Parthraj R. Kshirsagar

Highly efficient in vitro proliferation and genetic stability analysis of micropropagated Ceropegiaevansii by RAPD and ISSR markers: A critically endangered plant of Western Ghats

Ceropegiaevansii McCann (family: Asclepiadaceae), a critically endangered plant of Western Ghats has acquired significant importance due to its medicinal implications, edible tubers, and ornamental flowers. This study deals with the optimization of axillary bud proliferation using nodal explants followed by genetic stability analysis of regenerants. Maximum number of shoots (11.6 ± 1.1) was observed on the Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (4.0 mg/l) and indole-3-acetic acid (0.3 mg/l) with 85% shoot multiplication frequency. In vitro-grown shoots were rooted best in 1/2 MS medium supplemented with indole-3-butyric acid (1.0 mg/l) with an average of 10.3 ± 0.9 roots per shoot and 92% rooting frequency. Plantlets were acclimatized best (90%) in a mixture of sterile soil, sand, and coco peat (1:2:1). Micropropagated plants were subjected to random amplified polymorphic DNA and inter simple sequence repeat markers analyses. Collectively, 759 bands were generated which were monomorphic and similar to the mother plant. Findings of this study are the first report on micropropagation and assessment of genetic stability of micropropagated plantlets in C. evansii which suggests that axillary shoot proliferation can safely be used as an effective tool for propagation and conservation of C. evansii.

Phytochemical composition, antioxidant activity and HPLC profiles of Swertia species from Western Ghats.

Swertia chirayita is one of the potential medicinal plants of the family Gentianaceae in traditional medicine. Due to its high demand and scarcity, trade of chirayita is affected by adulterants. Swertia species from Western Ghats were compared with S. chirayita for phytochemical characterisation and antioxidant activities by using different extracts. This study revealed that acetone is the best extraction solvent of phenolic and flavonoid compounds with antioxidant properties as compared with other extracts. S. chirayita showed better antioxidant activity than other species with highest content of phenolics and flavonoids. Among the species from Western Ghats, Swertia minor has better antioxidant properties with higher content of phenolics and flavonoids when compared with S. chirayita. Gallic acid was detected in all species under study by using HPLC analysis. The Swertia species under study showed similar phytochemical properties and antioxidant potential and hence their use as substitute to S. chirayita needs to be further investigated.

Optimization of Extraction Techniques and RP-HPLC Analysis of Antidiabetic and Anticancer Drug Mangiferin from Roots of ‘Saptarangi’ (Salacia chinensis L.)

Saptarangi (Salacia chinensis L.) is a chief source of mangiferin, which has antidiabetic and anticancer properties. Extraction using continuous shaking extraction (CSE), ultrasonic extraction (UE), microwave assisted extraction (MAE), and steam bath assisted extraction (SBAE) were evaluated for extraction of mangiferin (MF) from roots of S. chinensis. The extracts were analyzed by reverse phase-high performance liquid chromatography (RP-HPLC) and Fourier transform-infra red spectroscopy (FT-IR) analysis. Operational parameters such as different solvents and exposure period were also evaluated. N,N-Dimethyl formamide (DMF 30%, v/v) extracted higher quantity of MF compared to other solvents. The results shows that the extraction of MF by SBAE was more efficient followed by MAE, CSE, and UE methods. Maximum quantity of MF (391.80 ppm) was obtained by SBAE technique and DMF (30%) as a solvent system. SBAE is the easiest and best method for screening of MF from S. chinensis. Furthermore, use of FT-IR validates the RP-HPLC technique for identification and quantification.

Seasonal variation in content of camptothecin from the bark of Nothapodytes nimmoniana (Grah.) Mabb., using HPLC analysis

Objective: To study and compare seasonal variation in camptothecin (CPT) content from bark samples of Nothapodytes nimmoniana obtained from geographically and climatologically isolated populations. Methods: A standard High Performance Liquid Chromatography methodology was used to analyze and quantify CPT from bark samples of N. nimmoniana. Results: Sample collected from Amboli yielded highest CPT content 1.337 g/100 g dry bark powder during the monsoon compared to other localities in study. Monsoon (August) showed to accumulate higher levels of CPT in barks of N. nimmoniana as compared to summer (May). Amboli averaged highest accumulation of CPT compared to other localities under study. Conclusion: These findings indicate season to have control over accumulation of CPT. Locality Amboli has highest CPT content in all seasons and were the elite population during the study. The study also suggests the need for further investigation in lights of biosynthesis in the plant.

Highly efficient in vitro regeneration, establishment of callus and cell suspension cultures and RAPD analysis of regenerants of Swertia lawii Burkill

Graphical abstract

Quantitative determination of three pentacyclic triterpenes from five Swertia L. species endemic to Western Ghats, India, using RP-HPLC analysis

Aim of this study was to identify pentacyclic triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from five Swertia species endemic to Western Ghats, which are used as substitutes as well as adulterants to Swertia chirayita. Our results indicate that the concentration of active compound vary largely among and within the species from different localities. OA was found in a higher amount in all species compared to BA and UA. From the results, it was clear that BA, OA and UA are present in the endemic species collected from Western Ghats, thus advocating the use of these species as alternate sources to S. chirayita. This in due course may release pressure of exploitation from natural resources of S. chirayita and help to bring it out from an endangered category from conservation point of view.

Extraction efficiency, phytochemical profiles and antioxidative properties of different parts of Saptarangi (Salacia chinensis L.) – An important underutilized plant

The study aimed to evaluate extraction efficiency, detection and quantification of phytochemicals, minerals and antioxidative capacity of different parts of Salacia chinensis L. Continuous shaking extraction, steam bath assisted extraction, ultrasonic extraction and microwave assisted extraction with varied time intervals were employed for extraction of phenolics, flavonoids, and antioxidants. Preliminary screening revealed the presence of wide array of metabolites along with carbohydrates and starch. Steam bath assisted extraction for 10 min exposure was found most suitable for extraction phenolics (46.02 ± 2.30 mg of gallic acid equivalent per gram of dry weight and 48.57 ± 2.42 mg of tannic acid equivalent per gram of dry weight) and flavonoids (35.26 ± 1.61 mg of quercetin equivalent per gram of dry weight and 51.60 ± 2.58 mg of ellagic acid equivalent per gram of dry weight). In support, reverse phase-high performance liquid chromatography- diode array detector confirmed the presence of seven pharmaceutically important phenolic acids. Antioxidant capacity was measured by 1, 1- diphenyl-1-picryl hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), 2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) scavenging (ABTS) and N, N-dimethyl-p-phenylenediamine (DMPD) assays and represented as trolox equivalent antioxidant capacity (TEAC) and ascorbic acid equivalent antioxidant capacity (AEAC). Antioxidant capacity ranged from 121.02 ± 6.05 to 1567.28 ± 78.36 µM trolox equivalent antioxidant capacity and 56.62 ± 2.83 to 972.48 ± 48.62 µM ascorbic acid equivalent antioxidant capacity. Roots showed higher yields of illustrated biochemical parameters, however fresh fruit pulp was found a chief source of minerals. Gas chromatography-mass spectroscopic analysis revealed the presence of a vast array of phytoconstituents associated with different plant parts. The present study revealed the amounts of minerals and diverse phytoconstituents in various parts of S. chinensis and confirmed its medicinal and nutritional implications.

Molecular authentication of the traditional medicinal plant “Lakshman Booti” (Smithia conferta Sm.) and its adulterants through DNA barcoding

Background: Smithia conferta Sm. is an annual herb widely used in Indian traditional medical practice and commonly known as “Lakshman booti” in Sanskrit. Morphological resemblance among the species of genus Smithia Aiton. leads to inaccurate identification and adulteration. This causes inconsistent therapeutic effects and also affects the quality of herbal medicine. Aim: This study aimed to generate potential barcode for authentication of S. conferta and its adulterants through DNA barcoding technique. Materials and Methods: Genomic DNA extracted from S. conferta and its adulterants was used as templates for polymerase chain reaction amplification of the barcoding regions. The amplicons were directed for sequencing, and species identification was conducted using BLASTn and unweighted pair-group method with arithmetic mean trees. In addition, the secondary structures of internal transcribed spacer (ITS) 2 region were predicted. Results: The nucleotide sequence of ITS provides species-specific single nucleotide polymorphisms and sequence divergence (22%) than psbA-trnH (10.9%) and rbcL (3.1%) sequences. The ITS barcode indicates that S. conferta and Smithia sensitiva are closely related compared to other species. Conclusion: ITS is the most applicable barcode for molecular authentication of S. conferta, and further chloroplast barcodes should be tested for phylogenetic analysis of genus Smithia. Abbreviations used: BLASTn: Basic Local Alignment Search Tool for Nucleotide; MEGA: Molecular Evolutionary Genetic Analysis; EMBL: European Molecular Biology Laboratory; psbA-trnH: Photosystem II protein D1- stuctural RNA: His tRNA gene; rbcL: Ribulose 1,5 bi-phosphate carboxylase/ oxygenase large subunit gene.

Vigna khandalensis (Santapau) Raghavan et Wadhwa: a promising underutilized, wild, endemic legume of the Northern Western Ghats, India

Vigna khandalensis (Santapau) Raghavan et Wadhwa, is an endemic legume belonging to subgenus Ceratotropis (Asian Vigna), genus Vigna of the family Fabaceae. It is commonly known as ‘Ranmung’, ‘Jungli mung’ or ‘Badamung’. V. khandalensis is sporadically distributed at high altitude areas in the Northern Western Ghats, India. It is an annual, erect, tall herb growing along roadsides, hilly slopes, pond sides and near human settlements. The legume seeds are utilized by human beings at the time of famine. Surprisingly, the plant remains neglected and underutilized although its high nutritional value. The scant information on V. khandalensis leads us to investigate this underexploited and underutilized legume as a food source for human. The present paper deals with geographical distribution, taxonomy, ecology, agro-morphological characterization, nutritional content and phytochemical analysis. The results obtained in the present investigation provide information on agro-morphological traits and nutritional potential of V. khandalensis. Furthermore, it is promoted as a legume ‘Crop for the Future’ for food security.