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This study aimed at investigating the chemical composition and the hepatoprotective activities of Plumbago indica L. and P. auriculata Lam. LC–MS/MS analyses for the hydroalcoholic extracts of the aerial parts of the two Plumbago species allowed the tentative identification of thirty and twenty-five compounds from P. indica and P. auriculata, respectively. The biochemical and histopathological alterations associated with thioacetamide (TAA)-induced liver fibrosis in rats were evaluated in vivo where rats received the two extracts at three different dose levels (100, 200 and 400 mg/kg p.o, daily) for 15 consecutive days with induction of hepatotoxicity by TAA (200 mg/kg/day, i.p.) at 14th and 15th days. Results of the present study showed a significant restoration in liver function biomarkers viz. alanine transaminase (ALT), aspartate transaminase (AST), gamma glutamyl transferase and total bilirubin. The liver homogenates exhibited increased levels of antioxidant biomarkers: reduced glutathione (GSH) and catalase (CAT), accompanied with decline in malondialdehyde (MDA). Furthermore, treated groups exhibited a significant suppression in liver inflammatory cytokines: tumor necrosis factor-α (TNF-α) and interlukin-6 (IL-6), and fibrotic biomarker: alpha smooth muscle relaxant. Histopathological examination of the liver showed normality of hepatocytes. Noteworthy, P. indica extract showed better hepatoprotective activity than P. auriculata, particularly at 200 mg/kg. To sum up, all these results indicated the hepatoprotective properties of both extracts, as well as their antifibrotic effect was evidenced by reduction in hepatic collagen deposition. However, additional experiments are required to isolate their individual secondary metabolites, assess the toxicity of the extracts and explore the involved mechanism of action. [ABSTRACT FROM AUTHOR]

Key message: The present study provides comparative transcriptome analysis, besides identifying functional secondary metabolite genes of Plumbago zeylanica with pharmacological potential for future functional genomics, and metabolomic engineering of secondary metabolites from this plant towards diversified biomedical applications. Plumbago zeylanica is a widely used medicinal plant of the traditional Indian system of medicine with wide pharmacological potential to treat several disorders. The present study aimed to carry out comparative transcriptome analysis in leaf and root tissue of P. zeylanica using Illumina paired end sequencing to identify tissue-specific functional genes involved in the biosynthesis of secondary metabolites, contributing to its therapeutic efficacy. De novo sequencing assembly resulted in the identification of 62,321 "Unigenes" transcripts with an average size of 1325 bp. Functional annotation using BLAST2GO resulted in the identification of 50,301 annotated transcripts (80.71%) and GO assigned to 18,814 transcripts. KEGG pathway annotation of the "Unigenes" revealed that 2465 transcripts could be assigned to 242 KEGG pathway maps wherein the number of transcripts involved in secondary metabolism was distinct in root and leaf transcriptome. Among the secondary metabolite biosynthesis pathways, the cluster of "Unigenes" encoding enzymes of 'Phenylpropanoid biosynthesis pathway' represents the largest group (84 transcripts) followed by 'Terpenoid Backbone biosynthesis' (48 transcripts). The transcript levels of the candidate unigenes encoding key enzymes of phenylpropanoid (PAL, TAL) and flavanoid biosynthesis (CHS, ANS, FLS) pathways were up-regulated in root, while the expression levels of candidate "Unigenes" transcript for monoterpenoid (DXS, ISPF), diterpenoid biosynthesis (SPS, SDS) and indole alkaloid pathways (STR) were significantly higher in leaf of P. zeylanica. Interestingly, validation of differential gene expression profile by qRT-PCR also confirmed that candidate "Unigenes" enzymes of phenylpropanoid and flavonoid biosynthesis were highly expressed in the root, while the key regulatory enzymes of terpenoid and indole alkaloid compounds were up-regulated in the leaf, suggesting that (differences in) the levels of these functional genes could be attributed to the (differential) pharmacological activity (between root and leaf) in tissues of P. zeylanica. [ABSTRACT FROM AUTHOR]

Background: Plumbagin is one of the pharmaceutically important biomolecule with anticancer potential. Among the plants reported to produce plumbagin, P. zeylanica topped the list. The plumbagin production is very slow with low yield and maximum 0.5% (of dry weight) was reported in P. zeylanica. To meet the increasing demand of the plumbagin at global level, the P. zeylanica are exploited at commercial level, which may pose serious threat on the germplasm of the plant populations. So, it is needed to enhance the contents of plumbagin in P. zeylanica using biotechnological approaches. Among the various methods used to enhance the contents of plumbagin in P. zeylanica, utilization of fungal endophytes to enhance the plumbagin contents is a widely accepted approach. As fungal endophytes have the potential to synthesize various secondary metabolites and also reported to influence the synthesis of the secondary metabolites in plants. In the present study, an attempt was made to assess the effect of fungal endophytes of the Plumbago zeylanica L. on enhancement of plumbagin contents at in vivo level. Results: Total 3 fungal endophytes were recorded from the roots of P. zeylanica collected from Khadki, Pune. The fungal endophytes were identified at morphological and molecular level. After 1 year of the treatment with fungal endophytes, significant enhancement of plumbagin was recorded in the roots of the P. zeylanica. Plumbagin contents in each were quantified against the standard plumbagin by employing LCMS-MS technique. Among the three fungal endophytes, the maximum enhancement of plumbagin content (122.67%) was reported with the treatment of Alternaria sp. (Isolate-3) in the roots of the P. zeylanica compared to control. Conclusion: Among the three fungal endophytes, the maximum enhancement of plumbagin content (122.67%) was reported with Alternaria sp. (Isolate 3) in the roots of the pot-grown plants of P. zeylanica at in vivo level. [ABSTRACT FROM AUTHOR]

The fermented foods have been a part of the human diet in both traditional and modern cultures. Fermented food products are those produced through the addition of microbes such as yeast and bacteria. Each fermented food has distinct population of microorganisms, acts as probiotics, and produces a variety of metabolites and bioactive compounds. The physical and chemical changes in fermented food are due to the presence of probiotics and prebiotics, resulting in improved quality and long-term stability of the product. The positive health benefits have been investigated intensively, and the consolidated data are presented in this review. Once ingested, the beneficial bacteria colonize in the gut microbiome, produce bioactive compounds, and protect the human body from pathogenic microorganisms in various ways. The purpose of this paper is to present various types of fermented foods and beverages, probiotics, and prebiotics present in the foods, bioprocess technologies used for processing of fermented foods, nutritional quality of the fermented foods, and the influence of fermented foods on gut microbiome and health. Therefore, the benefits of fermented foods and beverages on gut microbiome should be studied and pursued to promote good health. [ABSTRACT FROM AUTHOR]

Cactus (Opuntia ficus-indica) is a drought-resistant plant and the source of food and income for many farmers during the rainy season, and the only option to feed livestock during the dry seasons, especially in the southern part of the Tigray region of Ethiopia. The region is currently suffering from a damaging parasitoid of cactus called Dactylopius opuntiae- ficus biotype. An experimental study was conducted from August 2022 to February 2023 to investigate the efficacy of herbal extracts against cochineal insects. The experiment was conducted on the insecticidal property of methanol extracts of Solanum linnaenum, Euphorbia tirucalli, Nerium oleander, Tephrosia vogelii, Calpurnia aurea, Argemone Mexicana, Datura stramonium and Ricinus communis. Herbs were extracted, and their efficacy and LC50 were determined. Phytochemical screening for secondary metabolites was performed. There was a statistically significant difference in the mean mortality of insects across the levels of the herbs (F (8, 71) = 57.812, p = 0.0001, ƞ2 = 0.867) and the experimental extract concentrations (F (1, 71) = 4198.3, p = 0.0001, ƞ2 = 0.983). The individual extracts of S. linnaenum and N. oleander caused average mortality rates of 99.3% and 97.4%, respectively, at 5% extract concentration. Tannins and saponins were the major metabolites found in the extracts. The present study demonstrated the promising insecticidal efficacy of herbal extracts against the damaging cacti pest D. opuntia-ficus biotype, providing insights into sustainable pest control strategies for protecting vital cactus resources in the Tigray region. [ABSTRACT FROM AUTHOR]

The genus Verbascum, belonging to the family Scrophulariaceae, has a significant center of diversity in Iran. Two of its species, V. erianthum and V. stachydiforme, originate in the Iranian-Turanian region, but no studies have been conducted on the induction of their hairy roots. This genus is a valuable source of biologically active compounds such as iridoid glycosides and flavonoids. Hairy root culture is a suitable technique for the production and accumulation of secondary metabolites. Three different studies were conducted to optimize the induction and establishment of hairy roots. In the first experiment, the influence of explant type (leaf and hypocotyl), six infection methods, and co-culture time (48 and 72 h) on the efficiency of hairy root induction was investigated. The results showed that the highest hairy root induction (68.18%) was observed in the leaf explants inoculated by direct infection with three wounds within 72 h co-culture time. In the second experiment, the effect of four Agrobacterium rhizogenes strains (ATCC 15834, A4, A7, and A13) and leaf age (14, 21, and 28 days) on transformation efficiency and some morphological traits examined in both species were studied. The high transformation efficiency of hairy root (80.55%) was detected in the 21-day-old leaf explant of V. erianthum species that was inoculated with the A13 strain. The transformed hairy root colons were confirmed by PCR using rolB gene-specific primers. To optimize hairy root growth and avoid tissue browning, hairy roots were cultured in various media containing different antioxidants and improver agents (including ascorbic acid, citric acid, and NAA). The results showed that the highest fresh growth index (20.42) and the lowest tissue browning (9%) as well as total phenol (8.51 mg GA/g DW), and total flavonoid content (4.42 mg QUE/g DW) were obtained in medium B5 with 1.5 mg/l NAA. [ABSTRACT FROM AUTHOR]

Endophytes are microorganisms that inhabit various plant parts and cause no damage to the host plants. During the last few years, a number of novel endophytic fungi have been isolated and identified from medicinal plants and were found to be utilized as bio-stimulants and bio fertilizers. In lieu of this, the present study aims to isolate and identify endophytic fungi associated with the leaves of Anisomeles indica L. an important medicinal plant of the Terai-Duars region of West Bengal. A total of ten endophytic fungi were isolated from the leaves of A. indica and five were identified using ITS1/ITS4 sequencing based on their ability for plant growth promotion, secondary metabolite production, and extracellular enzyme production. Endophytic fungal isolates were identified as Colletotrichum yulongense Ai1, Colletotrichum cobbittiense Ai2, Colletotrichum alienum Ai2.1, Colletotrichum cobbittiense Ai3, and Fusarium equiseti. Five isolates tested positive for their plant growth promotion potential, while isolates Ai4. Ai1, Ai2, and Ai2.1 showed significant production of secondary metabolites viz. alkaloids, phenolics, flavonoids, saponins, etc. Isolate Ai2 showed maximum total phenolic concentration (25.98 mg g−1), while isolate Ai4 showed maximum total flavonoid concentration (20.10 mg g−1). Significant results were observed for the production of extracellular enzymes such as cellulases, amylases, laccases, lipases, etc. The isolates significantly influenced the seed germination percentage of tomato seedlings and augmented their growth and development under in vitro assay. The present work comprehensively tested these isolates and ascertained their huge application for the commercial utilization of these isolates both in the agricultural and industrial sectors. [ABSTRACT FROM AUTHOR]

Phyllanthus emblica (Aonla, Indian Gooseberry) is known to have various medicinal properties, but studies to understand its genetic structure are limited. Among the various secondary metabolites, ascorbic acid, flavonoids, terpenoids, phenols and tannins possess great potential for its pharmacological applications. Keeping this consideration, we assembled the transcriptome using the Illumina RNASeq500 platform, generating 39,933,248 high-quality paired-end reads assembled into 1,26,606 transcripts. A total of 87,771 unigenes were recovered after isoforms and unambiguous sequences deletion. Functional annotation of 43,377 coding sequences against the NCBI non-redundant (Nr) database search using BlastX yielded 38,692 sequences containing blast hits and found 4685 coding sequences to be unique. The transcript showed maximum similarity to Hevea brasilensis (16%), followed by to Jatropha curcas (12%). Considering key genes involved in the biosynthesis of flavonoids and various classes of terpenoid compounds, thirty EST-SSR primer sequences were designed based on transcriptomic data. Of which, 12 were found to be highly polymorphic with an average of 86.38%. The average value for marker index (MI), effective multiplicity ratio (EMR), resolution power (Rp) and polymorphic information content (PIC) was 7.20, 8.34, 8.64 and 0.80, respectively. Thus, from this study, we developed newly EST-SSRs linked to important genes involved in the secondary metabolites biosynthesis that will be serving as an invaluable genetic resource for crop improvement including the selection of elite genotypes in P. emblica and its closely related Phyllanthaceae species. [ABSTRACT FROM AUTHOR]

Plants that produce bioactive chemicals provide a viable in vitro method for producing key nutraceutical substances, especially in the presence of appropriate elicitors. The goal of this study is to use elicitors in Hassawi rice cell suspension culture to enhance production of phenolic compounds. Salicylic acid (SA), yeast extract (YE), and pectin were utilized at 50, 100, and 200 mg L−1. Impacts of elicitors on cell culture were measured in terms of biomass packed cell volume (PCV), dry and fresh weight, total phenolic, total flavonoid, anthocyanin, tannins content, antioxidant activity, and phenolic compounds using ultra performance liquid chromatography mass spectrometry (UPLC-mass spec). Except for high doses of pectin (200 mg L−1), the 50, 100, and 200 mg L−1 SA–, YE–, and pectin–treated cell culture produced increased PCV, dry (DW), and fresh weight (FW), and all of the aforementioned contents. The cell suspension supplied with 50 mg L−1 SA demonstrated greatest increases in PCV, FW, DW, all total secondary metabolites, and antioxidant activities as compared to other treatments. The highest concentrations of epigallocatechin and chlorogenic acid were found in cell cultures supplied with 100 mg L−1 SA and 50 mg L−1 pectin. Rutin and quercetin-3-O-α-L-rhamnopyranosyl (1–2)-β-D-glucopyranoside-7-O-α-L-rhamnopyranoside were detected at highest concentrations in cultures supplied with 50, 100 mg L−1 YE and 200 mg L−1 SA, respectively. These findings would facilitate in vitro mass production of nutraceuticals using rice cell cultures to assist commercial agronomic sectors. [ABSTRACT FROM AUTHOR]

Dendrobium hybrids have been cultivated as commercially important ornamental plants in the floriculture industry. However, the potential of Dendrobium hybrids as a source of valuable secondary metabolites has not been fully exploited. In this study, the effect of various elicitors on the proliferation of protocorm-like bodies (PLBs) of Dendrobium Sabin Blue and the corresponding production of dendrobine and anthocyanin was investigated. Yeast extract (YE) and glutamine (both at 200 mg L−1) increased dendrobine content in PLBs by 2.7- and 6.5-fold (54.874 ± 2.553 and 894.481 ± 26.185 ng mg−1dry weight (DW)) compared to controls (20.051 ± 0.182 and 136.956 ± 7.397 ng mg−1 DW). Glutamine (200 mg L−1) also significantly enhanced PLB biomass (0.190 ± 0.009 g) with respect to the control (0.144 ± 0.003 g). Chitosan and salicylic acid (SA) were not effective in enhancing the dendrobine and anthocyanin contents in PLBs. A combination of 4 mg L−1 NAA and 100 mg L−1 arginine increased dendrobine content by 8-fold (123.793 ± 5.446 ng mg−1 DW) compared to the control (15.554 ± 0.406 ng mg−1 DW). Anthocyanin content was increased by 1.7-fold (0.548 ± 0.019 Color Value (CV) g−1 DW) when PLBs treated with 4 mg L−1 TDZ were exposed to 1.5 h of UV irradiation as compared to those without UV treatment (0.331 ± 0.019 CV g−1 DW). Despite the low dendrobine content, PLBs as a source of dendrobine remains appealing due to its short culture duration (6 wk) compared to mother plants (2 yr). [ABSTRACT FROM AUTHOR]

This study reported the first-ever de novo transcriptome analysis of Operculina turpethum, a high valued endangered medicinal plant, using the Illumina HiSeq 2500 platform. The de novo assembly generated a total of 64,259 unigenes and 20,870 CDS (coding sequence) with a mean length of 449 bp and 571 bp respectively. Further, 20,218 and 16,458 unigenes showed significant similarity with identified proteins of NR (non-redundant) and UniProt database respectively. The homology search carried out against publicly available database found the best match with Ipomoea nil sequences (82.6%). The KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis identified 6538 unigenes functionally assigned to 378 modules with phenylpropanoid biosynthesis pathway as the most enriched among the secondary metabolite biosynthesis pathway followed by terpenoid biosynthesis. A total of 17,444 DEGs were identified among which majority of the DEGs (Differentially Expressed Gene) involved in secondary metabolite biosynthesis were found to be significantly upregulated in stem as compared to root tissues. The qRT-PCR validation of 9 unigenes involved in phenylpropanoid and terpenoid biosynthesis also showed a similar expression pattern. This finding suggests that stem tissues, rather than root tissues, could be used to prevent uprooting of O. turpethum in the wild, paving the way for the plant's effective conservation. Moreover, the study formed a valuable repository of genetic information which will provide a baseline for further molecular research. [ABSTRACT FROM AUTHOR]

Neem is a medicinal plant used as antimalarial, antibacterial, antiviral, insecticide, and antimicrobial drug. This study aimed to investigate and predict the effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression by response surface methodology. The highest fresh and dry cell weights were 580.25 g/L and 21.01 g/L, respectively obtained 6 days after using 100 mg/L yeast extract. The highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained 2 and 4 days, respectively after using 25 mg/L yeast extract. Maximum mevalonic acid accumulation (1.75 mg/g DW) and production (23.77 mg/L) were observed 2 days after application of 50 mg/L yeast extract. The highest amount of squalene accumulation (0.22 mg/g DW) and production (4.53 mg/L) were achieved 4 days after using 50 mg/L yeast extract. Prediction results exhibited the highest azadirachtin accumulation (13.61 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.50 mg/g DW) and production (5.57 mg/L), and squalene accumulation (0.30 mg/g DW) by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, without yeast extract for 6.54 days and 4 days, respectively. Also, it was predicted that the highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. The qRT-PCR analysis displayed the maximum relative gene expression of SQS1 and MOF1 by using 150 and 25 mg/L yeast extract for 4 and 2 days treatment. [ABSTRACT FROM AUTHOR]

In contrast to understanding spinescence in savanna woody species, little is known about the functions of plant secondary metabolites (PSM). Negative effects of PSMs on individual animal performance potentially translate into negative effects on herbivore population growth. Hence, understanding PSM functions is important for the conservation of savanna megafauna. We tested the view that black rhinoceros (Diceros bicornis) diet preference is not affected by spinescence or total phenolic abundance. We hypothesized that the composition of phenolic mixtures, however, would affect preference. Furthermore, we tested our data from 71 woody species for a trade-off between structural and chemical defenses. Spinescence type, and spinescence generally, did not deter black rhino feeding. Using eco-metabolomic data, we found that total abundance of phenolics did not affect preference, but mixture composition did and that the probability of spinescence trading off against phenolics depended on the mixture. We note that our study was restricted to black rhino and that diet preferences of other mammal herbivores might be influenced by subtle differences in phenolic mixtures. However, our results did support a previous, more detailed study of phenolic profiles of six species showing the same patterns in relation to preference generalised across mammal herbivore species in savannas. Our results represent substantial advancement in the understanding of the roles of PSMs, especially flavonoid compounds, in the functioning of savanna ecosystems, and highlight the need to dig deeper into broad groups of traits such as spinescence or total phenolics to improve understanding of woody plant defenses in savannas. [ABSTRACT FROM AUTHOR]

Hairy root (HR) culture, a successful biotechnology combining in vitro tissue culture with recombinant DNA machinery, is intended for the genetic improvement of plants. This technology has been put to use since the last three decades for genetic advancement of medicinal and aromatic plants and also to harvest the economical products in the form of secondary metabolites that are significantly important for their ethnobotanical and pharmacological properties. It also provides an efficient way out for the quicker extraction and quantification of the valuable phytochemicals. The current review provides an account of the in vitro HR culture technology and its wide-scale applications in the field of research as well as in pharmaceutical industries. Different facets of HR with respect to the culture establishment, phytochemical production as well as research investigations concerning the areas of gene manipulation, biotransformation of the secondary metabolites, phytoremediation, their industrial utilisations and different problems encountered during the application of this technology have been covered in this appraisal. Eventually, an idea has been provided on HR about the recent trends on the progress of this technology that may open up newer prospects in near future and calls for further research and explorations in this field. Key points: • Genetic engineering–based HR culture aims towards enhanced secondary metabolite production. • This review explores an insight in the HR technology and its multi-faceted approaches. • Up-to-date ground-breaking research applications and constraints of HR culture are discussed. [ABSTRACT FROM AUTHOR]

Morus alba is one of the most important cultivated crop in Indian sub-continent contributing towards production of silk fibre that carries economic importance worldwide. Post harvest preservation of M. alba leaves is a challenging factor as decrease in concentration of essential metabolites that needed for silk gland development takes place. Decrease in chlorophyll, protein, sugar concentration and increase in accumulation of free radicals and ROS takes place at post harvest stage of preservation, putting negative impact on larval development indicated by high mortality rate. Silver nitrate and nanosilver solution acts as an effective preservative, enhances the activity of enzymatic and non-enzymatic antioxidants thereby reducing the harmful effect of accumulated free radicals and ROS. The effectiveness of nanosilver solution was found to be on the upper site without any significant difference than silver nitrate, as higher retention of primary metabolites like pigments, proteins, and sugar takes place. The impact of feeding nanosilver preserved leaves on silkworm was found on the positive trend as larval growth rate, cocoon weight, shell weight, effective rate of rearing was observed almost same to the larvae fed with fresh leaves. [ABSTRACT FROM AUTHOR]

Main conclusion: Plant tissue culture as an important tool for the continuous production of active compounds including secondary metabolites and engineered molecules. Novel methods (gene editing, abiotic stress) can improve the technique.Humans have a long history of reliance on plants for a supply of food, shelter and, most importantly, medicine. Current-day pharmaceuticals are typically based on plant-derived metabolites, with new products being discovered constantly. Nevertheless, the consistent and uniform supply of plant pharmaceuticals has often been compromised. One alternative for the production of important plant active compounds is in vitro plant tissue culture, as it assures independence from geographical conditions by eliminating the need to rely on wild plants. Plant transformation also allows the further use of plants for the production of engineered compounds, such as vaccines and multiple pharmaceuticals. This review summarizes the important bioactive compounds currently produced by plant tissue culture and the fundamental methods and plants employed for their production. [ABSTRACT FROM AUTHOR]

The tea shot-hole borer beetle (TSHB, Euwallacea fornicatus) causes serious damage in plantations of tea, Camellia sinensis var. assamica, in Sri Lanka and South India. TSHB is found in symbiotic association with the ambrosia fungus, Monacrosporium ambrosium (syn. Fusarium ambrosium), in galleries located within stems of tea bushes. M. ambrosium is known to be the sole food source of TSHB. Six naphthoquinones produced during spore germination in a laboratory culture broth of M. ambrosium were isolated and identified as dihydroanhydrojavanicin, anhydrojavanicin, javanicin, 5,8-dihydroxy-2-methyl-3-(2-oxopropyl)naphthalene-1,4-dione, anhydrofusarubin and solaniol. Chloroform extracts of tea stems with red-colored galleries occupied by TSHB contained UV active compounds similar to the above naphthoquinones. Laboratory assays demonstrated that the combined ethyl acetate extracts of the fungal culture broth and mycelium inhibited the growth of endophytic fungi Pestalotiopsis camelliae and Phoma multirostrata, which were also isolated from tea stems. Thus, pigmented naphthoquinones secreted by M. ambrosium during spore germination may prevent other fungi from invading TSHB galleries in tea stems. The antifungal nature of the naphthoquinone extract suggests that it protects the habitat of TSHB. We propose that the TSHB fungal ectosymbiont M. ambrosium provides not only the food and sterol skeleton necessary for the development of the beetle during its larval stages, but also serves as a producer of fungal inhibitors that help to preserve the purity of the fungal garden of TSHB. [ABSTRACT FROM AUTHOR]

Abiotic stress in the form of gamma irradiation has been shown a potent inducer of oxidative stress in plant cell cultures which produce higher amounts of commercially important secondary metabolites. In the present study, the impact of low doses of gamma irradiation on growth and accumulation of 20-hydroxyecdysone of Sesuvium portulacastrum was investigated. Shoot cultures were established on Murashige and Skoog medium supplemented with indole-3-acetic acid (0.5 mg L) and N -benzylaminopurine (2.0 mg L). Mutations were induced in tissue culture by treating multiple shoots at low doses of gamma irradiation in the range from 5 to 40 Gy. In vitro cell growth and 20-hydroxyecdysone were assessed during MT, MT, MT, and MT generations. A gamma radiation dose of 20 Gy was calculated as 50% of the lethal value (LD). The survival rates of multiple shoot cultures exposed to high doses were gradually reduced in the course of increased order of generations. High-gamma irradiation doses were harmful to growth and 20-hydroxyecdysone production. The accumulation of 20-hydroxyecdysone of 0.139 mg/plant dry weight was significantly two-fold higher than non-irradiated shoot cultures. The stressed shoots increased 13-fold 20-hydroxyecdysone at 20 Gy during the MT generation compared to the yield of the MT generation. The ex vitro plants produced 0.321 mg/plant dry weight of 20-hydroxyecdysone which was remarkably greater than the untreated control. The present study postulated that gamma radiation induced metabolic changes and easy-to-achieve putative mutant comprised with the high amount of 20-hydroxyecdysone. [ABSTRACT FROM AUTHOR]

Evolvulus alsinoides L. is used for preparation of 'Shankhapushpi', an important popular ayurvedic drug that contributes considerably to the improvement of memory power. The improvement is attributed to the presence of furanocoumarin scopoletin, a metabolite with a wide range of biological activities. This report describes, for the first time, an in vitro culture system for propagation and enhanced production of scopoletin. Different concentrations of auxins and cytokinins individually and in combination were used in Murashige and Skoog (MS) medium to induce shoot regeneration in cotyledonary nodal explants and callus formation in leaf explants. The best response was achieved in MS medium fortified with 5.0 μM 6-benzyladenine (BA) in which 96 % of cultures produced 7.6 ± 0.6 shoots per explant. Regenerated shoots were rooted on MS medium with 5.0 μM indole-3-acetic acid (IAA). Plantlets were successfully acclimatized and established in soil. MS medium fortified with 10 μM BA + 5.0 μM IAA showed maximum growth and accumulation of scopoletin in cell cultures. Cell cultures could be maintained over 24 months. The influences of auxins, cytokinins, organic acids, amino acids, and fungal-derived elicitors on production of scopoletin were studied. Presence of either l-arginine, sodium pyruvate, or yeast extract highly promoted scopoletin production as compared with control and achieved 75.02-, 72.13-, and 57.98-fold higher accumulation, respectively. The results presented herein have laid solid foundation for large-scale production of scopoletin and further investigation of its purification and utilization as a novel pharmaceutical drug. [ABSTRACT FROM AUTHOR]

Bacopa monnieri (Linn.) Wettst. (family Scrophulariaceae), a therapeutically important perennial herb, was transformed to study the stability of the effects associated with the insertion of a gene encoding β-cryptogein, a proteinaceous elicitor, either alone or in addition to the T-DNA genes in long-term in vitro culture as well as after transfer to the greenhouse. Plant lines BmAr-IXcrypt obtained via transformation by LBA 9402-crypt (pRi1855+pBin19++ crypt) showed integration and expression of rolA, rolB, rolC, and rolD genes after 4 years of maintenance in vitro. The plant line BmAt-ncrypt obtained via transformation with A. tumefaciens strain LBA4404- crypt (harboring pBin19++ crypt) as well as plant line BmAr-IXcrypt showed stable integration and expression of nptII gene and crypt gene even after transfer to greenhouse. The clones of Ri-crypt-transformed plants differed morphologically from Ri-transformed plants in that the typical 'hairy root syndrome' was not observed in plants expressing crypt gene in the presence of rol genes. Except plant line BmAr-IXcrypt, none of the other transgenic plant lines showed any alteration in floral morphology and onset of flowering. The crypt-transformed plant lines (BmAr-IXcrypt and BmAt-ncrypt), maintained under long-term culture, showed significantly higher ( p ≤ 0.05) amount of bacoside production in vitro (1.66- to 2.05-fold, with respect to non-transformed plants). In addition, accumulation of all the four bacosides was significantly higher ( p ≤ 0.05) in crypt-transformed plants as compared to non-transformed plants grown for 1 year in greenhouse. Thus, the crypt-transformed plant lines of B. monnieri may be considered as the potential source for sustainable bioproduction of bacosides. [ABSTRACT FROM AUTHOR]

Tagetes erecta, L. an asteraceous plant of industrial and medicinal value, contains important compounds like pyrethrins, thiophenes and lutein, possessing immense potential for insecticidal, nematicidal and nutraceutical activities. Considering the importance and demand for these natural compounds, genetic manipulation of this crop for better productivity of secondary metabolites holds great significance. A rapid and reproducible direct regeneration and genetic transformation system is the prerequisite for genetic manipulation of any crop. This paper elucidates the establishment of an efficient direct regeneration and transformation protocol of T. erecta using Agrobacterium tumefaciens. Investigation of the effects of different types of explants (Hypocotyls, cotyledonary leaves, rachis and leaf sections) and different BAP and GA combinations on the regeneration frequency of T. erecta suggested that the best regeneration frequency (66 %) with an average of 5.08 ± 0.09 shoot buds/explant was observed from hypocotyl explants cultured on media containing 1.5 mg/l BAP and 5 mg/l GA The transformation protocol was established using A. tumefaciens strain LBA4404, containing the binary vector pBI121, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the neomycin phosphotransferase II ( nptII) gene as a kanamycin-resistant plant-selectable marker. Various parameters like optimization of kanamycin concentration (200 mg/l) for selection, standardization of cocultivation time (45 min) and acetosyringone concentration (150 μM) for obtaining higher transformation frequency were established using hypocotyl explants. The selected putative transgenic shoots were subsequently rooted on the Murashige and Skoog medium and transferred to the green house successfully. The plants were characterised by analysing the gus expression, amplification of 600 bp npt II fragment and Southern blot hybridization using the PCR-amplified gusA fragment as probe. The standardised protocol established during the study will open new vistas for genetic manipulation and introduction of desired genes for genetic improvement of T. erecta. [ABSTRACT FROM AUTHOR]

This review addresses methods of obtaining secondary metabolites from plant cell suspension and hairy root cultures and their exudates, particularly the physiological mechanisms of secondary metabolites release and trafficking. The efficiency for product recovery of metabolites can be increased by various methods, based on the principle of continuous product release into the cultivation medium. The most common methods for metabolite recovery are elicitation, influencing membrane permeability, and in situ product removal. The biosynthetic pathways can be influenced by cultivation conditions, transformation, or application of elicitors. The membrane permeability can be altered through the application of chemical or physical treatments. Product removal can be greatly increased through a two-phase system and the introduction of absorbents into the cultivation medium. In this review, we describe some improved approaches that have proven useful in these efforts. [ABSTRACT FROM AUTHOR]

Agrobacterium rhizogenes is a natural plant genetic engineer. It is a gram-negative soil bacterium that induces hairy root formation. Success has been obtained in exploring the molecular mechanisms of transferred DNA (T-DNA) transfer, interaction with host plant proteins, plant defense signaling and integration to plant genome for successful plant genetic transformation. T-DNA and corresponding expression of rol genes alter morphology and plant host secondary metabolism. During transformation, there is a differential loss of a few T-DNA genes. Loss of a few ORFs drastically affect the growth and morphological patterns of hairy roots, expression pattern of biosynthetic pathway genes and accumulation of specific secondary metabolites. [ABSTRACT FROM AUTHOR]

The carnivorous plant Drosera gigantea contains naphthoquinones among which plumbagin is known to have an antibacterial activity. The aims of this study were (i) to optimize conditions for micropropagation of D. gigantea and (ii) to evaluate the antibacterial activity of extracts of D. gigantea tissues and plumbagin against the bacterial fruit tree pathogens Pseudomonas syringae pv. syringae and P. syringae pv. morsprunorum. A number of media with different growth regulators [1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), thidiazuron (TDZ) and 6-benzylaminopurine (BAP)] or activated charcoal (CA) were tested for D. gigantea micropropagation. The best medium proved to be a halfstrenght Murashige and Skoog medium (1/2 MS) with 0.4% CA. All of the tested growth regulators resulted in changed phenotype of daughter plants and RAPD analysis showed that some of the regulators caused somaclonal variation. The minimal inhibitory concentrations and the minimal bactericidal concentrations (MBCs) of methanol and chloroform extracts of D. gigantea as well as pure plumbagin were determined and were in the range of 375-1875 μg dry weight (dw)/ml for methanol extract, 240-1425 μg dw/ml for chloroform extract and 5-150 μg dw/ml for plumbagin. The lowest MBCs (750-1875 μg dw/ml) were obtained with methanol extract of D. gigantea. These results suggest that in vitro-grown D. gigantea could be a potential source of antibacterial compounds, useful for the control of bacterial fruit tree pathogens but it should be further tested under in vivo conditions. [ABSTRACT FROM AUTHOR]

Plants are endowed with a large pool of structurally diverse small molecules known as secondary metabolites. The present study aims to virtually screen these plant secondary metabolites (PSM) for their possible anti-SARS-CoV-2 properties targeting four proteins/ enzymes which govern viral pathogenesis. Results of molecular docking with 4,704 ligands against four target proteins, and data analysis revealed a unique pattern of structurally similar PSM interacting with the target proteins. Among the top-ranked PSM which recorded lower binding energy (BE), > 50% were triterpenoids which interacted strongly with viral spike protein—receptor binding domain, > 32% molecules which showed better interaction with the active site of human transmembrane serine protease were belongs to flavonoids and their glycosides, > 16% of flavonol glycosides and > 16% anthocyanidins recorded lower BE against active site of viral main protease and > 13% flavonol glycoside strongly interacted with active site of viral RNA-dependent RNA polymerase. The primary concern about these PSM is their bioavailability. However, several PSM recorded higher bioavailability score and found fulfilling most of the drug-likeness characters as per Lipinski's rule (Coagulin K, Kamalachalcone C, Ginkgetin, Isoginkgetin, 3,3′-Biplumbagin, Chrysophanein, Aromoline, etc.). Natural occurrence, bio-transformation, bioavailability of selected PSM and their interaction with the target site of selected proteins were discussed in detail. Present study provides a platform for researchers to explore the possible use of selected PSM to prevent/ cure the COVID-19 by subjecting them for thorough in vitro and in vivo evaluation for the capabilities to interfering with the process of viral host cell recognition, entry and replication. [ABSTRACT FROM AUTHOR]

The nutgrass, Cyperus rotundus L. (Family: Cyperaceae), is a colonial, perennial herb considered to have originated in India 2000 years ago and widely used in Ayurveda to treat several ailments. In addition to its prehistoric uses, it is used in several systems of medicine for treating variety of diseases. The synergistic actions of the Cyperus’ compounds have added advantage over that of a single constituent. In the past decade, numerous studies proved analgesic, anti-allergic, anti-arthritic, anti-candida, anti-cariogenic, anti-convulsant, anti-diarrheal, anti-emetic, anti-helminthic, anti-histamine, anti-hyperglycemic, anti-hypertensive, anti-inflammatory, anti-malarial, anti-obesity, antioxidant, anti-platelet, anti-pyretic, anti-ulcer, anti-viral, cardioprotective, cytoprotective, cytotoxic, gastroprotective, hepatoprotective, neuroprotective, ovicidal, and larvicidal, wound healing and inhibition of brain Na+ K+ ATPase activities of C. rotundus and its chemical constituents. However, the exact the mechanism of action is not very clear and requires further evaluation. These properties strongly suggest an extensive use of C. rotundus for clinical applications. In this review, we attempted to provide information about the pharmacological effects of Cyperus and its proposed mechanisms of actions. [ABSTRACT FROM AUTHOR]