Accumulation Of Phenolic Acids Research Articles

Changes in secondary metabolites contents and stress responses in Salvia miltiorrhiza via ScWRKY35 overexpression: Insights from a wild relative Salvia castanea

Salvia castanea Diels, a close wild relative to the medicinal plant, Salvia miltiorrhiza Bunge, primarily grows in high-altitude regions. While the two species share similar active compounds, their content varies significantly. WRKY transcription factors are key proteins, which regulate plant growth, stress response, and secondary metabolism. We identified 46 ScWRKY genes in S. castanea and found that ScWRKY35 was a highly expressed gene associated with secondary metabolites accumulation. This study aimed to explore the role of ScWRKY35 gene in regulating the accumulation of secondary metabolites and its response to UV and cadmium (Cd) exposure in S. miltiorrhiza. It was found that transgenic S. miltiorrhiza hairy roots overexpressing ScWRKY35 displayed upregulated expression of genes related to phenolic acid synthesis, resulting in increased salvianolic acid B (SAB) and rosmarinic acid (RA) contents. Conversely, tanshinone pathway gene expression decreased, leading to lower tanshinone levels. Further, overexpression of ScWRKY35 upregulated Cd transport protein HMA3 in root tissues inducing Cd sequestration. In contrast, the Cd uptake gene NRAMP1 was downregulated, reducing Cd absorption. In response to UV radiation, ScWRKY35 overexpression led to an increase in the accumulation of phenolic acid and tanshinone contents, including upregulation of genes associated with salicylic acid (SA) and jasmonic acid (JA) synthesis. Altogether, these findings highlight the role of ScWRKY35 in enhancing secondary metabolites accumulation, as well as in Cd and UV stress modulation in S. miltiorrhiza, which offers a novel insight into its phytochemistry and provides a new option for the genetic improvement of the plants.

Effects of light-emitting diodes on the morphology and accumulation of glucosinolates, carotenoids and phenolic acids in red kale sprouts

Kale (Brassica oleracea var. acephala) has gained popularity as a nutritious and phytochemical-rich vegetable. This study has investigated the effects of three LED treatments (white, blue, and red) on the accumulation of secondary metabolites in kale sprouts. Ten DAS, the kale sprouts were harvested, and growth measurements were measured. Furthermore, selected sprouts were stored at -80 °C for further biochemical analysis, namely, phenolic acids, glucosinolates, and carotenoids. Sprouts irradiated with red LED light showed the best SL, RT, and FW values. For total GSLs, we found that kale sprouts irradiated with white LED lights showed the best results (41.59±0.41 μmol/g DW). Among the aliphatic GSLs, we found that progoitrin presented the best results under blue LED light (17.93±0.49 µmol/g DW), and among the indolic GSLs, glucobrassicin showed the best results under white and red LED light. The highest concentration of total carotenoids was found in kale sprouts under white LED light exposure (3341.27±206.96 μg/g DW). Individually, β-carotene was observed in high concentration under white LED light (1935.13±87.21 μg/g DW). Among the PAs, chlorogenic acid was found in the highest concentration in the treatments under white LED light (45.78±0.73 µg/g DW). In general, kale sprouts irradiated with white LED light showed high contents of GSLs, carotenoids, and PAs. Regarding morphological characteristics, kale sprouts under red LED light showed the most promise. This research offers a valuable approach to enhancing the phytochemicals found in kale sprouting.

Widely Targeted Metabolomic Analysis Provides New Insights into the Effect of Rootstocks on Citrus Fruit Quality.

The use of different rootstocks has a significant effect on the content of flavor components and overall fruit quality. However, little information is available about the metabolic basis of the nutritional value of citrus plants. In this study, UPLC-MS/MS (ultra-performance liquid chromatography-tandem mass spectrometry) was performed to analyze the metabolites of three late-maturing hybrid mandarin varieties ('Gold Nugget', 'Tango' and 'Orah') grafted on four rootstocks ('Trifoliate orange', 'Carrizo citrange', 'Red tangerine' and 'Ziyang Xiangcheng'). A total of 1006 metabolites were identified through OPLS-DA (Orthogonal Partial Least Squares-Discriminant Analysis) analysis. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis revealed the most critical pathways among the different pathways associated with genes grafted on the four rootstocks that were differentially activated, including tryptophan metabolism and sphingolipid metabolism in 'Gold Nugget'; tryptophan metabolism, phenylpropanoid biosynthesis and sphingolipid metabolism in 'Tango'; and pantothenate and CoA biosynthesis- and photosynthesis-related biosynthesis in 'Orah'. A considerable difference between the different rootstocks was also observed in the accumulation of lipids, phenolic acids and flavonoids; further analysis revealed that the rootstocks regulated specific metabolites, including deacetylnomylinic acid, sudachinoid A, amoenin evodol, rutaevin, cyclo (phenylalanine-glutamic acid), cyclo (proline-phenylalanine), 2-hydroxyisocaproic acid, and 2-hydroxy-3-phenylpropanoic acid. The results of this study provide a useful foundation for further investigation of rootstock selection for late-maturation hybrid mandarin varieties.

Constructed the metabolic compound accumulation profiles of daylily at different main producing areas using multi-omics techniques

Daylily flower buds (DFB) are widely cultivated in China. The discernible disparities exist in the nutritional composition of daylilies across different primary production areas. However, limited studies about the chemical composition and biosynthetic mechanisms of the principal constituents of DFB. This study systematically investigated daylilies from seven distinct growing regions by integrated metabolomics and transcriptomic sequencing to elucidate the environmental impact on DFB quality and metabolite biosynthesis mechanisms. In DFB, flavonoids constituting the majority, followed by phenolic acids and lipids. The total flavonoid and polyphenol content exhibited the highest levels in northern China (Ningxia, Gansu), and the lowest levels in southern regions (Hunan, Fujian). Correspondingly, antioxidant enzyme activity and free radical scavenging capacity mirrored these regional disparities. Thus, northern-producing areas demonstrated a propitious environment for the accumulation of secondary metabolites, particularly flavonoids and phenolic acids. In contrast, southern production areas exhibited a less conducive environment for the accumulation of flavonoid glycosides in daylilies. Furthermore, distinctive variations in the key metabolites were observed in FJQZ, a region characterized by diverse flower colors. Key enzymes and transcription factors were identified and validated through RT-qPCR. These results contribute valuable insights into the daylily quality formation mechanisms, offering a foundation for high-quality breeding strategies.

Withania somnifera (L.) Dunal: Enhanced production of withanolides and phenolic acids from hairy root culture after application of elicitors

Withania somnifera (L.) Dunal is an important indigenous medicinal plant with extensive pharmaceutical potential. The root is the main source of major bioactive compounds of this plant species including withanolides, withanine, phenolic acids, etc. Hairy root culture (HRC) is a crucial method for low-cost production of active compounds on a large scale. Four different Agrobacterium rhizogenes strains have been used for the hairy root induction. Maximum transformation efficiency (87.34 ± 2.13%) was achieved with A4 bacterial strain-mediated transformed culture. The genetic transformation was confirmed by using specific primers of seven different genes. Seven HR (Hairy root) lines were selected after screening 29 HR lines based on their fast growth rate and high accumulation of withanolides and phenolic acids content. Two biotic and three abiotic elicitors were applied to the elite root line to trigger more accumulation of withanolides and phenolic acids. While all the elicitors effectively increased withanolides and phenolic acids production, among the five different elicitors, salicylic acid (4.14 mg l–1) induced 11.49 -fold increase in withanolides (89.07 ± 2.75 mg g–1 DW) and 5.34- fold increase in phenolic acids (83.69 ± 3.11 mg g− 1 DW) after 5 days of elicitation compared to the non-elicited culture (7.75 ± 0.63 mg g–1 DW of withanolides and 15.66 ± 0.92 mg g–1 DW of phenolic acids). These results suggest that elicitors can tremendously increase the biosynthesis of active compounds in this system; thus, the HRC of W. somnifera is cost-effective and can be efficiently used for the industrial production of withanolides and phenolic acids.

Multi-omics analysis provides new insights into the changes of important nutrients and fructose metabolism in loquat bud sport mutant.

Bud sport is a common and stable somatic variation in perennial fruit trees, and often leads to significant modification of fruit traits and affects the breeding value. To investigate the impact of bud sport on the main metabolites in the fruit of white-fleshed loquat, we conducted a multi-omics analysis of loquat fruits at different developmental stages of a white-fleshed bud sport mutant of Dongting loquat (TBW) and its wild type (TBY). The findings from the detection of main fruit quality indices and metabolites suggested that bud sport resulted in a reduction in the accumulation of carotenoids, fructose, titratable acid and terpenoids at the mature stage of TBW, while leading to the accumulation of flavonoids, phenolic acids, amino acids and lipids. The comparably low content of titratable acid further enhances the balanced and pleasent taste profile of TBW. Expression patterns of differentially expressed genes involved in fructose metabolism exhibited a significant increase in the expression level of S6PDH (EVM0006243, EVM0044405) prior to fruit maturation. The comparison of protein sequences and promoter region of S6PDH between TBY and TBW revealed no structural variations that would impact gene function or expression, indicating that transcription factors may be responsible for the rapid up-regulation of S6PDH before maturation. Furthermore, correlation analysis helped to construct a comprehensive regulatory network of fructose metabolism in loquat, including 23 transcription factors, six structural genes, and nine saccharides. Based on the regulatory network and existing studies, it could be inferred that transcription factors such as ERF, NAC, MYB, GRAS, and bZIP may promote fructose accumulation in loquat flesh by positively regulating S6PDH. These findings improve our understanding of the nutritional value and breeding potential of white-fleshed loquat bud sport mutant, as well as serve as a foundation for exploring the genes and transcription factors that regulate fructose metabolism in loquat.

Waterlogging in soil restricts the growth of Gleditsia sinensis seedlings and inhibits the accumulation of lignans and phenolic acids in thorns.

Gleditsia sinensis, commonly known as Chinese Zaojiao, has important economic value and medicinal compounds in its fruits and thorns, making it widely cultivated artificially in China. However, the available literature on the impact of waterlogging on the growth of G. sinensis seedlings and the accumulation of metabolite compounds in its thorns is limited. To address this knowledge gap, G. sinensis seedlings were planted in soil supplemented with pindstrup substrate, which enhances the water-holding capacity of the soil. The analyses of morphological traits and nutrient elements in one-year-old G. sinensis seedlings grown naturally under ambient conditions and metabolite accumulation in its thorns were conducted. The results showed that the waterlogged soil significantly diminished the height, fresh weight, and dry weight of seedling roots and stems (P<0.05). Furthermore, waterlogging hindered the uptake of iron (Fe) and manganese (Mn), as well as the transport of potassium (K). The identified metabolites within the thorns were categorized into 16 distinct groups. Relative to the control soil, fatty acids and derivatives were the most down-regulated metabolites in the waterlogged soil, accounting for 40.58% of the total metabolites, followed by lignans (38.71%), phenolic acids (34.48%), saccharides and alcohols (34.15%), steroids (16.67%), alkaloids (12.24%), flavonoids (9.28%), and glycerophospholipids (7.41%). Conversely, nucleotides and derivatives experienced the greatest up-regulation in the waterlogged soil, accounting for 50.00% of the total metabolites. In conclusion, waterlogging negatively impacted the growth of G. sinensis seedlings and inhibited the accumulation of metabolites. Hence, when considering the accumulation of secondary metabolites such as lignans and phenolic acids, appropriate management of soil moisture levels should be taken into account.

Beneficial endophytic fungi improve the yield and quality of Salvia miltiorrhiza by performing different ecological functions.

Endophytic fungi can enhance the growth and synthesis of secondary metabolites in medicinal plants. Salvia miltiorrhiza Bunge is frequently employed for treating cardiovascular and cerebrovascular ailments, with the primary bioactive components being salvianolic acid and tanshinone. However, their levels in cultivated S. miltiorrhiza are inferior to that of the wild herbs, so the production of high-quality medicinal herbs is sharply declining. Consequently, the utilization of beneficial endophytic fungi to improve the yield and quality of S. miltiorrhiza holds great significance for the cultivation of medicinal plants. In this study, nine non-pathogenic, endophytic fungal strains were introduced into sterile S. miltiorrhiza seedlings and cultivated both in vitro and in situ (the greenhouse). The effects of these strains on the growth indices, C and N metabolism, antioxidant activity, photosynthesis, and content of bioactive ingredients in S. miltiorrhiza were then evaluated. The results showed that the different genera, species, or strains of endophytic fungi regulated the growth and metabolism of S. miltiorrhiza in unique ways. These endophytic fungi primarily exerted their growth-promoting effects by increasing the net photosynthetic rate, intercellular CO2 concentration, and the activities of sucrose synthase, sucrose phosphate synthase, nitrate reductase, and glutamine synthetase. They also enhanced the adaptability and resistance to environmental stresses by improving the synthesis of osmoregulatory compounds and the activity of antioxidant enzymes. However, their regulatory effects on the growth and development of S. miltiorrhiza were affected by environmental changes. Moreover, the strains that significantly promoted the synthesis and accumulation of phenolic acids inhibited the accumulation of tanshinones components, and vice versa. The endophytic fungal strains Penicillium meloforme DS8, Berkeleyomyces basicola DS10, and Acremonium sclerotigenum DS12 enhanced the bioaccumulation of tanshinones. Fusarium solani DS16 elevated the rosmarinic acid content and yields in S. miltiorrhiza. The strain Penicillium javanicum DS5 improved the contents of dihydrotanshinone, salvianolic acid B, and rosmarinic acid. The strains P. meloforme DS8 and B. basicola DS10 improved resistance. Various endophytic fungi affected the quality and yield of S. miltiorrhiza by regulating different physiological and metabolic pathways. This study also provides a novel and effective method to maximize the effects of beneficial endophytic fungi by selecting specific strains to design microbial communities based on the different ecological functions of endophytic fungi under varying environments and for specific production goals.

Abscisic Acid Affects Phenolic Acid Content to Increase Tolerance to UV-B Stress in Rhododendron chrysanthum Pall.

The presence of the ozone hole increases the amount of UV radiation reaching a plant's surface, and UV-B radiation is an abiotic stress capable of affecting plant growth. Rhododendron chrysanthum Pall. (R. chrysanthum) grows in alpine regions, where strong UV-B radiation is present, and has been able to adapt to strong UV-B radiation over a long period of evolution. We investigated the response of R. chrysanthum leaves to UV-B radiation using widely targeted metabolomics and transcriptomics. Although phytohormones have been studied for many years in plant growth and development and adaptation to environmental stresses, this paper is innovative in terms of the species studied and the methods used. Using unique species and the latest research methods, this paper was able to add information to this topic for the species R. chrysanthum. We treated R. chrysanthum grown in a simulated alpine environment, with group M receiving no UV-B radiation and groups N and Q (externally applied abscisic acid treatment) receiving UV-B radiation for 2 days (8 h per day). The results of the MN group showed significant changes in phenolic acid accumulation and differential expression of genes related to phenolic acid synthesis in leaves of R. chrysanthum after UV-B radiation. We combined transcriptomics and metabolomics data to map the metabolic regulatory network of phenolic acids under UV-B stress in order to investigate the response of such secondary metabolites to stress. L-phenylalanine, L-tyrosine and phenylpyruvic acid contents in R. chrysanthum were significantly increased after UV-B radiation. Simultaneously, the levels of 3-hydroxyphenylacetic acid, 2-phenylethanol, anthranilate, 2-hydroxycinnamic acid, 3-hydroxycinnamic acid, α-hydroxycinnamic acid and 2-hydroxy-3-phenylpropanoic acid in this pathway were elevated in response to UV-B stress. In contrast, the study in the NQ group found that externally applied abscisic acid (ABA) in R. chrysanthum had greater tolerance to UV-B radiation, and phenolic acid accumulation under the influence of ABA also showed greater differences. The contents of 2-phenylethanol, 1-o-p-coumaroyl-β-d-glucose, 2-hydroxy-3-phenylpropanoic acid, 3-(4-hydroxyphenyl)-propionic acid and 3-o-feruloylquinic ac-id-o-glucoside were significantly elevated in R. chrysanthum after external application of ABA to protect against UV-B stress. Taken together, these studies of the three groups indicated that ABA can influence phenolic acid production to promote the response of R. chrysanthum to UV-B stress, which provided a theoretical reference for the study of its complex molecular regulatory mechanism.

Integrative effects of phytohormones in the phenolic acids production in Salvia verticillata L. under multi-walled carbon nanotubes and methyl jasmonate elicitation

Salvia verticillata L. is a well-known herb rich in rosmarinic acid (RA) and with therapeutic values. To better understand the possible roles of phytohormones in the production of phenolic acids in S. verticillata, in this work, we investigated some physiological and biochemical responses of the species to methyl jasmonate (MJ) and multi-walled carbon nanotubes (MWCNTs) as two effective elicitors. The leaves were sprayed with aqueous solutions containing 100 mg L−1 MWCNTs and 100 µM MJ and then harvested during interval times of exposure up to 96 h. The level of abscisic acid, as the first effective phytohormone, was altered in the leaves in response to MJ and MWCNTs elicitation (2.26- and 3.06-fold more than the control, respectively), followed by significant increases (P ˂ 0.05) detected in jasmonic acid and salicylic acid contents up to 8 h after exposure. Obtained data revealed that simultaneously with changes in phytohormone profiles, significant (P ˂ 0.05) rises were observed in the content of H2O2 (8.85- and 9.74-folds of control), and the amount of lipid peroxidation (10.18- and 17.01-folds of control) during the initial times after exposure to MJ and MWCNTs, respectively. Later, the content of phenolic acids increased in the elicited leaves due to changes in the transcription levels of key enzymes involved in their biosynthesis pathways, so 2.71- and 11.52-fold enhances observed in the RA content of the leaves after exposure to MJ and MWCNTs, respectively. It is reasonable to conclude that putative linkages between changes in some phytohormone pools lead to the accumulation of phenolic acids in the leaves of S. verticillata under elicitation. Overall, the current findings help us improve our understanding of the signal transduction pathways of the applied stimuli that led to enhanced secondary metabolite production in medicinal plants.

Transcriptome Analysis of Salvia miltiorrhiza under Drought Stress.

Phenolic acids are one of the major secondary metabolites accumulated in Salvia miltiorrhiza with various pharmacological activities. Moderate drought stress can promote the accumulation of phenolic acids in S. miltiorrhiza, while the mechanism remains unclear. Therefore, we performed transcriptome sequencing of S. miltiorrhiza under drought treatment. A total of 47,169 unigenes were successfully annotated in at least one of the six major databases. Key enzyme genes involved in the phenolic acid biosynthetic pathway, including SmPAL, SmC4H, Sm4CL, SmTAT, SmHPPR, SmRAS and SmCYP98A14, were induced. Unigenes annotated as laccase correlated with SmRAS and SmCYP98A14 were analyzed, and seven candidates that may be involved in the key step of SalB biosynthesis by RA were obtained. A total of 15 transcription factors significantly up-regulated at 2 h and 4 h potentially regulating phenolic acid biosynthesis were screened out. TRINITY_DN14213_c0_g1 (AP2/ERF) significantly transactivated the expression of SmC4H and SmRAS, suggesting its role in the regulation of phenolic acid biosynthesis. GO and KEGG enrichment analysis of differential expression genes showed that phenylpropanoid biosynthesis and plant hormone signal transduction were significantly higher. The ABA-dependent pathway is essential for resistance to drought and phenolic acid accumulation. Expression patterns in drought and ABA databases showed that four PYLs respond to both drought and ABA, and three potential SnRK2 family members were annotated and analyzed. The present study presented a comprehensive transcriptome analysis of S. miltiorrhiza affected by drought, which provides a rich source for understanding the molecular mechanism facing abiotic stress in S. miltiorrhiza.

Metabolic adaptations of Pinus massoniana to low phosphorus and acidic aluminium with or without ectomycorrhization

Phosphorus (P) deficiency and aluminium toxicity often coexist and are the major constraints hampering plant growth in acidic soils. Plants have evolved several strategies to cope with these adverse environments, including the recruitment of beneficial microbes, especially mycorrhizal fungi. By employing microscopic observation, histochemical staining, and UPLC-MS/MS techniques, we analyzed the aluminium deposition and cell viability of root, symbiotic process with ECM fungus Suillus bovinus, as well as the metabolic adjustments in Pinus massoniana under low P and/or acidic aluminium conditions. The formation process of mycorrhizae between P. massoniana and S. bovinus was not affected by low P and acidic aluminium treatments, but low P postponed the development of mantle and Hartig nets. The colonization rate increased in the presence of acidic aluminium, and S. bovinus inoculation improved root cell viability and promoted the growth of P. massoniana under low P and/or acidic aluminium conditions. The metabolic responses of nonmycorrhizal and mycorrhizal P. massoniana to low P and/or acidic aluminium were different. Low P and/or acidic aluminium significantly increased the accumulation of phenolic acids and altered starch and sucrose metabolism in roots of nonmycorrhizal P. massoniana. However, for mycorrhizal P. massoniana, low P and/or acidic aluminium treatments increased the organic acids content of shared differentially accumulated metabolites (DAMs). Furthermore, acidic aluminium mainly altered arginine and proline metabolism, and phenylalanine metabolism was affected under low P conditions, linoleic acid metabolism was changed under the combined treatment of low P and acidic aluminium in mycorrhizal P. massoniana. These findings enhance our comprehension of how ECM fungi assist P. massoniana in coping with P deficiency and aluminium toxicity in acidic environments and provide a theoretical basis for ECM fungal application during seedling cultivation in acidic soils.

COI1 dependent jasmonic acid signalling positively modulates ROS scavenging system in transgenic hairy root culture of tomato

Reactive oxygen species (ROS) production is a routine event in plants. ROS function as signalling molecules in regulating plant development and defence. However, their accumulation beyond threshold leads to toxicity. Hence, plants are evolved with specialized ROS scavenging system involving phytohormones (synthesis and signalling), enzymes and metabolites. To understand the role of phytohormone jasmonic acid (JA) signalling in ROS scavenging, tomato coronatine insensitive 1 (SlCOI1), a key gene in JA signalling, was silenced and overexpressed in tomato transgenic hairy roots (HR) under the constitutive promoter. Targeted metabolomics of transgenic HR revealed accumulation of phenolic acids including ferulic acid, coumaric acid, vanillic acid, and flavonoid catechin in SlCOI1 overexpressed line. Moreover, osmolyte amino acids proline, asparagine, and glutamine showed a positive co-relation with transgenic overexpression of SlCOI1. Ascorbic acid-glutathione, a crucial antioxidant system was found to be influenced by COI1-mediated JA signalling. The expression of genes encoding enzymes superoxide dismutase 1, ascorbate peroxidase 1, and dehydroascorbate reductase 2 was found to be down and upregulated in SlCOI1 silenced and overexpressed lines, respectively. Methyl jasmonate and Fusarium oxysporum f.sp. lycopersici crude extract treatment further confirmed the regulatory role of COI1-mediated JA signalling in regulation of enzymatic components involved in ROS scavenging. The COI1-mediated JA signalling could also elevate the expression of RESPIRATORY BURST OXIDASE HOMOLOG-B gene which is involved in ROS wave signal generation. The present study underscores the role of COI1-mediated JA signalling in modulating enzymatic and non-enzymatic components of ROS scavenging system and pathogen associated molecular pattern triggered immunity.

Biochar preparation and its effects with reduced compound fertilizer on nutrients, phenolic acid and fungal community in tobacco rhizosphere soil

Biochar is a carbon-rich soil conditioner produced from pyrolysis of biomass, it has been widely used to enhance soil quality because of its physical adsorption as well as water and fertilizer conservation functions. This work aimed to improve the soil quality of continuously cropped flue-cured tobacco fields using biochar prepared from agricultural waste. To explore the impact of reduced compound fertilizer with biochar application on nutrients, phenolic acid contents and fungi diversity in the rhizosphere soil, 4 treatments were set: regular compound fertilizer application (T1), and biochar with reduced compound fertilizer in different proportions (T2, T3, T4, with 100, 75, 50% of compound fertilizer, respectively). The physicochemical properties of the prepared biochar were characterized and observed using electron microscopy. The results indicated that, a noticeable increase in the content of soil organic matter (SOM) and soil organic carbon (SOC) in T2 treatment compared to T1 treatment. Moreover, T2 treatment demonstrated a significant improvement in the contents of alkali-hydrolyzable nitrogen (AN), available phosphorus (AP), and available potassium (AK), with increases of 9.29%, 15.85% and 25.42% compared to T1 treatment, respectively. While a gradual decrease in soil AN, AP, and AK content was observed with the reduction of compound fertilizer application (T2–T4), there was no obvious difference between T3 and T1. For total phenolic acid, the content in T2 treatment significantly decreased by 35.99% compared to T1 treatment, while T3 and T4 treatments showed significant reductions of 15.30 and 18.73% respectively, compared to T2 treatment. Biochar application could enhance the fungal community’s abundance and diversity in the rhizosphere soil. Fungal community exhibited the highest richness under T3 treatment, while the relative abundance ofFusariumandMortierellareduced as the decrease of compound fertilizer (T2–T4). In conclusion, the reduced compound fertilizer with biochar application could reduce nutrient loss, phenolic acids accumulation, and improving the abundance of fungal community in the rhizosphere soil. This paper provides a reference for biochar combined with compound fertilizer to improve soil from the regulation of allelochemicals and soil fungi.

DNA methylation regulates biosynthesis of tanshinones and phenolic acids during growth of Salvia miltiorrhiza.

DNA methylation plays a crucial role in the regulation of plant growth and the biosynthesis of secondary metabolites. Danshen (Salvia miltiorrhiza) is a valuable Chinese herbal medicine commonly used to treat cardiovascular diseases; its active ingredients are tanshinones and phenolic acids, which primarily accumulate in roots. Here, we conducted a targeted metabolic analysis of S. miltiorrhiza roots at 3 distinct growth stages: 40 d old (r40), 60 d old (r60), and 90 d old (r90). The contents of tanshinones (cryptotanshinone, tanshinone I, tanshinone IIA, and rosmariquinone) and phenolic acids (rosmarinic acid and salvianolic acid B) gradually increased during plant development. Whole-genome bisulfite sequencing and transcriptome sequencing of roots at the 3 growth stages revealed an increased level of DNA methylation in the CHH context (H represents A, T, or C) context at r90 compared with r40 and r60. Increased DNA methylation levels were associated with elevated expression of various genes linked to epigenetic regulations, including CHROMOMETHYLASE2 (SmCMT2), Decrease in DNA Methylation 1 (SmDDM1), Argonaute 4 (SmAGO4), and DOMAINS REARRANGED METHYLTRANSFERASE 1 (SmDRM1). Moreover, expression levels of many genes involved in tanshinone and salvianolic acid biosynthesis, such as copalyldiphosphate synthase 5 (SmCPS5), cytochrome P450-related enzyme (SmCYP71D464), geranylgeranyl diphosphate synthase (SmGGPPS1), geranyl diphosphate synthase (SmGPPS), hydroxyphenylpyruvate reductase (SmHPPR), and hydroxyphenylpyruvate dioxygenase (SmHPPD), were altered owing to hyper-methylation, indicating that DNA methylation plays an important role in regulating tanshinone and phenolic acid accumulation. Our data shed light on the epigenetic regulation of root growth and the biosynthesis of active ingredients in S. miltiorrhiza, providing crucial clues for further improvement of active compound production via molecular breeding in S. miltiorrhiza.

Elucidating the Differentiation Synthesis Mechanisms of Differently Colored Resistance Quinoa Seedings Using Metabolite Profiling and Transcriptome Analysis.

Quinoa (Chenopodium quinoa wild.), a dicotyledonous plant native to the Andes, is an increasingly popular pseudograin owing to its high nutritional value, stress resistance capabilities, and gluten-free properties. In this study, we aimed to explore the dynamic changes in different varieties of quinoa at the seedling stage and their regulatory networks. Here, we found that the leaves of quinoa showed obvious coloration after 45 days, and four quinoa seedling types (red, white, yellow, and black) were subjected to ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and transcriptome sequencing to identify their differentially expressed genes and metabolites. A total of 29 differential metabolites and 19 genes (14 structural and 5 regulatory genes) were identified, and consistent differences were observed in the flavonoid, phenolic acid, and alkaloid metabolites in the different quinoa types. These differential metabolites were significantly enriched in flavonoid and flavonol biosynthesis, flavonoid biosynthesis, and phenylpropanoid biosynthesis pathways. In addition, real-time fluorescence quantitative PCR (RT-qPCR) technology was used to detect the expression of four structural genes involved in the flavonoid biosynthesis pathway and four regulatory genes (interaction network). The results revealed that the structural and regulatory gene transcript levels in the flavonoid pathway were higher in the red quinoa cultivars than in the white, yellow, and black. Additionally, the differences in the leaves of these four quinoa cultivars were mainly due to differences in flavonoid, phenolic acid, and alkaloid accumulation. Our findings provide a basis for understanding the accumulation and coloration mechanisms of flavonoids, phenolic acids, and alkaloids in quinoa seedlings of different colors and also provide a theoretical basis for future investigations.

Cultivar-dependent and drought-induced modulation of secondary metabolites, adaptative defense in Fagopyrum esculentum L

The present study investigates the biochemical responses of buckwheat to drought stress, particularly focusing on phenolic acids and flavonoids, abundant in this crop. We hypothesize that distinct genotypic responses to drought stress will lead to variations in phenolic acid accumulation. Two common buckwheat cultivars, Panda (East European origin) and PI 482597 (originating from Zimbabwe), were subjected to drought treatment, with biochemical traits, relative water content, and photosynthetic pigments regularly assessed. While chlorophyll content remained unaffected by dehydration, total carotenoid content decreased. The unique increase in the chlorophyll to carotenoid ratio suggests a specific role of carotenoids in buckwheat's metabolic stress response. While most phenolic acids and flavonoids exhibited increasing trends during progressive dehydration, their dynamics differed. Notably, rutin content increased early in drought stress, while chlorogenic acid and kaempferol showed enhanced levels only under severe dehydration. Genotypic differences were observed in chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, 4-hydroxybenzoic acid, and quercetin. Conversely, trans-p-coumaric acid, trans-ferulic acid, vanillic acid, rutin, and kaempferol showed similar trends in both cultivars. By aligning observed drought-induced changes in phenolic compound contents with biosynthesis pathways, trade-offs between individual compounds were identified, contributing to the mechanistic understanding of varied stress responses.

Salt elicitation to enhance phytochemicals in durum wheat seedlings.

Salt has been identified as an elicitor that can increase the accumulation of phytochemicals in seedlings during the germination process. However, the salinity level required to maximize the yield of phytochemicals, particularly phenolic compounds, needs further investigation for several plant species. To address this issue, we imposed increasing levels of salinity (NaCl solutions) on the sprouting substrate of Triticum durum (var. Platone) grains, at concentrations of 0, 50, 100, 150, 200, 250, and 300 mM (0_S, 50_S, 100_S, 150_S, 200_S, 250_S, and 300_S, respectively). The highest NaCl doses (250_S and 300_S) significantly impacted germination performance and were excluded from further analysis. The seedlings harvested at 8 days after sowing exhibited different growth stages depending on the salinity level: wheatgrass for 0_S, early wheatgrass for 50_S, intermediate between sprout and wheatgrass for 100_S, sprout for 150_S, and very early sprout for 200_S. Furthermore, salinity induced the concentration of phenolic compounds (PhCs) in the seedlings' tissues (i.e., both roots and shoots) in a salinity-dependent manner. The highest values were observed at 200_S, with an increase of 187% of the total investigated PhCs in comparison with 0_S, averaged over shoots and roots. In particular, in 200_S, the accumulation of phenolic acids was up to fourfold higher in roots, and that of flavonoids was up to twofold higher in shoots. Our findings suggest that the use of 200 mM NaCl applied to the sprouting substrate is excessive for producing edible sprouts but may be suitable for phytochemical extraction purposes. © 2023 Society of Chemical Industry.

Mycorrhizal colonization and Streptomyces viridosporus HH1 synergistically up-regulate the polyphenol biosynthesis genes in wheat against stripe rust

BackgroundStripe rust is considered one of the most devastating diseases of wheat all over the world, resulting in a high loss in its production. In this study, time-course changes in expression of the polyphenol biosynthesis pathways genes in wheat against stripe rust were investigated. The defense mechanisms triggered by mycorrhizal colonization and/or spraying with Streptomyces viridosporus HH1 against this disease were also investigated.ResultsResults obtained revealed that C3H, which is considered the key gene in lignin biosynthesis, was the most expressed gene. Furthermore, most of the chlorogenic acid and flavonoid biosynthesis genes were also overexpressed. Volcano plots of the studied genes reveal that the dual treatment led to a high significant overexpression of 10 out of the 13 studied genes. Heatmap of these genes showed that the most frequent expressed gene in response to all applied treatments along the study period was DFR, the key gene in the biosynthesis of anthocyanidins. Gene co-expression network of the studied genes showed that HQT was the most central gene with respect to the other genes, followed by AN2 and DFR, respectively. Accumulation of different flavonoids and phenolic acids were detected in response to the dual treatment, in particular, cinnamic acid, coumarin, and esculetin, which recorded the highest elevation level recording 1000, 488.23, and 329.5% respectively. Furthermore, results from the greenhouse experiment showed that application of the dual treatment led to an 82.8% reduction in the disease severity, compared with the control treatment.ConclusionsWe can conclude that the biosynthesis of lignin, chlorogenic acid, and flavonoids contributed to the synergistic triggering effect of the dual treatment on wheat resistance to stripe rust.

Mechanisms governing the impact of nitrogen stress on the formation of secondary metabolites in Artemisia argyi leaves

Nitrogen is a key factor in various physiological and metabolic processes in plants. Providing an adequate supply of nitrogen is essential for improving the total yield and quality of the medicinal plant Artemisia argyi (A. argyi), but the underlying mechanisms of how this nutrient alters the crop remains unclear. In this study, we conducted a series of pot experiments to investigate the agronomic traits and active components in the leaves of A. argyi plants under low and high nitrogen stress. Additionally, we used transcriptome analysis and RT-qPCR to explore the molecular pathways associated with nitrogen stress. Our results demonstrate a dramatic increase in the accumulation of phenolic acids and flavonoids in the low nitrogen (LN) stress group compared to the control (CK), with increases of 40.00% and 79.49%, respectively. Interestingly, plants in the high nitrogen (HN) stress group exhibited enhanced plant growth with larger leaves, thicker stems, and a 3% increase in volatile oil content compared to the CK. Moreover, A. argyi in the HN group displayed a 66% increase in volatile oil concentration compared to the LN group. Our combined transcriptome and q-PCR results indicate that LN stress promotes the expression of genes involved in flavonoid synthesis, while HN stress promotes the expression of genes related to terpene skeleton production and photosynthesis. Taken together, these findings suggest that different gene expression levels under LN and HN stress contribute to the photosynthesis capacity and the accumulation of active ingredients in A. argyi leaves. Our results elucidate the physiological and molecular mechanisms of nitrogen stress on A. argyi secondary metabolites and guide fertilization strategies for plant cultivation.