Secondary Metabolite Pathways Research Articles

Knockout of OsHMA3 in an indica rice increases cadmium sensitivity and inhibits plant growth

AbstractCadmium (Cd) is a heavy metal that poses harm to both plants and humans. OsHMA3, a member of the heavy metal ATPase (HMA) family, plays a crucial role in sequestering Cd into the vacuoles of roots, thereby limiting its accumulation in rice grains. However, the response of rice plants to Cd under complete loss-of-function of OsHMA3 remains unclear. In this study, we successfully generated OsHMA3 null mutants in an indica variety 93 − 11 using CRISPR/Cas9 technology. A Cd resistance experiment revealed that the Oshma3 mutants exhibited increased sensitivity to Cd compared to the wild-type at a tested concentration of 10 µM CdCl2. Furthermore, the seedlings of Oshma3 mutant lines displayed inhibited plant growth in the presence of 1 µM Cd, specifically suppressing aboveground growth. As expected, knockout lines of OsHMA3 showed lower Cd accumulation in roots but higher concentrations in shoots compared to wild-type plants, highlighting the role of OsHMA3 in root-to-shoot Cd translocation. We further performed RNA sequencing analysis on wild-type and Oshma3 plants under control and Cd treatment conditions and found that differentially expressed genes were mainly enriched in metal ion binding, integral component of the membranes, and biosynthesis pathways for secondary metabolites triggered by exposure to Cd. When grown in a paddy field, the Oshma3 mutants exhibited shorter plant height, lower seed setting rate, and higher Cd accumulation in grains compared to wild-type plants. Our results indicate that knockout of OsHMA3 in the 93 − 11 variety increases sensitivity to Cd and inhibits plant growth.

Integrated metabolome and transcriptome analyses reveal the molecular mechanism underlying dynamic metabolic processes during taproot development of Panax notoginseng

BackgroundPanax notoginseng (Burk) F. H. Chen is one of the most famous Chinese traditional medicinal plants. The taproot is the main organ producing triterpenoid saponins, and its development is directly linked to the quality and yield of the harvested P. notoginseng. However, the mechanisms underlying the dynamic metabolic changes occurring during taproot development of P. notoginseng are unknown.ResultsWe carried out metabolomic and transcriptomic analyses to investigate metabolites and gene expression during the development of P. notoginseng taproots. The differentially accumulated metabolites included amino acids and derivatives, nucleotides and derivatives, and lipids in 1-year-old taproots, flavonoids and terpenoids in 2- and 3-year-old taproots, and phenolic acids in 3-year-old taproots. The differentially expressed genes (DEGs) are related to phenylpropanoid biosynthesis, metabolic pathway and biosynthesis of secondary metabolites at all three developmental stages. Integrative analysis revealed that the phenylpropanoid biosynthesis pathway was involved in not only the development of but also metabolic changes in P. notoginseng taproots. Moreover, significant accumulation of triterpenoid saponins in 2- and 3-year-old taproots was highly correlated with the up-regulated expression of cytochrome P450s and uridine diphosphate-dependent glycosyltransferases genes. Additionally, a gene encoding RNase-like major storage protein was identified to play a dual role in the development of P. notoginseng taproots and their triterpenoid saponins synthesis.ConclusionsThese results elucidate the molecular mechanism underlying the accumulation of and change relationship between primary and secondary metabolites in P. notoginseng taproots, and provide a basis for the quality control and genetic improvement of P. notoginseng.

Genome sequencing and analysis of penicillin V producing Penicillium rubens strain BIONCL P45 isolated from India.

A filamentous fungus Penicillium rubens is widely recognized for producing industrially important antibiotic, penicillin at industrial scale. To better comprehend, the genetic blueprint of the wild-type P. rubens was isolated from India to identify the genetic/biosynthetic pathways for phenoxymethylpenicillin (penicillin V, PenV) and other secondary metabolites. Genomic DNA (gDNA) was isolated, and library was prepared as per Illumina platform. Whole genome sequencing (WGS) was performed according to Illumina NovoSeq platform. Further, SOAPdenovo was used to assemble the short reads validated by Bowtie-2 and SAMtools packages. Glimmer and GeneMark were used to dig out total genes in genome. Functional annotation of predicted proteins was performed by NCBI non-redundant (NR), UniProt, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO) databases. Moreover, secretome analysis was performed by SignalP 4.1 and TargetP v1.1 and carbohydrate-active enzymes (CAZymes) and protease families by CAZy database. Comparative genome analysis was performed by Mauve 2.4.0. software to find genomic correlation between P. rubens BIONCL P45 and Penicillium chrysogenum Wisconsin 54-1255; also phylogeny was prepared with known penicillin producing strains by ParSNP tool. Penicillium rubens BIONCL P45 strain was isolated from India and is producing excess PenV. The 31.09Mb genome was assembled with 95.6% coverage of the reference genome P. chrysogenum Wis 54-1255 with 10687 protein coding genes, 3502 genes had homologs in NR, UniProt, KEGG, and GO databases. Additionally, 358 CAZymes and 911 transporter coding genes were found in genome. Genome contains complete pathways for penicillin, homogentisate pathway of phenyl acetic acid (PAA) catabolism, Andrastin A, Sorbicillin, Roquefortine C, and Meleagrin. Comparative genome analysis of BIONCL P45 and Wis 54-1255 revealed 99.89% coverage with 2952 common KEGG orthologous protein-coding genes. Phylogenetic analysis revealed that BIONCL P45 was clustered with Fleming's original isolate P. rubens IMI 15378. This genome can be a helpful resource for further research in developing fermentation processes and strain engineering approaches for high titer penicillin production.

Genome-wide identification of Uncaria rhynchophylla bHLH transcription factors and in-vitro validation of UrbHLH1 through interaction with terpenoid indole alkaloid synthesis pathway members

Uncaria rhynchophylla (Gouteng), as an evergreen woody vine belong to Rubiaceae family, is a traditional medicinal herb in China. Its terpenoid indole alkaloids (TIAs), which have good antidepressant and combined therapeutic effects on Alzheimer's disease, have attracted widespread attention. However, the content of TIAs is relatively low in U.rhynchophylla, which is unable to meet the growing market demand. The basic helix loop-helix (bHLH) transcription factor family exists in all three eukaryotic kingdoms and can participate in regulating secondary metabolite pathways. So far, there has been no comprehensive analysis of the bHLH gene in U. rhynchophylla, and their role in TIAs is almost unknown. In this study, a total of 171 UrbHLH genes (UrbHLHs) were unevenly distributed on 22 chromosomes and divided into 23 subfamilies. In addition, the physicochemical properties of UrbHLHs were analyzed. Most UrbHLHs in each subgroup had similar gene structures and conserved motifs. Intraspecific collinearity analysis showed that UrbHLH1 may be related to the biosynthesis of TIAs. Subcellular localization experiments revealed that UrbHLH1 is located in the nucleus; Dual luciferase reporter gene analysis (Dual-LUC) showed that UrbHLH1 could activate the expression of UrG10H and Ur10HGO in the TIAs synthesis pathway of U. rhynchophylla. Finally, using yeast one hybrid (Y1H) it was found that the promoter regions of these two genes both have E-box binding elements, which can be bound by UrbHLH1 and produced strong interactions. Therefore, UrbHLH1 may participate in the synthesis of TIAs pathway. In conclusion, this study provides foundation data on the role of UrbHLH transcription factors in regulating TIAs of U. rhynchophylla.

Molecular mechanism of salinity and waterlogging tolerance in mangrove Kandelia obovata.

Mangrove forests are colloquially referred to as "Earth's kidneys" and serve many important ecological and commercial functions. Salinity and waterlogging stress are the most important abiotic stressors restricting the growth and development of mangroves. Kandelia obovata (K. obovata) is the greatest latitudinally-distributed salt mangrove species in China.Here, morphology and transcriptomics were used to study the response of K. obovata to salt and waterlogging stress. In addition, weighted gene co-expression network analysis of the combined gene expression and phenotypic datasets was used to identify core salinity- and waterlogging-responsive modules. In this study, we observed that both high salinity and waterlogging significantly inhibited growth and development in K. obovata. Notably, growth was negatively correlated with salt concentration and positively correlated with waterlogging duration, and high salinity was significantly more inhibitive than waterlogging. A total of 7, 591 salt-responsive and 228 waterlogging-responsive differentially expressed genes were identified by RNA sequencing. Long-term salt stress was highly correlated with the measured physiological parameters while long-term waterlogging was poorly correlated with these traits. At the same time, 45 salinity-responsive and 16 waterlogging-responsive core genes were identified. All 61 core genes were mainly involved in metabolic and biosynthesis of secondary metabolites pathways. This study provides valuable insight into the molecular mechanisms of salinity and waterlogging tolerance in K. obovata, as well as a useful genetic resource for the improvement of mangrove stress tolerance using molecular breeding techniques.

Melatonin Promotes Accumulation of Resveratrol and Its Derivatives through Upregulation of PAL, 4CL, C4H, and STS in Grape Seeds

Melatonin’s role in regulating the biosynthesis of phenolic compounds, particularly stilbenes, remains unknown in grape seeds. In this study, widely targeted metabolomics analysis revealed that 82 phenolic compounds were differentially accumulated in grape seeds from the berries treated with melatonin. The accumulation of resveratrol and its 11 derivatives was largely increased in melatonin-treated berry seeds. Additionally, melatonin treatment of preveraison grape berries increased the resveratrol content in seeds during berry ripening. The results of RNA-Seq showed that a total of 399 genes were differentially expressed, which were mainly involved in the plant hormone signal transduction, metabolic, and biosynthesis of secondary metabolites pathways, in melatonin-treated berry seeds compared to control seeds. Additionally, five PALs, four 4CLs, one C4H, and four STSs were elucidated to be induced by melatonin in seeds at different time points after treatment. Therefore, melatonin promotes resveratrol accumulation and its derivatives, probably by upregulating the genes in the phenylpropanoid pathway, which provides precursors for resveratrol biosynthesis, and the STS genes in grape seeds.

Unveiling Si’s shield: A holistic examination of Cd stress alleviation in maize through physiological and transcriptomic insights

Cadmium (Cd), present in agricultural soil, poses a substantial threat to public health through the food chain, adversely affecting food crops and yield. This study examined the role of silicon (Si) in alleviating Cd stress in maize (Zea mays L.). Germinated uniform seedlings were transplanted into 20 L pots filled with a low-concentration nutrient solution. At the three fully expanded leaves stage (21 days after transplanting), plants were treated with Si (1000 μΜ as K2SO3) and Cd (10 μM as CdCl2). Plants were assessed 0, 1, and 28 days after treatments. The results demonstrate that Si application leads to a reduction in Cd concentration and content in maize shoot tissue. Additionally, Si treatment positively influences the activity of antioxidant enzymes (peroxidase, superoxidase dismutase, catalase) and plant hormones (abscisic acid, gibberellic acid, salicylic acid) in maize leaves. Furthermore, Si application enhances plant growth, increases levels of antioxidant substances, improves gas exchange parameters, boosts chlorophyll content, and enhances fluorescence. Weighted gene co-expression network analysis (WGCNA) and identification of transcription factors (TFs) and structural genes among differentially expressed genes (DEGs) reveal physiological correlations and enriched signaling pathways, particularly those related to metabolism and biosynthesis. Si actively modulates the formation of metabolites such as arginine and proline, starch and sucrose, as well as biosynthetic pathways for secondary metabolites like benzoxazinoid and carotenoid. Additionally, Si exerts regulatory influence over pathways such as the MAPK signaling pathway and plant hormone signal transduction in plants subjected to Cd stress. These findings unveil the molecular mechanisms underpinning the alleviation of Cd-induced stress in maize leaves due to Si application. Consequently, Si application not only mitigates Cd stress in maize but also reduces the risk of Cd entering the food chain.

Widely targeted metabolomics combined with E-tongue and E-nose reveal dynamic changes of tender coconut water in responses to the infection of Ceratocystis paradoxa

Ceratocystis paradoxa is a major cause of postharvest disease in tender coconuts worldwide. We conducted a comprehensive study using widely targeted metabolomics, electronic tongue (E-tongue), and electronic nose (E-nose) analyses to investigate the impacts of C. paradoxa invasion on the quality of tender coconut water (TCW) from fresh control (FC), uninoculated (UN), skin-inoculated (SI), and deep-inoculated (DI) nuts. DI exhibited significantly higher taste indicators associated with bitterness, saltiness, astringency aftertaste, and bitter aftertaste, as well as odor sensor values related to various compounds such as long-chain alkanes, hydrides, methane, organic sulfides, etc. Invasion of C. paradoxa into the endosperm altered the flavor characteristics of TCW mainly through the modulation of carbohydrate and secondary metabolite pathways. Furthermore, significant correlations were observed between the differentially expressed flavorful metabolites and the sensor indicators of the E-nose and E-tongue. These findings offer valuable insights into understanding the impact of C. paradoxa infection on coconuts.

Genome-Wide Comparative Profiles of Triterpenoid Biosynthesis Genes in Ginseng and Pseudo Ginseng Medicinal Plants.

Saponin-rich medicinal plants, particularly ginseng and Pseudo ginseng, are valuable in traditional medical practice due to the presence of different saponins. These plants benefit from natural saponins/triterpenoids drugs, such as Ginsenosides, Gypenosides, Platycodins, and Lancemasides. Ginsenosides are highly required for research and functional materials preparation in industrial practices, and some compounds, like Compound-K, have been taken to human trials for various therapeutic applications. To elucidate the genes/transcripts profiles responsible for secondary metabolites and ginsenoside biosynthesis in Ginseng and Pseudo ginseng plant genomes, a comparative analysis was conducted in this study. Nine plant genomes with a 99% BUSCO completeness score were used, resulting in 49 KEGG secondary metabolite pathways, 571 cytochromes genes with 42 families, and 3529 carbohydrate genes with 103 superfamilies. The comparative analysis revealed 24 genes/transcripts belonging to the CYP716 family, which is involved in the ginsenoside biosynthesis pathway. Additionally, it found that various ginsenosides demonstrated strong binding affinity with twelve targets, with ginsenoside Rg3, Rg2, Rh1, Rh5, F3, Rh9, Panaxadione, Protopanaxatriol, Floral ginsenoside C, and Floral ginsenoside E exhibiting the highest binding affinities with the tested enzymes. Since these groups of enzymes are not yet fully characterized for Pseudo ginseng plants in the interconversion of triterpenoids, this comparative bioinformatics analysis could aid experimentalists in selecting and conducting characterization with practical knowledge.

Soybean-SCN Battle: Novel Insight into Soybean's Defense Strategies against Heterodera glycines.

Soybean cyst nematode (SCN, Heterodera glycines, Ichinohe) poses a significant threat to global soybean production, necessitating a comprehensive understanding of soybean plants' response to SCN to ensure effective management practices. In this study, we conducted dual RNA-seq analysis on SCN-resistant Plant Introduction (PI) 437654, 548402, and 88788 as well as a susceptible line (Lee 74) under exposure to SCN HG type 1.2.5.7. We aimed to elucidate resistant mechanisms in soybean and identify SCN virulence genes contributing to resistance breakdown. Transcriptomic and pathway analyses identified the phenylpropanoid, MAPK signaling, plant hormone signal transduction, and secondary metabolite pathways as key players in resistance mechanisms. Notably, PI 437654 exhibited complete resistance and displayed distinctive gene expression related to cell wall strengthening, oxidative enzymes, ROS scavengers, and Ca2+ sensors governing salicylic acid biosynthesis. Additionally, host studies with varying immunity levels and a susceptible line shed light on SCN pathogenesis and its modulation of virulence genes to evade host immunity. These novel findings provide insights into the molecular mechanisms underlying soybean-SCN interactions and offer potential targets for nematode disease management.

Proteome reveals the mechanism of selenium-sulfur interaction in regulating isothiocyanate biosynthesis and the physiological metabolism of broccoli sprouts.

Broccoli sprouts have a strong ability to accumulate isothiocyanate and selenium. In this study, the isothiocyanate content increased significantly as a result of ZnSO4 stress. Particularly, based on the isothiocyanate content is not affected, the combined ZnSO4 and Na2SeO3 treatment alleviated the inhibition of ZnSO4 and induced selenium content. Gene transcription and protein expression analyses revealed the changes in isothiocyanate and selenium metabolite levels in broccoli sprouts. ZnSO4 combined with Na2SeO3 was proven to activate a series of isothiocyanate metabolite genes (UGT74B1, OX1, and ST5b) and selenium metabolite genes (BoSultr1;1, BoCOQ5-2, and BoHMT1). The relative abundance of the total 317 and 203 proteins, respectively, in 4-day-old broccoli sprouts varied, and the metabolic and biosynthetic pathways for secondary metabolites were significantly enriched in ZnSO4/control and ZnSO4 combined Na2SeO3/ZnSO4 comparisons. The findings demonstrated how ZnSO4 combined with Na2SeO3 treatment reduced stress inhibition and the accumulation of encouraged selenium and isothiocyanates during the growth of broccoli sprouts.

Transcriptome Analysis of Diploid and Autotetraploid Hemerocallis Response to Drought Stress

Chromosome doubling in ornamental plants, as shown by our study in daylilies (Hemerocallis spp.), has great potential to increase tolerance to abiotic stress. Drought is the most critical growth-limiting factor in a changing climate. Drought tolerance is one of the decisive factors for the survival, productivity, and appearance of perennial ornamental plants. Understanding and elucidating the molecular mechanisms that determine plant response to abiotic stress is essential. De novo transcriptome assembly of diploid and autotetraploid Hemerocallis spp. cv. Trahlyta was performed under artificially induced stress to elucidate the molecular mechanisms related to plant response to drought. In daylily mRNA, 237,886 transcripts were detected, and 42.4% of them were identified as annotated unigenes. In the experiment, diploid plants were more stressed, with 2871 upregulated or downregulated DEGs (differentially expressed genes) responding to drought, while tetraploid plants had 1599 DEGs. The proportion of upregulated DEGs differed by 1.3 times between diploid and autotetraploid genotypes, whereas the proportion of downregulated DEGs was 1.8 times greater in diploid plants. Signaling pathways related to the drought response were activated in daylilies, and key candidate genes were identified in both ploidy genotypes. In autotetraploid plants, more drought-related pathways were activated than in diploids—43 and 19, respectively. The most abundant DEGs in both cases were KEGG (Kyoto Encyclopedia of Genes and Genomes), metabolic (ko01100), and biosynthesis of secondary metabolites (ko01110) pathways. Summarizing the data, it was found that autotetraploid plants of the daylily have a wider potential for adaptation to drought stress. Therefore, they adapt faster and better to adverse drought conditions by activating alternative signaling pathways. The comparative transcriptome analysis of diploid and autotetraploid plants allows us to understand the molecular mechanisms of drought resistance and it is also essential for daylily breeding programs to develop drought-resistant genotypes in the future.

Effects of topping and non-topping on growth-regulating hormones of flue-cured tobacco (Nicotiana tabacum L.)-a proteomic analysis.

Until now, the mechanism underlying the impact of topping on hormone regulation in tobacco plants remains unclear, and most studies investigating the hormone signaling pathways in plants rely on genes or transcriptional pathways. This study examines the regulatory mechanisms of hormones in the roots and leaves of tobacco plants with and without topping at the protein level. The results demonstrate that, compared with non-topped plants, topping leads to a decrease in the levels of IAA (auxin), ABA (abscisic acid), and GA (gibberellin) hormones in the leaves, whereas the content of the JA (jasmonic acid) hormone increases. Furthermore, in the roots, topping results in an increase in the levels of IAA, ABA, and JA hormones, along with a decrease in GA content. In the leaves, a total of 258 significantly different proteins were identified before and after topping, with 128 proteins upregulated and 130 proteins downregulated. In the roots, there were 439 proteins with significantly different quantities before and after topping, consisting of 211 upregulated proteins and 228 downregulated proteins. Notably, these proteins were closely associated with the metabolic and biosynthetic pathways of secondary metabolites, as indicated by functional categorization. When integrating the hormone changes and the proteomics results, it is evident that topping leads to increased metabolic activity and enhanced hormone synthesis in the root system. This research provides a theoretical foundation for further investigations into the regulation and signaling mechanisms of hormones at the protein level before and after topping in plants.

Two chromosome-level genome assemblies of Rhodiola shed new light on genome evolution in rapid radiation and evolution of the biosynthetic pathway of salidroside.

Rhodiola L. is a genus that has undergone rapid radiation in the mid-Miocene and may represent a typic case of adaptive radiation. Many species of Rhodiola have also been widely used as an important adaptogen in traditional medicines for centuries. However, a lack of high-quality chromosome-level genomes hinders in-depth study of its evolution and biosynthetic pathway of secondary metabolites. Here, we assembled two chromosome-level genomes for two Rhodiola species with different chromosome number and sexual system. The assembled genome size of R. chrysanthemifolia (2n = 14; hermaphrodite) and R. kirilowii (2n = 22; dioecious) were of 402.67 and 653.62 Mb, respectively, with approximately 57.60% and 69.22% of transposable elements (TEs). The size difference between the two genomes was mostly due to proliferation of long terminal repeat-retrotransposons (LTR-RTs) in the R. kirilowii genome. Comparative genomic analysis revealed possible gene families responsible for high-altitude adaptation of Rhodiola, including a homolog of plant cysteine oxidase 2 gene of Arabidopsis thaliana (AtPCO2), which is part of the core molecular reaction to hypoxia and contributes to the stability of Group VII ethylene response factors (ERF-VII). We found extensive chromosome fusion/fission events and structural variations between the two genomes, which might have facilitated the initial rapid radiation of Rhodiola. We also identified candidate genes in the biosynthetic pathway of salidroside. Overall, our results provide important insights into genome evolution in plant rapid radiations, and possible roles of chromosome fusion/fission and structure variation played in rapid speciation.

A Pseudomonas Plant Growth Promoting Rhizobacterium and Arbuscular Mycorrhiza differentially modulate the growth, photosynthetic performance, nutrients allocation, and stress response mechanisms triggered by a mild Zinc and Cadmium stress in tomato

This study aimed to assess the effectiveness of plant growth-promoting rhizobacteria (PGPR; Pseudomonas strain So_08) and arbuscular mycorrhizal fungi (AMF; Rhizoglomus irregulare BEG72 and Funneliformis mosseae BEG234) in mitigating the detrimental effects of cadmium (Cd) and zinc (Zn) stress in tomato plants. Plant biomass, root morphology, leaf relative water content, membrane stability, photosynthetic performance, chlorophyll content, and heavy metals (HMs) accumulation were determined. Furthermore, an ionomic profile was conducted to investigate whether microbial inoculants affected the uptake and allocation of macro- and micronutrients. Metabolomics with pathway analysis of both roots and leaves was performed to unravel the mechanisms underlying the differential responses to HMs stress. The findings revealed that the levels of HMs did not significantly affect plant growth parameters; however, they affected membrane stability, photosynthetic performance, nutrient allocation, and chlorophyll content. Cadmium was mainly accumulated in roots, whilst Zn exhibited accumulation in various plant organs. Our findings demonstrate the beneficial effects of PGPR and AMF in mitigating Cd and Zn stress in tomato plants. The microbial inoculations improved physiological parameters and induced differential accumulation of macro- and micronutrients, modulating nutrient uptake balance. These results provide insights into the mechanisms underlying the plant-microbe interactions and highlight the differential modulation of the biosynthetic pathways of secondary metabolites related to oxidative stress response, membrane lipids stability, and phytohormone crosstalk.

Secondary Metabolites and Their Role in Strawberry Defense.

Strawberry is a high-value commercial crop and a model for the economically important Rosaceae family. Strawberry is vulnerable to attack by many pathogens that can affect different parts of the plant, including the shoot, root, flowers, and berries. To restrict pathogen growth, strawberry produce a repertoire of secondary metabolites that have an important role in defense against diseases. Terpenes, allergen-like pathogenesis-related proteins, and flavonoids are three of the most important metabolites involved in strawberry defense. Genes involved in the biosynthesis of secondary metabolites are induced upon pathogen attack in strawberry, suggesting their transcriptional activation leads to a higher accumulation of the final compounds. The production of secondary metabolites is also influenced by the beneficial microbes associated with the plant and its environmental factors. Given the importance of the secondary metabolite pathways in strawberry defense, we provide a comprehensive overview of their literature and their role in the defense responses of strawberry. We focus on terpenoids, allergens, and flavonoids, and discuss their involvement in the strawberry microbiome in the context of defense responses. We discuss how the biosynthetic genes of these metabolites could be potential targets for gene editing through CRISPR-Cas9 techniques for strawberry crop improvement.

Multilayer omics landscape analyses reveal the regulatory responses of tea plants to drought stress

Adverse environments, especially drought conditions, deeply influence plant development and growth in all aspects, and the yield and quality of tea plants are largely dependent on favorable growth conditions. Although tea plant responses to drought stress (DS) have been studied, a comprehensive multilayer epigenetic, transcriptomic, and proteomic investigation of how tea responds to DS is lacking. In this study, we generated DNA methylome, transcriptome, proteome, and phosphoproteome data to explore multiple regulatory landscapes in the tea plant response to DS. An integrated multiomics analysis revealed the response of tea plants to DS at multiple regulatory levels. Furthermore, a set of DS-responsive genes involved in photosynthesis, transmembrane transportation, phytohormone metabolism and signaling, secondary metabolite pathways, transcription factors, protein kinases, posttranslational and epigenetic modification, and other key stress-responsive genes were identified for further functional investigation. These results reveal the multilayer regulatory landscape of the tea plant response to DS and provide insight into the mechanisms of these DS responses.

Extreme environmental adaptation mechanisms of Antarctic bryophytes are mainly the activation of antioxidants, secondary metabolites and photosynthetic pathways

The environment in Antarctica is characterized by low temperature, intense UVB and few vegetation types. The Pohlia nutans M211 are bryophytes, which are the primary plants in Antarctica and can thrive well in the Antarctic harsh environment. The transcriptional profiling of Pohlia nutans M211 under low temperature and high UVB conditions was analyzed to explore their polar adaptation mechanism in the extreme Antarctic environment by third-generation sequencing and second-generation sequencing. In comparison to earlier second-generation sequencing techniques, a total of 43,101 non-redundant transcripts and 10,532 lncRNA transcripts were obtained, which were longer and more accurate. The analysis results of GO, KEGG, AS (alternative splicing), and WGCNA (weighted gene co-expression network analysis) of DEGs (differentially expressed genes), combined with the biochemical kits revealed that antioxidant, secondary metabolites pathways and photosynthesis were the key adaptive pathways for Pohlia nutans M211 to the Antarctic extreme environment. Furthermore, the low temperature and strong UVB are closely linked for the first time by the gene HY5 (hlongated hypocotyl 5) to form a protein interaction network through the PPI (protein–protein interaction networks) analysis method. The UVR8 module, photosynthetic module, secondary metabolites synthesis module, and temperature response module were the key components of the PPI network. In conclusion, this study will help to further explore the polar adaptation mechanism of Antarctic plants represented by bryophytes and to enrich the polar gene resources.

Establishment of Agrobacterium rhizogenes-mediated RNAi of Phryma leptostachya accelerates the functional identification of key genes of the furofuran lignan biosynthetic pathway

Phryma leptostachya is a well-known medicinal plant that produces furofuran lignans with various biological activities. However, the biosynthetic pathways of these natural products in P. leptostachya remain unclear. Hairy root cultures provide a novel method for elucidating the biosynthetic pathways of secondary metabolites. Here, we established a hairy root system for P. leptostachya by optimizing five parameters: Agrobacterium strains, explant types, infection time, A. rhizogenes cell density, and acetosyringone (AS) concentration. The highest transformation frequency (74.51 %) was achieved when the stem explants were infected with ATCC15834 (OD600 = 0.8) strain of A. rhizogenes for 30 min on 1/2 MS medium supplemented with 200 µM AS. Based on the optimal protocol, RNAi hairy roots were induced by silencing the cytochrome P450 gene, PlCYP81Q38, which converts (+)-pinoresinol to (+)-sesamin by means of (+)-piperitol in P. leptostachya, to determine its in vivo role in the lignan biosynthetic pathway and verify the availability of this system. PlCYP81Q38 silencing resulted in significant down-regulation of the PlCYP81Q38 gene at the transcriptional level, consistent with a remarkable decrease in the content of leptostachyol acetate and 6-desmethoxy-leptostachyol acetate, demonstrating its significance in the production of lignans. In conclusion, the approaches and results presented here provide a solid foundation for elucidating the role of key genes involved in the lignan biosynthesis pathway in P. leptostachya and for the industrial production of lignan through plant biotechnology.

Identification of altered metabolic functional components using metabolomics to analyze the different ages of fruiting bodies of Sanghuangporus vaninii cultivated on cut log substrates.

Sanghuangporus vaninii is a profitable traditional and medicinal edible fungus with uncommon therapeutic properties and medicinal value. The accumulation of active ingredients in this fungus that is used in traditional Chinese medicine is affected by its years of growth, and their pharmacological activities are also affected. However, the effects of age on the medicinal value of fruiting bodies of S. vaninii cultivated on cut log substrate remain unclear. In this study, an untargeted liquid chromatography mass spectrometry (LC-MS)-based metabolomics approach was performed to characterize the profiles of metabolites from 1-, 2- and 3-year-old fruiting bodies of S. vaninii. A total of, 156 differentially accumulated metabolites (DAMs) were screened based on the criterion of a variable importance projection greater than 1.0 and p < 0.01, including 75% up regulated and 25% down regulated. The results of enrichment of metabolic pathways showed that the metabolites involved the biosynthesis of plant secondary metabolites, biosynthesis of amino acids, central carbon metabolism in cancer, steroid hormone biosynthesis, linoleic acid metabolism, prolactin signaling pathway, and arginine biosynthesis, and so on. The biosynthesis of plant secondary metabolites pathway was significantly activated. Five metabolites were significantly elevated within the growth of fruiting bodies, including 15-keto-prostaglandin F2a, (4S, 5R)-4,5,6-trihydroxy-2-iminohexanoate, adenylsuccinic acid, piplartine, and chenodeoxycholic acid. 15-keto-prostaglandin F2a is related to the pathway of arachidonic acid metabolism and was significantly increased up to 1,320- and 535-fold in the 2- and 3-year-old fruiting bodies, respectively, compared with those in the 1-year-old group. The presence of these bioactive natural products in S. vaninii is consistent with the traditional use of Sanghuang, which prompted an exploration of its use as a source of natural prostaglandin in the form of foods and nutraceuticals. These findings may provide insight into the functional components of S. vaninii to develop therapeutic strategies.