Chalcone-flavanone Isomerase Research Articles

Consensus QTL map deciphered genes and pathways regulating tolerance to post‐flowering diseases in maize

AbstractPost‐flowering diseases (PFDs), such as ear rot, stalk rot and smut, affect maize yield and quality by damaging the reproductive organs, stalks and seeds. We hypothesized that quantitative trait loci (QTL) associated with different PFDs colocalize and share similar defence mechanisms. Hence, to find a stable consensus meta‐QTL (MQTL) for single or multiple PFDs, MQTL analysis was performed. QTL conferring resistance to PFD reported in 31 independent studies were collated to develop a consensus map. As many as 49 MQTL conferring PFD resistance were projected using appropriate algorithms. Most MQTL regions encompass genes encoding a wide range of defence‐related proteins. MQTL1.1 and MQTL10.5 included QTL/genes for resistance to all PFDs, which supported our hypothesis. Candidate genes for PFDs in MQTL7.1 were associated with pathogenesis‐related 1 protein and mitogen‐activated protein kinase (MAPK) signalling. MQTL5.2 encompassed chalcone flavanone isomerase and cinnamoyl coenzyme A (CoA) reductase genes involved in flavonoid and phenylpropanoid biosynthesis, respectively. Furthermore, MQTL10.4 was found to harbour genes encoding E3 ubiquitin ligase, WRKY‐TF11, calcium‐binding domains and zinc finger motifs. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis reiterated the role of genes within MQTL7.1 in the MAPK signalling pathway, phytohormone signal transduction and plant–pathogen interaction. Hence, we propose that these genes are potential candidates for PFD resistance. Furthermore, 75% of the genes within the MQTL showed orthology with sorghum and rice, indicating that these genes were conserved across species. The role of 27 MQTL, including the six most significant MQTL, was confirmed with reported genome‐wide association study (GWAS) results. Thus, the hotspots associated with PFDs identified in our study could be reliably used in marker‐assisted breeding for PFD resistance.

1H-NMR-based metabolomic profiling and proteomic analysis of soybean (Glycine max L.) in response to dicarboxylic acids (photon) application as a stress priming agent

Soybean (Glycinemax L.) serves not only as food for humans, animals, and industrial purposes, but is also a plant that can be used to comprehend molecular mechanisms occurring in stress response to various development techniques. To reveal the effect of applying dicarboxylic acids as stress priming agents on a metabolic level in soybean leaf extracts, the chemical profile of methanolic extracts were collected at different time points (1 h, 2 h, 12 h, 24 h, 1 week, 2 weeks and 3 weeks) after spraying were analyzed using 1H-NMR based metabolomics by way of PCA and OPLS-DA. The OPLS-DA revealed several metabolites, including organic acids (fumarate, citrate and malate) and amino acids (asparagine, alanine and GABA), which accumulated in higher amounts, with fumarate accumulating the highest in Glycinemax L. leaf extracts compared to untreated leaves. Denaturing 1DE gels were prepared for MS-based protein analysis and the presence of fatty acids (linolenic, oleic and α-linolenic acid) were confirmed by gas chromatography coupled with mass spectrometry (GC-MS), which served as jasmonic acid precursors. The MS-based profiling of proteins on the denaturing 1DE gels revealed several proteins that were differentiated between the treated and untreated leaf extracts. These proteins included ferritins, CaM, ferredoxin-thioredoxin reductase and chalcone-flavanone isomerase 1A. Following the treatment, fumarate was significantly elevated at 12 h to 3 weeks, compared to other compounds. It is, therefore, proposed that elevated quantities of fumarate could be related to the KEAP1-NRF2 metabolic pathway. This study represents the initial investigation of the effect of dicarboxylic acid application as a stress priming agent on Glycinemax L. using 1H-NMR metabolomic analysis, GC-MS and proteomic analysis.

Comparative Analysis of the Chalcone-Flavanone Isomerase Genes in Six Citrus Species and Their Expression Analysis in Sweet Orange (Citrus sinensis).

Citrus fruit contains rich nutrients which is edible and of officinal value. Citrus flavanones are widely used in the treatment of cardiovascular and other diseases, and they are a foundational material of Chinese medicine. The chalcone-flavanone isomerase (CHI) plays a key role in flavanone synthesis. Therefore, we comprehensively analyzed CHI genes in Citrus species. Here, thirty CHI genes were identified for the first time in six Citrus species, which were divided into CHI and FAP groups. Evolutionary analysis showed that CHI gene members were highly conserved and were an ancient family. All CsCHI genes showed the highest expression level after the second physiological fruit-falling period in C. sinensis. CsCHI1 and CsCHI3 were highly expressed at 50 days after the flowering (DAF) stage in albedo. The expression of CsFAP2 and CsCHI3 genes at the 50 DAF stage was 16.5 and 24.3 times higher than that at the 220 DAF stage, respectively. The expression of CsCHI1, CsCHI3, and CsFAP2 genes in the peel was higher than that in the pulp, especially in common sweet orange. The CsCHI3 gene maintained a high expression level in the epicarp and juice sac at all periods. The members of CHIs interacted with chalcone synthase (CHS), flavonol synthase/flavanone 3-hydroxylase (FLS) and naringenin, and 2-oxoglutarate 3-dioxygenase (F3H) to form heterodimers, which might together play a regulatory role and participate in the flavonoid pathway. This study will provide the basis for the selection of flavonoids in plant tissues and periods and fundamental information for further functional studies.

Metabolite Profiling and Transcriptome Analysis Explains Difference in Accumulation of Bioactive Constituents in Licorice (Glycyrrhiza uralensis) Under Salt Stress.

Salinity stress significantly affects the contents of bioactive constituents in licorice Glycyrrhiza uralensis. To elucidate the molecular mechanism underlying the difference in the accumulation of these constituents under sodium chloride (NaCl, salt) stress, licorice seedlings were treated with NaCl and then subjected to an integrated transcriptomic and metabolite profiling analysis. The transcriptomic analysis results identified 3,664 differentially expressed genes (DEGs) including transcription factor family MYB and basic helix-loop-helix (bHLH). Most DEGs were involved in flavonoid and terpenoid biosynthesis pathways. In addition, 121 compounds including a triterpenoid and five classes of flavonoids (isoflavone, flavone, flavanone, isoflavan, and chalcone) were identified, and their relative levels were compared between the stressed and control groups using data from the ultrafast liquid chromatography (UFLC)–triple quadrupole–time of flight–tandem mass spectrometry (TOF–MS/MS) analysis. Putative biosynthesis networks of the flavonoids and triterpenoids were created and combined with structural DEGs such as phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase [4CL], cinnamate 4-hydroxylase [C4H], chalcone synthase [CHS], chalcone-flavanone isomerase [CHI], and flavonoid-3′,5′ hydroxylase (F3′,5′H) for flavonoids, and CYP88D6 and CYP72A154 for glycyrrhizin biosynthesis. Notably, significant upregulation of UDP-glycosyltransferase genes (UGT) in salt-stressed licorice indicated that postmodification of glycosyltransferase may participate in downstream biosynthesis of flavonoid glycosides and triterpenoid saponins. Accordingly, the expression trend of the DEGs is positively correlated with the accumulation of glycosides. Our study findings indicate that key DEGs and crucial UGT genes co-regulate flavonoid and saponin biosynthesis in licorice under salt stress.

Nitrogen fixation and crop productivity enhancements co‐driven by intercrop root exudates and key rhizosphere bacteria

Abstract Crop diversity management is widely used to increase agricultural productivity and sustainability. Recent studies have demonstrated that maize root exudates can drive interspecific facilitation to enhance N2 fixation of bean in intercropping systems. However, the mechanisms of N2 fixation enhancement stimulated by root exudates in the intercropping systems remain unclear. Four experiments were designed to provide a progressively deeper understanding of how root exudates stimulate microbial‐mediated N2 fixation. First, the effects of faba bean/maize intercropping on yields and soil microbial communities were determined in a field experiment. Second, root‐derived interspecific facilitation was evaluated using a root partitioning approach. Third, the key microbial taxa in the faba bean rhizosphere were traced using 13C‐labelled maize root exudates. Fourth, the co‐driven mechanism of maize root exudates and micro‐organisms in the faba bean rhizosphere were explored. Faba bean/maize intercropping with maize residue return increased the yields of faba bean (26%), maize (27%) and broccoli (9.1%) compared to that under monocropping. Nodulin‐like 4 (NODL4), chalcone‐flavanone isomerase (CFI) and early nodulin‐like (ENODL2) gene expression in faba bean roots intercropped with maize increased by 1.5–2.3‐fold compared to that observed under monoculture. More than half of the N2 fixation of faba bean increase under intercropping was due to interactions with micro‐organisms. Nine key bacterial genera in the faba bean rhizosphere were identified by 13C‐DNA‐based stable isotope probing analysis. Among them Agromyces, Arthrobacter, Bacillus, Lysobacter and Paenibacillus directly fix N2, while Gemmatimonas, Heliobacillus, Natronocella and Sorangium increase the N2 fixation capacity of Azotobacter by providing additional carbon sources. These key bacteria triggered by maize root exudates played an important role in the rhizosphere facilitation of intercropping. Synthesis and applications. We demonstrated a novel root–root facilitation of N2 fixation and increased crop yields co‐driven by root exudates and rhizosphere bacteria under faba bean/maize intercropping, and nine key bacteria associated with this process were identified by 13C‐DNA‐based stable isotope probing. We recommend the adoption and optimization of intercropping systems with residue return to reduce the shortcomings of continuous cropping and to increase the sustainability of crop production.

Advances in the mechanism of cereal/legume intercropping promotion of symbiotic nitrogen fixation

Biological nitrogen fixation by rhizobia in symbiotic association with legumes is a valuable N source for agroecosystems, and thus it is crucial for increased sustainability of agricultural production. Previous studies found that legume and cereal intercropping can promote symbiotic nitrogen fixation mainly due to rhizosphere nitrogen depletion by the cereal and that soil mineral nitrogen is absorbed more by cereals than legumes, which forces the legume to increase its reliance on symbiotic nitrogen fixation. However, in addition to the nitrogen depletion theory in the rhizosphere, stoichiometry (such as nitrogen and phosphorus coupling) and molecular regulation mechanisms are also important in understanding legume nitrogen fixation. The legume usually maintains a stable stoichiometric relationship between nitrogen and phosphorus. Symbiotic nitrogen fixation of legumes tends to have a high demand for phosphorus, mainly because nitrogen fixation by legumes is a high energy-consuming process, and the required energy is directly provided by ATP. Therefore, when the phosphorus obtained by the intercropped legume is insufficient to meet the increase in symbiotic nitrogen fixation, legume growth is limited by phosphorus. Under phosphorus limitation, the phosphorus uptake strategy of the legume can be stimulated, i.e. by increasing the infection of arbuscular mycorrhizal fungi (AMF) and secretion of root exudates. The AMF mycelia of legumes can extend beyond the rhizosphere, linking the roots to the surrounding soil microhabitats and expanding the area of phosphorus absorbed by the roots. On the other hand, mobilizing rhizosphere insoluble phosphorus through root exudates is also an important pathway for improving phosphorus utilization. Increased soil phosphorus availability promotes plant growth and increased allocation of carbon (C) sources to roots and nodules, resulting in a larger root system or greater nodule formation, finally higher symbiotic nitrogen fixation. In addition, the formation of root nodules is influenced by rhizosphere talk between the cereal and legume, which are directly related to the level of nitrogen. A recent study found that under low N concentration conditions, cereal root exudates stimulated flavonoid biosynthesis and secretion partly through upregulating expression of the Chalcone-flavanone isomerase gene ( CFI ) in legumes. Meanwhile, an auxin-responsive GH3 family gene ( GH3.1 ) and the nodulation gene ENODL2 displayed significant increases, suggesting that nodulation continues under the regulation of auxin signaling and the nodulin-like protein. At nodule maturity, cereal root exudates continue to up-regulate key nodulation genes, such as NODL4 , ENOD93 and the N fixation gene FixI3 , finally promoting symbiotic nitrogen fixation. Moreover, a high concentration of soil mineral N significantly suppressed expression of the early nodulin gene NIN in Lotus corniculatus , which inhibited nodulation and symbiotic nitrogen fixation. In the cereal/legume intercropping system, rhizosphere nitrogen depletion can relieve the inhibition of nitrate on the expression of key nodulation and N fixation genes, thus promoting nodulation and symbiotic nitrogen fixation. In conclusion, intercropping can promote nitrogen uptake and optimize nitrogen use efficiency through the coupling effects of rhizosphere nitrogen depletion, phosphorus mobilization and the regulation of nitrogen fixation genes. This review will help improve our understanding of the mechanism of symbiotic nitrogen fixation in the cereal/legume intercropping system.

A Convenient Plant-Based Detection System to Monitor Androgenic Compound in the Environment.

Environmental androgen analogues act as endocrine disruptors, which inhibit the normal function of androgen in animals. In the present work, through the expression of a chimeric gene specified for the production of the anthocyanin in response to androgen DHT (dihydrotestosterone), we generated an indicator Arabidopsis that displays a red color in leaves in the presence of androgen compounds. This construct consists of a ligand-binding domain of the human androgen receptor gene and the poplar transcription factor gene PtrMYB119, which is involved in anthocyanin biosynthesis in poplar and Arabidopsis. The transgenic Arabidopsis XVA-PtrMYB119 displayed a red color in leaves in response to 10 ppm DHT, whereas it did not react in the presence of other androgenic compounds. The transcript level of PtrMYB119 peaked at day 13 of DHT exposure on agar media and then declined to its normal level at day 15. Expressions of anthocyanin biosynthesis genes including chalcone flavanone isomerase, chalcone synthase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, UFGT (UGT78D2), and anthocyanidin synthase were similar to that of PtrMYB119. It is assumed that this transgenic plant can be used by nonscientists for the detection of androgen DHT in the environment and samples such as food solution without any experimental procedures.

Organ-Specific Analysis of Morus alba Using a Gel-Free/Label-Free Proteomic Technique.

Morus alba is an important medicinal plant that is used to treat human diseases. The leaf, branch, and root of Morus can be applied as antidiabetic, antioxidant, and anti-inflammatory medicines, respectively. To explore the molecular mechanisms underlying the various pharmacological functions within different parts of Morus, organ-specific proteomics were performed. Protein profiles of the Morus leaf, branch, and root were determined using a gel-free/label-free proteomic technique. In the Morus leaf, branch, and root, a total of 492, 414, and 355 proteins were identified, respectively, including 84 common proteins. In leaf, the main function was related to protein degradation, photosynthesis, and redox ascorbate/glutathione metabolism. In branch, the main function was related to protein synthesis/degradation, stress, and redox ascorbate/glutathione metabolism. In root, the main function was related to protein synthesis/degradation, stress, and cell wall. Additionally, organ-specific metabolites and antioxidant activities were analyzed. These results revealed that flavonoids were highly accumulated in Morus root compared with the branch and leaf. Accordingly, two root-specific proteins named chalcone flavanone isomerase and flavonoid 3,5-hydroxylase were accumulated in the flavonoid pathway. Consistent with this finding, the content of the total flavonoids was higher in root compared to those detected in branch and leaf. These results suggest that the flavonoids in Morus root might be responsible for its biological activity and the root is the main part for flavonoid biosynthesis in Morus.

Development of bisphenol A (BPA)-sensing indicator Arabidopsis thaliana which synthesizes anthocyanin in response to BPA in leaves

Bisphenol A (BPA) is an estrogenic endocrine disruptor which disturbs a normal animal development. We generated an indicator plant that senses and provides a clear visual indicator of an estrogen-like compound BPA in the environment. We developed transgenic Arabidopsis lines expressing a construct designed to synthesize anthocyanin (thus showing a red color) in response to BPA. We transformed Arabidopsis with a recombinant vector containing the chimeric estrogen receptor (XVE region), LAP and coding region of PtrMYB119 (transcription factor involved in anthocyanin biosynthesis in poplar and Arabidopsis). Upon binding of the estrogen compound to the ligand-binding domain of E (estrogen receptor) in XVE, the XV domain binds to LAP promoter and triggering the transcription of PtrMYB119 with a subsequent enhancement of anthocyanin biosynthetic gene expression, resulting in anthocyanin synthesis. The leaves of the transgenic Arabidopsis line XVE-PtrMYB119 turned red in the presence of 10 ppm BPA. The transcript level of PtrMYB119 peaked at day 3 of BPA exposure, then decreased to its minimal level at day 5. Similar expression patterns to that of PtrMYB119 were detected for genes encoding the anthocyanin biosynthetic enzymes chalcone synthase, chalcone flavanone isomerase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, anthocyanidin synthase, and UFGT (UGT78D2). The leaves of transgenic plants did not turn red in response to BPA at concentrations below 10 ppm, but PtrMYB119 expression was induced by BPA at concentrations as low as 1 ppt BPA. Since this transgenic plant turns red in the presence of BPA without any experimental procedures, this line can be easily used by non-scientists.

Flavonoid biosynthesis controls fiber color in naturally colored cotton.

The existence of only natural brown and green cotton fibers (BCF and GCF, respectively), as well as poor fiber quality, limits the use of naturally colored cotton (Gossypium hirsutum L.). A better understanding of fiber pigment regulation is needed to surmount these obstacles. In this work, transcriptome analysis and quantitative reverse transcription PCR revealed that 13 and 9 phenylpropanoid (metabolic) pathway genes were enriched during pigment synthesis, while the differential expression of phenylpropanoid (metabolic) and flavonoid metabolic pathway genes occurred among BCF, GCF, and white cotton fibers (WCF). Silencing the chalcone flavanone isomerase gene in a BCF line resulted in three fiber phenotypes among offspring of the RNAi lines: BCF, almost WCF, and GCF. The lines with almost WCF suppressed chalcone flavanone isomerase, while the lines with GCF highly expressed the glucosyl transferase (3GT) gene. Overexpression of the Gh3GT or Arabidopsis thaliana 3GT gene in BCF lines resulted in GCF. Additionally, the phenylpropanoid and flavonoid metabolites of BCF and GCF were significantly higher than those of WCF as assessed by a metabolomics analysis. Thus, the flavonoid biosynthetic pathway controls both brown and green pigmentation processes. Like natural colored fibers, the transgenic colored fibers were weaker and shorter than WCF. This study shows the potential of flavonoid pathway modifications to alter cotton fibers’ color and quality.

Engineering co-culture system for production of apigetrin in Escherichia coli.

Microbial cells have extensively been utilized to produce value-added bioactive compounds. Based on advancement in protein engineering, DNA recombinant technology, genome engineering, and metabolic remodeling, the microbes can be re-engineered to produce industrially and medicinally important platform chemicals. The emergence of co-culture system which reduces the metabolic burden and allows parallel optimization of the engineered pathway in a modular fashion restricting the formation of undesired byproducts has become an alternative way to synthesize and produce bioactive compounds. In this study, we present genetically engineered E. coli-based co-culture system to the de novo synthesis of apigetrin (APG), an apigenin-7-O-β-D-glucopyranoside of apigenin. The culture system consists of an upstream module including 4-coumarate: CoA ligase (4CL), chalcone synthase, chalcone flavanone isomerase (CHS, CHI), and flavone synthase I (FNSI) to synthesize apigenin (API) from p-coumaric acid (PCA). Whereas, the downstream system contains a metabolizing module to enhance the production of UDP-glucose and expression of glycosyltransferase (PaGT3) to convert API into APG. To accomplish this improvement in titer, the initial inoculum ratio of strains for making the co-culture system, temperature, and media component was optimized. Following large-scale production, a yield of 38.5µM (16.6mg/L) of APG was achieved. In overall, this study provided an efficient tool to synthesize bioactive compounds in microbial cells.

The potassium channel FaTPK1 plays a critical role in fruit quality formation in strawberry (Fragaria × ananassa).

SummaryPotassium (K+), an abundant cation in plant cells, is important in fruit development and plant resistance. However, how cellular K+ is directed by potassium channels in fruit development and quality formation of strawberry (Fragaria × ananassa) is not yet fully clear. Here, a two‐pore K+ (TPK) channel gene in strawberry, FaTPK1, was cloned using reverse transcription–PCR. A green fluorescent protein subcellular localization analysis showed that FaTPK1 localized in the vacuole membrane. A transcription analysis indicated that the mRNA expression level of FaTPK1 increased rapidly and was maintained at a high level in ripened fruit, which was coupled with the fruit's red colour development, suggesting that FaTPK1 is related to fruit quality formation. The down‐ and up‐regulation of the FaTPK1 mRNA expression levels using RNA interference and overexpression, respectively, inhibited and promoted fruit ripening, respectively, as demonstrated by consistent changes in firmness and the contents of soluble sugars, anthocyanin and abscisic acid, as well as the transcript levels of ripening‐regulated genes PG1 (polygalacturonase), GAL6 (beta‐galactosidase), XYL2 (D‐xylulose reductase), SUT1 (sucrose transporter), CHS (chalcone synthase) and CHI (chalcone flavanone isomerase). Additionally, the regulatory changes influenced fruit resistance to Botrytis cinerea. An isothermal calorimetry analysis showed that the Escherichia coli‐expressed FaTPK1 recombinant protein could bind K+ with a binding constant of 2.1 × 10–3 m −1 and a dissociation constant of 476 μm. Thus, the strawberry TPK1 is a ubiquitously expressed, tonoplast‐localized two‐pore potassium channel that plays important roles in fruit ripening and quality formation.

Proteomic Characterization of Differential Abundant Proteins Accumulated between Lower and Upper Epidermises of Fleshy Scales in Onion (Allium cepa L.) Bulbs.

The onion (Allium cepa L.) is widely planted worldwide as a valuable vegetable crop. The scales of an onion bulb are a modified type of leaf. The one-layer-cell epidermis of onion scales is commonly used as a model experimental material in botany and molecular biology. The lower epidermis (LE) and upper epidermis (UE) of onion scales display obvious differences in microscopic structure, cell differentiation and pigment synthesis; however, associated proteomic differences are unclear. LE and UE can be easily sampled as single-layer-cell tissues for comparative proteomic analysis. In this study, a proteomic approach based on 2-DE and mass spectrometry (MS) was applied to compare LE and UE of fleshy scales from yellow and red onions. We identified 47 differential abundant protein spots (representing 31 unique proteins) between LE and UE in red and yellow onions. These proteins are mainly involved in pigment synthesis, stress response, and cell division. Particularly, the differentially accumulated chalcone-flavanone isomerase and flavone O-methyltransferase 1-like in LE may result in the differences in the onion scale color between red and yellow onions. Moreover, stress-related proteins abundantly accumulated in both LE and UE. In addition, the differential accumulation of UDP-arabinopyranose mutase 1-like protein and β-1,3-glucanase in the LE may be related to the different cell sizes between LE and UE of the two types of onion. The data derived from this study provides new insight into the differences in differentiation and developmental processes between onion epidermises. This study may also make a contribution to onion breeding, such as improving resistances and changing colors.

Root exudates drive interspecific facilitation by enhancing nodulation and N2 fixation

Plant diversity in experimental systems often enhances ecosystem productivity, but the mechanisms causing this overyielding are only partly understood. Intercropping faba beans (Vicia faba L.) and maize (Zea mays L.) result in overyielding and also, enhanced nodulation by faba beans. By using permeable and impermeable root barriers in a 2-y field experiment, we show that root-root interactions between faba bean and maize significantly increase both nodulation and symbiotic N2 fixation in intercropped faba bean. Furthermore, root exudates from maize promote faba bean nodulation, whereas root exudates from wheat and barley do not. Thus, a decline of soil nitrate concentrations caused by intercropped cereals is not the sole mechanism for maize promoting faba bean nodulation. Intercropped maize also caused a twofold increase in exudation of flavonoids (signaling compounds for rhizobia) in the systems. Roots of faba bean treated with maize root exudates exhibited an immediate 11-fold increase in the expression of chalcone-flavanone isomerase (involved in flavonoid synthesis) gene together with a significantly increased expression of genes mediating nodulation and auxin response. After 35 d, faba beans treated with maize root exudate continued to show up-regulation of key nodulation genes, such as early nodulin 93 (ENOD93), and promoted nitrogen fixation. Our results reveal a mechanism for how intercropped maize promotes nitrogen fixation of faba bean, where maize root exudates promote flavonoid synthesis in faba bean, increase nodulation, and stimulate nitrogen fixation after enhanced gene expression. These results indicate facilitative root-root interactions and provide a mechanism for a positive relationship between species diversity and ecosystem productivity.

The B-, G- and S-genomic Chi genes in family Triticeae

As result of a close evolutionary relationship between Triticeae B, G, and S genomes, the exchange of genetic material between them is possible and may be beneficial for broadening the genetic diversity of cultivated bread wheat. However, the extent to which regulatory networks are conserved remains poorly researched. Here, the structural organization and transcriptional activity of the B, S, and G genome copies of a gene encoding flavonoid biosynthesis enzyme chalcone-flavanone isomerase (CHI) were explored using introgression lines which differ from the wild type by carrying a non-bread wheat Chi-1 gene. Chi-S1, Chi-G1, and Chi-B1 all mapped to a comparable region of chromosomes 5S, 5G, and 5B, respectively. Nucleotide sequences of Aegilops speltoides Chi-S1 and Triticum timopheevii Chi-G1 were determined and compared with T. aestivum Chi-B1 sequences. The enzymes encoded by these three genes shared the same predicted tertiary structure and active sites. However, the replacement of Chi-B1 by Chi-S1 or Chi-G1 in a wheat background resulted in a significant decrease in the global amount of the Chi-1 transcript present in the seedling shoot indicating divergence in regulation of expression of the orthologous Chi-1 genes among Triticeae ssp.

Defence strategies adopted by the medicinal plant Coleus forskohlii against supplemental ultraviolet-B radiation: Augmentation of secondary metabolites and antioxidants

Supplementary ultraviolet-B (ambient+3.6 kJ m−2 day−1) induced changes on morphological, physiological, and biochemical characteristics (specifically the defence strategies: UV-B protective compounds and antioxidants) of Coleus forskohlii were investigated under field conditions at 30, 60, and 90 days after transplantation. Levels of secondary metabolites increased under s-UV-B stress; flavonoids and phenolics (primary UV-B screening agents) were recorded to be higher in leaves which are directly exposed to s-UV-B. This was also verified by enhanced activities of phenylpropanoid pathway enzymes: phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), 4-coumarate-CoA ligase (4CL), chalcone–flavanone isomerase (CHI), and dihydroflavonol reductase (DFR). Antioxidants, both enzymatic (ascorbate peroxidase, catalase, glutathione reductase, peroxidase, polyphenol oxidase, and superoxide dismutase) and non-enzymatic (ascorbic acid and α-tocopherol) also increased in the treated organs of the test plant, higher contents being recorded in roots except for ascorbic acid. On the contrary, protein and chlorophyll content (directly implicated in regulating plant growth and development) declined under s-UV-B. These alterations in plant biochemistry led the plant to compromise on its photosynthate allocation towards growth and biomass production as evidenced by a reduction in its height and biomass. The study concludes that s-UV-B is a potent stimulating factor in increasing the concentrations of defense compounds and antioxidants in C. forskohlii to optimize its performance under stress.

Polysaccharide elicitors enhance phenylpropanoid and naphtodianthrone production in cell suspension cultures of Hypericum perforatum

Hypericum perforatum cell suspensions were evaluated for their growth, phenylpropanoid and naphtodianthrone productions, and antioxidant activity after treatments with polysaccharide elicitors chitin (CHI), pectin (PEC) and dextran (DEX). Polysaccharide elicitation of cell cultures showed a considerable improving effect on phenylpropanoid production (phenolics, flavonoids, flavanols and anthocyanins) during the post-elicitation period. Even that enhanced phenylpropanoid production was related to growth suppression of CHI elicited cells, PEC and DEX were presented as effective elicitors without loss of cell biomass. Phenylpropanoid accumulation in elicited cells was correlated to markedly higher enzyme activities of phenylalanine ammonia lyase and chalcone-flavanone isomerase. Cell cultures were found to respond rapidly towards the applied elicitors through an early stimulation of both naphtodiantrones, hypericin (HYP) and pseudohypericin (PHYP). The CHI was proposed as a much more prominent elicitor in stimulation of HYP and PHYP levels in cell suspensions. All tested polysaccharide elicitors stimulated non-enzymatic antioxidant properties in cell extracts indicating the potential role of phenolic compounds as effective antioxidants. With regards to the antioxidant enzymes, an up-regulation of peroxidase and catalase activities was observed in PEC and DEX elicited cells, whereas CHI did not give any stimulatory effect on enzymatic activities. These findings suggest the involvement of an efficient antioxidant defense system in the adaptive response of cells to polysaccharide elicitation. Altogether, these results indicated that H. perforatum cells elicited with polysaccharides represent a promising experimental system for enhanced production of phenylpropanoids and naphtodianthrones.

Marker-assisted development of bread wheat near-isogenic lines carrying various combinations of purple pericarp (Pp) alleles

The commercial interest in pigmented wheat grain flows from an understanding that they are nutritionally superior to white kernels. The pigment of purple coloured bread and durum wheat grains results from the accumulation of anthocyanins in the pericarp; its genetic basis is the action of Pp-1 and Pp3 genes. Here, the development of a set of bread wheat near isogenic lines (NILs) carrying various combinations of Pp alleles is described, along with a demonstration of their utility for the genetic dissection of the purple pericarp trait. A marker-assisted backcrossing strategy was based on the use of microsatellite markers linked to Pp3 (chromosome 2A), Pp-A1 (7A) and Pp-D1 (7D). Pp-A1 is a newly uncovered gene of weak effect. A qRT-PCR-based analysis of the anthocyanin synthesis structural genes [Chi (chalcone-flavanone isomerase) and F3h (flavanone 3-hydroxylase)] transcript abundance in the pericarp of the NILs suggested that the Pp genes up-regulate their transcription in contrasting ways. These NILs represent a resource for studying the effect of grain pigmentation on other wheat traits and end products.

Transcriptome profiling of the UV-B stress response in the desert shrub Lycium ruthenicum.

Ultraviolet-B (UV-B) is a natural component of the solar radiation. Due to its high energy, low dosages of UV-B can bring huge potential damage effect to organisms. Despite much research that has analyzed the gene expression changes of plants that under UV-B radiation, the transcriptome response of Lycium ruthenicum under the UV-B induction is still un-available. The aim of our study was to identify UV-B responsive genes and gain an insight into the underlying genetic basis of the pathobiology of UV-B related damage. We collected leaf samples from L. ruthenicum with and without UV-B exposure, and then performed a transcriptome profiling to comprehensively investigate their expression signatures. By employing the high throughput RNA-sequencing analysis of samples with and without UV-B exposure, we identified 1,913 up-regulated and 536 down-regulated genes at least by twofold changes. The activity of antioxidant enzyme related genes, including the superoxide dismutase, was decreased, genes related to the synthesis of secondary metabolites and defense responses, such as cinnamyl alcohol dehydrogenase, chalcone-flavanone isomerase and dihydroflavonol reductase were also downregulated. The expression patterns of 14 randomly selected genes resulted from quantitative real-time PCR were basically consistent with their transcript abundance changes identified by RNA-sequencing. We found that several biological pathways related to biotic and abiotic stresses, including cell defense, photosynthesis processes, energy metabolism, were involved in the process of UV-B stress response. A genome-wide screening of gene deregulation under UV-B induction would provide an insight into the understanding of the molecular bases and pathogenesis of UV-B responses.

Secondary metabolites and phenylpropanoid pathway enzymes as influenced under supplemental ultraviolet-B radiation in Withania somnifera Dunal, an indigenous medicinal plant

The present study aims to investigate the effects of supplemental ultraviolet B (3.6kJm−2day−1 above ambient) radiation on secondary metabolites and phenylpropanoid pathway enzymes of Withania somnifera under field conditions at 40, 70, and 100days after transplantation. Secondary metabolites’ (alkaloids, anthocyanins, carotenoids, flavonoids, lignin, phytosterols, saponins, and tannins) concentrations were analysed at the end of the treatments. Activities of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), 4-coumarate-CoA ligase (4CL), chalcone–flavanone isomerase (CHI), and dihydroflavonol reductase (DFR) were also determined. In treated plants, secondary metabolite-concentrations generally increased (higher concentrations being recorded in roots compared to leaves). Anomalies were recorded for lycopene in roots and phytosterols in leaves (all sampling ages); β-carotene declined in leaves at third sampling age. s-UV-B-treated plants depicted decrease in withanolide A content with concomitant increase in withaferin A (two major alkaloids analysed by HPLC) compared to their respective controls. Phenylpropanoid pathway enzyme-activities increased in leaves and roots under s-UV-B treatment, the latter showing greater increase. The study concludes that s-UV-B is a potent factor in increasing the concentrations of secondary metabolites and their biosynthetic pathway enzymes in W. somnifera.