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The present study illustrates the transformation ability of two wild-type bacterial strains of Rhizobium rhizogenes (MTCC 532 and MTCC 2364) on the embryogenic callus and callus-derived plantlets of a threatened Indian orchid, Dendrobium ovatum. Co-culture of the bacterium with the explants gave marginal hairy root phenotype that failed to multiply in the culture medium. Some primary and secondary metabolites were subdued in infected explants. Moscatilin, the stilbenoid active principle in D. ovatum, was found below the detection limit. The presence of two metabolites viz., Laudanosine, a benzyltetrahydroisoquinoline alkaloid and Lyciumin B, a cyclic peptide, were detected exclusively in the infected explants. The subjugated amino acids and phenolics in the infected plantlets were routed to produce phytoanticipins, and phenanthrenes, strengthening the defence mechanism in infected tissues. This research implies that the plant's defence mechanism activation could have prevented the extensive hairy root formation in the explants, even though nodulations and phenotype transitions were witnessed. Moscatilin has a structural resemblance with Resveratrol, a phytoalexin that combats bacterial and fungal pathogens. The study favours the possibility of Moscatlin being a precursor for phenanthrene compounds, thereby serving as a 'phytoanticipin' during the infection phase. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03180-9.

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Moscatilin, a bibenzyl derivative (stilbenoid), mostly found in one of the largest genera of Orchidaceae; Dendrobium has many therapeutic benefits. Its function as an anticancer agent has been widely demonstrated through many research investigations. However, the compound has not been produced in vitro to date. The present study highlights the development of cultures viz., seedling generation, callus induction and callus regeneration (transformation of callus into plantlets). These cultures were devised to conserve the threatened tropical epiphytic orchid species, Dendrobium ovatum and identify their potential towards moscatilin bioproduction in vitro. Among the three culture platforms, callus-derived plantlets could yield high moscatilin when treated with l-Phenylalanine as a precursor. Tissue differentiation was found to be indispensable for the high production of this polyphenol. These cultures also offer potential commercial benefits as they can serve as appropriate platforms to decode moscatilin biosynthesis and other significant bibenzyl derivatives. Elicitors, such as chitosan, salicylic acid, and methyl jasmonate, were found, causing an enhancement in moscatilin content in the cultures. The seedlings obtained can serve towards ecorestoration and preservation of the studied species. Callogenesis was useful in plantlet regeneration, as callus-derived plantlets could be utilized for the enrichment and commercial scale-up of moscatilin-like chemicals.

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BACKGROUND: Moscatilin is a bibenzyl derivative (stilbenoid), mainly found in Dendrobium species. This plant-derived chemical is a potential cytotoxic anticancer drug that acts against different cancer types. The present study compared the structural interactions of Moscatilin along with five clinically relevant drugs against two target proteins, viz., Anaphase-Promoting Complex subunit 10/Death of Cyclase 1 and Pyruvate Kinase Muscle isozyme M2 in silico. Out of five clinical ligands, four were plant-derived compounds, viz., Resveratrol, Paclitaxel, Shikonin, and Colchicine. The synthetic chemotherapeutic agent, Mitomycin-C, was used as a ligand to compare the mechanistic insights. The objective of the study was to determine the anticancer potency of Moscatilin in silico. RESULTS: Moscatilin was found to have an advantage over other drugs of interest due to its structural simplicity and folding bridge connecting the bibenzyl structures. Moscatilin exhibited dual function by exclusively affecting the cancer cells, creating instabilities in biochemical and molecular cascades. CONCLUSIONS: The study demonstrates that Moscatilin is has a multi-antimetastatic function. Moscatilin interaction with APC10/DOC1 indicated that the drug is involved with post-replicative inhibition, and with PKM2 showed glycolytic pathway inhibition in cancer cells. Moscatilin can function as an effective cell cycle inhibitor.

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Moscatilin (stilbenoid) is a plant-derived anticancer compound, and it has mostly been isolated from threatened wild Dendrobium species. The present study attempts to evaluate the cytotoxicity of Moscatilin on several cancer cell lines through MTT assay. Additionally, it also aims towards estimating and comparing the radiosensitivity, cell-cycle progression, and apoptotic/necrotic effect induced by Moscatilin on different cell lines. The effects of Moscatilin was compared with another significant stilbenoid anticancer agent, Resveratrol (a structural analog of Moscatilin), whose presence has also been reported in Dendrobiums. Considering the threatened nature of this genus, crude extracts of a tropical and epiphytic Dendrobium species, viz., Dendrobium ovatum, prepared from in vitro seedlings were also tested towards cytotoxicity and radiosensitization efficacy. Moscatilin functioned as an effective radiosensitizer at 5 µg/ml along with 1 Gy X-ray and 200 J/m2 UV-C radiations. It was also able to perturb cell cycle both at replicative and post-replicative phases with the aforementioned combination. Moscatilin, in unison with radiation, triggered immunogenic death specifically on cancer cells starting from Pyroptosis, terminating in Necroptosis. Moscatilin, when used singly, could evoke immunogenic cell death. Analyses of Damage-Associated Molecular Patterns released during radiation and Moscatilin treatment would aid in ascertaining the mode of cell death. Moscatilin is a potential radiosensitizer and must be tested for preclinical and clinical trials to combat cancer. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02827-3.

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Metabolic pathway reconstruction and gene edits for native natural product synthesis in single plant cells are considered to be less complicated when compared to the production of non-native metabolites. Being an efficient eukaryotic system, plants encompass suitable post-translational modifications. However, slow cell division rate and heterogeneous nature is an impediment for consistent product retrieval from plant cells. Plant cell synchrony can be attained in cultures developed in vitro. Isolated plant protoplasts capable of division, can potentially enhance the unimpaired yield of target bioactives, similar to microbes and unicellular eukaryotes. Evidence from yeast experiments suggests that 'critical cell size' and division rates for enhancement machinery, primarily depend on culture conditions and nutrient availability. The cell size control mechanisms in Arabidopsis shoot apical meristem is analogous to yeast notably, fission yeast. If protoplasts isolated from plants are subjected to cell size studies and cell cycle progression in culture, it will answer the underlying molecular mechanisms such as, unicellular to multicellular transition states, longevity, senescence, 'cell-size resetting' during organogenesis, and adaptation to external cues.

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The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4% pectinase together with 2% cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 104/ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.

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In the present study, the seeds of brinjal (Solanum melongena L.) var. Mattu Gulla were irradiated with single exposure of He-Ne laser at different doses of 5-40 J cm(-2) and germinated aseptically. Thirty day old seedlings were harvested and the germination, growth, physiological and biochemical parameters were estimated and compared with un-irradiated control seedlings. A significant enhancement in growth characters were noted with respect to length, fresh and dry weight of shoots and roots. In addition, chlorophyll (a and b), carotenoid content, anthocyanin and amylases (α and ß) activities were found to be altered. Significant alterations in percentage of seed germination (P < 0.001) and time to 50% germination (P < 0.001) were observed in the irradiated seeds compared with the un-irradiated controls. In conclusion, the results of the present study demonstrated that low dose (5-30 J cm(-2) ) of He-Ne laser irradiation enhanced the germination process and altered growth, by positively influencing physiological and biochemical parameters of the brinjal seedlings compared with un-irradiated control under in vitro conditions.

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There has been a resurgence and prevalence of fever with symptoms of Chikungunya (CHIK) and increased death toll in Kerala, the southern-most state of India. The objective of this study was to develop a rapid detection method to determine the presence of CHIK- virus in the serum samples collected from febrile patients in Kerala, India. Serum specimens were analyzed for CHIK viral RNA by RT-PCR using primers specific for nsP1 and E1 genes. Five out of twenty clinical samples were positive for CHIK virus. The partial sequences of the E1 and nsP1 genes of the strain, IndKL01 were highly similar to the Reunion strains and the recently isolated Indian strains. A novel substitution, A148V, was detected in the E1 gene of the isolate, IndKL02. The detection procedure used in this study was simple, sensitive and rapid (less than 4 hr). This result suggests that CHIK viruses similar to the Reunion strains, which had resulted in high morbidity and mortality rates, may have caused the recent Chikungunya outbreak in India. The effect of the variant, E1-A148V, in the virulence and the rate of transmission of the virus deserves further investigation.

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