved in these pathways by treating 40-day-old E. purpurea seedlings with 100 μM methyl jasmonate (MeJA). Subsequently, RNA-sequencing was performed, identifying 509 differentially expressed transcripts for co-expression network analysis. The correlation between differentially expressed lncRNAs and coding transcripts was calculated, constructing a gene network to identify hub lncRNAs and their associated genes. Gene ontology and KEGG enrichment analyses were conducted to gain insights into the functions of these hub differentially expressed lncRNAs (DElncRNAs). Results revealed that these hub DElncRNAs regulate key genes involved in various pathways. For instance, CSE and BEBT are associated with the conversion of caffeoylshikimate to caffeate and shikimate, respectively. Additionally, SAT and NEC3 contribute to alkaloid biosynthesis, while NES1 and CFAT are involved in the biosynthesis of aromatic compounds, phenylpropanoids, and terpenoids. HST and SILD are associated with lignin biosynthesis. Furthermore, these lncRNAs mediate the acyl group transfer on both shikimate and quinate in the phenylpropanoid and flavonoid biosynthesis pathways. In summary, our findings offer comprehensive insights into the transcriptomic and metabolomic changes induced by methyl jasmonate treatment. We provide novel insights into the regulatory mechanisms of secondary metabolite biosynthesis, specifically highlighting the role of lncRNAs in E. purpurea.

High-intensity green light potentially activates the actinorhodin biosynthetic pathway in Streptomyces coelicolor A3(2).

The development of practices that enhance the potential of actinomycetes as major antibiotic producers is a challenge in discovering new secondary metabolites. Light, an essential external stimulus for most microorganisms, could be exploited to manipulate their physiological processes. However, the effects of monochromatic green light on the production of secondary metabolites in actinomycetes have not yet been reported. In this paper, we report a novel and simple method that uses high-intensity monochromatic green light to potentially induce the production of cryptic secondary metabolites in the model actinomycete Streptomyces coelicolor A3(2). Using actinorhodin (ACT), a blue-pigmented antibiotic, and undecylprodigiosin (RED), a red-pigmented antibiotic, as indicators, we found that irradiation with high-intensity monochromatic green light-emitting diodes promoted sporulation, significantly decreased RED production, and increased ACT production. Semi-quantitative reverse transcription-polymerase chain reaction and western blot analyses revealed, for the first time, that stimulation with green light accelerated the expression of ActII-ORF4, a pathway-specific regulator of ACT biosynthesis in S. coelicolor A3(2). This approach of stimulating secondary metabolite biosynthesis pathways in actinomycetes by irradiation with high-intensity monochromatic green light is expected to facilitate the discovery of cryptic antibiotics that are not typically produced under conventional dark culture conditions. However, the effective intensity and duration of irradiation with green light that are required to activate these metabolite pathways may vary markedly among actinomycetes.

Herbgenomics meets Papaveraceae: a promising -omics perspective on medicinal plant research.

Herbal medicines were widely used in ancient and modern societies as remedies for human ailments. Notably, the Papaveraceae family includes well-known species, such as Papaver somniferum and Chelidonium majus, which possess medicinal properties due to their latex content. Latex-bearing plants are a rich source of diverse bioactive compounds, with applications ranging from narcotics to analgesics and relaxants. With the advent of high-throughput technologies and advancements in sequencing tools, an opportunity exists to bridge the knowledge gap between the genetic information of herbs and the regulatory networks underlying their medicinal activities. This emerging discipline, known as herbgenomics, combines genomic information with other -omics studies to unravel the genetic foundations, including essential gene functions and secondary metabolite biosynthesis pathways. Furthermore, exploring the genomes of various medicinal plants enables the utilization of modern genetic manipulation techniques, such as Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR/Cas9) or RNA interference. This technological revolution has facilitated systematic studies of model herbs, targeted breeding of medicinal plants, the establishment of gene banks and the adoption of synthetic biology approaches. In this article, we provide a comprehensive overview of the recent advances in genomic, transcriptomic, proteomic and metabolomic research on species within the Papaveraceae family. Additionally, it briefly explores the potential applications and key opportunities offered by the -omics perspective in the pharmaceutical industry and the agrobiotechnology field.

Comparative transcriptome profiling of fruit tissue provides novel insights into piperine biosynthesis in black pepper (Piper nigrum L.)

Black pepper (Piper nigrum L.) is a storehouse of numerous bioactive compounds, most important being the alkaloid piperine. This metabolite has immense applications in both food and pharmaceutical industries. Therefore, studying the pathways involved in piperine biosynthesis is of paramount importance for improving the piperine content in black pepper. In the present study, we have estimated the piperine content across different fruit/berry developmental stages (DS1 to DS8) and leaf tissue of two different traditional landraces of black pepper, using the high-performance liquid chromatography (HPLC) method. Based on the results obtained, the landrace Thottumuriyan was identified to possess higher piperine content amongst the two landraces with high piperine content detected at the mid-developmental stages. The fruit tissues spanning the different developmental stages (DS2, DS4, DS6 and DS8) were selected for comparative transcriptome profiling. An average of 150 million read pairs were generated for the five different samples (4 berry development stages and the leaf tissue). These were mapped against the black pepper reference genome and differentially expressed genes (DEGs) were identified for the four different combinations. Further transcript profiling of twenty candidate genes potentially involved in secondary metabolite biosynthesis pathways, including piperine synthesis, was performed using Real time PCR in the eight different berry developmental stages. Highest expression for the genes associated with piperine biosynthesis was observed at mid-developmental stages. This included the genes coding for piperine synthase and piperic acid-CoA ligase. Our data provides critical information to understand the molecular mechanisms underlying piperine biosynthesis and other pathways in black pepper.

Comparative analysis of the molecular response characteristics in Platycodon grandiflorus irradiated with heavy ion beams and X-rays

The response of plants to radiation is an essential topic in both space plant cultivation and mutation breeding by radiation. In this study, heavy ion beams (HIB) generated by the ground accelerator and X-rays (XR) were used as models of high linear energy transfer (LET) and low LET radiation to study the molecular response mechanism of Platycodon grandiflorus (P. grandiflorus) seedlings after irradiation. The gene and protein expression profiles of P. grandiflorus after 15 Gy HIB and 20 Gy XR radiation were analyzed by transcriptome and proteome. The results showed that the number of differentially expressed genes (DEGs) induced by HIB radiation was less than that of XR group, but HIB radiation induced more differentially expressed proteins (DEPs). Both HIB and XR radiation activated genes of RNA silencing, double-strand break repair and cell catabolic process. DNA replication and cell cycle related genes were down-regulated. The genes of cell wall and external encapsulating structure were up-regulated after HIB radiation. The gene expression of protein folding and glucan biosynthesis increased after XR radiation. Protein enrichment analysis indicated that HIB radiation resulted in differential protein enriched in photosynthesis and secondary metabolite biosynthesis pathways, while XR radiation induced differential protein of glyoxylate and dicarboxylate metabolism and carbon metabolism. After HIB and XR radiation, the genes of antioxidant system and terpenoid and polyketide metabolic pathways presented different expression patterns. HIB radiation led to the enrichment of non-homologous end-joining pathway. The results will contribute to understanding the biological effects of plants under space radiation.

Amorphophallus muelleri activates ferulic acid and phenylpropane biosynthesis pathways to defend against Fusarium solani infection.

Amorphophallus sp. is an economically important crop for rural revitalization in southwest China. However, Fusarium solani often infects Amorphophallus sp. corms during storage, damaging the corm quality and affecting leaf elongation and flowering in the subsequent crop. In this study, the mechanism of resistance to F. solani was investigated in the leaf bud and flower bud corms of Amorphophallus muelleri through transcriptome and metabolome analyses. A total of 42.52 Gb clean reads and 1,525 metabolites were detected in a total of 12 samples including 3 samples each of disease-free leaf bud corms (LC), leaf bud corms inoculated with F. solani for three days (LD), disease-free flower bud corms (FC), and flower bud corms inoculated with F. solani for three days (FD). Transcriptome, metabolome, and conjoint analyses showed that 'MAPK signal transduction', 'plant-pathogen interaction', 'plant hormone signal transduction', and other secondary metabolite biosynthesis pathways, including 'phenylpropane biosynthesis', 'arachidonic acid metabolism', 'stilbene, diarylheptane and gingerolin biosynthesis', and 'isoquinoline alkaloids biosynthesis', among others, were involved in the defense response of A. muelleri to F. solani. Ultimately, the expression of six genes of interest (AmCDPK20, AmRBOH, AmWRKY33, Am4CL, Am POD and AmCYP73A1) was validated by real-time fluorescence quantitative polymerase chain reaction, and the results indicated that these genes were involved in the response of A. muelleri to F. solani. Ferulic acid inhibited the growth of F. solani, reducing the harm caused by F. solani to A. muelleri corms to a certain extent. Overall, this study lays a strong foundation for further investigation of the interaction between A. muelleri and F. solani, and provides a list of genes for the future breeding of F. solani-resistant A. muelleri cultivars.

Transcriptome profiling, physiological, and biochemical analyses provide new insights towards drought stress response in sugar maple (Acer saccharum Marshall) saplings.

Sugar maple (Acer saccharum Marshall) is a temperate tree species in the northeastern parts of the United States and is economically important for its hardwood and syrup production. Sugar maple trees are highly vulnerable to changing climatic conditions, especially drought, so understanding the physiological, biochemical, and molecular responses is critical. The sugar maple saplings were subjected to drought stress for 7, 14, and 21 days and physiological data collected at 7, 14, and 21 days after stress (DAS) showed significantly reduced chlorophyll and Normalized Difference Vegetation Index with increasing drought stress time. The drought stress-induced biochemical changes revealed a higher accumulation of malondialdehyde, proline, and peroxidase activity in response to drought stress. Transcriptome analysis identified a total of 14,099 differentially expressed genes (DEGs); 328 were common among all stress periods. Among the DEGs, transcription factors (including NAC, HSF, ZFPs, GRFs, and ERF), chloroplast-related and stress-responsive genes such as peroxidases, membrane transporters, kinases, and protein detoxifiers were predominant. GO enrichment and KEGG pathway analysis revealed significantly enriched processes related to protein phosphorylation, transmembrane transport, nucleic acids, and metabolic, secondary metabolite biosynthesis pathways, circadian rhythm-plant, and carotenoid biosynthesis in response to drought stress. Time-series transcriptomic analysis revealed changes in gene regulation patterns in eight different clusters, and pathway analysis by individual clusters revealed a hub of stress-responsive pathways. In addition, qRT-PCR validation of selected DEGs revealed that the expression patterns were consistent with transcriptome analysis. The results from this study provide insights into the dynamics of physiological, biochemical, and gene responses to progressive drought stress and reveal the important stress-adaptive mechanisms of sugar maple saplings in response to drought stress.

Pathogenesis and defense mechanism while Beauveria bassiana JEF-410 infects poultry red mite, Dermanyssus gallinae.

The poultry red mite, Dermanyssus gallinae (Mesostigmata: Dermanyssidae), is a major pest that causes great damage to chicken egg production. In one of our previous studies, the management of red mites using entomopathogenic fungi was evaluated, and the acaricidal fungus Beauveria bassiana JEF-410 was selected for further research. In this study, we tried to elucidate the pathogenesis of B. bassiana JEF-410 and the defense mechanisms of red mites at a transcriptome level. Red mites collected from a chicken farm were treated with B. bassiana JEF-410. When the mortality of infected red mites reached 50%, transcriptome analyses were performed to determine the interaction between B. bassiana JEF-410 and red mites. Uninfected red mites and non-infecting fungus served as controls. In B. bassiana JEF-410, up-regulated gene expression was observed in tryptophan metabolism and secondary metabolite biosynthesis pathways. Genes related to acetyl-CoA synthesis were up-regulated in tryptophan metabolism, suggesting that energy metabolism and stress management were strongly activated. Secondary metabolites associated with fungal up-regulated DEGs were related to the production of substances toxic to insects such as beauvericin and beauveriolide, efflux pump of metabolites, energy production, and resistance to stress. In red mites, physical and immune responses that strengthen the cuticle against fungal infection were highly up-regulated. From these gene expression analyses, we identified essential factors for fungal infection and subsequent defenses of red mites. These results will serve as a strong platform for explaining the interaction between B. bassiana JEF-410 and red mites in the stage of active infection.

UDP-Glycosyltransferases in Edible Fungi: Function, Structure, and Catalytic Mechanism

UDP-glycosyltransferases (UGTs) are the most studied glycosyltransferases, and belong to large GT1 family performing the key roles in antibiotic synthesis, the development of bacterial glycosyltransferase inhibitors, and in animal inflammation. They transfer the glycosyl groups from nucleotide UDP-sugars (UDP-glucose, UDP-galactose, UDP-xylose, and UDP-rhamnose) to the acceptors including saccharides, proteins, lipids, and secondary metabolites. The present review summarized the recent of UDP-glycosyltransferases, including their structures, functions, and catalytic mechanism, especially in edible fungi. The future perspectives and new challenges were also summarized to understand of their structure–function relationships in the future. The outputs in this field could provide a reference to recognize function, structure, and catalytic mechanism of UDP-glycosyltransferases for understanding the biosynthesis pathways of secondary metabolites, such as hydrocarbons, monoterpenes, sesquiterpene, and polysaccharides in edible fungi.

Metabolomics provide a novel interpretation of the changes in flavonoids during sea buckthorn (Hippophae rhamnoides L.) drying

Flavonoids are major nutrients in sea buckthorn berries. However, the effects of drying methods on flavonoids in sea buckthorn berries are unclear. In this study, ultra-performance liquid chromatography and metabolomics were adopted to analyse the effects of hot air drying (HAD) and infrared drying (IRD) on flavonoid compounds and antioxidant capacity in sea buckthorn berries. In total, 97 metabolites belonging to 12 classes were identified, including 26 flavones, 23 flavonols, and 11 flavanones. Additionally, 32 differential metabolites were identified among groups. Isorhamnetin and quercetin contents increased in response to HAD and IRD, while (-)-epigallocatechin and (-)-gallocatechin contents decreased. Differential metabolism of flavonoid compounds occurred mainly via the flavonoid biosynthesis and secondary metabolite biosynthesis pathways. Flavonoid compound degradation might be associated with antioxidant activity during drying. This study elucidated the effect of drying on nutritional components of sea buckthorn berries and may guide the improvement of quality during food processing.

Genome-Wide Identification of the Cytochrome P450 Superfamily Genes and Targeted Editing of BnCYP704B1 Confers Male Sterility in Rapeseed.

The cytochrome P450 (CYP450) monooxygenase superfamily, which is involved in the biosynthesis pathways of many primary and secondary metabolites, plays prominent roles in plant growth and development. However, systemic information about CYP450s in Brassica napus (BnCYP450) was previously undiscovered and their biological significance are far from understood. Members of clan 86 CYP450s, such as CYP704Bs, are essential for the formation of pollen exine in plant male reproduction, and the targeted mutagenesis of CYP704B genes has been used to create new male sterile lines in many crops. In the present study, a total of 687 BnCYP450 genes were identified in Brassica napus cultivar "Zhongshuang 11" (ZS11), which has nearly 2.8-fold as many CYP450 members as in Arabidopsis thaliana. It is rationally estimated since Brassica napus is a tetraploid oil plant with a larger genome compared with Arabidopsis thaliana. The BnCYP450 genes were divided into 47 subfamilies and clustered into nine clans. Phylogenetic relationship analysis reveals that CYP86 clan consists of four subfamilies and 109 BnCYP450s. Members of CYP86 clan genes display specific expression profiles in different tissues and in response to ABA and abiotic stresses. Two BnCYP450s within the CYP704 subfamily from CYP86 clan, BnCYP704B1a and BnCYP704B1b, display high similarity to MS26 (Male Sterility 26, also known as CYP704B1). These two BnCYP704B1 genes were specifically expressed in young buds. We then simultaneously knocked-out these two BnCYP704B1 genes through a clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) genome engineering system. The edited plants displayed a pollenless, sterile phenotype in mature anthers, suggesting that we successfully reproduced genic male sterility (GMS, also known as nuclear male sterility) lines in Brassica napus. This study provides a systemic view of BnCYP450s and offers a strategy to facilitate the commercial utility of the CRISPR/Cas9 system for the rapid generation of GMS in rapeseed via knocking-out GMS controlling genes.