Isoflavone Biosynthesis Research Articles

Melatonin mediates isoflavone accumulation in germinated soybeans (Glycine max L.) under ultraviolet-B stress

Soybean germination under ultraviolet-B (UV–B) radiation stress is a common and effective way to enrich the isoflavone content of sprouts. However, the growth and biomass of germinated soybeans are significantly suppressed using this method. Melatonin (MT), a novel plant biostimulant, not only plays a vital protective role in responses to various abiotic stresses but also regulates the accumulation of secondary metabolites. In the present study, the effects of exogenous MT on the growth and isoflavone metabolism of germinating soybeans exposed to UV-B stress were investigated. Compared to UV-B stress, the application of exogenous MT (25 μM) significantly increased sprout length, fresh weight, Ca2+ influx, and peroxidase activity; markedly decreased the content of malondialdehyde and H2O2 and the fluorescence intensity of H2O2 and O2•-; but had no noticeable effect on the activity of superoxide dismutase and catalase during germination. Moreover, the content of total flavonoids and isoflavone monomers (including daidzein, genistein, daidzin, glycitin and genistin) in 4-day-old germinated soybeans was significantly enhanced by MT application under UV-B stress and was not only companied by dramatically increased phenylalanine ammonia lyase activity, but also by markedly increased relative expression levels of phenylalanine ammonia lyase1, chalcone synthase, isoflavone reductase and flavanone 3-hydroxylase that are involved in the isoflavone biosynthesis pathway. The inhibitory effects of UV-B stress on the growth and biomass of germinated soybeans were alleviated with exogenous MT. MT enhanced the content of total flavonoids and isoflavone monomers under UV-B stress by increasing the activity and relative gene expression level of critical isoflavone biosynthesis-related enzymes.

Field mold stress induced catabolism of storage reserves in soybean seed and the resulting deterioration of seed quality in the field

Excessive rainfall provides a favorable condition for field mold infection of plants, which triggers field mold (FM) stress. If FM stress occurs during the late maturation stage of soybean seed, it negatively affects seed yield and quality. To investigate the responses of soybean seed against FM stress and identify the underlying biochemical pathways involved, a greenhouse was equipped with an artificial rain producing system to allow the induction of mold growth on soybean seed. The induced quality changes and stress responses were revealed on the levels of both transcriptome and metabolome. The results showed that soybean seeds produced under FM stress conditions had an abnormal and inferior appearance, and also contained less storage reserves, such as protein and polysaccharide. Transcriptional analysis demonstrated that genes involved in amino acid metabolism, glycolysis, tricarboxylic acid, β-oxidation of fatty acids, and isoflavone biosynthesis were induced by FM stress. These results were supported by a multiple metabolic analysis which exhibited increases in the concentrations of a variety of amino acids, sugars, organic acids, and isoflavones, as well as reductions of several fatty acids. Reprogramming of these metabolic pathways mobilized and consumed stored protein, sugar and fatty acid reserves in the soybean seed in order to meet the energy and substrate demand on the defense system, but led to deterioration of seed quality. In general, FM stress induced catabolism of storage reserves and diminished the quality of soybean seed in the field. This study provides a more profound insight into seed deterioration caused by FM stress.

Novel QTL and Meta-QTL Mapping for Major Quality Traits in Soybean.

Isoflavone, protein, and oil are the most important quality traits in soybean. Since these phenotypes are typically quantitative traits, quantitative trait locus (QTL) mapping has been an efficient way to clarify their complex and unclear genetic background. However, the low-density genetic map and the absence of QTL integration limited the accurate and efficient QTL mapping in previous researches. This paper adopted a recombinant inbred lines (RIL) population derived from ‘Zhongdou27’and ‘Hefeng25’ and a high-density linkage map based on whole-genome resequencing to map novel QTL and used meta-analysis methods to integrate the stable and consentaneous QTL. The candidate genes were obtained from gene functional annotation and expression analysis based on the public database. A total of 41 QTL with a high logarithm of odd (LOD) scores were identified through composite interval mapping (CIM), including 38 novel QTL and 2 Stable QTL. A total of 660 candidate genes were predicted according to the results of the gene annotation and public transcriptome data. A total of 212 meta-QTL containing 122 stable and consentaneous QTL were mapped based on 1,034 QTL collected from previous studies. For the first time, 70 meta-QTL associated with isoflavones were mapped in this study. Meanwhile, 69 and 73 meta-QTL, respectively, related to oil and protein were obtained as well. The results promote the understanding of the biosynthesis and regulation of isoflavones, protein, and oil at molecular levels, and facilitate the construction of molecular modular for great quality traits in soybean.

MATE-Type Proteins Are Responsible for Isoflavone Transportation and Accumulation in Soybean Seeds.

Soybeans are nutritionally important as human food and animal feed. Apart from the macronutrients such as proteins and oils, soybeans are also high in health-beneficial secondary metabolites and are uniquely enriched in isoflavones among food crops. Isoflavone biosynthesis has been relatively well characterized, but the mechanism of their transportation in soybean cells is largely unknown. Using the yeast model, we showed that GmMATE1 and GmMATE2 promoted the accumulation of isoflavones, mainly in the aglycone forms. Using the tobacco BrightYellow-2 (BY-2) cell model, GmMATE1 and GmMATE2 were found to be localized in the vacuolar membrane. Such subcellular localization supports the notion that GmMATE1 and GmMATE2 function by compartmentalizing isoflavones in the vacuole. Expression analyses showed that GmMATE1 was mainly expressed in the developing soybean pod. Soybean mutants defective in GmMATE1 had significantly reduced total seed isoflavone contents, whereas the overexpression of GmMATE1 in transgenic soybean promoted the accumulation of seed isoflavones. Our results showed that GmMATE1, and possibly also GmMATE2, are bona fide isoflavone transporters that promote the accumulation of isoflavones in soybean seeds.

Transcriptome analysis revealed key genes involved in flavonoid metabolism in response to jasmonic acid in pigeon pea (Cajanus cajan (L.) Millsp.)

Flavonoids are important metabolites of pigeon pea in relation to its stress resistance. However, the molecular basis and regulatory mechanisms of flavonoids in pigeon pea remain unclear. Methyl jasmonate (MeJA) is a signaling molecule associated with biosynthesis of flavonoids. In this study, after exogenous treatment of 10 mg/L MeJA, infection of pathogenic fungi to pigeon pea was alleviated and the content of flavonoids was increased. Results of gene expression and metabolic changes that were respectively analyzed by transcriptome sequencing and high performance liquid chromatography (HPLC) showed that (1) concentrations of various flavonoids, such as genistein, apigenin, vitexin and biochanin A were significantly up-regulated; (2) 13675 differentially expressed genes were produced, mainly enriched in signal transduction and isoflavone biosynthesis pathways: (3) the expression levels of key synthase genes (CcI2′H, CcHIDH, Cc7-IOMT) in the flavonoid biosynthesis pathway were significantly up-regulated; (4) Overexpression of CcbHLH35 significantly induced upregulation of flavonoid synthase genes and accumulation of genistein, vitexin and apigenin. Our findings reveals the pivotal roles of MeJA in synthesis and functioning of flavonoids in pigeon pea, which provide a basis for further studies on flavonoid-mediated defense responses.

Isolation, sequencing of the HvnHID gene and its role in the purple-grain colour development in Tibetan hulless barley

2-hydroxyisoflavanone dehydratase (HID) plays an important role in isoflavone biosynthesis. In this study, HID was isolated from the seeds of the purple-grained Tibetan hulless barley variety Nerumuzha and the white-grained variety Kunlun 10. The HvnHID gene includes the 981 bp open reading frame and encodes a protein of 327 amino acids. It has a typical Abhydrolase_3 domain (78–306) and belongs to the carboxylesterase (CXE) family of the Abhydrolase_3 (α/β hydrolase) superfamily. There are eight nucleotide differences in the HvnHID coding sequence and two amino acid differences (one in the Abhydrolase_3 domain) between Nerumuzha and Kunlun 10. The HvnHID of hulless barley has the closest relationship with the HID in Hordeum vulgare, and the most distant relationship in Panicum hallii. At the early-mid stage of the seed colour development, the HvnHID expression levels in the purple and black seeds were significantly higher than in the white and blue ones (P < 0.01). During the seed colour development of purple-grained hulless barley, the expression of the key genes (HvnF3'H, HvnDRF, HvnANT1, and HvnGT) in the anthocyanidin biosynthetic pathway increased significantly, while the HvnHID expression decreased significantly (P < 0.01). Thus, it is likely that HvnHID negatively regulates the anthocyanidin biosynthesis. This result provides an important basis for further study of the biological functions of HvnHID in the anthocyanidin biosynthetic pathway.

Cover Image

The cover image is based on the Original Article Changing light promotes isoflavone biosynthesis in soybean pods and enhances their resistance to mildew infection by Xiaoman Li et al., https://doi.org/10.1111/pce.14128.

The Induction of the Isoflavone Biosynthesis Pathway Is Associated with Resistance to Common Bacterial Blight in Phaseolus vulgaris L.

Xanthomonas axonopodis infects common bean (Phaseolus vulgaris L.) causing the disease common bacterial blight (CBB). The aim of this study was to investigate the molecular and metabolic mechanisms underlying CBB resistance in P. vulgaris. Trifoliate leaves of plants of a CBB-resistant P. vulgaris recombinant inbred line (RIL) and a CBB-susceptible RIL were inoculated with X. axonopodis or water (mock treatment). Leaves sampled at defined intervals over a 48-h post-inoculation (PI) period were monitored for alterations in global transcript profiles. A total of 800 genes were differentially expressed between pathogen and mock treatments across both RILs; approximately half were differentially expressed in the CBB-resistant RIL at 48 h PI. Notably, there was a 4- to 32-fold increased transcript abundance for isoflavone biosynthesis genes, including several isoflavone synthases, isoflavone 2′-hydroxylases and isoflavone reductases. Ultra-high performance liquid chromatography-tandem mass spectrometry assessed leaf metabolite levels as a function of the PI period. The concentrations of the isoflavones daidzein and genistein and related metabolites coumestrol and phaseollinisoflavan were increased in CBB-resistant RIL plant leaves after exposure to the pathogen. Isoflavone pathway transcripts and metabolite profiles were unaffected in the CBB-susceptible RIL. Thus, induction of the isoflavone pathway is associated with CBB-resistance in P. vulgaris.

Changing light promotes isoflavone biosynthesis in soybean pods and enhances their resistance to mildew infection.

Mildew severely reduces soybean yield and quality, and pods are the first line of defence against pathogens. Maize-soybean intercropping (MSI) reduces mildew incidence on soybean pods; however, the mechanism remains unclear. Changing light (CL) from maize shading is the most important environmental feature in MSI. We hypothesized that CL affects isoflavone accumulation in soybean pods, affecting their disease resistance. In the present study, shading treatments were applied to soybean plants during different developmental stages according to various CL environments under MSI. Chlorophyll fluorescence imaging (CFI) and classical evaluation methods confirmed that CL, especially vegetative stage shading (VS), enhanced pod resistance to mildew. Further metabolomic analyses and exogenous jasmonic acid (JA) and biosynthesis inhibitor experiments revealed the important relationship between JA and isoflavone biosynthesis, which had a synergistic effect on the enhanced resistance of CL-treated pods to mildew. VS promoted the biosynthesis and accumulation of constitutive isoflavones upstream of the isoflavone pathway, such as aglycones and glycosides, in soybean pods. When mildew infects pods, endogenous JA signalling stimulated the biosynthesis of downstream inducible malonyl isoflavone (MIF) and glyceollin to improve pod resistance.

Metabolic Engineering of Isoflavones: An Updated Overview

Isoflavones are ecophysiologically active secondary metabolites derived from the phenylpropanoid pathway. They were mostly found in leguminous plants, especially in the pea family. Isoflavones play a key role in plant–environment interactions and act as phytoalexins also having an array of health benefits to the humans. According to epidemiological studies, a high intake of isoflavones-rich diets linked to a lower risk of hormone-related cancers, osteoporosis, menopausal symptoms, and cardiovascular diseases. These characteristics lead to the significant advancement in the studies on genetic and metabolic engineering of isoflavones in plants. As a result, a number of structural and regulatory genes involved in isoflavone biosynthesis in plants have been identified and characterized. Subsequently, they were engineered in various crop plants for the increased production of isoflavones. Furthermore, with the advent of high-throughput technologies, the regulation of isoflavone biosynthesis gains attention to increase or decrease the level of isoflavones in the crop plants. In the review, we begin with the role of isoflavones in plants, environment, and its benefits in human health. Besides, the main theme is to discuss the updated research progress in metabolic engineering of isoflavones in other plants species and regulation of production of isoflavones in soybeans.

Iris domestica (iso)flavone 7- and 3'-O-Glycosyltransferases Can Be Induced by CuCl2.

In many plants, isoflavones are the main secondary metabolites that have various pharmacological activities, but the low water solubility of aglycones limits their usage. The O-glycosylation of (iso)flavones is a promising way to overcome this barrier. O-glycosyltransferases (UGTs) are key enzymes in the biosynthesis of (iso)flavonoid O-glycosides in plants. However, limited investigations on isoflavonoid O-UGTs have been reported, and they mainly focused on legumes. Iris domestica (L.) Goldblatt et Mabberley is a non-legume plant rich in various isoflavonoid glycosides. However, there are no reports regarding its glycosylation mechanism, despite the I. domestica transcriptome previously being annotated as having non-active isoflavone 7-O-UGTs. Our previous experiments indicated that isoflavonoid glycosides were induced by CuCl2 in I. domestica calli; therefore, we hypothesized that isoflavone O-UGTs may be induced by Cu2+. Thus, a comparative transcriptome analysis was performed using I. domestica seedlings treated with CuCl2, and eight new active BcUGTs were obtained. Biochemical analyses showed that most of the active BcUGTs had broad substrate spectra; however, substrates lacking 5-OH were rarely catalyzed. Real-time quantitative PCR results further indicated that the transcriptional levels of BcUGTs were remarkably induced by Cu2+. Our study increases the understanding of UGTs and isoflavone biosynthesis in non-legume plants.

Exogenous Serotonin and Melatonin Regulate Dietary Isoflavones Profoundly through Ethylene Biosynthesis in Soybean [Glycine max (L.) Merr.

Serotonin and melatonin are important signaling and stress mitigating molecules. However, their role and molecular mechanism in the accumulation of isoflavones are not clearly defined. To elucidate their functions, serotonin and melatonin were applied to in vitro cultures of soybean at different concentrations and analyzed to assess the accumulation of isoflavone content followed by transcript levels of biosynthesis genes at different time intervals. Increased total phenolics, total flavonoids, and different forms of isoflavone content were observed in the treatments. Expression levels of critical genes in isoflavone, ethylene, jasmonic acid, abscisic acid, and melatonin biosynthesis and related transcription factor were quantified. A correlation was observed between the expression of ethylene biosynthesis genes (S-adenosylmethionine synthase and 1-aminocyclopropane-1-carboxylate oxidase) and isoflavone biosynthesis genes (chalcone synthase, chalcone reductase, and isoflavone synthase). We hypothesize that, under serotonin and melatonin treatments, ethylene biosynthesis may play a role in the increase/decrease in isoflavone content in soybean culture.

Isoflavone levels, nodulation and gene expression profiles of a CRISPR/Cas9 deletion mutant in the isoflavone synthase gene of red clover.

Isoflavones are not involved in rhizobial signaling in red clover, but likely play a role in defense in the rhizosphere. Red clover (Trifolium pratense) is a high-quality forage legume, well suited for grazing and hay production in the temperate regions of the world. Like many legumes, red clover produces a number of phenylpropanoid compounds including anthocyanidins, flavan-3-ols, flavanols, flavanones, flavones, and isoflavones. The study of isoflavone biosynthesis and accumulation in legumes has come into the forefront of biomedical and agricultural research due to potential for medicinal, antimicrobial, and environmental implications. CRISPR/Cas9 was used to knock out the function of a key enzyme in the biosynthesis of isoflavones, isoflavone synthase (IFS1). A hemizygous plant carrying a 9-bp deletion in the IFS1 gene was recovered and was intercrossed to obtain homozygous mutant plants. Levels of the isoflavones formononetin, biochanin A and genistein were significantly reduced in the mutant plants. Wild-type and mutant plants were inoculated with rhizobia to test the effect of the mutation on nodulation, but no significant differences were observed, suggesting that these isoflavones do not play important roles in nodulation. Gene expression profiling revealed an increase in expression of the upstream genes producing the precursors for IFS1, namely, phenylalanine ammonium lyase and chalcone synthase, but there were no significant differences in IFS1 gene expression or in the downstream genes in the production of specific isoflavones. Higher expression in genes involved in ethylene response was observed in the mutant plants. This response is normally associated with biotic stress, suggesting that the plants may have been responding to cues in the surrounding rhizosphere due to lower levels of isoflavones.

Chromosome-level reference genome assembly provides insights into aroma biosynthesis in passion fruit (Passiflora edulis).

Passion fruit, native to tropical America, is an agriculturally, economically and ornamentally important fruit plant that is well known for its acid pulp, rich aroma and distinctive flavour. Here, we present a chromosome-level genome assembly of passion fruit by incorporating PacBio long HiFi reads and Hi-C technology. The assembled reference genome is 1.28Gb size with a scaffold N50 of 126.4Mb and 99.22% sequences anchored onto nine pseudochromosomes. This genome is highly repetitive, accounting for 86.61% of the assembled genome. A total of 39,309 protein-coding genes were predicted with 93.48% of those being functionally annotated in the public databases. Genome evolution analysis revealed a core eudicot-common γ whole-genome triplication event and a more recent whole-genome duplication event, possibly contributing to the expansion of certain gene families. The 33 rapidly expanded gene families were significantly enriched in the pathways of isoflavone biosynthesis, galactose metabolism, diterpene biosynthesis and fatty acid metabolism, which might be responsible for the formation of featured flavours in the passion fruit. Transcriptome analysis revealed that genes related to ester and ethylene biosynthesis were significantly upregulated in the mature fruit and the expression levels of those genes were consistent with the accumulation of volatile lipid compounds. The passion fruit genome analysis improves our understanding of the genome evolution of this species and sheds new lights into the molecular mechanism of aroma biosynthesis in passion fruit.

Regulatory protein genes and microRNAs in response to selenium stimuli in Pueraria lobata (Willd.) Ohwi

Abstract Regulatory protein genes and microRNAs (miRNAs) play important roles in response to abiotic and biotic stress, and the biosynthesis of secondary metabolites in plants. However, their responses to selenium (Se) stimuli have not been comprehensively studied in Pueraria lobata (Willd.) Ohwi, a selenocompound-rich medicinal and edible plant. In this study, we identified a total of 436/556/1161/624 transcription factors, 134/157/308/172 transcriptional regulators, and 341/456/250/518 protein kinases, which were co-expressed with at least one selenocompound-related structural gene/sulfate transporter or phosphate transporter/reactive oxygen species (ROS) scavenging structural gene/isoflavone-related structural gene, respectively. Then, we identified a total of 87 expressed miRNAs by Se disposure, in which 11 miRNAs, including miR171f-3p, miR390b-3P, miR-N111b, miR-N118, miR-N30, miR-N38-3P, miR-N61a, miR-N61b, miR-N80-3p, miR-N84-3P, and miR-N90.2-3P, were significantly upregulated. We also identified a total of 1172 target genes for the 87 expressed miRNAs. Gene Ontology enrichment analysis of these target genes showed that regulation of transcription, DNA-templated, integral component of membrane, nucleus, ATP binding, and plasma membrane are the top five subclassifications. Finally, we revealed that 5 miRNAs targeted 10 regulatory protein genes, which are highly correlated with at least one selenocompound-related structural gene or transporter gene; 5 miRNAs targeted 10 regulatory protein genes, which are highly correlated with at least one ROS scavenging structural gene; and 5 miRNAs targeted 9 regulatory protein genes, which are potentially involved in the isoflavone biosynthesis. Overall, the study provides us the comprehensive insight into the roles of regulatory proteins and miRNAs in response to Se stimuli in P. lobata.

Diurnal metabolic regulation of isoflavones and soyasaponins in soybean roots.

Isoflavones and soyasaponins are major specialized metabolites accumulated in soybean roots and secreted into the rhizosphere. Unlike the biosynthetic pathway, the transporters involved in metabolite secretion remain unknown. The developmental regulation of isoflavone and soyasaponin secretions has been recently reported, but the diurnal regulation of their biosynthesis and secretion still needs to be further studied. To address these challenges, we conducted transcriptome and metabolite analysis using hydroponically grown soybean plants at 6‐hr intervals for 48 hr in a 12‐hr‐light/12‐hr‐dark condition. Isoflavone and soyasaponin biosynthetic genes showed opposite patterns in the root tissues; that is, the former genes are highly expressed in the daytime, while the latter ones are strongly induced at nighttime. GmMYB176 encoding a transcription factor of isoflavone biosynthesis was upregulated from ZT0 (6:00 a.m.) to ZT6 (12:00 a.m.), followed by the induction of isoflavone biosynthetic genes at ZT6. The isoflavone aglycone content in the roots accordingly increased from ZT6 to ZT18 (0:00 a.m.). The isoflavone aglycone content in root exudates was kept consistent throughout the day, whereas that of glucosides increased at ZT6, which reflected the decreased expression of the gene encoding beta‐glucosidase involved in the hydrolysis of apoplast‐localized isoflavone conjugates. Co‐expression analysis revealed that those isoflavone and soyasaponin biosynthetic genes formed separate clusters, which exhibited a correlation to ABC and MATE transporter genes. In summary, the results in this study indicated the diurnal regulation of isoflavone biosynthesis in soybean roots and the putative transporter genes responsible for isoflavone and soyasaponin transport.

Isoflavonoid biosynthesis in cultivated and wild soybeans grown in the field under adverse climate conditions

The cultivation of soybean plants is one of the most important crop production sectors in the world. Isoflavones are an important defence against pathogens in soybeans. The aim of the present study was to analyse isoflavone biosynthesis in wild and cultivated soybeans grown in the field conditions in an unfavourable climate. We analysed by LCMS-IT-TOF the composition and content of isoflavonoids, productivity and fungal disease resistance of wild and cultivated. The Hefeng25 and Sfera varieties have the highest isoflavonoid content and fungal tolerance. We have shown a 3-fold increase of total isoflavonoids in Sfera, comparing with wild type, and 4- and 7-fold increases of total isoflavone aglycones in Hefeng25 and Sfera, respectively. Accordingly, the expression of genes encoding enzymes of the isoflavonoid biosynthetic pathway was also maximal in these cultivars. Thus, biosynthetic status is an important indicator of soybean productivity and resistance to pathogens in adverse climates.

GABA mediates NO/cGMP/GSK-3-induced isoflavone accumulation in soybean sprouts

The effects of glycogen synthase kinase-3 (GSK-3) on nitric oxide (NO)/guanosine 3′,5′-cyclic monophosphate (cGMP)-activated γ-aminobutiric acid (GABA) accumulation and the role of GABA in isoflavone biosynthesis in response to UV-B exposure in soybean sprouts were investigated. The sprouts were sprayed using donors and inhibitors of cGMP and NO as well as inhibitors of GSK-3 and GABA. It turned out that GSK-3, as a second messenger, mediated NO/cGMP/PKG-induced GABA accumulation by activating diamine oxidase (DAO) under UV-B treatment. GABA mediated NO/cGMP/PKG/GSK-3-stimulated the increment of gene and protein expression and activity of chalcone synthase (CHS) and isoflavone synthase (IFS), and consequently induced the elevation in isoflavone content. Overall, GABA mediates NO/cGMP/PKG/GSK-3-stimulated isoflavone production in response to UV-B exposure in soybean sprouts.

Transcriptome sequencing and analysis reveals the molecular response to selenium stimuli in Pueraria lobata (willd.) Ohwi.

Pueraria lobata (willd.) Ohwi is a consumable selenium-enriched plant used for medicinal purposes. The molecular response to selenium (Se) stimuli in P. lobata is currently unknown. We used RNA-Seq to identify potential genes involved in selenite metabolism and analyzed their expression profiles. We obtained a total of 150,567 unigenes, of which 90,961 were annotated, including 16 structural genes, 14 sulfate transporters, and 13 phosphate transporters that may be involved in Se metabolism, and 33 candidate structural genes involved in isoflavone biosynthesis. The genes with a —foldchange— >2 and q value <0.05 after sodium selenite treatment were identified as differentially expressed genes (DEGs). We obtained a total of 4,246 DEGs, which were enriched in GO terms that included “response to stimulus”, “response to stress”, “signal transduction”, “response to abiotic stimulus”, and “response to chemical”. Of the 4,246 DEGs, one sulfate transporter and five phosphate transporter genes involved Se metabolism, and nine structural genes involved in isoflavone biosynthesis were up-regulated. The expression patterns of 10 DEGs were selected randomly and validated using qRT-PCR. The Pearson Correlation Coefficient (r) was 0.86, indicating the reliability of RNA-Seq results. 22 Reactive Oxygen Species (ROS) scavenging DEGs were found, 11 of which were up-regulated. 436, 624 transcription factors (TFs) correlated with structural genes were identified that may be involved in Se and isoflavone biosynthesis, respectively, using r (r > 0.7 or r < − 0.7). 556 TFs were related to at least one sulfate and phosphate transporter. Our results provided a comprehensive description of gene expression and regulation in response to Se stimuli in P. lobata.

Effect of biotic and abiotic elicitors on isoflavone biosynthesis during seed development and in suspension cultures of soybean (Glycine max L.).

The present investigation aimed to look at the effects of biotic and abiotic elicitors during Soybean seed development and cell suspension culture in isoflavones accumulation. The expression levels of four major genes viz., CHS7, CHS8, IFS2, and IFS1 involved on isoflavones biosynthesis during seed developmental stages from R5L-R7 was seen in both MAUS-2 and JS-335 Soybean varieties. The R7 stage showed 1.24-fold upregulation of IFS1transcript level and considered as the control for Soybean seed development. Both varieties during R6-R8 stages responded differently to the foliar application of 10µM SA, 10µMMJ and 0.1% Aspergillus niger. The IFS2 transcripts were upregulated by SA at the R7 stage with 5.21- and 4.68-fold in JS-335 and MAUS-2, respectively. IFS1 expression was significantly increased by A. niger treatment at R7 stage with 3.98- and 3.21-fold in MAUS-2 and JS-335, respectively. The expression of CHS7 and CHS8 by 10μM SA at R7 level revealed maximum up-regulation of 0.51- and 1.01-fold in MAUS-2; 0.37- and 0.82-fold in JS-335, respectively. In the soybean callus suspension culture, biosynthetic genes were used to validate the effects of elicitor on isoflavones. Both biotic and abiotic treatments contribute to the upregulation of IFS1 and IFS2 expression, that in turn, leads to the accumulation of isoflavone in seed development as well as in suspension cultures. These data further suggested that the IFS2 is the key gene responsible for the isoflavone accumulation during elicitor treatment.