Nishi Mishra

In this study, we have discriminated and identified the Genus of Curcuma and related species Zingiberaceae using rbcL and trnL DNA barcode primers. Curcuma genus related to the family Zingiberaceae comprises a significant number of medicinal plants renowned for their use in ethnomedicine, playing a pivotal role in the medical, health, and pharmaceutical sectors. Traditionally, morphological methods alone have proven insufficient for accurately identifying species within this family. However, DNA barcoding technology provides a contemporary solution by utilizing plant DNA sequences for species identification, thus enabling effective conservation efforts. We used DNA barcoding techniques and for analysis used the Maximum Parsimony tree in MEGA 11 with the Kimura 2-parameter (KP model) to analyse the genetic relationships between species. Out of the 13 accessions that were studied, 12 accessions belonged to Curcuma caesia and 1 accession belonged to Curcuma aeruginosa. The genetic relationships observed were correlated with the geographical distributions of these species. It was determined that C. aeruginosa is a mutated species of C. caesia. Additionally, 1 specimen of Alpinia galanga, a plant species related to the Zingiberaceae. Barcode primer trnL primer demonstrated a 92% efficiency during the investigation. The rbcL and trnL loci are recommended as potential barcode markers for discriminating between different plant species. This study developed a comprehensive DNA barcoding database that can confidently differentiate between species by combining morphological and molecular data. This database has the potential to identify adulteration in herbal products, combat illegal trade and adulteration of plant species, and assist in germplasm conservation efforts.

Curcuma caesia belongs to the genus Curcuma and the family of Zingiberaceae, which is a very important but unexplored medicinal plant. It is locally known as black turmeric or kali halide and is mainly used by the local tribal community as traditional medicine for the health sector. The rhizome of black turmeric has wide applications in the economic pharma sector due to essential active ingredients. This research aims to standardize a rapid, simple and efficient protocol for DNA extraction in Curcuma caesia which can be used for another genus of Zingiberaceae to obtain DNA from leaf samples. Changes in the concentration of components of DNA extraction buffer have improved the quantity of DNA from leaf samples compared to rhizomes. Extracted DNA samples also proved more efficient in PCR amplification of DNA barcode primers. The protocol developed in the present study is more efficient for leaf samples of Curcuma caesia compared to rhizome samples.

Santalum album (L.) is a prized tropical tree species of high therapeutic and industrial importance. The wood of these naturally grown plants is extensively harvested to acquire therapeutically important metabolite santalol and be used for additional functions such as in wood statuette industries. Due to high demand, it is crucial to maintain a sufficient plant population. An easy protocol for establishing cell suspension culture initiated from the loose embryogenic callus mass of sandalwood was realized by shifting 6–8-week-old morphogenic calli acquired from the mature embryonic axis and cotyledon explant cultures in fluid media. The asynchronous embryogenic cultures were sloughed with clumps of flourishing cell clumps and embryos of various progressive phases along with diffident non-embryogenic tissues. The frequency of embryo proliferation was evidenced to determinethe expansion pace of embryogenic masses under diverse conditions. The intonation of initiation and creation of cell suspension was under the directive of the influence of exogenous plant growth regulators amended in the nutrient medium at different concentrations and combinations. Maximum relative growth rate (386%) and clumps/embryoids in elevated integers (321.44) were accomplished on MS nutrient medium fortified with 2.0 mg L⁻¹ 2,4-D in association with 0.5 mg L⁻¹ BA and 30.0 g L⁻¹ sucrose raised from mature embryonic axis-derived calli. Plantlet regeneration in higher frequency (84.43%) was evidenced on MS medium amended with 1.0 mg L⁻¹ each of TDZ and GA₃ in conjunction with 0.5 mg L⁻¹ NAA and 20.0 g L⁻¹ sucrose. Mature embryonic axis-derived calli were found to be constantly better than mature cotyledon-derived calli for raising profitable and reproducible cell suspension cultures. Regenerants displayed normal growth and morphology and were founded successfully in the external environment after hardening.

Soybean (Glycine max (L) Merrill) is used in India mostly as a substantial fund of protein and oil, which makes the crop significantly important. Somaclonal variation has been researched as a base of additional variability for drought in soybean. In the present experiment calli/cell clumps/embryoids rose from immature and mature embryonic axis and cotyledons explants were exposed to different concentrations of polyethylene glycol (PEG6000). A discontinuous method proved to be superior as it permitted the calli/embryoids/cell clumps to regain their regeneration competence. A total of 64 (12.21%) plantlets of genotype JS335 and 78 (13.13%) of genotype JS93-05 were regenerated after four consequent subcultures on the selection medium with an effective lethal concentration of 20% PEG6000, and proliferated calli/embryoids/cell clumps were further subcultured on Murashige and Skoog regeneration medium supplemented with 0.5 mgL⁻¹ each of α-napthalene acetic acid (NAA), 6-benzyladenine (BA) and Kinetin (Kn), 20.0 gL⁻¹ sucrose and 7.5 gL⁻¹ agar. Putative drought-tolerant plantlets were acquired from genotype JS93-05 (38) in more numbers compared to genotype JS335 (26). Random decamer primers confirmed the presence of variability between mother plants and regenerated plants from both the genotypes. Since these plantlets recovered from tolerant calli/embryoids/cell clumps selected from the medium supplemented with PEG6000, the possibility exists that these plants may prove to be tolerant against drought stress.