Flavonoid Biosynthesis In Plants Research Articles

A R2R3-MYB transcription factor, GmMYB12B2, affects the expression levels of flavonoid biosynthesis genes encoding key enzymes in transgenic Arabidopsis plants

Isoflavones play diverse roles in plant–microbe interactions and are potentially important for human nutrition and health. To study the regulation of isoflavonoid synthesis in soybean, the R2R3-MYB transcription factor GmMYB12B2 was isolated and characterized. Yeast expression experiments demonstrated that GmMYB12B2 showed transcriptional activity. GmMYB12B2 was localized in the nucleus when it was transiently expressed in onion epidermal cells. Real-time quantitative PCR analysis revealed that GmMYB12B2 transcription was increased in roots and mature seeds compared with other organs. The gene expression level in immature embryos was consistent with the accumulation of isoflavones. CHS8 is a key enzyme in plant flavonoid biosynthesis. Transient expression experiments in soybean calli demonstrated that CHS8 was regulated by GmMYB12B2 and produced more fluorescence. The expression levels of some key enzymes in flavonoid biosynthesis were examined in transgenic Arabidopsis lines. The results showed that the expression levels of PAL1, CHS and FLS in transgenic plants were significantly higher than those in wild type plants. However, the expression level of DFR was lower, and the expression levels of CHI, F3H and F3′H were the same in all lines. GmMYB12B2 expression caused a constitutive increase in the accumulation of flavonoids in transgenic Arabidopsis lines compared with wild type plants.

Increasing Plant Flavonoid Biomaterials in Response to UV-A Light

Flavonoid biomaterials have a protecting function from various stresses. We examined the flavonoid biosynthesis in plant treated under visible light (VL) and additional UV-A light. The transgenic tobacco containing PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) cDNA, involved in flavonoid biosynthesis from Arabidopsis thaliana, were used for studying the flavonoid biosynthesis under both light conditions comparing to non transgenic tobacco. The flavonoid biomaterials were extracted with acidic methanol and water solvent from treated plant leaves. The absorbance of each biomaterial in the extract was measured under specific wavelength using a spectrophotometer. Additional UV-A radiated to non transgenic and transgenic tobacco affect the increasing of p-coumaric acid, naringenin, apigenin and kaempherol biomaterials from themselves grown under VL (approximately 120-130%). However, PAP1 transgenic tobaccos under additional UV-A radiation enhance the accumulation of these biomaterials up to160-180% higher than non transgenic tobaccos grown under VL condition. Moreover, PAP1 transgenic tobacco radiated with UVA light also significantly increased pelargonidin biomaterial. PAP1 transgenic tobaccos had a similar phenotype with non transgenic tobaccos but the color of fully expanding flower was more pink intensity than non transgenic.

Metabolic engineering of flavonoids in plants and microorganisms

Over 9,000 flavonoid compounds have been found in various plants, comprising one of the largest families of natural products. Flavonoids are an essential factor in plant interactions with the environment, often serving as the first line of defense against UV irradiation and pathogen attacks. Flavonoids are also major nutritional compounds in foods and beverages, with demonstrated health benefits. Some flavonoids are potent antioxidants, and specific flavonoid compounds are beneficial in many physiological and pharmacological processes. Therefore, engineering of flavonoid biosynthesis in plants or in microorganisms has significant scientific and economical importance. Construction of biosynthetic pathways in heterologous systems offers promising results for large-scale flavonoid production by fermentation or bioconversion. Genomics and metabolomics now offer unprecedented tools for detailed understanding of the engineered transgenic organism and for developing novel technologies to further increase flavonoid production yields. We summarize some of the recent metabolic engineering strategies in plants and microorganisms, with a focus on applications of metabolic flux analysis. We are confident that these engineering approaches will lead to successful industrial flavonoid production in the near future.

Effect of latitude on flavonoid biosynthesis in plants

The growth conditions in different latitudes vary markedly with season, day length, light quality and temperature. Many plant species have adapted well to the distinct environments through different strategies, one of which is the production of additional secondary metabolites. Flavonoids are a widely spread group of plant secondary metabolites that are involved in many crucial functions of plants. Our understanding of the biosynthesis, occurrence and function of flavonoids has increased rapidly in recent decades. Numerous studies have been published on the influence of environmental factors on the biosynthesis of flavonoids. However, extensive long-term studies that examine the effect of the characteristics of northern climates on flavonoid biosynthesis are still scarce. This review focuses on the current knowledge about the effect of light intensity, photoperiod and temperature on the gene-environment interaction related to flavonoid biosynthesis in plants.

Relationship between homoeologous regulatory and structural genes in allopolyploid genome – A case study in bread wheat

BackgroundThe patterns of expression of homoeologous genes in hexaploid bread wheat have been intensively studied in recent years, but the interaction between structural genes and their homoeologous regulatory genes remained unclear. The question was as to whether, in an allopolyploid, this interaction is genome-specific, or whether regulation cuts across genomes. The aim of the present study was cloning, sequence analysis, mapping and expression analysis of F3H (flavanone 3-hydroxylase – one of the key enzymes in the plant flavonoid biosynthesis pathway) homoeologues in bread wheat and study of the interaction between F3H and their regulatory genes homoeologues – Rc (red coleoptiles).ResultsPCR-based cloning of F3H sequences from hexaploid bread wheat (Triticum aestivum L.), a wild tetraploid wheat (T. timopheevii) and their putative diploid progenitors was employed to localize, physically map and analyse the expression of four distinct bread wheat F3H copies. Three of these form a homoeologous set, mapping to the chromosomes of homoeologous group 2; they are highly similar to one another at the structural and functional levels. However, the fourth copy is less homologous, and was not expressed in anthocyanin pigmented coleoptiles. The presence of dominant alleles at the Rc-1 homoeologous loci, which are responsible for anthocyanin pigmentation in the coleoptile, was correlated with F3H expression in pigmented coleoptiles. Each dominant Rc-1 allele affected the expression of the three F3H homoeologues equally, but the level of F3H expression was dependent on the identity of the dominant Rc-1 allele present. Thus, the homoeologous Rc-1 genes contribute more to functional divergence than do the structural F3H genes.ConclusionThe lack of any genome-specific relationship between F3H-1 and Rc-1 implies an integrative evolutionary process among the three diploid genomes, following the formation of hexaploid wheat. Regulatory genes probably contribute more to the functional divergence between the wheat genomes than do the structural genes themselves. This is in line with the growing consensus which suggests that although heritable morphological traits are determined by the expression of structural genes, it is the regulatory genes which are the prime determinants of allelic identity.

Functional analysis of soybean genes involved in flavonoid biosynthesis by virus‐induced gene silencing

Virus-induced gene silencing (VIGS) is a powerful tool for functional analysis of genes in plants. A wide-host-range VIGS vector, which was developed based on the Cucumber mosaic virus (CMV), was tested for its ability to silence endogenous genes involved in flavonoid biosynthesis in soybean. Symptomless infection was established using a pseudorecombinant virus, which enabled detection of specific changes in metabolite content by VIGS. It has been demonstrated that the yellow seed coat phenotype of various cultivated soybean lines that lack anthocyanin pigmentation is induced by natural degradation of chalcone synthase (CHS) mRNA. When soybean plants with brown seed coats were infected with a virus that contains the CHS gene sequence, the colour of the seed coats changed to yellow, which indicates that the naturally occurring RNA silencing is reproduced by VIGS. In addition, CHS VIGS consequently led to a decrease in isoflavone content in seeds. VIGS was also tested on the putative flavonoid 3'-hydroxylase (F3'H) gene in the pathway. This experiment resulted in a decrease in the content of quercetin relative to kaempferol in the upper leaves after viral infection, which suggests that the putative gene actually encodes the F3'H protein. In both experiments, a marked decrease in the target mRNA and accumulation of short interfering RNAs were detected, indicating that sequence-specific mRNA degradation was induced. The present report is a successful demonstration of the application of VIGS for genes involved in flavonoid biosynthesis in plants; the CMV-based VIGS system provides an efficient tool for functional analysis of soybean genes.

Role of anthocyanidin reductase, encoded by BANYULS in plant flavonoid biosynthesis.

Condensed tannins (CTs) are flavonoid oligomers, many of which have beneficial effects on animal and human health. The flavanol (-)-epicatechin is a component of many CTs and contributes to flavor and astringency in tea and wine. We show that the BANYULS (BAN) genes from Arabidopsis thaliana and Medicago truncatula encode anthocyanidin reductase, which converts anthocyanidins to their corresponding 2,3-cis-flavan-3-ols. Ectopic expression of BAN in tobacco flower petals and Arabidopsis leaves results in loss of anthocyanins and accumulation of CTs.

Localized Changes in Flavonoid Biosynthesis in Roots of Lotus pedunculatus after Infection by Rhizobium loti

Two-dimensional paper chromatography in four solvent systems, high-sensitivity spray reagents, and UV absorption spectroscopy were used to separate and characterize flavonoids and isoflavonoids in roots and root nodules of 20-d-old Lotus pedunculatus Cav. Seedlings were grown either under sterile conditions or after inoculation with Fix(+) or Fix(-) strains of Rhizobium loti. Flavonoids rather than isoflavonoids predominated in all tissues. Flavonoid profiles in sterile and denodulated root tissues were remarkably similar, both qualitatively and quantitatively. At least 14 partially purified flavonoid aglycones and conjugates were found in root extracts; denodulated root tissues contained no compounds that were not also present in sterile roots. Fix(+) rhizobia were responsible for major postinfection shifts in plant flavonoid biosynthesis at the sites of nodule morphogenesis. Polymeric flavolans were absent from Fix(+) nodules but present in all root tissues and in Fix(-) nodules. Catechin was detected only in Fix(+) nodules.