Ghansham B. Dixit

Free amino acid profiling in grain amaranth using LC-MS/MS.

Reliable and reproducible techniques for identification and quantification of amino acids usually require derivatization. However, techniques such as LC-MS/MS may perhaps sideline the derivatization with significant accuracy. Understanding the interaction between free amino acids and several physiological processes apparently requires that the pool be studied in the seed itself. Data on free amino acid content of grain amaranth is very inadequate. The present analysis revealed a higher concentration of essential amino acids especially Threonine and Tryptophan than non essential amino acids. The study constitutes first report of use of LC-MS/MS method for analysing the free amino acid composition of amaranth grain.

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.

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

Phytochemical constituents and antioxidant potential of some underused fruits

Phenols, a major group of antioxidant phytochemicals, have profound importance due to their biological and free radical scavenging activities. To identify their potential sources, extracts of some underused fruits ( Muntingia calabura , Averrhoea bilimbi and Artocarpus altilis ) were assessed with regard to their total phenolics and flavonoid content, as well as antioxidant activity in different solvent systems (methanol, ethanol and acetone) and distilled water. The phenolics and flavonoid content of M. calabura was 1.356 to 3.872 mg tannic acid equivalents/g fresh weight (TAE/g fw) and 0.026 to 0.068 mg rutin equivalents/g fresh weight (RE/g fw), respectively. That of A. bilimbi was 0.581 to 1.334 mg TAE/g fw and 0.021 to 0.037 mg RE/g fw, and that of A. altilis was 0.689 to 1.723 mg TAE/g fw and 0.013 to 0.043 mg RE/g fw. The extracts were also found to have significantly different levels of antioxidant activities. A correlation between antioxidant activity and phenolic content was observed, which shows that the tested plant extracts are having potential antioxidants of natural origin.

Biosystematics and evolutionary studies in Indian Drimia species

Abstract  The taxonomic position and genetic relationship within Indian Drimia species is controversial due to their morphological similarities and genomic complexities. The present work gives an insight on the genetic relationship between Indian Drimia species on the basis of their karyotype, pollen morphology, flower opening characteristics, hybridization behavior, and by use of DNA sequence of two molecular markers (internal transcribed spacers [ITS] and maturase K [matK]). The karyotypic studies of Indian Drimia species revealed various polyploid forms making their identification and delimitation more difficult. The five species of Indian Drimia are grouped into two complexes, indica complex and wightii complex on the basis of their pollen morphology, karyotype, and hybridization behavior. These two groups were found to be evolving separately. The cytomorphological studies of wightii complex revealed that it is evolving through polyploid and chromosome repatterning, while indica complex have adapted polyploid as well as hybridization for evolution. Phylogeny obtained from DNA sequences of molecular markers (ITS and matK) confirmed that the indica complex and wightii complex are evolving parallely, by grouping them in two clusters. Thus, a combination of conventional and molecular methods proved to be of great use for delimiting a small but complex group of Indian Drimia species.

First report of Aspergillus niger as a plant pathogen on Zingiber officinale from India.

Zingiber officinale Rosc., belonging to the Zingiberaceae family, is medicinally important. It is commonly known as ginger and has been extensively cultivated for many centuries for use as a spice and traditional medicine in India. During a survey in September of 2007, leaves of Z. officinale showed a necrotic leaf spot disease on a plant growing in Kolhapur, India. Symptoms of the disease appeared as small (10 to 15 mm), rectangular to irregular, yellow spots on the leaves that covered a major area of the leaf when severe. The infection caused defoliation. Symptomatic leaves were collected and isolations from infected leaves were made on Czapeks Dox agar supplemented with streptomycin sulfate (30 mg/l). Plates were incubated at 28 to 30°C for 6 to 7 days. The fungal colonies were colorless to pale on the reverse side and covered with a dense layer of dark brown-to-black conidial heads. Conidia were globose to subglobose (3.5 to 5.0 μm in diameter), dark brown to black, and rough walled. 16S rRNA of isolates was amplified and sequenced (EMBL Accession No. AM941157) and compared with sequences of known Aspergillus species obtained from GenBank. The closest matches (99% identity) were with A. niger. On the basis of morphological and molecular approaches, the pathogen was confirmed as A. niger. Of five isolates, AN-5 was used for pathogenicity study. Kochs postulates were satisfied after reisolating the fungus from leaves inoculated with a conidial suspension that showed symptoms (6 days after inoculation) similar to the lesions observed on the leaves collected from Z. officinale. The tests were repeated three times in the greenhouse. Healthy, potted Z. officinale plants were grown in isolation after artificial inoculation with a suspension of spores (10,000 conidia per ml), which was prepared in 100 ml of sterile distilled water and then sprayed on the abaxial and adaxial surface of leaves. Noninoculated plants served as controls. These results confirmed the pathogenicity of A. niger on Z. officinale. To our knowledge, this is the first report of A. niger causing a disease on ginger (1). Reference: (1) G. N. Dake. J. Spices Aromatic Crops. 4:40, 1995.

A new species of Brachystelma (Asclepiadaceae) from India

Erect, perennial herb, 9-30 cm in height; roots subglobose, tuberous; tubers 2-5 cm in diameter. Stem terete, 0-2-0-4 cm in diam., usually unbranched, hispid. Leaves sessile or subsessile, lanceolate or narrowly elliptic, 4-9 x 05-2 5 cm, acute. Flowering axis with internodes 1-3 cm long bearing flowers in whorls at each node. Flowering nodes with two opposite, subulate, reduced leaves, flowers 2 on either side of each reduced leaf, nodes 3-5 but usually 4-flowered; flowering nodes 3-7. Flowers small, brown-black, bracteate. Pedicels 0 -3-0 5 cm long, puberulous; bracts subulate, 12 mm long. Calyx 5-partite, sepals 1-2 mm long, puberulous. Corolla divided almost to base, dark purple and variously variegated within, greenish-purple outside, corolla-tube short, c. 1.5 mm long; corolla-lobes broad at base, tapering at apex, connate or free,

RP-HPLC Analysis of an Alkaloid–Methyllycaconitine from Mutagenic Delphinium malabaricum (Huth) Munz

A systematic search of some phytochemicals in Delphinium malabaricum will be undertaken to evaluate the therapeutic quality of D. malabaricum. The populations of D. malabaricum were subjected to analysis of alkaloids by Reversed phase HPLC technique. Among the various alkaloids present in Delphinium species methyllycaconitine is a principle alkaloid. Therefore, the populations of D. malabaricum were assessed for Methyllycaconitine (MLA) content in the mutants and their control. The results revealed that the MLA content was highest with 0.01% concentration of EMS (0.78 mg/g plant material) as compared to the control (0.76 mg/g plant material). The concentration of MLA decreased with an increase in dose/concentration of the mutagens. Less concentration of MLA in the roots of D. malabaricum indicates that it is less toxic and a further decrease in concentration after mutagenic treatment minimizes its toxic effects. As these populations are growing in the same environmental and edaphic conditions, observed variability can be related to genetic makeup altered due to mutation. The RP-HPLC analysis results could form the basis of a more detailed study of root alkaloids and any marked similarity/dissimilarity in their alkaloid pattern may draw reasonable conclusions of the difference between the mutants and the parent cultivar.