Sesquiterpenoid Biosynthesis Research Articles

Exploring the Genetic Basis of Calonectria spp. Resistance in Eucalypts.

Selecting high-quality varieties with disease resistance by artificial crossbreeding is the most fundamental way to address the damage caused by Calonectria spp. in eucalypt plantations. However, understanding the mechanism of disease-resistant heterosis occurrence in eucalypts is crucial for successful crossbreeding. Two eucalypt hybrids, the susceptible EC333 (H1522 × unknown) and the resistant EC338 (W1767 × P9060), were screened through infection with Calonectria isolates, a pathogen that causes eucalypt leaf blight. RNA-Seq was performed on the susceptible hybrid, the disease-resistant hybrid, and their parents. The gene differential expression analysis showed that there were 3912 differentially expressed genes between EC333 and EC338, with 1631 up-regulated and 2281 down-regulated genes. The expression trends of the differential gene sets in P9060 and EC338 were similar. However, the expression trend of W1767 was opposite that of EC338. The similarity of the expression and the advantage of stress resistance in E. pellita suggested that genes with significant differences in expression likely relate to disease resistance. A GSEA based on GO annotations revealed that the carbohydrate binding pathway genes were differentially expressed between EC338 and EC333. The gene pathways that were differentially expressed between EC338 and EC333 revealed by the GSEA based on KEGG annotations were the sesquiterpenoid and triterpenoid biosynthesis pathways. The alternative splicing analysis demonstrated that an AS event between EC338 and EC333 occurred in LOC104426602. According to our SNP analysis, EC338 had 626 more high-impact mutation loci than the male parent P9060 and 396 more than the female parent W1767; W1767 had 259 more mutation loci in the downstream region than EC338, while P9060 had 3107 fewer mutation loci in the downstream region than EC338. Additionally, EC338 had 9631 more mutation loci in the exon region than EC333. Modules were found via WGCNA that were strongly and oppositely correlated with EC338 and EC333, such as module MEsaddlebrown, likely associated with leaf blight resistance. The present study provides a detailed explanation of the genetic basis of eucalypt leaf blight resistance, providing the foundation for exploring genes related to this phenomenon.

Efficient biosynthesis of β-caryophyllene in Saccharomyces cerevisiae by β-caryophyllene synthase from Artemisia argyi

Artemisia argyi H. Lév. & Vaniot is an important traditional Chinese medicinal plant known for its volatile oils, which are the main active components of A. argyi, including monoterpenes, sesquiterpenes and their derivatives. Despite its medicinal significance, the biosynthesis of sesquiterpenoids in A. argyi remains underexplored. In this study, we identified four β-caryophyllene synthases from A. argyi. A high-yield β-caryophyllene engineered Saccharomyces cerevisiae cell factory has been built in this study. By fusing ERG20 and AarTPS88 with a flexible linker (GGGS)2 and enhancing metabolic flux in the MVA pathway (HIF-1, tHMGR, and UPC2-1), we achieved a titer of β-caryophyllene reached 15.6 g/L by fed-batch fermentation in a 5 L bioreactor. To our knowledge, this represents the highest reported titer of β-caryophyllene in yeast to date. This study provides a valuable tool for the industrial-scale production of β-caryophyllene.

Enhancement of Sesquiterpenoid Production by Methyl Jasmonate in Atractylodes chinensis Adventitious Root Culture and its Transcriptional Regulation.

The adventitious root (AR) culture of Atractylodes chinensis is an efficient platform for sustainable production of its sesquiterpenoid compounds (atractylon and β-eudesmol). However, their limited accumulation levels need an effective elicitation approach, and the present study solved this problem using methyl jasmonate (MeJA) as an elicitor. The effects of its treatment concentration and duration on metabolite production were investigated. The ARs treated with 100 µM MeJA for seven days increased atractylon and β-eudesmol by 3.64- and 1.90-fold, respectively, compared with the control. This study further performed transcriptome analysis to explore the transcriptional regulation mechanism of the MeJA elicitation. A total of 124,464 unigenes were identified in A. chinensis ARs, of which 3,568 genes were upregulated and 3,864 genes were downregulated under the MeJA treatment. The MeJA treatment activated the endogenous JA biosynthesis and signaling pathways and sesquiterpenoid biosynthesis. The MeJA treatment more significantly activated the MEP pathway than the MVA pathway. In addition, 14 genes encoding terpene synthase were identified to be significantly upregulated. A total of 2,700 transcription factors (TFs) were identified in A. chinensis ARs, of which Tify, MYB, and MADS were significantly enriched under the MeJA treatment. We predicted a new antagonistic interaction between MYC2 and CPP TFs, which was significantly regulated by the MeJA treatment. The results of real-time quantitative PCR and enzyme activity assays proved the reliability of the transcriptome data. This study will help improve the in vitro production system of A. chinensis sesquiterpenoids and understand the transcriptional regulation mechanism of MeJA elicitation.

Exploration of diverse secondary metabolites from Penicillium brasilianum by co-culturing with Armillaria mellea

Bioinformatic analysis revealed that the genomes of ubiquitous Penicillium spp. might carry dozens of biosynthetic gene clusters (BGCs), yet many clusters have remained uncharacterized. In this study, a detailed investigation of co-culture fermentation including the basidiomycete Armillaria mellea CPCC 400891 and the P. brasilianum CGMCC 3.4402 enabled the isolation of five new compounds including two bisabolene-type sesquiterpenes (arpenibisabolanes A and B), two carotane-type sesquiterpenes (arpenicarotanes A and B), and one polyketide (arpenichorismite A) along with seven known compounds. The assignments of their structures were deduced by the extensive analyses of detailed spectroscopic data, electronic circular dichroism spectra, together with delimitation of the biogenesis. Most new compounds were not detected in monocultures under the same fermentation conditions. Arpenibisabolane A represents the first example of a 6/5-fused bicyclic bisabolene. The bioassay of these five new compounds exhibited no cytotoxic activities in vitro against three human cancer cell lines (A549, MCF-7, and HepG2). Moreover, sequence alignments and bioinformatic analysis to other metabolic pathways, two BGCs including Pb-bis and Pb-car, responsible for generating sesquiterpenoids from co-culture were identified, respectively. Furthermore, based on the chemical structures and deduced gene functions of the two clusters, a hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposed. These results demonstrated that the co-culture approach would facilitate bioprospecting for new metabolites even from the well-studied microbes. Our findings would provide opportunities for further understanding of the biosynthesis of intriguing sesquiterpenoids via metabolic engineering strategies.Key points• Penicillium and Armillaria co-culture facilitates the production of diverse secondary metabolites• Arpenibisabolane A represents the first example of 6/5-fused bicyclic bisabolenes• A hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposedGraphical

Transcriptomic and Metabolomic Analysis Reveals the Potential Roles of Polyphenols and Flavonoids in Response to Sunburn Stress in Chinese Olive (Canarium album).

Sunburn stress is one of the main environmental stress factors that seriously affects the fruit development and quality of Chinese olive, a tropical and subtropical fruit in south China. Therefore, the understanding of the changes in physiological, biochemical, metabolic, and gene expression in response to sunburn stress is of great significance for the industry and breeding of Chinese olive. In this study, the different stress degrees of Chinese olive fruits, including serious sunburn injury (SSI), mild sunburn injury (MSI), and ordinary (control check, CK) samples, were used to identify the physiological and biochemical changes and explore the differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) by using transcriptomics and metabolomics. Compared with CK, the phenotypes, antioxidant capacity, and antioxidant-related enzyme activities of sunburn stress samples changed significantly. Based on DEG-based KEGG metabolic pathway analysis of transcriptomics, the polyphenol and flavonoid-related pathways, including phenylpropanoid biosynthesis, sesquiterpenoid, and triterpenoid biosynthesis, monoterpene biosynthesis, carotenoid biosynthesis, isoflavonoid biosynthesis, flavonoid biosynthesis, were enriched under sunburn stress of Chinese olive. Meanwhile, 33 differentially accumulated polyphenols and 99 differentially accumulated flavonoids were identified using metabolomics. According to the integration of transcriptome and metabolome, 15 and 8 DEGs were predicted to regulate polyphenol and flavonoid biosynthesis in Chinese olive, including 4-coumarate-CoA ligase (4CL), cinnamoyl-CoA reductase (CCR), cinnamoyl-alcohol dehydrogenase (CAD), chalcone synthase (CHS), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS). Additionally, the content of total polyphenols and flavonoids was found to be significantly increased in MSI and SSI samples compared with CK. Our research suggested that the sunburn stress probably activates the transcription of the structural genes involved in polyphenol and flavonoid biosynthesis in Chinese olive fruits to affect the antioxidant capacity and increase the accumulation of polyphenols and flavonoids, thereby responding to this abiotic stress.

Temporal transcriptome and metabolome study revealed molecular mechanisms underlying rose responses to red spider mite infestation and predatory mite antagonism.

Red spider mite (Tetranychus urticae) infestation (SMI) is a detrimental factor for roses grown indoors. Although predatory mite (Neoseiulus californicus) antagonism (PMA) is often utilized to alleviate SMI damage, little is known about the defensive response of greenhouse-grown roses to SMI and the molecular mechanism by which PMA protects roses. To determine the transcriptome and metabolome responses of roses to SMI and PMA, the leaves of a rose cultivar ("Fairy Zixia/Nightingale") were infested with T. urticae, followed by the introduction of predator mite. Leaf samples were collected at various time points and subjected to transcriptome and metabolome analyses. We found that 24 h of SMI exerted the most changes in the expression of defense-related genes and metabolites in rose leaves. KEGG pathway analysis of differentially expressed genes (DEGs) and metabolites revealed that rose responses to SMI and PMA were primarily enriched in pathways such as sesquiterpenoid and triterpenoid biosynthesis, benzoxazinoid biosynthesis, stilbenoid, diarylheptanoid and gingerol biosynthesis, phytosterol biosynthesis, MAPK signaling pathway, phenylpropanoid biosynthesis, and other pathways associated with resistance to biotic stress. Rose reacted to SMI and PMA by increasing the expression of structural genes and metabolite levels in phytosterol biosynthesis, mevalonate (MVA) pathway, benzoxazinoid biosynthesis, and stilbenoid biosynthesis. In addition, PMA caused a progressive recover from SMI, allowing rose to revert to its normal growth state. PMA restored the expression of 190 essential genes damaged by SMI in rose leaves, including transcription factors DRE1C, BH035, MYB14, EF110, WRKY24, NAC71, and MY108. However, after 144 h of PMA treatment, rose responsiveness to stimulation was diminished, and after 192 h, the metabolic levels of organic acids and lipids were recovered in large measure. In conclusion, our results offered insights on how roses coordinate their transcriptome and metabolome to react to SMI and PMA, therefore shedding light on how roses, T. urticae, and N.californicus interact.

Ultrasonic treatment can improve maize seed germination and abiotic stress resistance

Constant-frequency ultrasonic treatment helped to improve seed germination. However, variable-frequency ultrasonic treatment on maize seed germination were rarely reported. In this study, maize seeds were exposed to 20–40 kHz ultrasonic for 40 s. The germination percentage and radicle length of maize seeds increased by 10.4% and 230.5%. Ultrasonic treatment also significantly increased the acid protease, α-amylase, and β-amylase contents by 96.4%, 73.8%, and 49.1%, respectively. Transcriptome analysis showed that 11,475 differentially expressed genes (DEGs) were found in the ultrasonic treatment and control groups, including 5,695 upregulated and 5,780 downregulated. Metabolic pathways and transcription factors (TFs) were significantly enriched among DEGs after ultrasonic treatment. This included metabolism and genetic information processing, that is, ribosome, proteasome, and pyruvate metabolism, sesquiterpenoid, triterpenoid, and phenylpropanoid biosynthesis, and oxidative phosphorylation, as well as transcription factors in the NAC, MYB, bHLH, WRKY, AP2, bZIP, and ARF families. Variable-frequency ultrasonic treatment increased auxin, gibberellin, and salicylic acid by 5.5%, 37.3%, and 28.9%, respectively. Abscisic acid significantly decreased by 33.2%. The related DEGs were upregulated and downregulated to varying degrees. Seed germination under the abiotic stress conditions of salt stress (NaCl solution), drought (PEG solution), and waterlogging (water-saturated sand bed) under ultrasonic treatment were promoted, radicle length was significantly increased by 30.2%, 30.5%, and 27.3%, respectively; and germination percentage by 14.8%, 20.1%, and 21.6%, respectively. These findings provide new insight into the mechanisms through ultrasonic to promote maize seed germination.

Coronatine-treated seedlings increase the tolerance of cotton to low-temperature stress

Coronatine, an analog of Jasmonic acid (JA), has been shown to enhance crop tolerance to abiotic stresses, including chilling stress. However, the underlying molecular mechanism remains largely unknown. In this study, we investigated the effect of Coronatine on cotton seedlings under low temperature using transcriptomic and metabolomics analysis. Twelve cDNA libraries from cotton seedlings were constructed, and pairwise comparisons revealed a total of 48,322 differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified the involvement of these unigenes in various metabolic pathways, including Starch and sucrose metabolism, Sesquiterpenoid and triterpenoid biosynthesis, Phenylpropanoid biosynthesis, alpha-Linolenic acid metabolism, ABC transporters, and Plant hormone signal transduction. Additionally, substantial accumulations of jasmonates (JAs), abscisic acid and major cell wall metabolites were observed. Transcriptome analysis revealed differential expression of regulatory genes, and qRT-PCR analysis confirmed the expression patterns of 9 selected genes. Co-expression analysis showed that the JA-responsive genes might form a network module with ABA biosynthesis genes or cell wall biosynthesis genes, suggesting the existence of a COR-JA-cellulose and COR-JA-ABA-cellulose regulatory pathway in cotton seedlings. Collectively, our findings uncover new insights into the molecular basis of coronatine--associated cold tolerance in cotton seedlings.

Mikania micrantha Kunth and its derived biochar impacts on heavy metal bioavailability and siderophore-related genes during chicken manure composting

Biochar can potentially reduce heavy metals (HMs) mobility and bioavailability during composting. However, siderophores secreted by functional microbes might lead to the re-mobilization of metals like Cu and Zn. Therefore, this study intended to explore the impacts of Mikania micrantha Kunth (MM) and MM-derived biochar (MMB) in the reduction of Cu and Zn bioavailability, and siderophore-related gene abundances during composting. Compared with MM and corn straw (CS) composts, a significant decline was noticed in the extractable and reducible Cu [(2.3 mg kg−1 + 12.1 mg kg−1), and (3.3 mg kg−1 + 14.6 mg kg−1)], and Zn [(103.1 mg kg−1 + 110.1 mg kg−1), and (109.6 mg kg−1 + 117.2 mg kg−1)] in MMB and corn straw biochar (CSB) composts, respectively. Besides, the lowest relative abundance of HMs-resistant bacteria particularly Corynebacterium (0.40%), Pseudomonas (0.46%), and Enterobacter (0.47%), was noted in MMB compost. Also, a significant increase in sesquiterpenoid and triterpenoid biosynthesis abundance (5.77%) accompanied by a reduction in the abundance of clusters related to siderophore transport, and siderophore transmembrane transporter activity was detected in MMB compost. Multivariate analysis labeled temperature, moisture content, total organic carbon, Corynebacterium, and Bacillus as the primary factors significantly correlated with the Cu and Zn bioavailability (− 0.90 ≤ r ≤ 0.90, P < 0.05). The structural equation model revealed that physicochemical parameters, microbial abundance, and siderophores exert a substantial influence on Cu and Zn bioavailability. Accordingly, MM and its derived biochar are recommended as an effective approach for accelerating Cu and Zn bioavailability reduction and managing the growth and distribution of invasive plants.Graphical

Comparative Transcriptomics of Soybean Genotypes with Partial Resistance Toward Phytophthora sojae, Conrad, and M92-220 to Moderately Susceptible Fast Neutron Mutant Soybeans and Sloan.

The breeding of disease-resistant soybeans cultivars to manage Phytophthora root and stem rot caused by the pathogen Phytophthora sojae involves combining quantitative disease resistance (QDR) and Rps gene-mediated resistance. To identify and confirm potential mechanisms of QDR toward P. sojae, we conducted a time course study comparing changes in gene expression among Conrad and M92-220 with high QDR to susceptible genotypes, Sloan, and three mutants derived from fast neutron irradiation of M92-220. Differentially expressed genes from Conrad and M92-220 indicated several shared defense-related pathways at the transcriptomic level but also defense pathways unique to each cultivar, such as stilbenoid, diarylheptanoid, and gingerol biosynthesis and monobactam biosynthesis. Gene Ontology pathway analysis showed that the susceptible fast neutron mutants lacked enrichment of three terpenoid-related pathways and two cell wall-related pathways at either one or both time points, in contrast to M92-220. The susceptible mutants also lacked enrichment of potentially important Kyoto Encyclopedia of Genes and Genomes pathways at either one or both time points, including sesquiterpenoid and triterpenoid biosynthesis; thiamine metabolism; arachidonic acid; stilbenoid, diarylheptanoid, and gingerol biosynthesis; and monobactam biosynthesis. Additionally, 31 genes that were differentially expressed in M92-220 following P. sojae infection were not expressed in the mutants. These 31 genes have annotations related to unknown proteins; valine, leucine, and isoleucine biosynthesis; and protein and lipid metabolic processes. The results of this study confirm previously proposed mechanisms of QDR, provide evidence for potential novel QDR pathways in M92-220, and further our understanding of the complex network associated with QDR mechanisms in soybean toward P. sojae.

Effects of Lactobacillus-fermented low-protein diets on the growth performance, nitrogen excretion, fecal microbiota and metabolomic profiles of finishing pigs

This study investigated the effects of Lactobacillus-fermented low-protein diet on the growth performance, nitrogen balance, fecal microbiota, and metabolomic profiles of finishing pigs. A total of 90 finishing pigs were assigned to one of three dietary treatments including a normal protein diet (CON) as well as two experimental diets in which a low-protein diet supplemented with 0 (LP) or 1% Lactobacillus-fermented low-protein feed (FLP). In comparison with CON, the LP and FLP significantly increased average daily gain (P = 0.044), significantly decreased feed to gain ratio (P = 0.021), fecal nitrogen (P < 0.01), urine nitrogen (P < 0.01), and total nitrogen (P < 0.01), respectively. The LP group exhibited increased abundances of unclassified_f_Selenomonadaceae, Coprococcus, Faecalibacterium, and Butyricicoccus, while the abundances of Verrucomicrobiae, Verrucomicrobiales, Akkermansiaceae, and Akkermansia were enriched in the FLP group. Low-protein diet-induced metabolic changes were enriched in sesquiterpenoid and triterpenoid biosynthesis and Lactobacillus-fermented low-protein feed-induced metabolic changes were enriched in phenylpropanoid biosynthesis and arginine biosynthesis. Overall, low-protein diet and Lactobacillus-fermented low-protein diet improved the growth performance and reduce nitrogen excretion, possibly via altering the fecal microbiota and metabolites in the finishing pigs. The present study provides novel ideas regarding the application of the low-protein diet and Lactobacillus-fermented low-protein diet in swine production.

Elucidating the ecophysiology of soybean pod-sucking stinkbug Riptortus pedestris (Hemiptera: Alydidae) based on de novo genome assembly and transcriptome analysis

Food security is important for the ever-growing global population. Soybean, Glycine max (L.) Merr., is cultivated worldwide providing a key source of food, protein and oil. Hence, it is imperative to maintain or to increase its yield under different conditions including challenges caused by abiotic and biotic stresses. In recent years, the soybean pod-sucking stinkbug Riptortus pedestris has emerged as an important agricultural insect pest in East, South and Southeast Asia. Here, we present a genomics resource for R. pedestris including its genome assembly, messenger RNA (mRNA) and microRNA (miRNA) transcriptomes at different developmental stages and from different organs. As insect hormone biosynthesis genes (genes involved in metamorphosis) and their regulators such as miRNAs are potential targets for pest control, we analyzed the sesquiterpenoid (juvenile) and ecdysteroid (molting) hormone biosynthesis pathway genes including their miRNAs and relevant neuropeptides. Temporal gene expression changes of these insect hormone biosynthesis pathways were observed at different developmental stages. Similarly, a diet-specific response in gene expression was also observed in both head and salivary glands. Furthermore, we observed that microRNAs (bantam, miR-14, miR-316, and miR-263) of R. pedestris fed with different types of soybeans were differentially expressed in the salivary glands indicating a diet-specific response. Interestingly, the opposite arms of miR-281 (-5p and -3p), a miRNA involved in regulating development, were predicted to target Hmgs genes of R. pedestris and soybean, respectively. These observations among others highlight stinkbug’s responses as a function of its interaction with soybean. In brief, the results of this study not only present salient findings that could be of potential use in pest management and mitigation but also provide an invaluable resource for R. pedestris as an insect model to facilitate studies on plant-pest interactions.

Divergent Biosynthesis of Bridged Polycyclic Sesquiterpenoids by a Minimal Fungal Biosynthetic Gene Cluster.

The bridged polycyclic sesquiterpenoids derived from sativene, isosativene, and longifolene have unique structures, and many chemical synthesis approaches with at least 10 steps have been reported. However, their biosynthetic pathway remains undescribed. A minimal biosynthetic gene cluster (BGC), named bip, encoding a sesquiterpene cyclase (BipA) and a cytochrome P450 (BipB) is characterized to produce such complex sesquiterpenoids with multiple carbon skeletons based on enzymatic assays, heterologous expression, and precursor experiments. BipA is demonstrated as a versatile cyclase with (-)-sativene as the dominant product and (-)-isosativene and (-)-longifolene as minor ones. BipB is capable of hydroxylating different enantiomeric sesquiterpenes, such as (-)-longifolene and (+)-longifolene, at C-15 and C-14 in turn. The C-15- or both C-15- and C-14-hydroxylated products are then further oxidized by unclustered oxidases, resulting in a structurally diverse array of sesquiterpenoids. Bioinformatic analysis reveals the BipB homologues as a discrete clade of fungal sesquiterpene P450s. These findings elucidate the concise and divergent biosynthesis of such intricate bridged polycyclic sesquiterpenoids, offer valuable biocatalysts for biotransformation, and highlight the distinct biosynthetic strategy employed by nature compared to chemical synthesis.

Transcriptomic and physiological analysis of atractylodes chinensis in response to drought stress reveals the putative genes related to sesquiterpenoid biosynthesis

BackgroundAtractylodes chinensis (DC) Koidz., a dicotyledonous and hypogeal germination species, is an important medicinal plant because its rhizome is enriched in sesquiterpenes. The development and production of A. chinensis are negatively affected by drought stress, especially at the seedling stage. Understanding the molecular mechanism of A. chinensis drought stress response plays an important role in ensuring medicinal plant production and quality. In this study, A. chinensis seedlings were subjected to drought stress treatment for 0 (control), 3 (D3), and 9 days (D9). For the control, the sample was watered every two days and collected on the second morning after watering. The integration of physiological and transcriptomic analyses was carried out to investigate the effects of drought stress on A. chinensis seedlings and to reveal the molecular mechanism of its drought stress response.ResultsThe malondialdehyde, proline, soluble sugar, and crude protein contents and antioxidative enzyme (superoxide dismutase, peroxidase, and catalase) activity were significantly increased under drought stress compared with the control. Transcriptomic analysis indicated a total of 215,665 unigenes with an average length of 759.09 bp and an N50 of 1140 bp. A total of 29,449 differentially expressed genes (DEGs) were detected between the control and D3, and 14,538 DEGs were detected between the control and D9. Under drought stress, terpenoid backbone biosynthesis had the highest number of unigenes in the metabolism of terpenoids and polyketides. To identify candidate genes involved in the sesquiterpenoid and triterpenoid biosynthetic pathways, we observed 22 unigene-encoding enzymes in the terpenoid backbone biosynthetic pathway and 15 unigene-encoding enzymes in the sesquiterpenoid and triterpenoid biosynthetic pathways under drought stress.ConclusionOur study provides transcriptome profiles and candidate genes involved in sesquiterpenoid and triterpenoid biosynthesis in A. chinensis in response to drought stress. Our results improve our understanding of how drought stress might affect sesquiterpenoid and triterpenoid biosynthetic pathways in A. chinensis.

Proteomic and metabolomic revealed the effect of shading treatment on cigar tobacco.

Shading or low light conditions are essential cultivation techniques for cigar wrapper tobacco leaves production, yet their impact on protein and metabolic regulatory networks is not well understood. In this study, we integrated proteomic and metabolomic analyses to uncover the potential molecular mechanisms affecting cigar tobacco leaves under shading treatment. Our findings include: (1) Identification of 780 significantly differentially expressed proteins (DEPs) in the cigar wrapper tobacco leaves, comprising 560 up-regulated and 220 down-regulated proteins, predominantly located in the chloroplast, cytoplasm, and nucleus, collectively accounting for 50.01%. (2) Discovery of 254 significantly differentially expressed metabolites (DEMs), including 148 up-regulated and 106 down-regulated metabolites. (3) KEGG pathway enrichment analysis revealed that the mevalonate (MVA) pathway within 'Terpenoid backbone biosynthesis' was inhibited, leading to a down-regulation of 'Sesquiterpenoid and triterpenoid biosynthesis'. Conversely, the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway was enhanced, resulting in an up-regulation of 'Monoterpenoid biosynthesis', 'Diterpenoid biosynthesis', and 'Carotenoid biosynthesis', thereby promoting the synthesis of terpenoids such as carotenoids and chlorophylls. Simultaneously, the Calvin cycle in 'Carbon fixation in photosynthetic organisms' was amplified, increasing photosynthetic efficiency. These results suggest that under low light conditions, cigar tobacco optimizes photosynthetic efficiency by reconfiguring its energy metabolism and terpenoid biosynthesis. This study contributes valuable insights into protein and metabolic analyses, paving the way for future functional studies on plant responses to low light.

Plant-microbial interactions inspired the discovery of novel sesquiterpenoid dimeric skeletons of hidden natural products from Hibiscus tiliaceus

Hidden natural products are representative of defensive strategies produced in vivo in diseased plants, a process that is induced by the plant immune system. The first transcriptome library of uninfected and pathogen infected Hibiscus tiliaceus stems was constructed by transcriptome sequencing technology, genes related to cadinene-type sesquiterpenoid biosynthesis were screened and combined with ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF-MS) analysis data, which indicated pathological tissue had potential to produce novel carbon skeletons of cadinane sesquiterpenoid dimers. Successfully, two cadinane-derived sesquiterpenoid dimers with unprecedented carbon skeletons, hibisceusanols A (1) and B (2) were isolated for the first time from the stems of H. tiliaceus induced by plant-microbial interactions. Their structures and absolute configurations were unambiguously established by spectroscopy, advanced chemistry development (ACD) and electronic circular dichroism (ECD) methods. Compounds 1 and 2 exhibited significant antitumor activity in vitro with half maximal inhibitory concentration (IC50) values of 2.3–7.2 µmol/L. The anticancer effect was generated via the induction of HepG2 cell apoptosis by inhibiting the phosphatidylinositol 3-kinase (PI3K) pathway.

Physiological and Transcriptional Analysis Provides Insights into Tea Saponin Biosynthesis and Regulation in Response to SA in Camellia vietnamensis Huang

Camellia vietnamensis Huang is an important and famous woody oil crop with high economic value in China because of its high-quality, edible, and medicinal oil. As one of its major active components, tea saponin (triterpenoid saponin) has shown anticancer, antioxidant, bacteriostatic, and other pharmacological activities. In this study, C. vietnamensis was used as an experimental material to determine the tea saponin content and physiological activity indicators after salicylic acid (SA) treatment and to analyze the differential expression genes of key metabolic pathways in response to SA by combining transcriptome data. The results showed that SA treatment increased the content of tea saponin and total phenols in leaves; effectively promoted the activity of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX); and decreased the content of malondialdehyde (MDA). A total of 60,038 genes, including 5871 new genes, were obtained by the RNA-seq. There were 6609 significantly differential expression genes mainly enriched in pathways such as sesquiterpenoid and triterpenoid biosynthesis, terpenoid backbone biosynthesis, diterpenoid biosynthesis, and flavonoid biosynthesis. The SA-induced key structural genes (SQS, SQE, bAS, CYP450, and UGT) and transcription factors related to the tea saponin biosynthetic pathway were screened by weighted gene co-expression network analysis (WGCNA). The results of this study could provide a theoretical basis and a new technical method to improve the content of tea saponin, with its excellent anticancer activity, in C. vietnamensis.

Multiomics Analysis Reveals the Genetic Basis of Volatile Terpenoid Formation in Oolong Tea.

Oolong tea has gained great popularity in China due to its pleasant floral and fruity aromas. Although numerous studies have investigated the aroma differences across various tea cultivars, the genetic mechanism is unclear. This study performed multiomics analysis of three varieties suitable for oolong tea and three others with different processing suitability. Our analysis revealed that oolong tea varieties contained higher levels of cadinane sesquiterpenoids. PanTFBS was developed to identify variants of transcription factor binding sites (TFBSs). We found that the CsDCS gene had two TFBS variants in the promoter sequence and a single nucleotide polymorphism (SNP) in the coding sequence. Integrating data on genetic variations, gene expression, and protein-binding sites indicated that CsDCS might be a pivotal gene involved in the biosynthesis of cadinane sesquiterpenoids. These findings advance our understanding of the genetic factors involved in the aroma formation of oolong tea and offer insights into the enhancement of tea aroma.

Transcriptome analysis of resistant and susceptible grapes reveals molecular mechanisms underlying resistance of white rot disease

Grape production in China is significantly impacted by white rot disease, which is caused by Coniella diplodiella (Speg.) Sacc. This study analyzes the differences in leaf transcriptomes and phenotypes of two grape species, ‘Manicure Finger (Vitis vinifera L.)’ and ‘0940 (Vitis davidii Foex)’, following inoculation with C. diplodiella. Leaf anatomy and H2O2 content confirm the greater resistance of '0940' to C. diplodiella compared to 'Manicure Finger.' Comparative transcriptome analysis reveals that the defense mechanism of '0940' against C. diplodiella involves sesquiterpenoid and triterpenoid biosynthesis, plant-pathogen interactions, sulfur relay systems, suberin and wax biosynthesis, monoterpenoid biosynthesis, as well as flavonoid and flavonol biosynthesis pathways. Using Weighted Gene Co-expression Network Analysis (WGCNA), we identified three modules highly correlated with C. diplodiella resistance and 125 candidate genes, including resistant genes (R genes), pattern-recognition receptors (PRRs), and pathogenesis-related proteins genes (PR genes), which may play important roles in grape resistance to this disease.

Transcriptomic Analysis of the Effect of Pruning on Growth, Quality, and Yield of Wuyi Rock Tea.

Pruning is an important agronomic measure in tea plantation management. In this study, we analyzed the effect of pruning on gene expression in tea leaves from a transcriptomics perspective and verified the results of a transcriptomic analysis in terms of changes in physiological indicators of tea leaves. The results showed that pruning enhanced the gene expression of nine metabolic pathways in tea leaves, including fatty acid synthesis and carbohydrate metabolism, nitrogen metabolism, protein processing in the endoplasmic reticulum, and plant hormone signal transduction, thereby promoting the growth of tea plants and increasing tea yield. However, pruning reduced the gene expression of nine metabolic pathways, including secondary metabolites biosynthesis, flavonoid biosynthesis, phenylpropanoid biosynthesis, and sesquiterpenoid and triterpenoid biosynthesis, and lowered the content of caffeine, flavonoids, and free amino acids in tea plant leaves. In conclusion, pruning could promote the growth of tea plants and increase the yield of tea, but it was not conducive to the accumulation of some quality indicators in tea leaves, especially caffeine, flavonoids, and free amino acids, which, in turn, reduced the quality of tea. This study provides an important theoretical reference for the management of agronomic measures in tea plantations.