Plants as Food and Medicine: An Ethnobotanical Survey among Kanikaran Community in Southern India.

The study mainly focused on the plants used as food as well as medicine by Kani tribals in Kalakad Mundanthurai Tiger Reserve in Southern Western Ghats, India. An ethnobotanical survey was carried out among the Kani tribals through questionnaires and consultations with age-old and knowledgeable tribals during June 2007 to September 2009. A total of 59 species of plants were recorded in this study with their vernacular names, parts used as food either raw or cooked and medicinal uses. Among all the edible plants, unripe fruits of Artocarpus heterophyllus and tubers of Manihot esculenta are favorite edibles and these are the main food for the Kani tribal community. Due to indiscriminate exploitation, destruction of forests and changing scenario of rural life, the oral folklore of plants is on the way of extinction. The present investigation underlines the potential of ethnobotanical research and the need for documentation of traditional knowledge pertaining to the utilization of plants for greater benefit of mankind.

Development of transgenic finger millet (Eleusine coracana (L.) Gaertn.) resistant to leaf blast disease.

Finger millet plants conferring resistance to leaf blast disease have been developed by inserting a rice chitinase (chi11) gene through Agrobacterium-mediated transformation. Plasmid pHyg-Chi.11 harbouring the rice chitinase gene under the control of maize ubiquitin promoter was introduced into finger millet using Agrobacterium strain LBA4404 (pSB1). Transformed plants were selected and regenerated on hygromycin-supplemented medium. Transient expression of transgene was confirmed by GUS histochemical staining. The incorporation of rice chitinase gene in R0 and R1 progenies was confirmed by PCR and Southern blot analyses. Expression of chitinase gene in finger millet was confirmed by Western blot analysis with a barley chitinase antibody. A leaf blast assay was also performed by challenging the transgenic plants with spores of Pyricularia grisea. The frequency of transient expression was 16.3% to 19.3%. Stable frequency was 3.5% to 3.9%. Southern blot analysis confirmed the integration of 3.1 kb chitinase gene. Western blot analysis detected the presence of 35 kDa chitinase enzyme. Chitinase activity ranged from 19.4 to 24.8. In segregation analysis, the transgenic R1 lines produced three resistant and one sensitive for hygromycin, confirming the normal Mendelian pattern of transgene segregation. Transgenic plants showed high level of resistance to leaf blast disease compared to control plants. This is the first study reporting the introduction of rice chitinase gene into finger millet for leaf blast resistance.

Antimycobacterial activity of two natural alkaloids, vasicine acetate and 2-acetyl benzylamine, isolated from Indian shrub Adhatoda vasica Ness. leaves.

In folk medicine, Adhatoda vasica Ness. (Acanthaceae) is used to treat asthma and cough. The leaves of A.vasica were powdered and extracted with hexane, ethyl acetate and methanol. The hexane extract showed 97% reduction in colony-forming units (CFU) at 100 μg/ml. The hexane extract was subjected to column chromatography. Two natural compounds, vasicine acetate and 2-acetyl benzylamine, were isolated from it. They were bioassayed against Mycobacterium tuberculosis. The two compounds showed strong antimycobacterial activity. Vasicine acetate and 2- acetyl benzylamine isolated from hexane extract of A.vasica leaves, signifi cantly inhibited M. tuberculosis and one multi-drug-resistant (MDR) strain and one sensitive strain at 200 and 50 μg/ml, respectively. Our study demonstrated that both the compounds, vasicine acetate and 2-acetyl benzylamine, could be evaluated further for developing a drug to control M. tuberculosis.

Buffering capacity and membrane H+ conductance of protease producing facultative alkaliphilic bacterium Bacillus flexus from mangrove soil. Indian Journal of Biochemistry & Biophysics. 2009 June; 46(3): 261-265.

A facultative alkaliphilic protease-producing gram-positive rod-shaped bacteria (EMGA 5) was isolated from mangrove soil and confirmed as Bacillus flexus by the 16S rDNA sequence. Buffering capacity and membrane H+ conductance of this alkaliphilic isolate were investigated for the cells grown at pH 7.2 and 10.5 using acid pulse technique. Suspensions of B. flexus cells grown in poly peptone yeast glucose medium at pH 10.5 exhibited higher cytoplasmic membrane buffering capacity values (70 µmol H+/pH unit/mg protein at pH 9.9) than the cells grown at pH 7.2 (41 µmol H+/pH unit/mg protein at pH 9.9). B. flexus grown aerobically at pH 7.2 showed higher H+ conductance values than the cells grown at pH 10.5 (0.032 µmol H+/s/pH unit/mg protein at pH 9.9 and 0.028 µmol H+/s/pH unit/mg protein at pH 9.8, respectively). The present study revealed that the buffering capacity and membrane H+ conductance of the B. flexus isolates were influenced by pH of the medium.

Facultative alkalophilic bacteria from mangrove soil with varying buffering capacity and H+ conductance.

Facultative alkalophilic bacteria Planococcus sp. (EMGA-26), Bacillus sp. (EMGA-29) and Corynebacterium spp. (EMGA-33 and 130) were isolated from mangrove soil samples. Neutrophiles were predominant than alkalophiles. Buffering capacity and membrane H+ conductance were investigated for the strains grown in PPYG medium at pH 10.5 using acid pulse technique. Bacillus sp. showed higher buffering capacity than Planococcus sp. and Corynebacterium spp. Buffering capacity was two-fold higher in Corynebacterium sp. EMGA-33 than in EMGA-130. The membrane H+ conductance was high in Bacillus sp. and was directly proportional to the buffering capacity values. The Bacillus sp. (EMGA-29) had higher cell membrane adaptability in high pH environment than the Planococcus sp. and Corynebacterium spp.

Agrobacterium-mediated transformation of chickpea with alpha-amylase inhibitor gene for insect resistance.

Chickpea is the world's third most important pulse crop and India produces 75% of the world's supply. Chickpea seeds are attacked by Callosobruchus maculatus and C. chinensis which cause extensive damage. The alpha-amylase inhibitor gene isolated from Phaseolus vulgaris seeds was introduced into chickpea cultivar K850 through Agrobacterium-mediated transformation. A total of 288 kanamycin resistant plants were regenerated. Only 0.3% of these were true transformants. Polymerase chain reaction (PCR) analysis and Southern hybridization confirmed the presence of 4.9 kb alpha-amylase inhibitor gene in the transformed plants. Western blot confirmed the presence of alpha-amylase inhibitor protein. The results of bioassay study revealed a significant reduction in the survival rate of bruchid weevil C. maculatus reared on transgenic chickpea seeds. All the transgenic plants exhibited a segregation ratio of 3:1.

Efficient protocols for in vitro regeneration of Pennisetum glaucum (L) Br.

A system was developed for in vitro regeneration of Pennisetum glaucum through organogenesis and somatic embryogenesis. Mature embryo and leaf base explants of Pennisetum glaucum (L) Br. cv HH B60 (Poaceae) were cultured on Murashige and Skoog agar medium supplemented with 11.3 microM of 2,4-D for callus induction. Embryogenic calli were induced within eight weeks. Percentage of callus induction and somatic embryogenesis was significantly higher in mature embryo than leaf base explants. Maximum shoot regeneration was obtained via organogenesis on MS medium supplemented with 4.43 microM of BAP and 4.64 microM of kinetin from the calli of both the explants. The frequency of plant regeneration through somatic embryogenesis was comparatively lower than organogenesis. Regeneration frequency was higher in mature embryo explants than leaf base explants. The shoots regenerated via organogenesis were elongated and rooted efficiently on MS medium supplemented with IBA (0.49 microM). The rooted plantlets were hardened and transferred to soil.

Use of RAPD in assessing the genetic variability in Spodoptera litura.

A polymerase chain reaction based assay to distinguish six different ecotypes of the Spodoptera litura, a sporadic insect pest and a predominant defoliator of various crops was carried out. A total of 40 random primers were screened to reveal the existence of polymorphism between the populations. Among them eight showed scorable banding patterns and three primers (OPA-01, OPA-05, OPM-01) exhibited distinguishable banding patterns. However, Chengalpattu and Chennai populations revealed their closed relatedness and Coimbatore population stood distantly from others.

Hepatoprotective effect of coumestans isolated from the leaves of Wedelia calendulacea Less. in paracetamol induced liver damage.

Effect of coumestans isolated form the leaves of W. calendulacea was evaluated in paracetamol induced liver damage. The increased serum enzyme levels (lactate dehydrogenase, alanine and aspartate transaminase and alkaline phophatase) by paracetamol induction were significantly lowered due to coumestans treatment. Results of this study revealed that coumestans of W. calendulacea afforded a significant protective action in the alleviation of paracetamol induced hepatocellular injury.

Influence of growth regulators and explant type on in vitro shoot propagation and rooting of red sandal wood (Pterocarpus santalinus L.).

In vitro shoot regeneration in Pterocarpus santalinus L. was achieved when detached cotyledons from in vitro germinated seedlings were cultured on MS medium containing NAA (0.1 mg/L), BA (1 mg/L) and kinetin (1 mg/L). The regenerated shoots rooted on 1/4 strength MS medium with IAA (1 mg/L) and the fully developed plantlets were successfully established in the soil.

Agrobacterium mediated transformation of Vigna sesquipedalis Koern (asparagus bean).

Agrobacterium mediated transformation of Vigna sesquipedalis was achieved using cotyledonary node explants prepared from 5 days old seedlings germinated on B5 basal medium, and transformed using Agrobacterium tumefaciens strain EHA101, carrying the phosphinothricin-N-acetyltransferase gene and neomycin-3-phosphotransferase-II gene as selectable markers and GUS gene as a screenable marker. Gene transfer was achieved by inoculation of cotyledonary node explants with a bacterial suspension and a further cocultivation with Agrobacterium suspension for 3 days on B5 basal medium. Only 10% of the explants were transformed with EHA101 and exhibited transient expression of GUS genes, while 2% of shoots exhibited stable integration of genes and developed into plants. Transgenic character of tissues was confirmed by GUS assay and Southern analysis. Histological analysis of GUS gene expression directly after cocultivation revealed a high competence of subepidermal cell layers of cotyledonary node and associated cotyledons for transformation with Agrobacterium.

Resistance of legume seeds to the bruchid, Callosobruchus maculatus: metabolites relationship.

A study was initiated to categorize the seeds of various wild and cultivar legume varieties on the basis of their relative resistance to the bruchid, C. maculatus, and to correlate the important primary and secondary metabolites (non-protein anti-metabolites) in these seeds to the developmental parameters of the bruchid. In general, the wild seed varities showed greater amount of resistance to the bruchid attack when compared to that of the cultivar varieties. All the cultivar varieties studied showed higher amounts of primary metabolites, namely, proteins, carbohydrates, lipids and free amino acids thus showing a positive correlation between the primary metabolites content and the infestation rate. The wild varieties, however, showed significantly lower amounts of these primary metabolites and consequently a lower level of infestation. The non-protein anti-metabolites such as total phenols, ortho- dihydroxy phenols and tannis were significantly lower in the cultivars. The wild varieties, in contrast, revealed higher amounts of these secondary metabolites showing a negative correlation between these secondary metabolites content and the infestation rate. The study revealed that these non-protein anti-metabolites are important in conferring resistance to the seeds.

Changes of haemolymph protein profile in the larva of Pericallia ricini (Fabricius) parasitised by the Braconid Wasp, Apanteles taragamae Viereck (Hymenoptera: Braconidae).

Parasitism by the braconid wasp, A. taragamae caused alterations in the haemolymph polypeptides of woolly bear larvae of P. ricini. Analysis of haemolymph proteins by SDS-PAGE and densitometry showed that the quantities of haemolymph proteins were reduced dramatically in the parasitised larvae. Simultaneously, parasitism induced large amount of 95 kDa polypeptides in the haemolymph of the parasitised larvae. Also, a remarkable induction of 43 and 45 kDa polypeptides which are not detectable in non-parasitised larvae appeared in the parasitised larvae.

Effect of solvent residues of Vitex negundo Linn. and Cassia fistula Linn. on pulse beetle, Callosobruchus maculatus Fab. and its larval parasitoid, Dinarmus vagabundus (Timberlake).

Effect of solvent residues of Vitex negundo L. and Cassia fistula L. leaves (0.5 and 1%) was studied on egg laying and adult emergence of Callosobruchus maculatus Fab. and on percentage of larval parasitism by Dinarmus vagabundus (Timberlake). Cowpea seeds treated with methanol and aqueous extract of Vitex, at these concentrations (0.5 and 1%), significantly reduced the number of eggs and emergence of F1 adults of C. maculatus. Both Vitex and Cassia extracts did not affect the percentage of parasitism by D. vagabundus on C. maculatus grubs.

Ribosomal DNA repeat unit polymorphism in some pulses.

Restriction fragment length polymorphism (RFLP) of nuclear ribosomal RNA genes (rDNA) was carried out in some pulse species such as Lablab purpureus, Dolichos tribolus, Vigna bournii, V. grahmiana, V. unguiculata and V. wightii. Restriction endonuclease digestion and Southern blot analysis with wheat probe pTa 71 revealed a bewildering diversity. The fragment patterns produced by different enzymes such as Dpn II, Hind III, EcoRI and Bam H1 already illustrate considerable polymorphism.

Hypoglycemic activity of Cajanus cajan (seeds) in mice.

The effect of roasted and unroasted seeds of C. cajan on serum glucose levels of normal and alloxan diabetic mice were studied. Single doses of unroasted seeds (60% and 80%) on administration to normal as well as alloxanized mice showed significant reduction in the serum glucose levels after 1-2 hr and a significant rise at 3 hr. In case of roasted seeds, on other hand there was a significant increase in serum glucose levels during 3 hr experimental period. It is therefore concluded that roasting of seeds at high temperature for an half hour period resulted in the total loss of hypoglycemic principle but not the hyperglycemic principle present in the seeds.