Enhanced Flavonoid Accumulation Reduces Combined Salt and Heat Stress Through Regulation of Transcriptional and Hormonal Mechanisms

Abiotic stresses, such as salt and heat stress, coexist in some regions of the world and can have a significant impact on agricultural plant biomass and production. Rice is a valuable crop that is susceptible to salt and high temperatures. Here, we studied the role of flavanol 3-hydroxylase in response to combined salt and heat stress with the aim of better understanding the defensive mechanism of rice. We found that, compared with wild-type plants, the growth and development of transgenic plants were improved due to higher biosynthesis of kaempferol and quercetin. Furthermore, we observed that oxidative stress was decreased in transgenic plants compared with that in wild-type plants due to the reactive oxygen species scavenging activity of kaempferol and quercetin as well as the modulation of glutathione peroxidase and lipid peroxidase activity. The expression of high-affinity potassium transporter (HKT) and salt overly sensitive (SOS) genes was significantly increased in transgenic plants compared with in control plants after 12 and 24 h, whereas sodium-hydrogen exchanger (NHX) gene expression was significantly reduced in transgenic plants compared with in control plants. The expression of heat stress transcription factors (HSFs) and heat shock proteins (HSPs) in the transgenic line increased significantly after 6 and 12 h, although our understanding of the mechanisms by which the F3H gene regulates HKT, SOS, NHX, HSF, and HSP genes is limited. In addition, transgenic plants showed higher levels of abscisic acid (ABA) and lower levels of salicylic acid (SA) than were found in control plants. However, antagonistic cross talk was identified between these hormones when the duration of stress increased; SA accumulation increased, whereas ABA levels decreased. Although transgenic lines showed significantly increased Na+ ion accumulation, K+ ion accumulation was similar in transgenic and control plants, suggesting that increased flavonoid accumulation is crucial for balancing Na+/K+ ions. Overall, this study suggests that flavonoid accumulation increases the tolerance of rice plants to combined salt and heat stress by regulating physiological, biochemical, and molecular mechanisms.

The Aconitum carmichaelii F3′5′H Gene Overexpression Increases Flavonoid Accumulation in Transgenic Tobacco Plants

Aconitum carmichaelii Debx. is a herbal species that contains many precious bioactive substances, which are alkaloids, flavonoids, steroids, and glycosides. Flavonoids, which are major secondary compounds, play an important role in maintaining redox balance in the cells of the plant body. Many flavonoids have antibacterial, antioxidant, and anticancer properties. However, studies have mainly focused on aconitine, which is a highly toxic group A poison belonging to the alkaloid group, but with little mention of flavonoids. The flavonoids in A. carmichaelii are a group of substances with high content, concentrated in leaves and flowers, including quercetin and kaempferol. F3′5′H (Flavonoid 3′5′-hydroxylase) has been identified as the key enzyme involved in the final steps of flavonoid biosynthesis in plants in general and in A. carmichaelii specifically. This study offers the first report, and demonstrates that the overexpression of the F3′5′H gene from a herbal plant, A. carmichaelii, increases flavonoid content in genetically modified tobacco plants. The A. carmichaelii gene was transformed into tobacco leaf tissue to create transgenic tobacco plants. The AcF3′5′H gene was incorporated into the tobacco genome and was expressed in four transgenic tobacco lines (T01, T03, T05, and T014). The F3′5′H content increased from 20.33% to 32.00% compared with that in non-transformed plants (P < 0.001). Therefore, the flavonoid content of four transgenic tobacco lines increased compared to the WT, from 69.23% to 122.23% (P < 0.001). The results of the successful expression of the AcF3′5′H gene in model tobacco plants are the basis for using the AcF3′5′H gene for improving flavonoid content in other medicinal plants. Thus, the AcF3′5′H gene considered in this work could be a candidate for gene technology to enhance flavonoid accumulation in plants.

Carbon and nitrogen metabolism under nitrogen variation affects flavonoid accumulation in the leaves of Coreopsis tinctoria

Flavonoids are phytochemicals present in medicinal plants and contribute to human health. Coreopsis tinctoria, a species rich in flavonoids, has long been used in traditional medicine and as a food resource. N (nitrogen) fertilization can reduce flavonoid accumulation in C. tinctoria. However, there is limited knowledge regarding N regulatory mechanisms. The aim of this study was to determine the effect of N availability on flavonoid biosynthesis in C. tinctoria and to investigate the relationship between C (carbon) and N metabolism coupled with flavonoid synthesis under controlled conditions. C. tinctoria seedlings were grown hydroponically under five different N levels (0, 0.625, 1.250, 2.500 and 5.000 mM). The related indexes of C, N and flavonoid metabolism of C. tinctoria under N variation were measured and analysed. N availability (low and moderate N levels) regulates enzyme activities related to C and N metabolism, promotes the accumulation of carbohydrates, reduces N metabolite levels, and enhances the internal C/N balance. The flavonoid content in roots and stalks remained relatively stable, while that in leaves peaked at low or intermediate N levels. Flavonoids are closely related to phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate: coenzyme A ligase (4CL), and chalcone-thioase (CHS) activity, significantly positively correlated with carbohydrates and negatively correlated with N metabolites. Thus, C and N metabolism can not only control the distribution of C in amino acid and carbohydrate biosynthesis pathways but also change the distribution in flavonoid biosynthesis pathways, which also provides meaningful information for maintaining high yields while ensuring the nutritional value of crop plants.

Morphological characteristics and flavonoid accumulation of Echinacea purpurea cultivated at various salinity

Choirunnisa JP, Widiyastuti Y, Sakya AT, Yunus A. 2021. Morphological characteristics and flavonoid accumulation of Echinacea purpurea cultivated at various salinity. Biodiversitas 22: 3716-3721. Purple coneflower (Echinacea purpurea (L.) Moench) is an introduced medicinal plant from North America. E. purpurea has high morphological characteristics on stems, leaves and flowers. This plant has not much cultivated as a raw material for traditional medicine in Indonesia due to not much information about flavonoid accumulation of E. purpurea in this country. The purpose of this research was to study morphological characteristics from three accessions of E. purpurea cultivated with various salinity and to select E. purpurea accessions that have high flavonoid accumulation. This study design using a factorial Completely Randomized Design (CRD). The first factor is 3 accessions of E. purpurea (E1; E2; E3). The second factor is 4 levels of CaCl2 (0 ppm; 2500 ppm; 5000 ppm; 10000 ppm). The study was conducted by observing the morphological characteristics of stems, leaves flowers, and herb extract and flavonoid accumulation were analyzed using SPSS. The results demonstrated that morphological characteristics are easy to observed on leaf shape and flower color. The highest herb extract with 10.043% and flavonoid accumulation with 0.510% were in accession 2 with the addition of CaCl2 concentration of 10000 ppm. This study concludes that there are morphological characteristics of E. purpurea cultivated at various salinity and the highest CaCl2 concentration can increase with significance to herb extract and flavonoid accumulation.

Leaf Age-Dependent Photosystem II Photochemistry and Oxidative Stress Responses to Drought Stress in Arabidopsis thaliana Are Modulated by Flavonoid Accumulation

We investigated flavonoid accumulation and lipid peroxidation in young leaves (YL) and mature leaves (ML) of Arabidopsis thaliana plants, whose watering stopped 24 h before sampling, characterized as onset of drought stress (OnDS), six days before sampling, characterized as mild drought stress (MiDS), and ten days before sampling, characterized as moderate drought stress (MoDS). The response to drought stress (DS) of photosystem II (PSII) photochemistry, in both leaf types, was evaluated by estimating the allocation of absorbed light to photochemistry (ΦPSII), to heat dissipation by regulated non-photochemical energy loss (ΦNPQ) and to non-regulated energy dissipated in PSII (ΦNO). Young leaves were better protected at MoDS than ML leaves, by having higher concentration of flavonoids that promote acclimation of YL PSII photochemistry to MoDS, showing lower lipid peroxidation and excitation pressure (1 − qp). Young leaves at MoDS possessed lower 1 − qp values and lower excess excitation energy (EXC), not only compared to MoDS ML, but even to MiDS YL. They also possessed a higher capacity to maintain low ΦNO, suggesting a lower singlet oxygen (1O2) generation. Our results highlight that leaves of different developmental stage may display different responses to DS, due to differential accumulation of metabolites, and imply that PSII photochemistry in Arabidopsis thaliana may not show a dose dependent DS response.

Nitrogen deficiency maintains the yield and improves the antioxidant activity of Coreopsis tinctoria Nutt

Nitrogen (N) deficiency levels were investigated for their potential to maintain the yield and improve antioxidant activity of Coreopsis tinctoria. Inflorescences and leaves at 0, 10, 20, 30, 40 and 50 d after flowering were frozen at −80 °C and plant growth, antioxidant activity, bioactive substance, enzyme activity and gene expression were evaluated. N deficiency maintained the total number of flowers, promoted phenol and flavonoid accumulation and enhanced antioxidant activity. Moreover, N deficiency stimulated activities of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate: coenzyme A ligase (4CL) and induced CtPAL, CtC4H and Ct4CL gene expression. The data also suggest that N deficiency-induced phenolic and flavonoid accumulation occurs due to the activation of biosynthetic pathways in C. tinctoria. We characterize the unique features of C. tinctoria under N deficiency conditions and provide valuable information for the cultivation of high-N use efficiency varieties with low input and high output.