Regulation Of Secondary Metabolites Research Articles

General Analysis of Heat Shock Factors in the Cymbidium ensifolium Genome Provided Insights into Their Evolution and Special Roles with Response to Temperature.

Heat shock factors (HSFs) are the key regulators of heat stress responses and play pivotal roles in tissue development and the temperature-induced regulation of secondary metabolites. In order to elucidate the roles of HSFs in Cymbidium ensifolium, we conducted a genome-wide identification of CeHSF genes and predicted their functions based on their structural features and splicing patterns. Our results revealed 22 HSF family members, with each gene containing more than one intron. According to phylogenetic analysis, 59.1% of HSFs were grouped into the A subfamily, while subfamily HSFC contained only two HSFs. And the HSF gene families were differentiated evolutionarily between plant species. Two tandem repeats were found on Chr02, and two segmental duplication pairs were observed on Chr12, Chr17, and Chr19; this provided evidence for whole-genome duplication (WGD) events in C. ensifolium. The core region of the promoter in most CeHSF genes contained cis-acting elements such as AP2/ERF and bHLH, which were associated with plant growth, development, and stress responses. Except for CeHSF11, 14, and 19, each of the remaining CeHSFs contained at least one miRNA binding site. This included binding sites for miR156, miR393, and miR319, which were responsive to temperature and other stresses. The HSF gene family exhibited significant tissue specificity in both vegetative and floral organs of C. ensifolium. CeHSF13 and CeHSF15 showed relatively significant expression in flowers compared to other genes. During flower development, CeHSF15 exhibited markedly elevated expression in the early stages of flower opening, implicating critical regulatory functions in organ development and floral scent-related regulations. During the poikilothermic treatment, CeHSF14 was upregulated over 200-fold after 6 h of heat treatment. CeHSF13 and CeHSF14 showed the highest expression at 6 h of low temperature, while the expression of CeHSF15 and CeHSF21 continuously decreased at a low temperature. The expression patterns of CeHSFs further confirmed their role in responding to temperature stress. Our study may help reveal the important roles of HSFs in plant development and metabolic regulation and show insight for the further molecular design breeding of C. ensifolium.

Whole genome sequencing and annotation of Aspergillus flavus JAM-JKB-B HA-GG20.

Groundnuts are mostly contaminated with the mold Aspergillus flavus which produces a carcinogenic mycotoxin called as aflatoxin. It is very important to understand the genetic factors underlying its pathogenicity, regulation, and biosynthesis of secondary metabolites and animal toxicities, but it still lacks useful information due to certain gaps in the era of modern technology. Therefore, the present study was considered to determine the key genes and metabolites involved in the biosynthesis of aflatoxin by using a molecular approach in a virulent strain of Aspergillus. The whole genome sequence of highly toxic and virulent Aspergillus isolates JAM-JKB-B HA-GG20 revealed 3,73,54,834bp genome size, 2, 26, 257 number of contigs with N50 value of 49,272bp, 12,400 genes and 48.1% of GC contained respectively. The genome sequence was compared with other known aflatoxin producing and non-producing genome of Aspergillus spp. and 61 secondary metabolite (SM) gene clusters were annotated with the toxic strain JAM-JKB-BHA-GG20 which showed similarity with other Aspergillus spp. A total number of eight genes (ver-1, AflR, pksA, uvm8, omt1, nor-1, Vha and aflP) were identified related to biosynthesis of aflatoxin and ochratoxin. Also, 69 SSR with forward and reverse primers and 137 di and tri nucleotide motifs were identified in the nucleotide sequence region related to aflatoxin gene pathway. The genes and putative metabolites identified in this study are potentially involved in host invasion and pathogenicity. As such, the genomic information obtained in this study is helpful in understanding aflatoxin gene producing pathway in comparison to other Aspergillus spp. and predicted presence of other secondary metabolites clusters viz. Nrps, T1pks etc. genes associated with a biosynthesis of OTA mycotoxin.

Gene Co-expression Network Analysis to Mine Genes Involved in Plant Secondary Metabolite Biosynthesis and Regulation.

Plants produce diverse specialized metabolites (SMs) that do not participate in plant growth and development but help them adapt to various environmental conditions. In addition to aiding in plant adaptation, different SMs serve as active ingredients for pharmaceutical and cosmetics products. However, despite their significant role in plant adaptation and industrial importance, the genes involved in the biosynthesis and regulation of many SMs remain largely unknown. This hinders deciphering the specific role of SMs in plant adaptation and limits their industrial utilization. Since many SMs pathway genes are expected to act in tight association with each other within a coexpression network, the network biology approach, such as weighted gene coexpression network analysis, could be used to identify the unknown genes. This chapter describes a workflow for constructing a gene coexpression network to identify genes that could be associated with the biosynthesis and regulation of SMs.

Genome-wide transcriptome analysis of toxigenic Fusarium verticillioides in response to variation of temperature and water activity on maize kernels

Fusarium verticillioides is one of the important mycotoxigenic pathogens of maize since it causes severe yield losses and produces fumonisins (FBs) to threaten human and animal health. Previous studies showed that temperature and water activity (aw) are two pivotal environmental factors affecting F. verticillioides growth and FBs production during maize storage. However, the genome-wide transcriptome analysis of differentially expressed genes (DEGs) in F. verticillioides under the stress combinations of temperature and aw has not been studied in detail. In this study, DEGs of F. verticillioides and their related regulatory pathways were analyzed in response to the stress of temperature and aw combinations using RNA-Seq. The results showed that the optimal growth conditions for F. verticillioides were 0.98 aw and 25 °C, whereas the highest per-unit yield of the fumonisin B1 (FB1) was observed at 0.98 aw and 15 °C. The RNA-seq analysis showed that 9648 DEGs were affected by temperature regardless of aw levels, whereas only 218 DEGs were affected by aw regardless of temperature variations. Gene Ontology (GO) analysis revealed that a decrease in temperature at both aw levels led to a significant upregulation of genes associated with 24 biological processes, while three biological processes were downregulated. Furthermore, when aw was decreased at both temperatures, seven biological processes were significantly upregulated and four were downregulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that the genes, whose expression was upregulated when the temperature decreased, were predominantly associated with the proteasome pathway, whereas the genes, whose expression was downregulated when the aw decreased, were mainly linked to amino acid metabolism. For the FB1, except for the FUM15 gene, the other 15 biosynthetic-related genes were highly expressed at 0.98 aw and 15 °C. In addition, the expression pattern analysis of other biosynthetic genes involved in secondary metabolite production and regulation of fumonisins production was conducted to explore how this fungus responds to the stress combinations of temperature and aw. Overall, this study primarily examines the impact of temperature and aw on the growth of F. verticillioides and its production of FB1 using transcriptome data. The findings presented here have the potential to contribute to the development of novel strategies for managing fungal diseases and offer valuable insights for preventing fumonisin contamination in food and feed storage.

Genome-Wide Identification and Characterization of the bHLH Gene Family and Its Response to Abiotic Stresses in Carthamus tinctorius.

The basic helix-loop-helix (bHLH) transcription factors possess DNA-binding and dimerization domains and are involved in various biological and physiological processes, such as growth and development, the regulation of secondary metabolites, and stress response. However, the bHLH gene family in C. tinctorius has not been investigated. In this study, we performed a genome-wide identification and analysis of bHLH transcription factors in C. tinctorius. A total of 120 CtbHLH genes were identified, distributed across all 12 chromosomes, and classified into 24 subfamilies based on their phylogenetic relationships. Moreover, the 120 CtbHLH genes were subjected to comprehensive analyses, including protein sequence alignment, evolutionary assessment, motif prediction, and the analysis of promoter cis-acting elements. The promoter region analysis revealed that CtbHLH genes encompass cis-acting elements and were associated with various aspects of plant growth and development, responses to phytohormones, as well as responses to both abiotic and biotic stresses. Expression profiles, sourced from transcriptome databases, indicated distinct expression patterns among these CtbHLH genes, which appeared to be either tissue-specific or specific to certain cultivars. To further explore their functionality, we determined the expression levels of fifteen CtbHLH genes known to harbor motifs related to abiotic and hormone responses. This investigation encompassed treatments with ABA, salt, drought, and MeJA. The results demonstrated substantial variations in the expression patterns of CtbHLH genes in response to these abiotic and hormonal treatments. In summary, our study establishes a solid foundation for future inquiries into the roles and regulatory mechanisms of the CtbHLH gene family.

THE OBTAINING RHODIOLA ROSEA L. BIOLOGICALLY ACTIVE COMPOUNDS BY BIOTECHNOLOGICAL METHODS (OVERVIEW)

Due to its rich chemical composition including unique biologically active compounds the gold root (Rhodiola rosea L.) is still of interest to researchers all over the world. However, deficiency of the raw material base and metabolome variability are determination the modern development ways of pharmacologically active substances in vitro obtaining. The article summarizes and systematizes the information about of Rhodiola rosea biologically active compounds obtaining, which are most often obtaining from callus tissues. Callus tissue is usually obtaining from leaf explants, and Murashige and Skoog medium with plant growth regulators in various combinations and concentrations is most commonly used for its induction. At the same time, the tissue growth efficiency and target compounds accumulation depends on the intact plant genotype, the precursors presence of these compounds in the medium, elicitors, as well as external influence factors. Recently, a promising direction is the transgenic hairy roots cultivation, which reveals new aspects of synthesis, accumulation and secondary metabolites production regulation. Biosynthesis genetic regulation of significant secondary metabolites is also a promising direction. Numerous studies have demonstrated that salidroside biosynthesis depends on the expression of the TyrDC gene encoding tyrosinedecarboxylase. Consequently, understanding the molecular and genetic mechanisms opens up opportunities for its regulation and metabolic engineering. In this regard, biotechnological methods may be a priority to obtaining salidroside, rosin and their derivatives at higher or at least comparable levels with wild type or cultivated plants.
 Among the various biotechnological strategies used to increase the accumulation of salidroside and glycosides of cinnamic alcohol in cells and cultures of Rhodiola organs, the approach with the addition of the metabolic precursors to the nutrient medium proved to be the most effective. In vitro cultures, a significant increase in the production of rosin and its derivatives was observed when precursors (cinnamic alcohol, cinnamic acid and cinnamic aldehyde) were added to the medium.

The role of signaling compounds in enhancing rice allelochemicals for sustainable agriculture: an overview.

Plant phytotoxin synthesis is influenced by intricate signaling networks like jasmonic acid (JA) and salicylic acid (SA). These compounds not only induce allelochemical production but also aid weed suppression and plant immunity. (-)-Loliolide, JA, SA, and their derivatives trigger rice allelochemical synthesis and gene expression. Enhancing allelochemical synthesis in crops offers an alternative, reducing reliance on traditional herbicides for effective weed management. Rice (Oryza sativa L.) serves as a crucial staple food crop, nourishing over half of the global population, particularly in South Asia. Within rice plants, various secondary metabolites are produced, contributing to its nutritional value and providing energy to consumers. Over the last 5 decades, researchers have investigated 276 distinct types of secondary metabolites found in rice plants. These metabolites predominantly include phenolic acids, flavonoids, steroids, alkaloids, terpenoids, and their derivatives. The role of these secondary metabolites is to regulate the growth and development of the rice plant. In this research paper, we have focused on the allelopathic potential of rice, which involves its active defense strategy to suppress other species in its vicinity. This defense mechanism is regulated by plant signaling compounds. These signaling compounds enable rice plants to recognize and detect competitors, pathogens, and herbivores in their environment. As a response, the rice plants elevate the production of defensive secondary metabolites. One crucial aspect of rice allelopathy is the phenomenon of neighbor detection. Rice plants can sense the presence of neighboring plants and respond accordingly to establish their competitive advantage and ensure their survival. This paper specifically highlights the impact of exogenously applied signaling compounds, namely Methyl salicylate (MeSA) and Methyl Jasmonate (MeJA), on paddy rice. The aim is to provide deeper insights into the signaling mechanisms involved in rice allelopathy and how the exogenous application of signaling compounds influence the induction and regulation of defensive secondary metabolites in rice plants. Comprehensive analysis of various researchers' studies clearly reveals that the application of these elicitor compounds noticeably augments the allelopathic potential of rice, resulting in heightened accumulation of phenolic acid compounds. Expansion in more enlistment of phenolics may be because of expansion in the activities of enzymes, such as cinnamate 4-hydroxylase (C4H) and phenylalanine ammonia-lyase (PAL), the two main enzymes of the phenylpropanoid pathway, which are associated with allelopathic crop plants, and along this, they recognize the presence of weeds and react by expanding allelochemical focuses. Consequently, substantial endeavors have been dedicated in recent times to discover and characterize plant-derived signaling molecules. In bioassays conducted by Patni et al. in 2019, both competitive and non-competitive rice genotypes exhibited elevated phytotoxicity against Echino colona following treatment with MeSA. MeSA-treated rice plants displayed accelerated growth, increased yield, and concurrently demonstrated weed-suppressing properties. Published studies from 1976 to 2021 are reviewed in this paper. The study indicates that signaling compounds induce allelochemical concentrations, enhancing allelopathic activity. This insight may lead to development of novel herbicides for effective sustainable weed management.

Genomic and transcriptomic analyses reveal polygenic architecture for ecologically important traits in aspen (Populustremuloides Michx.).

Intraspecific genetic variation in foundation species such as aspen (Populus tremuloides Michx.) shapes their impact on forest structure and function. Identifying genes underlying ecologically important traits is key to understanding that impact. Previous studies, using single-locus genome-wide association (GWA) analyses to identify candidate genes, have identified fewer genes than anticipated for highly heritable quantitative traits. Mounting evidence suggests that polygenic control of quantitative traits is largely responsible for this "missing heritability" phenomenon. Our research characterized the genetic architecture of 30 ecologically important traits using a common garden of aspen through genomic and transcriptomic analyses. A multilocus association model revealed that most traits displayed a highly polygenic architecture, with most variation explained by loci with small effects (likely below the detection levels of single-locus GWA methods). Consistent with a polygenic architecture, our single-locus GWA analyses found only 38 significant SNPs in 22 genes across 15 traits. Next, we used differential expression analysis on a subset of aspen genets with divergent concentrations of salicinoid phenolic glycosides (key defense traits). This complementary method to traditional GWA discovered 1243 differentially expressed genes for a polygenic trait. Soft clustering analysis revealed three gene clusters (241 candidate genes) involved in secondary metabolite biosynthesis and regulation. Our work reveals that ecologically important traits governing higher-order community- and ecosystem-level attributes of a foundation forest tree species have complex underlying genetic structures and will require methods beyond traditional GWA analyses to unravel.

Genome sequencing provides potential strategies for drug discovery and synthesis

Medicinal plants are renowned for their abundant production of secondary metabolites, which exhibit notable pharmacological activities and great potential for drug development. The biosynthesis of secondary metabolites is highly intricate and influenced by various intrinsic and extrinsic factors, resulting in substantial species diversity and content variation. Consequently, precise regulation of secondary metabolite synthesis is of utmost importance. In recent years, genome sequencing has emerged as a valuable tool for investigating the synthesis and regulation of secondary metabolites in medicinal plants, facilitated by the widespread use of high-throughput sequencing technologies. This review highlights the latest advancements in genome sequencing within this field and presents several strategies for studying secondary metabolites. Specifically, the article elucidates how genome sequencing can unravel the pathways for secondary metabolite synthesis in medicinal plants, offering insights into the functions and regulatory mechanisms of participating enzymes. Comparative analyses of plant genomes allow identification of shared pathways of metabolite synthesis among species, thereby providing novel avenues for obtaining cost-effective biosynthetic intermediates. By examining individual genomic variations, genes or gene clusters associated with the synthesis of specific compounds can be discovered, indicating potential targets and directions for drug development and the exploration of alternative compound sources. Moreover, the advent of gene-editing technology has enabled the precise modifications of medicinal plant genomes. Optimization of specific secondary metabolite synthesis pathways becomes thus feasible, enabling the precise editing of target genes to regulate secondary metabolite production within cells. These findings serve as valuable references and lessons for future drug development endeavors, conservation of rare resources, and the exploration of new resources.

The Transcriptome Response to Azole Compounds in Aspergillus fumigatus Shows Differential Gene Expression across Pathways Essential for Azole Resistance and Cell Survival.

The opportunistic pathogen Aspergillus fumigatus is found on all continents and thrives in soil and agricultural environments. Its ability to readily adapt to novel environments and to produce billions of spores led to the spread of azole-resistant A. fumigatus across the globe, posing a threat to many immunocompromised patients, including critically ill patients with severe influenza or COVID-19. In our study, we sought to compare the adaptational response to azoles from A. fumigatus isolates that differ in azole susceptibility and genetic background. To gain more insight into how short-term adaptation to stressful azole compounds is managed through gene expression, we conducted an RNA-sequencing study on the response of A. fumigatus to itraconazole and the newest clinically approved azole, isavuconazole. We observed many similarities in ergosterol biosynthesis up-regulation across isolates, with the exception of the pan-azole-resistant isolate, which showed very little differential regulation in comparison to other isolates. Additionally, we found differential regulation of membrane efflux transporters, secondary metabolites, iron metabolism, and various stress response and cell signaling mechanisms.

High hydrostatic pressure harnesses the biosynthesis of secondary metabolites via the regulation of polyketide synthesis genes of hadal sediment-derived fungi.

Deep-sea fungi have evolved extreme environmental adaptation and possess huge biosynthetic potential of bioactive compounds. However, not much is known about the biosynthesis and regulation of secondary metabolites of deep-sea fungi under extreme environments. Here, we presented the isolation of 15 individual fungal strains from the sediments of the Mariana Trench, which were identified by internal transcribed spacer (ITS) sequence analysis as belonging to 8 different fungal species. High hydrostatic pressure (HHP) assays were performed to identify the piezo-tolerance of the hadal fungi. Among these fungi, Aspergillus sydowii SYX6 was selected as the representative due to the excellent tolerance of HHP and biosynthetic potential of antimicrobial compounds. Vegetative growth and sporulation of A. sydowii SYX6 were affected by HHP. Natural product analysis with different pressure conditions was also performed. Based on bioactivity-guided fractionation, diorcinol was purified and characterized as the bioactive compound, showing significant antimicrobial and antitumor activity. The core functional gene associated with the biosynthetic gene cluster (BGC) of diorcinol was identified in A. sydowii SYX6, named as AspksD. The expression of AspksD was apparently regulated by the HHP treatment, correlated with the regulation of diorcinol production. Based on the effect of the HHP tested here, high pressure affected the fungal development and metabolite production, as well as the expression level of biosynthetic genes which revealed the adaptive relationship between the metabolic pathway and the high-pressure environment at the molecular level.

Exogenous Application of Moringa Leaf Extract Confers Salinity Tolerance in Sunflower by Concerted Regulation of Antioxidants and Secondary Metabolites

Exogenous Application of Moringa Leaf Extract Confers Salinity Tolerance in Sunflower by Concerted Regulation of Antioxidants and Secondary Metabolites

Lavandula Species, Their Bioactive Phytochemicals, and Their Biosynthetic Regulation.

Lavandula species are one of the most useful aromatic and medicinal plants and have great economic potential. The phytopharmaceutical contribution of the secondary metabolites of the species is unquestionable. Most recent studies have been focusing on the elucidation of the genetic background of secondary metabolite production in lavender species. Therefore, knowledge of not only genetic but especially epigenetic mechanisms for the regulation of secondary metabolites is necessary for the modification of those biosynthesis processes and the understanding of genotypic differences in the content and compositional variability of these products. The review discusses the genetic diversity of Lavandula species in relation to the geographic area, occurrence, and morphogenetic factors. The role of microRNAs in secondary-metabolites biosynthesis is described.

Novel players in organogenesis and flavonoid biosynthesis in cucumber glandular trichomes.

Glandular trichomes (GTs) are outgrowths of plant epidermal cells that secrete and store specialized secondary metabolites that protect plants against biotic and abiotic stresses and have economic importance for human use. While extensive work has been done to understand the molecular mechanisms of trichome organogenesis in Arabidopsis (Arabidopsis thaliana), which forms unicellular, nonglandular trichomes (NGTs), little is known about the mechanisms of GT development or regulation of secondary metabolites in plants with multicellular GTs. Here, we identified and functionally characterized genes associated with GT organogenesis and secondary metabolism in GTs of cucumber (Cucumis sativus). We developed a method for effective separation and isolation of cucumber GTs and NGTs. Transcriptomic and metabolomic analyses showed that flavonoid accumulation in cucumber GTs is positively associated with increased expression of related biosynthesis genes. We identified 67 GT development-related genes, the functions of 7 of which were validated by virus-induced gene silencing. We further validated the role of cucumber ECERIFERUM1 (CsCER1) in GT organogenesis by overexpression and RNA interference transgenic approaches. We further show that the transcription factor TINY BRANCHED HAIR (CsTBH) serves as a central regulator of flavonoid biosynthesis in cucumber GTs. Work from this study provides insight into the development of secondary metabolite biosynthesis in multicellular GTs.

Hormone and carbohydrate regulation of defense secondary metabolites in a Mediterranean forest during drought

Plants have evolved complex physiological mechanisms to regulate production of defense secondary metabolites (SMs) to cope with environmental stress such as drought. Yet, these mechanisms remain under-studied in mature trees, limiting our ability to understand effects of climate change on tree productivity and survival. We investigated defense SMs and their relations to growth, nonstructural carbohydrates and phytohormones across seasons, in a Mediterranean forest dominated by two tree species with different resistance to drought. Our results showed seasonality of leaf SMs, with a strong accumulation of SMs during cold and dry conditions in winter. The drought-resistant species Phillyrea latifolia had high levels of SMs in both leaves (ca. 10–20%) and stem phloem (4–6%), but experimental drought decreased SMs and soluble sugars in stem phloem in spring and summer. By contrast, the drought-susceptible species Quercus ilex had relatively low and constant levels of SMs, irrespective of drought. We further showed that P. latifolia leaves had generally higher levels of jasmonic acid and lower salicylic acid than Q. ilex leaves, potentially leading to differences in SMs and growth between the two co-occucrring species. Our study suggests that species- and organ-specific dynamics of defense SMs are driven by phytohormones and carbohydrates in mature trees, and that P. latifolia may become less resistant to abiotic and biotic stress due to reduced phloem defense SMs and carbohydrates in a drying climate.

Phylogenetic Analysis of R2R3-MYB Family Genes in Tetrastigma hemsleyanum Diels et Gilg and Roles of ThMYB4 and ThMYB7 in Flavonoid Biosynthesis.

Tetrastigma hemsleyanum Diels et Gilg (T. hemsleyanum) is an extensively used Chinese folk herb with multiple bioactivities. Among these bioactivities, flavonoids are recognized as the representative active ingredients. We previously found an elevated accumulation of flavonoids in T. hemsleyanum under water stress; however, the mechanism remains unclear. R2R3-MYB transcription factors play vital roles in the plant response to environmental stress and the regulation of secondary metabolites. Herein, a systematic transcriptome identification of R2R3-MYB family genes under water stress in T. hemsleyanum was performed to explore their potential function in the biosynthesis of flavonoids. A total of 26 R2R3-MYB genes were identified, most of which were clustered into functional branches of abiotic stress. ThMYB4 and ThMYB7 were then screened out to be associated with the biosynthesis of flavonoids through a protein-protein interaction prediction. An expression correlation analysis based on RNA-seq further confirmed that ThMYB4 and ThMYB7 were positively related to the flavonoid biosynthetic pathway genes of T. hemsleyanum. In ThMYB4- and ThMYB7-overexpression hairy roots, it was found that the expression of ThCHS and ThCHI was significantly increased, suggesting that ThMYB4 and ThMYB7 may act as regulators in flavonoid biosynthesis. This will shed new light on the promotion of flavonoid production and the medicinal value of T. hemsleyanum by manipulating transcription factors.

Ultraviolet-B and Heavy Metal-Induced Regulation of Secondary Metabolites in Medicinal Plants: A Review.

Despite a rich history and economic importance, the potential of medicinal plants has not been fully explored under different abiotic stress conditions. Penetration of UV-B radiation and contamination of heavy metals are two important environmental stress for plants with remarkable influence on the defense-related and pharmaceutically important secondary metabolites of medicinal plants. UV-B and heavy metal contamination may become a critical issue that either positively or negatively affects the quality and quantity of secondary metabolites. Such effects may result from changes in the expression level of genes that encode the corresponding enzymes or the inactivation and/or stimulation of specific enzymes involved in the different biosynthetic pathways of the secondary metabolites. Therefore, a comprehensive study of the impact of UV-B and heavy metals individually and in combination on the biosynthesis and accumulation of secondary metabolites in medicinal plants is discussed in the present review.

Genome-wide analysis of Panax MADS-box genes reveals role of PgMADS41 and PgMADS44 in modulation of root development and ginsenoside synthesis

Panax root is an important material used in food and medicine. Its cultivation and production usually depend on root shape and ginsenoside content. There is limited understanding about the synergistic regulatory mechanisms underlying root development and ginsenoside accumulation in Panax. MADS-box transcription factors possibly play a significant role in regulation of root growth and secondary metabolites. In this study, we identified MADS-box transcription factors of Panax, and found high expression levels of SVP, ANR1 and SOC1-like clade genes in its roots. We confirmed that two SOC1-like genes, PgMADS41 and PgMADS44, bind to expansion gene promoters (PgEXLB5 and PgEXPA13), which contribute to root growth, and to SE-4, CYP716A52v2-4, and β-AS-13 promoters, which participate in ginsenoside Ro biosynthesis. These two genes were found to increase lateral root number and main root length in transgenic Arabidopsis thaliana by improving AtEXLA1, AtEXLA3, AtEXPA5, and AtEXPA6 gene expression. As a non-phytohormone regulatory tool, Ro can stimulate adventitious root growth by influencing their expression and ginsenoside accumulation. Our study provides new insights into the coordinated regulatory function of SOC1-like clade genes in Panax root development and triterpenoid accumulation, paving the way towards understanding root formation and genetic improvement in Panax.

Palliating Salt Stress in Mustard through Plant-Growth-Promoting Rhizobacteria: Regulation of Secondary Metabolites, Osmolytes, Antioxidative Enzymes and Stress Ethylene.

The severity of salt stress is alarming for crop growth and production and it threatens food security. Strategies employed for the reduction in stress are not always eco-friendly or sustainable. Plant-growth-promoting rhizobacteria (PGPR) could provide an alternative sustainable stress reduction strategy owning to its role in various metabolic processes. In this study, we have used two strains of PGPR, Pseudomonas fluorescens (NAIMCC-B-00340) and Azotobacter chroococcum Beijerinck 1901 (MCC 2351), either singly or in combination, and studied their effect in the amelioration of salt toxicity in mustard cultivar Pusa Jagannath via its influence on plants' antioxidants' metabolism, photosynthesis and growth. Individually, the impact of Pseudomonas fluorescens was better in reducing stress ethylene, oxidative stress, photosynthesis and growth but maximal alleviation was observed with their combined application. MDA and H2O2 content as indicator of oxidative stress decreased by 27.86% and 45.18% and osmolytes content (proline and glycine-betaine) increased by 38.8% and 26.3%, respectively, while antioxidative enzymes (SOD, CAT, APX and GR) increased by 58.40, 25.65, 81.081 and 55.914%, respectively, over salt-treated plants through the application of Pseudomonas fluorescens. The combined application maximally resulted in more cell viability and less damage to the leaf with lesser superoxide generation due to higher antioxidative enzymes and reduced glutathione formation (GSH). Considering the obtained results, we can supplement the PGPR in combination to plants subjected to salt stress, prevent photosynthetic and growth reduction, and increase the yield of plants.

RADseq-based population genomic analysis and environmental adaptation of rare and endangered recretohalophyte Reaumuria trigyna.

Genetic diversity reflects the survival potential, history, and population dynamics of an organism. It underlies the adaptive potential of populations and their response to environmental change. Reaumuria trigyna is an endemic species in the Eastern Alxa and West Ordos desert regions in China. The species has been considered a good candidate to explore the unique survival strategies of plants that inhabit this area. In this study, we performed population genomic analyses based on restriction-site associated DNA sequencing to understand the genetic diversity, population genetic structure, and differentiation of the species. Analyses of 92,719 high-quality single-nucleotide polymorphisms (SNPs) indicated that overall genetic diversity of R. trigyna was low (HO=0.249 and HE=0.208). No significant genetic differentiation was observed among the investigated populations. However, a subtle population genetic structure was detected. We suggest that this might be explained by adaptive diversification reinforced by the geographical isolation of populations. Overall, 3513 outlier SNPs were located in 243 gene-coding sequences in the R. trigyna transcriptome. Potential sites under diversifying selection occurred in genes (e.g., AP2/EREBP, E3 ubiquitin-protein ligase, FLS, and 4CL) related to phytohormone regulation and synthesis of secondary metabolites which have roles in adaptation of species. Our genetic analyses provide scientific criteria for evaluating the evolutionary capacity of R. trigyna and the discovery of unique adaptions. Our findings extend knowledge of refugia, environmental adaption, and evolution of germplasm resources that survive in the Ordos area.