Blue LED light promoting the growth, accumulation of high-value isoflavonoids and astragalosides, antioxidant response, and biosynthesis gene expression in Astragalus membranaceus (Fisch.) Bunge hairy root cultures.

The root of Astragalus membranaceus (Fisch.) Bunge is one of the most frequently used herbs in traditional Chinese medicine (TCM) formulae for fighting COVID-19 infections, due to the presence of isoflavonoids and astragalosides associated with antiviral and immune-enhancing activities. For the first time, the exposure of A. membranaceus hairy root cultures (AMHRCs) to different colors of LED lights i.e., red, green, blue, red/green/blue (1/1/1, RGB), and white, was conducted to promote the root growth and accumulation of isoflavonoids and astragalosides. LED light treatment regardless of colors was found beneficial for root growth, which might be a result of the formation of more root hairs upon light stimulation. Blue LED light was found most effective for enhancing phytochemical accumulation. Results showed that the productivity of root biomass in blue-light grown AMHRCs with an initial inoculum size of 0.6% for 55 days was 1.40-fold higher than that in dark (control), and yields of high-value isoflavonoids and astragalosides including calycosin, formononetin, astragaloside IV, and astragaloside I increased by 3.17-fold, 2.66-fold, 1.78-fold, and 1.52-fold relative to control, respectively. Moreover, the photooxidative stress together with transcriptional activation of biosynthesis genes might contribute to the enhanced accumulation of isoflavonoids and astragalosides in blue-light grown AMHRCs. Overall, this work offered a feasible approach for obtaining higher yields of root biomass and medicinally important compounds in AMHRCs via the simple supplementation of blue LED light, which made blue-light grown AMHRCs industrially attractive as plant factory in controlled growing systems. Supplementary Information: The online version contains supplementary material available at 10.1007/s11240-023-02486-7.

Cocultivation of pigeon pea hairy root cultures and Aspergillus for the enhanced production of cajaninstilbene acid.

Pigeon pea hairy root cultures (PPHRCs) have been proven to be a promising alternative for the production of health-beneficial phenolic compounds, such as the most important health-promoting compound, i.e., cajaninstilbene acid (CSA). In this study, PPHRCs were cocultured with live Aspergillus fungi for further improving phenolic productivity via biological elicitation. Aspergillus oryzae CGMCC 3.951 (AO 3.951) was found to be the optimal fungus that could achieve the maximum increment of CSA (10.73-fold increase) in 42-day-old PPHRCs under the inoculum size of mycelia 0.50% and cocultivation time 36 h. More precisely, the contents of CSA in hairy roots and culture media after fungal elicitation increased by 9.87- and 62.18-fold over control, respectively. Meanwhile, the contents of flavonoid glycosides decreased, while aglycone yields increased upon AO 3.951 elicitation. Moreover, AO 3.951 could trigger the oxidative stress and pathogen defense response thus activating the expression of biosynthesis- and ABC transporter-related genes, which contributed to the intracellular accumulation and extracellular secretion of phenolic compounds (especially CSA) in PPHRCs. And PAL2, 4CL2, STS1, and I3'H were likely to be the potential key enzyme genes regulating the biosynthesis of CSA, and ABCB11X1-1, ABCB11, and ABCG24X2 were closely related to the transmembrane transport of CSA. Overall, the cocultivation approach could make PPHRCs more commercially attractive for the production of high-value phenolic compounds such as CSA and flavonoid aglycones in nutraceutical/medicinal fields. And the elucidation of crucial biosynthesis and transport genes was important for systematic metabolic engineering aimed at increasing CSA productivity. KEY POINTS: • Cocultivation of PPHRCs and live fungi was to enhance CSA production and secretion. • PPHRCs augmented CSA productivity 10.73-fold when cocultured with AO 3.951 mycelia. • Several biosynthesis and transport genes related to CSA production were clarified.

Health-Promoting Phenolic Compound Accumulation, Antioxidant Response, Endogenous Salicylic Acid Generation, and Biosynthesis Gene Expression in Germinated Pigeon Pea Seeds Treated with UV-B Radiation.

Germinated pigeon pea seeds (GPPSs) are good dietary supplements with satisfactory nutritional and medicinal values. In this study, UV-B treatment was used to promote the accumulation of health-promoting phenolic compounds (10 flavonoids and 1 stilbene) in GPPS. The total yield of 11 phenolic compounds (235 839.76 ± 17 118.24 ng/g DW) significantly improved (2.53-fold increase) in GPPSs exposed to UV-B radiation (3 W/m2) for 8 h, whereas free amino acid and reducing sugar contents exhibited a decreasing tendency during UV-B exposure. Meanwhile, the positive response in the antioxidant activities of enzymes and nonenzymatic extracts was noticed in UV-B-treated GPPSs. Moreover, UV-B radiation could cause tissue damages in hypocotyls and cotyledons of the GPPSs and enhance the generation of endogenous salicylic acid, thus activating the expression of biosynthesis genes (especially CHS and STS1). Overall, the simple UV-B supplementation strategy makes GPPSs more attractive as functional foods/nutraceuticals in diet for promoting human health.

High-potential accumulation and tolerance in the submerged hydrophyte Hydrilla verticillata (L.f.) Royle for nickel-contaminated water.

Water contamination by nickel (Ni) has become an increasing concern in recent decades. Hydrilla verticillata (L.f.) Royle has been recognized as a promising accumulator of several potentially toxic elements (PTEs) in phytoremediation, but its Ni-accumulation characteristics and its mechanisms of tolerance to Ni remain largely unknown. This research investigated the biochemical responses of leaves and stems of H. verticillata to various concentrations of Ni (5, 10, 15, 20, and 40 µM) over periods of 7, 14, or 21 days. Plants accumulated considerable Ni to a maximum amount of 1080 mg kg-1 dry weight (DW) with a maximum bioconcentration factor of 1100; thus, high Ni accumulation was detected in H. verticillata. Low concentrations (5-15 µM) or short durations (less than 14 days) of Ni exposure might promote plant growth without adversely affecting normal metabolism. After peaking at day 14, a decline in bioaccumulation was unexpectedly observed as a long-term effect of Ni toxicity. Malondialdehyde content and the activities of defense-related enzymes changed in a similar pattern after treatment with Ni, increasing with both Ni concentration and exposure time to a peak (often at 5-15 µM on day 14), followed by a decline. Through a comprehensive analysis of all the test parameters, the tolerance thresholds were determined to be > 40.0 µM, 24.0 µM, and 15.8 µM at days 7, 14, and 21, respectively. Hydrilla verticillata could be a "high-potential accumulator" capable of decontaminating aquatic bodies polluted by Ni within the threshold range.

Bioaccumulation and tolerance characteristics of a submerged plant (Ceratophyllum demersum L.) exposed to toxic metal lead.

A hydroponic study was conducted to investigate the lead bioaccumulation and tolerance characteristics of Ceratophyllum demersum L. exposed to various lead concentrations (5-80 µM) for 7, 14 or 21 days. Lead accumulation increased with increasing concentrations of metal in the solution, to a maximum accumulation of 4016.4 mg kg(-1) dw. Unexpectedly, the release of accumulated lead from the plants into solution was observed for all experimental groups except those exposed to 5 µM. Both the biomass and protein content of the plants responded significantly to lead stress. Malondialdehyde (MDA) levels increased substantially at lead concentrations below 20 µM, further indicating that this metal is toxic to the plants. To reveal the mechanism underlying the defense against lead stress, plants were also assayed for the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), as well as other relevant enzymes such as phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO). The activities of both SOD and CAT increased at lower lead concentrations and with shorter exposure times, followed by a decline, but the activities of POD and its isoenzymes continued to increase under all conditions. Moreover, increases in the activities of PAL and PPO were observed only for the 14-day treatment, and these two enzymes were not sensitive to lead concentration. These results suggest that C. demersum exhibits strong tolerance within a specific concentration range of lead in solution; according to regression analysis, 40 µM is suggested to be this plant's tolerance threshold for lead in water. Furthermore, the malfunction of this tolerance mechanism might accelerate the metal-release process. These attributes are likely to be beneficial for utilizing C. demersum in phytoremediation applications.