Isolation and molecular identification of β-carotene producing strains of Dunaliella salina and Dunaliella bardawil from salt soil samples by using species-specific primers and internal transcribed spacer (ITS) primers

Dunaliella salina and Dunaliella bardawil are unique species of the genus Dunaliella that produce large amounts of â-carotene when cultivated under appropriate conditions. These include high light intensity, high sodium chloride concentration, nitrate deficiency and extreme temperatures. Under these conditions, only D. salina and D. bardawil can accumulate â-carotene to as much as 10% of the cellular dry weight. Because the morphological characterization is based on the environmental factors, the Dunaliella will change the shape, so identification and differentiation of Dunaliella species by morphology is very difficult. In this research study, we isolated, identified and discriminated the different Dunaliella β-carotene producing strains from salt soil samples, by using 18S rDNA and internal transcribed spacer (ITS) gene sequences. The soil samples were collected from four different provinces of the North Eastern part of Thailand.namely: UdonThani, BuriRam, AmnartCharoen and Chaiyaphum. Among the four isolates, only BuriRam KU01 and UdonThani KU01 were D. salina and D. bardawil, respectively whereas AmnartCharoen KU01 and Chaiyaphum KU01 were not these Dunaliella species. At 4 M NaCl, with deficiency of nitrate (KNO3) and phosphate (KH2PO4) in the medium, the D. salina strain BuriRam KU01 produced β-carotene at the level of 56.25 ± 0.97 pg·cell-1 and D. bardawil strain UdonThani KU01 produced β-carotene at the level of 52.91 ± 0.29 pg·cell-1 at the 25th day after inoculation. The 18S rDNA and ITS sequences of D. salina strain BuriRam KU01 and D. bardawil strain UdonThani KU01 were submitted to the National Center for Biotechnology Information (NCBI) database with accession numbers of JN052202, JN052203, JN034031 and JN052204, respectively. By using the species-specific primers and ITS primers the â-carotene producing strains of Dunaliella was identified.Key words: 18S rDNA, β-carotene, carotenoid, Dunaliella bardawil, Dunaliella salina, internal transcribed spacer (ITS), salinity

Differential expression profiles of anthocyanidin biosynthesis gene during black rice seed development

The black rice (Oryza sativa cv. Heugjinju) is rich in anthocyanins which is beneficial to human health. To correlate the biosynthesis of the pigments with relevant genes, the mRNA level of genes involved in anthocyanin biosynthesis was monitored by quantitative real-time polymerase chain reaction (qRT-PCR) during seed development of black rice. The mRNA level of F3’H, DFR, and ANS, key enzymes in anthocyanidin biosynthesis, peaked at 10 days after flowering. In general, the absolute level of ANS was approximately one order higher than F3’H, F3’5’H, and DFR in 10 days after flowering. The transcript level of major seed protein gene GluA-3, taken as reference, was also at the highest on the 10 days after flowering. However, the level of CHS isogenes was highest at 15 or 20 days after flowering. The highest transcript level of the genes, except CHS, preceded the highest anthocyanidin content by 5 days. This pattern coincided with an increase of anthocyanin content between 10 and 15 days after flowering. From these findings, it is suggested that particular CHS isoforms might be responsible for the anthocyanin production in black rice

Effect of cytokinins on growth and phenylpropanoid accumulation in Tartary buckwheat sprouts (Fagopyrum esculentum Moench)

This study analyzed the effect of plant hormones, zeatin, 6-benzyl amino purine (BAP), kinetin, and thidiazuron (TDZ) on the growth of Tartary buckwheat sprouts and analyzed the fresh weight, shoot and root length, and production of phenolic compounds. All the hormone-treated plants at the lowest concentration (0.1 mg/L) showed the highest levels of growth parameters (fresh weight, shoot, and root length) when compared to the control. Among the various hormones treatment, the plant treated with 1 mg/L of BAP, kinetin, and zeatin showed the highest total phenolic level, whereas the TDZ showed the highest accumulation of total phenolic at the lowest concentration (0.1 mg/L). A total of 6 compounds were identified (4-hydroxybenzoic acid, caffeic acid, chlorogenic acid, p-coumaric acid, rutin, and trans-cinnamic acid) were quantified by high liquid performance chromatography (HPLC) after treatment of plant with different concentrations of hormones. Among these individual phenolic compounds, at the higher hormonal concentration (1 mg/L) the rutin showed the highest accumulation in BAP, zeatin, and kinetin treated sprout, whereas in the TDZ treated sprout the rutin content was highest at the lowest concentration (0.1 mg/L). From these results, it is suggested that BAP, zeatin, and kinetin at the lowest concentrations might positively enhance the growth of buckwheat sprouts, whereas at the highest hormonal treatment the accumulation of the phenolic compounds was higher. However, in TDZ treatment the growth and phenolic compound accumulation were highest at the lowest concentration. From these results, it is showed that suitable concentrations might enhance the growth and phenolic compound accumulation in Tatary buckwheat sprout

Biosynthesis of phenylpropanoids and their protective effect against heavy metals in nitrogen-fixing black locust (Robinia pseudoacacia)

Purpose: To examine the effect of various heavy metals (HMs) on phenylpropanoid pathway compounds in Robinia pseudoacacia.Methods: A series of pot culture experiments were performed to understand how the metabolic profile of phenylpropanoid compounds were affected by various HMs, such as redox-active HMs (AgNO3 and CuCl2), and non-redox-active HMs (HgCl2). Phenylpropanoid compound level was evaluated by high performance liquid chromatography.Results: The total phenylpropanoid level in leaves increased significantly in all the treated groups when compared to that in the untreated group (p < 0.05). However, a significant effect on the total phenylpropanoid levels was only found for redox-active HMs (p < 0.05), whereas non-redox-active HMs showed less accumulation. Chlorogenic acid and rutin were the two major phenylpropanoid compounds found after the plants were subjected to redox and non-redox-active HMs stress. However, when compared to these two compounds, the levels of catechin hydrate, epicatechin, p-coumaric acid, kaempferol, and quercetin were lower. Caffeic acid level was significantly decreased in both redox and non-redox-active HMs when compared to that in the control (p < 0.05). In addition, trans-cinnamic acid accumulation was altered based on the types and concentration of HMs.Conclusion: Phenylpropanoid metabolic pathway participated in the HM tolerance process for the protection of R. pseudoacacia from oxidative damage caused by HMs, thus allowing the species to grow in highly HMs-contaminated areas. Keywords: Heavy metals, Non-redox-active metals, Phenylpropanoid compounds, Redox-active metals, Robinia pseudoacaci

Expression analysis of diosgenin pathway genes and diosgenin accumulation in fenugreek sprouts after exposure to copper sulfate

Trigonella foenum-graecum L. is an annual herb belonging to the family Fabaceae commonly called Fenugreek. It is rich in various secondary metabolites such as alkaloids, flavonoids, phenolic compounds, and steroidal saponins. In recent years, diosgenin has much attention in the cosmetic, functional food, and pharmaceutical industries. In this study we aimed to examine the effect of different concentrations of copper sulfate (CuSO4) on growth, diosgenin biosynthetic (DB) gene expression, and diosgenin accumulation in T. foenum-graecum sprouts. Results showed that the seed germination, fresh weight, shoot length, and root length were gradually decreased with increasing the CuSO4 concentrations. In contrast, the expression level of DBGs i.e., TfSQS, TfSQLE, TfCAS, and TfSTRL were gradually upregulated with increasing the CuSO4 concentrations. Among all those tested concentrations, the expression levels of all those genes were significantly higher in 0.5 mM CuSO4 treated sprouts. The highest expression level was obtained in the TfCAS gene, which was 3.25-fold higher than the unexposed sprouts. The diosgenin content was significantly influenced in the CuSO4 exposed sprouts. The highest diosgenin content was achieved in the 5.0 mM followed by 1.0, 10.0, and 0.5 mM CuSO4 exposed concentrations, with a reduction of 41%, 39%, 36%, and 35%, respectively. From these results, it is shown that exposure of fenugreek sprout to CuSO4 is one of the suitable strategies to enhance the accumulation of diosgenin content

Effect of media and gelling agents on shoot organogenesis of Liriope platyphylla

Liriope platyphylla can be multiplied either by planting seeds or dividing its tuberous roots. In this study, a method for L. platyphylla plant shoot organogenesis from meristem explants was developed employing medium and gelling agents. For the selection of optimal medium conditions for shoot organogenesis, the effects of full- and half-strength B5, SH, and MS media were examined. Different concentrations of the gelling agents such as phytagar (6, 7, 8, and 9 g L-1) and gellan gum (2, 3, 4, and 5 g L-1) were examined for efficient shoot formation. The results revealed the superiority of half-strength MS basal medium in shoot organogenesis and growth of Liriope platyphylla. But the half-strength B5 media performed poorly. Compared to plant agar, gellan gum performed well in terms of shoot regeneration and shoot length. When gellan gum was used at 3 g L-1 the maximum number of shoots explant-1 (5.8) and longest shoot (45.8 mm) was observed but the lowest number of shoots explant-1 (3.2) and shortest shoot (21.4 mm) was registered with 5 g L-1. It is proposed from our study that half-strength MS media and gellan gun gelling agent at 3 g/l could be applied in shoot organogenesis and growth of L. platyphylla

An endophyte Paenibacillus dendritiformis strain APL3 promotes Amaranthus polygonoides L. sprout growth and their extract inhibits food-borne pathogens

Green leafy vegetables are rich sources of antioxidants and minerals, which prevent food-borne pathogen infections during our diet. This study was aimed to isolate and identify the plant growth-promoting endophytic bacterium from several plant species to enhance the growth of Amaranthus polygonoides L. and their antimicrobial potential against food-borne pathogens. Seven endophytic bacterial isolates were tested on two Amaranthus species to identify the suitable beneficial bacterium. The antioxidants capacity and antimicrobial activity of bacterial isolate (APL3) treated plants were analyzed. The bacterial isolate, APL3 showed a significantly higher growth of A. polygonoides L. than other isolates. It was identified as Paenibacillus dendritiformis strain APL3 by 16S rRNA gene sequencing and phylogenetic analysis. The endophyte (APL3) treated A. polygonoides L. sprouts had higher antioxidants potentials and significantly inhibited the growth of Escherichia coli, Salmonella sp., Staphylococcus sp. and Pseudomonas sp. The results of the present study suggest that utilization of P. dendritiformis strain APL3 triggers the growth of A. polygonoides L. and induces metabolic changes in plants to improve their antimicrobial properties to prevent foodborne pathogens

Production, antimicrobial, antioxidant, sensory, and therapeutic properties of herbal wine – A comprehensive review

Wine is a fermented beverage. Herbal-infused wine is beneficial to health due to its antimicrobial and anticancer properties. The constituents of these plants, including flowers, fruits, stems, roots, bark, and leaves, contain antioxidant activity. The herbs can be extracted through various methods such as maceration, decoction, infusion, crushing, grinding, and blending. Saccharomyces cerevisiae is the primary organism responsible for fermentation, converting glucose into metabolic energy. This review analyses the potential medicinal value of herbal wine in treating human diseases. Herbal wine is a recent development in culinary technology, as herbs possess antioxidant and antimicrobial properties that make them effective against cancer and diabetes. Polyphenols found in wine have been reported to be effective in treating human ailments such as coronary heart disease, diabetes, microbial infections, neurodegenerative diseases, and aging. Therefore, fortifying alcoholic beverages may increase health benefits and clinical applications. The qualities of these herbal extracts are comparable to those of fortified wines, making drinking fortified wines a healthier option than consuming conventional wines. However, the production of herbal wine from certain extracts may require the addition of taste enhancers. Our focus is on the fermentative production of wine from various herbal extracts, including physicochemical, antioxidant, antimicrobial, and sensory evaluation. We compare and describe the health benefits and harmful effects of fruit wine and herbal wine

Effect of heavy metals on phenylpropanoid biosynthesis in Euonymus alatus

The productivity of the phenylpropanoid biosynthesis pathway in plants varies depending on the type of stress. In this work, we looked into how different phenylpropanoid chemicals accumulated in Euonymus alatus following exposure to different concentrations of CuCl2 (0.1, 0.5, and 1 mM), HgCl2 (0.1, 0.5, and 1 mM), and NiSO4 (10, 50, and 100 mM). We analyzed some of the individual phenolic chemicals by high-performance liquid chromatography (HPLC). In nearly all cases, rutin showed the largest concentration among the phenylpropanoid chemicals, followed by epicatechin, sinapic acid, p-coumaric acid, trans-cinnamic acid, ferulic acid, and caffeic acid. However, due to the change in the concentration of the heavy metals, the amount of phenylpropanoid changed. The highest accumulation of phenylpropanoid was documented in 0.1 mM CuCl2, whereas it was reduced in 1 mM HgCl2 exposed plants. These findings unequivocally demonstrate that the phenylpropanoid metabolic pathway took part in the heavy metal tolerance process, which shielded E. alatus from the oxidative damage brought on by heavy metals. Thus, under a variety of environmental stress situations, this species with a high tolerance to heavy metals may survive

Analysis of triterpenoids, carotenoids, and phenylpropanoids in the flowers, leaves, roots, and stems of white bitter melon (Cucurbitaceae, Momordica charantia)

Purpose: To evaluate the contents of carotenoids, triterpenoids, and phenylpropanoids in different parts of white bitter melon.Methods: We evaluated the accumulation of 2 triterpenoids, 10 carotenoids, and 11 phenylpropanoids in different parts of white bitter melon, including fruits at four different developmental stages using HPLC.Results: Charantin, lutein, and rutin were the main triterpenoids, carotenoids, and phenylpropanoids, respectively. The accumulation of triterpenoids (momordicine and charantin), carotenoids (antheraxanthin, lutein, violaxanthin, α-carotene, and β-carotene), and phenylpropanoids (caffeic acid, chlorogenic acid, epicatechin, gallic acid, p-coumaric acid, rutin, and trans-cinnamic acid) was high inthe leaves and/or flowers, which are exposed to direct sunlight, but low in the roots.Conclusion: Most of the analyzed components were accumulated at high levels in the leaves and/or flowers. These results will help exploit the compounds in various parts of white bitter melon that are beneficial for human health. Keywords: Momordica charantia, Bitter melon, Triterpenoid, Carotenoid, Phenylpropanoi