M. B. Shivanna

Ethno-medicinal application of plants in the eastern region of Shimoga district, Karnataka, India

AIM OF THE STUDY The present paper aimed to document and study the role of traditional herbal drugs in the treatment of human and veterinary ailments by communities residing in the eastern part of Shimoga district of Karnataka in India and to determine the consensus of informants knowledge on different category of ailments and fidelity level of plant species in treating particular disease. METHODOLOGY The ethno-medico-botanical information was collected from herbal healers and knowledgeable elder people residing in 15 randomly selected villages using semi-structured interviews. The data were subjected to informant consensus factor (ICF) and fidelity level (FL) analyses. RESULTS Eighty-five plant species of 41 families used to treat 31 human and 10 veterinary ailments were documented. Those medicinal plants which are effective in the treatment of liver complaints category had high ICF (0.77) and the skin diseases and disorders category attracted low ICF (0.12). Certain medicinal plants like Cyclea peltata, Justicia adhatoda, Memecylon umbellatum, Phyllanthus amarus and Tabernaemontana alternifolia were assigned with high degree of FL (100%) value and certain species like Cassia fistula was assigned with very low FL (25%). Some of the noteworthy medicinal uses of plant species in the study area were not reported for such purposes in the previously published literature on ethnobotany. CONCLUSION The local communities residing in eastern part of Shimoga district depended on herbal formulations for the treatment of diseases and disorders that affected them and their animals, as well. A high ICF and FL values indicated the possible occurrence of valuable bioactive compounds in certain plants and such of these plants need scientific evaluation for their pharmacological activities.

Endophytic fungi from Miquelia dentata Bedd., produce the anti-cancer alkaloid, camptothecine

Camptothecine (Campothecin, CPT), a quinoline alkaloid, is a potent inhibitor of eukaryotic topoisomerase I. Several semi-synthetic derivatives of CPT are in clinical use against ovarian, small lung and refractory ovarian cancers. While CPT is produced by several plant species belonging to the Asterid clade, in recent years, efforts have been made to isolate endophytic fungi from some of these plants as possible alternative sources of CPT. In this study we report the isolation of three endophytic fungi from fruit and seed regions of Miquelia dentata (Icacinaceae), that produce CPT, 9-methoxy CPT (9-MeO-CPT) and 10-hydroxy CPT (10-OH-CPT). All the three fungi identified as, Fomitopsis sp. P. Karst (MTCC 10177), Alternaria alternata (Fr.) Keissl (MTCC 5477) and Phomposis sp. (Sacc.) produced CPT, 9-MeO-CPT and 10-OH-CPT in mycelial mats in shake flasks containing potato dextrose broth. Methanolic and ethyl acetate extracts of these fungal species were cytotoxic to colon and breast cancer cell lines. We discuss these results in the context of the recent interest in endophytic fungi as possible alternative sources of plant secondary metabolites.

Biological control of anthracnose of chilli with rhizosphere and rhizoplane fungal isolates from grasses

The fungal species from rhizosphere and rhizoplane of perennial grasses of the Western Ghats of India were studied for their pathogenicity, antagonism in vitro, substrate and root colonization abilities, rhizosphere competence, growth in different soil pH and inoculum shelf-life. Out of 138 non-pathogenic fungal isolates tested, 85 were antagonistic in vitro to chilli anthracnose pathogen Colletotrichum capsici. Fifteen isolates with >60% inhibition zone to pathogen culture had saprophytic and root and rhizosphere colonization abilities. The sorghum grain inocula of test antagonistic fungi- Fusarium oxysporum, Chaetomium globosum and Trichoderma harzianum had the shelf-life of 90 days at 20 ± 2 °C and required optimum soil pH of 6.5. The above fungal isolates when tested for biocontrol of anthracnose disease in greenhouse and field caused reduction in seedling mortality and decreased disease incidence and severity at various plant growth stages and significant reduction in chilli fruit and seed infection. The test antagonistic fungi promoted seedling and mature plant growth and increased fruit and seed yield. Populations of these antagonistic fungi were fairly high in chilli rhizosphere at harvest. The present study indicated that antagonistic fungi from grass rhizosphere and rhizoplane could be used to control anthracnose and promote plant growth, and increase yield of chilli in field.

Fungal foliar diseases of Rauwolfia serpentina in wild, its seasonal occurrence, seed transmission and disease management

Rauwolfia serpentina is an important medicinal herb. Studies were conducted to determine causal organisms of foliar diseases in R. serpentina in Bhadra Wildlife Sanctuary during 2006–2009. The foliar disease incidence and its distribution and disease severity in nine state forest regions of the sanctuary were determined. The seedborne nature and transmission of the causal organism was also determined. The management of seedborne inoculum was done by seed dressing with fungicides. The effect of foliage infection on secondary metabolite content was also determined. Results of the present study indicated that Cercospora rauwolfiae is major leaf spot disease causing pathogen. The disease is homogeneously distributed through the study area. The foliar disease severity was high in Kagemanegiri forest during October–November. The minor leaf spot disease caused by Alternaria alternata occurred occasionally. Alternaria alternata is seedborne and seed transmitted and could be managed by seed treatment with Captra or Hyzeb. The secondary metabolites like alkaloids and steroids decreased with increase in foliar infection by C. rauwolfiae, while phenol and flavonoid contents increased. The study suggested that R. serpentina is affected in wild by C. rauwolfiae and A. alternata. The latter pathogen is seedborne and seed transmitted and controlled by seed treatment.

Foliar disease of Clitorea ternatea due to Colletotrichum dematium and its effect on secondary metabolite production

Clitorea ternatea L. finds extensive application in therapeutic medicine. Forest regions of the Western Ghats, India were surveyed for foliar disease(s) caused in C. ternatea. The incidence and severity of foliar disease were determined during August 2006 to July 2009. The distribution of foliar disease incidence was determined by Taylor’s power law. The pathogen was tested for its seed-borne nature and seed transmission. Fungicides, like Bavistin, Antrocol, Hyzeb M-45 and Captra, were used for managing seed-borne pathogens in C. ternatea. Diseased foliages were tested for secondary metabolite content. Results indicated that the foliar disease in C. ternatea is caused by Colletotrichum dematium. The disease incidence and severity was moderately high. The disease is distributed homogeneously and heterogeneously in two forest regions. The pathogen is seedborne and transmitted from seed to plant. Among fungicides, Antrocol and Bavistin reduced seed mycoflora incidence. The disease caused increased production of alkaloid, flavonoid, phenol and sterol in foliages.

Fungal foliar diseases in Withania somnifera and its effect on secondary metabolites

Withania somnifera is a promising revitalizing medicinal herb. The plant is affected by foliar diseases in Lakkavalli forest region of Bhadra Wildlife Sanctuary. The symptomatology of foliar fungal disease incidence, severity and distribution in the study area was examined during 2006–2009. The seedborne nature and transmission of the causal pathogen and its management with seed dressing fungicides were studied. The results of the study indicated that Alternaria alternata caused severe leaf spot disease, while Myrothecium roridum and Fusarium oxysporum caused minor diseases. Based on the internal transcribed spacer (ITS1 and ITS2) regions of rDNA, the major pathogen was identified as A. alternata. The disease is homogeneously distributed in Lakkavalli forest region and high severity is recorded during November. Alternaria alternata and Fusarium oxysporum were the dominant seedborne pathogens that are transmitted to seedlings. Among the seed dressing fungicides used, Hyzeb was the most effective, followed by Captra, Antracol and Bavistin, in reducing the incidence of A. alternata and other seedborne fungi. The infected W. somnifera foliages had decreased steroids and alkaloids and increased phenolics and flavonoids. Analysis of alkaloids in diseased foliages by high performance thin layer chromatography indicated the occurrence of transformed compounds at Rf = 0.1, 0.77 (254 nm) and 0.2 (366 nm).

Pod and seed traits in candidate plus trees of Pongamia pinnata (L.) Pierre from southern peninsular India in relation to provenance variation and genetic variability

Provenance variation and genetic variability in pod and seed traits were carried out with 232 candidate plus trees of Pongamia pinnata collected from selected agro-ecological zones of southern peninsular India during February–March 2008 to 2010. Significant variation (P < 0.01) for pod and seed traits across zones and provenances within zones was recorded. The southern dry and transition zone of Karnataka showed the highest mean value for all the pod and seed traits. A significant negative (P < 0.05) correlation and positive correlation was recorded for latitude with 100-seed weight and seed germination, respectively. Altitude positively correlated with 100-pod weight and negatively correlated with oil content. Phenotypic co-efficient of variation was higher than the genotypic co-efficient of variation indicating the influence of environment. Central dry zone and eastern dry zone of Karnataka showed high heritability and genetic advance for 100-pod weight, 100-seed weight, and seed germination. Positive significant (P < 0.05) genotypic correlation of co-efficient was recorded between seed thickness and oil content (0.211). Germination percentage showed significant positive correlation with seed weight and oil content. Path analysis revealed that seed thickness contributed directly to oil content (0.272). Based on D2 analysis, pongamia accessions could be grouped into seven clusters. Maximum diverse Clusters I and VII were identified based on inter-cluster distance (40.19). Thus on the basis of the present findings, it might be suggested that accessions from these clusters could be most suitably used for hybridization and in situ germplasm conservation.

Serological diagnosis and host range studies of important viral diseases of a few cucurbitaceous crops in Maharashtra, India

Around 39 well characterised viruses affect cucurbits crops in developing countries and their viral diversity may be the consequence for genetic and ecological diversity of their hosts. Indeed, cucurbits are grown in variety of climatic, environmental and agricultural conditions, and this may provide more or less favourable conditions for the specific viruses or their hosts. The presence of various viral diseases caused by different viruses in Maharashtra was studied from the infected samples collected from cucurbits and melons during the survey conducted in 2009–2010 in different locations. The virus isolates collected from various cucurbitaceous crops were established and their host ranges were studied by sap transmission. The study revealed Cucumber Mosaic Virus (CMV), Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV) and Cucumber green mottle mosaic virus infections predominately found in Nashik region, and Watermelon bud necrosis virus (WBNV), CMV, ZYMV, WMV and Watermelon silver mottle virus (WSMoV) infections in Aurangabad and Paithan regions. In Sangamner region, the crop was mostly affected by WBNV, ZYMV and WSMoV, and CMV was found only in Sillod region. The protocols for performing sap transmission tests in assay hosts were standardised for ZYMV, CMV and WBNV. Using direct antigen-coating enzyme-linked immunosorbent assay, of all the plant parts, young leaves were found to have high concentration of virus and suitable for virus detection in screening programmes. CMV and ZYMV was found to have high concentration of virus and suitable for virus detection in screening programmes.

PCR-based RAPD technique to determine induced salinity tolerance in vitro in Acacia auriculiformis

Acacia auriculiformis is an agro-forestry species in India with application in paper industry. The species is saline intolerant and fails to establish. Experiments were conducted to induce saline tolerance in callus cultures of A. auriculiformis on Murashige & Skoogs medium supplemented with different concentrations of NaCl+CaCl2––– (2:1; 50, 100, 150, 200, 250 or 300 mM). Calli were trained for eight sub-culture generations. The genome from saline-tolerant calli was extracted by modified cetyl trimethyl ammonium bromide method and subjected to polymerase chain reaction-based random amplified polymorphic DNA technique. The polymorphic DNA bands were eluted and cloned to Escherichia coli with the help of pTZ57R/T. The transformed plasmid DNA was sequenced and subjected to BLAST at National Center for Biotechnology Information. Results indicated that saline training of calli resulted in the expression of tolerance to 100 and 150 mM saline concentration after eight sub-cultures. Both the saline concentrations produced conspicuous bands in trained calli. Three nucleotide sequences were similar to those of Bacillus amyloliquefaciens sub sp. plantarum and BLAST search result indicated high homologies to ThiT thiamine transporter, YuaG flottilin type band 7, and U32 class peptidase protein genes in bacterial genome. The annotation of these protein genes in saline tolerance was discussed. The study suggested that the endophytic B. amyloliquefaciens sub sp. plantarum might be inducing saline tolerance in A. auriculiformis.