Expression Of Phenylpropanoid Pathway Genes Research Articles

Expression Analysis of Phenylpropanoid Pathway Genes and Phenylpropanoid Accumulation in Different Organs of Chelidonium majus L.

Objective This study set out to examine the phenylpropanoid pathway gene expression and phenolic compound accumulation in different organs of Chelidonium majus (C. majus). Methods Total RNA was extracted and expression of phenylpropanoid biosynthesis genes was assessed using quantitative real-time polymerase chain reaction in different organs of C. majus. In addition, the phenylpropanoid compounds were analyzed from different organs of C. majus by using high-performance liquid chromatography. Results The results demonstrate the variance in the expression levels of phenylpropanoid pathway genes in the flowers, leaves, stems, and roots of C. majus. CmPAL and CmC4H were expressed at their greatest levels in the stem, while CmCHS and CmCHI were strongly expressed in the leaf, and CmF3H, CmFLS, Cm4CL, and CmDFR were more abundant in the flower. The total phenolic contents in the different organs also varied significantly. The highest value was recorded in the flowers (2152.89 ± 78.71 μg/g dry wt.), which was 54.26 times higher than the lowest values obtained from roots (39.68 ± 0.05 μg/g dry wt.). Except for roots, rutin showed the highest level in all other organs. According to a heat map, the majority of the phenolic compounds were highest in the flower. By using variable importance in projection analysis, the most significant metabolite was found to be chlorogenic acid, which had a positive association with all phenolic chemicals, but it showed a negative correlation with trans-cinnamic acid. Except for trans-cinnamic acid, all other phenolic compounds likewise showed positive correlation with other phenolic compounds. Conclusions This work offers pertinent data on the expression analysis of genes involved in the phenylpropanoid biosynthetic pathway and the presence of phenolic compounds in C. majus. These findings will be helpful for the researcher to understand the relationship between the expression level of phenylpropanoid pathway genes and phenylpropanoid compound accumulation in C. majus.

Global changes in metabolic pathways in endocarp of ‘Dayagan’ hybrid citrus fruit during segment drying revealed by widely targeted metabolomics and transcriptomics analysis

Segment drying is a severe physiological disorder of citrus fruit that occurs during on-tree and postharvest storage. Although many research have focused on the analysis of metabolic pathways in juice vesicles during segment drying, the associated changes in endocarp are poorly understood. Physiological analysis, widely targeted metabolomics, and transcriptomics were utilized to systematically compare the changes in endocarp between normal and disordered fruit affected by segment drying in ‘Dayagan’ hybrid citrus. Physiological analysis showed that segment drying was associated with modification of phenotype, increase in cell wall components contents, and decrease in pectin degradation enzymes activities. Metabolomic profiling identified a total of 951 differentially accumulated metabolites upon segment drying. These metabolites were enriched in metabolic pathways relating to the synthesis of amino acids and derivatives, lipids, and flavonoids. Transcriptomics analysis revealed 6103 genes that were differentially expressed during segment drying, with increased expression of phenylpropanoid pathway genes and cell wall synthesis genes, while decreased expression of genes involved in pectin degradation. The global metabolic pathways that have been revealed suggest that the alteration of endocarp may be related to the water deficit stress in the pulp during segment drying. Finally, we carried out in vitro and in vivo assays, which proved that the endocarp could impede water loss of the pulp during segment drying. The present study is the first to offer insights into the metabolic changes that occur in the endocarp during segment drying, further indicating that water loss may be a key early event of the segment drying process.

Temporal transcriptome of tomato elucidates the signaling pathways of induced systemic resistance and systemic acquired resistance activated by Chaetomium globosum.

C. globosum is an endophytic fungus, which is recorded effective against several fungal and bacterial diseases in plants. The exclusively induce defense as mechanism of biocontrol for C. globosum against phyto-pathogens is reported. Our pervious study states the effectiveness of induced defense by C. globosum (Cg), in tomato against Alternaria solani. In this study the temporal transcriptome analysis of tomato plants after treatment with C. globosum was performed for time points at 0 hpCi, 12 hpCi, 24 hpCi and 96 phCi. The temporal expression analysis of genes belonging to defense signaling pathways indicates the maximum expression of genes at 12h post Cg inoculation. The sequential progression in JA signaling pathway is marked by upregulation of downstream genes (Solyc10g011660, Solyc01g005440) of JA signaling at 24 hpCi and continued to express at same level upto 96 hpCi. However, the NPR1 (Solyc07g040690), the key regulator of SA signaling is activated at 12h and repressed in later stages. The sequential expression of phenylpropanoid pathway genes (Solyc09g007920, Solyc12g011330, Solyc05g047530) marks the activation of pathway with course of time after Cg treatment that results in lignin formation. The plant defense signaling progresses in sequential manner with time course after Cg treatment. The results revealed the involvement of signaling pathways of ISR and SAR in systemic resistance induced by Cg in tomato, but with temporal variation.

Increased phenylpropanoids production in UV-B irradiated Salvia verticillata as a consequence of altered genes expression in young leaves

Ultraviolet-B (UV–B) radiation as an environmental potential elicitor induces the synthesis of plant secondary metabolites. The effects of UV-B radiation on photosynthetic pigments and dry weight, biochemical and molecular features of old and young leaves of Salvia verticillata were investigated. Plants were exposed to 10.97 kJ m−2 day−1 of biologically effective UV-B radiation for up to 10 days. The sampling process was performed in four steps: 1, 5, 10, and 13 days (recovery time) after the start of irradiation. As a result of plant investment in primary and secondary metabolism, the production of phenolic compounds increased, while chlorophyll levels and leaf dry weight (%) declined. Under long-term UV-B exposure, young leaves exhibited the most significant reduction in chlorophyll a and b content and leaf dry weight. The highest level of total phenol (1.34-fold) and flavonoid concentration (2-fold) relative to the control was observed on the 5th day and recovery time, respectively. Young leaves demonstrated the highest amount of phenolic acids in recovery time. Young leaves on the 5th day of the experiment exerted the highest level of antioxidant activity when compared to the control. A positive correlation was observed between antioxidant activity and the amount of phenolic compounds. Regarding the expression of phenylpropanoid pathway genes, UV-B enhanced the expression of phenylalanine ammonia-lyase, tyrosine aminotransferase, and rosmarinic acid synthase with the highest level in young leaves on the 10th day. Overall, young leaves of S. verticillata indicated higher sensitivity to UV-B radiation and developed more tangible reactions to such radiation.

Postharvest UV-C irradiation increased the flavonoids and anthocyanins accumulation, phenylpropanoid pathway gene expression, and antioxidant activity in sweet cherries (Prunus avium L.)

Ultraviolet (UV) influences postharvest changes in secondary metabolites of fruit. In this study, sweet cherries were treated with UV-C irradiation (1.05, 2.10, and 4.20 kJ/m2) for different treatment periods, they were stored at room temperature (25.0 ± 2.0 ℃) for 6 days. The results showed that the total phenolics, flavonoids, and anthocyanins increased in response to UV-C during storage. UPLC-ESI-MS analysis indicated that the individual flavonoids and anthocyanins contents increased, including cyanidin 3-O-galactoside, cyanidin O-syringic acid, cyanidin 3-O-glucoside, pelargonidin 3-O-glucoside, tricin 5-O-hexoside, luteolin C-hexoside, Di-O-methylquercetin, and naringenin-7-O-glucoside. Furthermore, UV-C (2.10 kJ/m2) treatment upregulated the enzymes activity and genes expression of phenylpropanoid pathway. Highly significant correlations were found among flavonoids, anthocyanins, phenylalanine ammonia lyase (PAL) and genes expression of ANS, DFR, UFGT (r > 0.6, p < 0.01) and cinnamate 4-hydroxylase (C4H) and PAL, C4H, 4CL, CHI, ANS, DFR expressions (r> 0.6, p < 0.01). These results contribute to illustrating the molecular mechanism of flavonoids and anthocyanins biosynthesis under the UV-C irradiation in sweet cherries and promote the development of postharvest in agriculture.

Changes in phenylpropanoid pathway gene expression in roots and leaves of susceptible and resistant Brassica napus lines in response to Plasmodiophora brassicae inoculation

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is currently one of the most severe diseases of the Brassicaceae. As a soil-borne pathogen, P. brassicae infects the roots of host plants, inducing the formation of galls and negatively affecting plant growth and productivity. Roots and leaves of two clubroot susceptible and one resistant Brassica napus lines, inoculated with P. brassicae spores, were monitored for disease progression at four time points after inoculation. Quantitative gene expression analysis of phenylpropanoid pathway genes in these tissues showed increased expression of several genes involved in lignin biosynthesis in response to infection. In leaves, a higher expression of genes involved in flavonoid biosynthesis was also observed. In both roots and leaves of the resistant line, phenylpropanoid pathway genes were upregulated at more time points and at higher levels than in susceptible lines. The differential expression of phenylpropanoid pathway genes between resistant and susceptible lines, suggests that this pathway is instrumental in resistance to clubroot disease progression in the resistant line.

Aspen phenylpropanoid genes' expression levels correlate with genets' tannin richness and vary both in responses to soil nitrogen and associations with phenolic profiles.

Condensed tannin (CT) contents of European aspen (Populus tremula L.) vary among genotypes, and increases in nitrogen (N) availability generally reduce plants' tannin production in favor of growth, through poorly understood mechanisms. We hypothesized that intrinsic tannin production rates may co-vary with gene expression responses to soil N and resource allocation within the phenylpropanoid pathway (PPP). Thus, we examined correlations between soil N levels and both expression patterns of eight PPP genes (measured by quantitative-reverse transcription PCR) and foliar phenolic compounds (measured by liquid chromatography-mass spectrometry) in young aspen genets with intrinsically extreme CT levels. Monitored phenolics included salicinoids, lignins, flavones, flavonols, CT precursors and CTs. The PPP genes were consistently expressed more strongly in high-CT trees. Low N supplements reduced expression of genes throughout the PPP in all genets, while high N doses restored expression of genes at the beginning and end of the pathway. These PPP changes were not reflected in pools of tannin precursors, but varying correlations between gene expression and foliar phenolic pools were detected in young and mature leaves, suggesting that processes linking gene expression and the resulting phenolics vary spatially and temporally. Precursor fluxes suggested that CT-related metabolic rate or sink controls are linked to intrinsic carbon allocation strategies associated with N responses. Overall, we found more negative correlations (indicative of allocation trade-offs) between PPP gene expression and phenolic products following N additions in low-CT plants than in high-CT plants. The tannin-related expression dynamics suggest that, in addition to defense, relative tannin levels may also be indicative of intraspecific variations in the way aspen genets respond to soil fertility.

Elucidation of defense-related signaling responses to spot blotch infection in bread wheat (Triticum aestivum L.).

Spot blotch disease, caused by Bipolaris sorokiniana, is an important threat to wheat, causing an annual loss of ~17%. Under epidemic conditions, these losses may be 100%, yet the molecular responses of wheat to spot blotch remain almost uncharacterized. Moreover, defense-related phytohormone signaling genes have been poorly characterized in wheat. Here, we have identified 18 central components of salicylic acid (SA), jasmonic acid (JA), ethylene (ET), and enhanced disease susceptibility 1 (EDS1) signaling pathways as well as the genes of the phenylpropanoid pathway in wheat. In time-course experiments, we characterized the reprogramming of expression of these pathways in two contrasting genotypes: Yangmai #6 (resistant to spot blotch) and Sonalika (susceptible to spot blotch). We further evaluated the performance of a population of recombinant inbred lines (RILs) by crossing Yangmai#6 and Sonalika (parents) and subsequent selfing to F10 under field conditions in trials at multiple locations. We characterized the reprogramming of defense-related signaling in these RILs as a consequence of spot blotch attack. During resistance to spot blotch attack, wheat strongly elicits SA signaling (SA biogenesis as well as the NPR1-dependent signaling pathway), along with WRKY33 transcription factor, followed by an enhanced expression of phenylpropanoid pathway genes. These may lead to accumulation of phenolics-based defense metabolites that may render resistance against spot blotch. JA signaling may synergistically contribute to the resistance. Failure to elicit SA (and possibly JA) signaling may lead to susceptibility against spot blotch infection in wheat.

AtMYB12 expression in tomato leads to large scale differential modulation in transcriptome and flavonoid content in leaf and fruit tissues.

Plants synthesize secondary metabolites, including flavonoids, which play important role during various stresses for their survival. These metabolites are also considered as health-protective components in functional foods. Flavonols, one of the important groups of flavonoids, apart from performing several roles in plants have been recognized as potent phytoceuticals for human health. Tomato fruits are deficient in this group of flavonoids and have been an important target for enhancing the accumulation of flavonols through genetic manipulations. In the present study, AtMYB12 transcription factor of the Arabidopsis has been expressed under constitutive promoter in tomato. Transgenic tomato lines exhibited enhanced accumulation of flavonols and chlorogenic acid (CGA) in leaf and fruit accompanied with elevated expression of phenylpropanoid pathway genes involved in flavonol biosynthesis. In addition, global gene expression analysis in leaf and fruit suggested that AtMYB12 modulates number of molecular processes including aromatic amino acid biosynthesis, phytohormone signaling and stress responses. Besides this, a differential modulation of the genes in fruits and leaves is reported in this study. Taken together, results demonstrate that modulation of primary carbon metabolism and other pathways by AtMYB12 in tomato may lead to sufficient substrate supply for enhanced content of phenolics in general and flavonols in particular.

An R2R3-MYB Transcription Factor Regulates Eugenol Production in Ripe Strawberry Fruit Receptacles.

Eugenol is a volatile phenylpropanoid that contributes to flower and ripe fruit scent. In ripe strawberry (Fragaria × ananassa) fruit receptacles, eugenol is biosynthesized by eugenol synthase (FaEGS2). However, the transcriptional regulation of this process is still unknown. We have identified and functionally characterized an R2R3 MYB transcription factor (emission of benzenoid II [FaEOBII]) that seems to be the orthologous gene of PhEOBII from Petunia hybrida, which contributes to the regulation of eugenol biosynthesis in petals. The expression of FaEOBII was ripening related and fruit receptacle specific, although high expression values were also found in petals. This expression pattern of FaEOBII correlated with eugenol content in both fruit receptacle and petals. The expression of FaEOBII was repressed by auxins and activated by abscisic acid, in parallel to the ripening process. In ripe strawberry receptacles, where the expression of FaEOBII was silenced, the expression of cinnamyl alcohol dehydrogenase1 and FaEGS2, two structural genes involved in eugenol production, was down-regulated. A subsequent decrease in eugenol content in ripe receptacles was also observed, confirming the involvement of FaEOBII in eugenol metabolism. Additionally, the expression of FaEOBII was under the control of FaMYB10, another R2R3 MYB transcription factor that regulates the early and late biosynthetic genes from the flavonoid/phenylpropanoid pathway. In parallel, the amount of eugenol in FaMYB10-silenced receptacles was also diminished. Taken together, these data indicate that FaEOBII plays a regulating role in the volatile phenylpropanoid pathway gene expression that gives rise to eugenol production in ripe strawberry receptacles.

Novel Roles for the Polyphenol Oxidase Enzyme in Secondary Metabolism and the Regulation of Cell Death in Walnut

The enzyme polyphenol oxidase (PPO) catalyzes the oxidation of phenolic compounds into highly reactive quinones. Polymerization of PPO-derived quinones causes the postharvest browning of cut or bruised fruit, but the native physiological functions of PPOs in undamaged, intact plant cells are not well understood. Walnut (Juglans regia) produces a rich array of phenolic compounds and possesses a single PPO enzyme, rendering it an ideal model to study PPO. We generated a series of PPO-silenced transgenic walnut lines that display less than 5% of wild-type PPO activity. Strikingly, the PPO-silenced plants developed spontaneous necrotic lesions on their leaves in the absence of pathogen challenge (i.e. a lesion mimic phenotype). To gain a clearer perspective on the potential functions of PPO and its possible connection to cell death, we compared the leaf transcriptomes and metabolomes of wild-type and PPO-silenced plants. Silencing of PPO caused major alterations in the metabolism of phenolic compounds and their derivatives (e.g. coumaric acid and catechin) and in the expression of phenylpropanoid pathway genes. Several observed metabolic changes point to a direct role for PPO in the metabolism of tyrosine and in the biosynthesis of the hydroxycoumarin esculetin in vivo. In addition, PPO-silenced plants displayed massive (9-fold) increases in the tyrosine-derived metabolite tyramine, whose exogenous application elicits cell death in walnut and several other plant species. Overall, these results suggest that PPO plays a novel and fundamental role in secondary metabolism and acts as an indirect regulator of cell death in walnut.

Thiamine modulates metabolism of the phenylpropanoid pathway leading to enhanced resistance to Plasmopara viticola in grapevine

BackgroundPreviously, we have reported the ability of thiamine (vitamin B1) to induce resistance against Plasmopara viticola in a susceptible grapevine cv. Chardonnay. However, mechanisms underlying vitamins, especially, thiamine-induced disease resistance in grapevine are still largely unknown. Here, we assessed whether thiamine could modulate phenylpropanoid pathway-derived phytoalexins in grapevine plants, as well as, the role of such secondary metabolites in thiamine-induced resistance process to P. viticola.ResultsOur data show that thiamine treatment elicited the expression of phenylpropanoid pathway genes in grapevine plants. The expression of these genes correlated with an accumulation of stilbenes, phenolic compounds, flavonoids and lignin. Furthermore, the total anti-oxidant potential of thiamine-treaded plants was increased by 3.5-fold higher level as compared with untreated-control plants. Four phenolic compounds are responsible of 97% of the total anti-oxidant potential of thiamine-treated plants. Among these compounds, is the caftaric acid, belonging to the hydroxy-cinnamic acids family. This element contributed, by its own, by 20% of this total anti-oxidant potential. Epifluorescence microscopy analysis revealed a concomitant presence of unbranched-altered P. viticola mycelia and stilbenes production in the leaf mesophyll of thiamine-treated inoculated plants, suggesting that stilbenes are an important component of thiamine-induced resistance in grapevine.ConclusionThis work is the first to show the role of thiamine, as a vitamin, in the modulation of grapevine plant secondary metabolism contributing to an enhanced resistance to P. viticola, the most destructive fungal disease in vineyards.

Chemical and Biochemical Change of Healthy Phenolic Fractions in Winegrape by Means of Postharvest Dehydration

Clusters of Aleatico winegrape were picked at 18 degrees Brix and placed at 10, 20, or 30 degrees C, 45% relative humidity (RH) and 1.5 m/s of air flow to dehydrate the berries up to 40% of loss of initial fresh weight. Sampling was done at 0, 10, 20, 30, and 40% weight loss (wl). Selected polyphenols and sugar content (expressed as SSC = soluble solids content) both measured on dry weight basis, polyphenol oxidase (PPO), and phenylpropanoid pathway gene expression were analyzed. Phenolic acids increased significantly at 20% wl at 20 degrees C, while at 10 degrees C the increase was lower. Stilbenes (trans-resveratrol and trans-piceid) and catechins rose more than double to 100 mg/kg and more than 3-fold to 135 mg/kg at 20 degrees C and 10% wl. At 10 degrees C the increase of these compounds was less, but higher than initial values. At 30 degrees C, except for a significant rise at 10% wl for catechins and stilbenes, all the rest of the compounds diminished. Anthocyanins increased at 10 and 20 degrees C, but decreased at 30 degrees C. PPO rapidly increased at 20 and 30 degrees C at 10% wl and then declined, while at 10 degrees C the activity lasted longer. Relative gene expression of phenylalanine ammonia lyase (PAL), stilbene synthase (STS), chalcone isomerase (CHI), dihydroflavonol reductase (DFR) were upregulated at 10 degrees C more than at 20 degrees C, at 20% wl, while at 30 degrees C the gene expression was downregulated.

Expression of Phenylpropanoid Pathway Genes in Chickpea Defense Against Race 3 of Ascochyta rabiei

The fungal disease, ascochyta blight, caused by Ascochyta rabiei is a major yield limiting factor of chickpea (Cicer arietinum L.) around the world. Expression analysis of genes induced in general defense response can provide clues to elucidate major defense mechanisms against pathogen infection in chickpea plants. The role of key phenylpropanoid pathway enzymes response to Ascochyta rabiei in chickpea was studied under greenhouse conditions using a reverse transcription and semi-quantitative polymerase chain reaction (SQ-PCR). Transcript accumulation of four genes encoding phenylalanine ammonia-lyase (PAL), chalcon synthase (CHS), isoflavone reductase (IFR) and Flavanone 3-Hydroxylase (F3H) induced in response to race 3 of A. rabiei was compared in resistant and susceptible genotypes. Results obtained in this study showed that in resistant genotype all 4 phenylpropanoid pathway genes: PAL, CHS, IFR and F3H were rapidly up regulated 6 h after inoculation with race 3 of A. rabiei. However, transcripts of PAL and IFR genes were rapidly accumulated in both resistant and susceptible cultivars. Therefore, induction of key enzymes of phenylpropanoid pathway appeared to be an important defense mechanism of chickpea plants against A. rabiei.