Generation of Variability for Salt Tolerance in Rice Using Tissue Culture Techniques

The study deals with the generation of variability for salt tolerance in rice using tissue culture techniques. Rice is the staple food of more than half of the world’s population. The management of drought, salinity and acidity in soils are all energy intensive agricultural practices. The Genetic variability is the basis of crop improvement. Somaclonal and androclonal variation can be effectively used for this purpose. In the present study, eight isozymes were studied and esterase and isocitric dehydrogenase was found to have varietal specific, developmental stage specific and stress specific banding pattern in rice. Under salt stress thickness of bands and enzyme activity showed changes. Pokkali, a moderately salt tolerant variety, had a specific band 7, which was present only in this variety and showed slight changes under stress. This band was faint in tillering and flowering stage .Based on the results obtained in the present study it is suggested that esterase could possibly be used as an isozyme marker for salt tolerance in rice. Varietal differences and stage specific variations could be detected using esterase and isocitric dehydrogenase . Moreover somaclonal and androclonal variation could be effectively detected using isozyme markers.

Immune Response of Penaeus monodon to the Inactivated White Spot Syndrome Virus Preparation

National Centre for Aquatic Animal Health, Cochin University of Science and Technology

Paddy Field Cyanobacteria : Diversity and Pesticide Tolerance

School of Environmental Studies, Cochin University of Science and Technology

Probiotic effect of Bacillus NL110 and Vibrio NE17 on the survival, growth performance and immune response of Macrobrachium rosenbergii (de Man)

Eight hundred and eighty-¢ve strains of bacterial isolates fromvarious samples associatedwith the natural habitat ofMacrobrachiumrosenbergii were screened for their probiotic potential. Two putative probionts namely Bacillus NL110 and Vibrio NE17 isolated from the larvae and egg samples, respectively, were selected for experimental studies and were introduced to the juveniles of M. rosenbergii (0.080 0.001g) through di¡erent modes such as through feed, water and both. The probiotic potential of the above bacteria in terms of improvements inwater quality, growth, survival, speci¢c growth rate (SGR), feed conversion ratio and immune parameters was evaluated. The treatment groups showed a signi¢cant improvement in SGR and weight gain (Po0.001). Survival among di¡erent treatment groups was better than that in the control group. There were also signi¢cant improvements in the water quality parameters such as the concentration of nitrate and ammonia in the treatment groups (Po0.05). Improvements in immune parameters such as the total haemocyte count (Po0.05), phenoloxidase activity and respiratory burst were also signi¢cant (Po0.001). It is concluded that screening of the natural micro£ora of cultured ¢sh and shell¢sh for putative probionts might yield probiotic strains of bacteria that could be utilized for an environment-friendly and organic mode of aquaculture.

Heavy metal tolerance patterns of total heterotrophic bacteria isolated from the soils of Mahatma Gandhi University campus, Kottayam, Kerala

Industrialization of our society has led to an increased production and discharge of both xenobiotic and natural chemical substances. Many of these chemicals will end up in the soil. Pollution of soils with heavy metals is becoming one of the most severe ecological and human health hazards. Elevated levels of heavy metals decrease soil microbial activity and bacteria need to develop different mechanisms to confer resistances to these heavy metals. Bacteria develop heavy-metal resistance mostly for their survivals, especially a significant portion of the resistant phenomena was found in the environmental strains. Therefore, in the present work, we check the multiple metal tolerance patterns of bacterial strains isolated from the soils of MG University campus, Kottayam. A total of 46 bacterial strains were isolated from different locations of the campus and tested for their resistant to 5 common metals in use (lead, zinc, copper, cadmium and nickel) by agar dilution method. The results of the present work revealed that there was a spatial variation of bacterial metal resistance in the soils of MG University campus, this may be due to the difference in metal contamination in different sampling location. All of the isolates showed resistance to one or more heavy metals selected. Tolerance to lead was relatively high followed by zinc, nickel, copper and cadmium. About 33% of the isolates showed very high tolerance (>4000μg/ml) to lead. Tolerance to cadmium (65%) was rather low (<100 μg/ml). Resistance to zinc was in between 100μg/ml - 1000μg/ml and the majority of them shows resistance in between 200μg/ml - 500μg/ml. Nickel resistance was in between 100μg/ml - 1000μg/ml and a good number of them shows resistance in between 300μg/ml - 400μg/ml. Resistance to copper was in between <100μg/ml - 500μg/ml and most of them showed resistance in between 300μg/ml - 400μg/ml. From the results of this study, it was concluded that heavy metal-resistant bacteria are widely distributed in the soils of MG university campus and the tolerance of heavy metals varied among bacteria and between locations

Probiotic effect of Bacillus NL110 and Vibrio NE17 on the survival, growth performance and immune response of Macrobrachium rosenbergii (de Man)

Eight hundred and eighty-¢ve strains of bacterial isolates fromvarious samples associatedwith the natural habitat ofMacrobrachiumrosenbergii were screened for their probiotic potential. Two putative probionts namely Bacillus NL110 and Vibrio NE17 isolated from the larvae and egg samples, respectively, were selected for experimental studies and were introduced to the juveniles of M. rosenbergii (0.080 0.001g) through di¡erent modes such as through feed, water and both. The probiotic potential of the above bacteria in terms of improvements inwater quality, growth, survival, speci¢c growth rate (SGR), feed conversion ratio and immune parameters was evaluated. The treatment groups showed a signi¢cant improvement in SGR and weight gain (Po0.001). Survival among di¡erent treatment groups was better than that in the control group. There were also signi¢cant improvements in the water quality parameters such as the concentration of nitrate and ammonia in the treatment groups (Po0.05). Improvements in immune parameters such as the total haemocyte count (Po0.05), phenoloxidase activity and respiratory burst were also signi¢cant (Po0.001). It is concluded that screening of the natural micro£ora of cultured ¢sh and shell¢sh for putative probionts might yield probiotic strains of bacteria that could be utilized for an environment-friendly and organic mode of aquaculture

Primary hemocyte culture of Penaeus monodon as an in vitro model for white spot syndrome virus titration, viral and immune related gene expression and cytotoxicity assays

Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1 ), tryptose phosphate broth (2.95 g l 1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 lgml 1 chloramphenicol, 100 lgml 1 streptomycin and 100 IU ml 1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-20-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals

Lymphoid organ cell culture system from Penaeus monodon (Fabricius) as a platform for white spot syndrome virus and shrimp immune-related gene expression

Shrimp cell lines are yet to be reported and this restricts the prospects of investigating the associated viral pathogens, especially white spot syndrome virus (WSSV). In this context, development of primary cell cultures from lymphoid organs was standardized. Poly-l-lysine-coated culture vessels enhanced growth of lymphoid cells, while the application of vertebrate growth factors did not, except insulin-like growth factor-1 (IGF-1). Susceptibility of the lymphoid cells to WSSV was confirmed by immunofluoresence assay using monoclonal antibody against the 28 kDa envelope protein of WSSV. Expression of viral and immunerelated genes in WSSV-infected lymphoid cultures could be demonstrated by RT-PCR. This emphasizes the utility of lymphoid primary cell culture as a platform for research in virus–cell interaction, virus morphogenesis, up and downregulation of shrimp immune-related genes, and also for the discovery of novel drugs to combat WSSV in shrimp culture

Proline Accumulation and Salt Tolerance in Rice

The morphological and biochemical response of calli and seedlings of different rice cultivars were compared under acid saline conditions. Calli of both tolerant and sensitive varieties showed severe stress symptoms like browning and necrosis, but the onset of stress symptoms was delayed in Pokkali. Seedlings of Pokkali showed minimal stress symptoms in lower salinities, and curling and senescence of older leaves in higher salinities although plants revived on amelioration of stress. Seedlings of the other varieties showed severe stress symptoms even at low salinities and plant death at higher salinities. Salt stress induced accumulation of the putative osmoprotectant proline in calli and seedlings of all varieties. Proline accumulation was higher in sensitive varieties than in Pokkali. These results indicate that proline accumulation is not directly correlated with salt tolerance in rice.

Distribution, extracellular virulence factors and antibiogram of motile aeromonads in fresh water ornamental fishes and immune response of Cyprinus carpio against Aeromonas hydrophila infection

The thesis deals with the prevalence and distribution of motile aeromonads in selected ornamental fishes. The presence of motile aeromonads in ornamental fishes and associated carriage water is well documented. Though aeromonads are a part of autochthonous flora of natural waters, disease outbreak occurs as a result of environmental stress on the cultured species and virulence of the pathogens. While ornamental aquaculture in many parts of the world is highly organized and practiced scientifically, it is highly unorganized in India. The culture ponds/tanks are often maintained in very poor manner and the fishes are subjected to high degree of stress during transportation from the production facility to retail vendors. The situation is no better at retail outlets, where fishes are maintained in crowded condition without proper aeration or food. All these could result in high prevalence of diseases caused by motile aeromonads. No systematic study has been carried out to understand the prevalence of motile aeromonads in ornamental fishes and carriage water . It also gives an account of the production of extracellular virulence factors and the antibiogram of the different species of motile aeromonads isolated. The growth characteristics and virulence potential of a representative strain of Aeromonas hydrophila is also studied. The nucleotide sequencing of the strain was carried out and sequences deposited in Genbank. Survival and immune response of Cyprinus carpio under different stress conditions and on probiotic treatment with Bacillus NL110, when challenged with A. hydrophila is also dealt within this thesis.