Identification of quantitative trait loci for seedling cold tolerance using RILs derived from a cross between japonica and tropical japonica rice cultivars

Cold tolerance at the seedling stage of rice is an important phenotypic trait that causes normal plant growth and stable rice production in temperate regions as well as tropical high-lands in Asia and Africa. In order to find quantitative trait loci (QTLs)/genes associated with cold tolerance, we constructed a linkage map using 153 recombinant inbred lines (RILs) derived from a cross between a cold-tolerant temperate japonica cultivar, Geumobyeo, and a cold-sensitive tropical japonica breeding line, IR66160-121-4-4-2. The RILs were phenotyped for cold tolerance or sensitivity based on the degrees of cold tolerance as cold tolerance indices at the seedling stage. The seedlings for cold-tolerance/-sensitive traits were scored on the 7th day of the recovery period at 25°C after cold treatment at 10°C. Two QTLs (qCTS4a and qCTS4b) associated with cold tolerance at the seedling stage were identified on the long and short arms of chromosome 4 with an LOD score of 2.89 and 2.75, respectively, using composite interval mapping. The QTLs were flanked by simple sequence repeat (SSR) markers RM3648-RM2799 and RM3375a-RM558 that explained 8.3 and 7.8% of the total phenotypic variation, respectively. Seven of the selected RILs expressed cold tolerance at both the seedling and reproductive stages. The SSR markers associated with the QTLs will be useful for tracking favorable QTLs/genes into cold-sensitive elite cultivars and may have potential for pyramiding different QTLs for the improvement of cold tolerance in rice.     

Evaluation of genetic diversity among commercial cultivars, hybrids and local mango (Mangifera indica L.) genotypes of India using cumulative RAPD and ISSR markers

An assessment of genetic diversity studies was undertaken to understand the level and pattern of diversity in 65 mango (Mangifera indica L.) genotypes of India including 20 commercial cultivars, 18 hybrids, 25 local genotypes and two exotic cultivars based on qualitative and quantitative fruit characters as well as RAPD and ISSR profiles. A considerable variation was observed in respect of three important qualitative characters namely table quality, fruit attractiveness and storage life of ripe fruits and potentially superior genotypes for the above traits were identified. A wide variation was noticeable regarding metabolite composition of pulp of ripe mango fruit with respect to total soluble solids, total sugar, reducing sugar, acidity, sugar:acid ratio, ascorbic acid and phenolic content. Fifteen RAPD primers yielded 27 monomorphic and 129 polymorphic bands with percent polymorphism averaging 82.7%. Of a total 70 ISSR bands generated from eight ISSR primers, 60 bands (85.71%) were found to be polymorphic. Cumulative band data from these two methods precisely arranged accessions into eight clusters which correspond well with their pedigree relationship. UPGMA dendrograms drawn using RAPD, ISSR and cumulative data showed highly similar grouping of genotypes on the basis of their parental origin. No clear-cut geographical separation was revealed among East, West, North and South Indian mango cultivars by neither of these molecular markers nor their combinations. This supports the common gene pool origin of mango as well as operation of similar selection pressure as the cultivar preferences in these areas are largely similar.     

Nitrogen uptake efficiency of finger millet grown in hydroponic culture

The optimum nitrogen requirement along with nitrogen uptake efficiency of finger millet (Eleusine coracana) for obtaining plants of a high phenotypic quality was studied in this research using hydroponic culture. 20 days old seedlings were transferred in nutrient solution containing all the essential macro and micro nutrients. Plants were subjected to three treatments of nitrate viz. 0.05 mM, 0.1 mM and 0.5 mM by dissolving it in the nutrient solution. Plants grown in nutrient solution devoid of nitrate were treated as control. Plants grown in 0.5 mM nitrate conc. attained more height compared to other low nitrogen treatments (0.1 mM and 0.05 mM). Similarly plants of 0.5 mM nitrate treatments possessed more number of lateral roots, surface area of leaves, dry weight of plants and chlorophyll content compared to other low nitrogen treatments. Specific activity of nitrate reductase in plants treated with 0.5 mM concentration was found nearly 3 times higher than that of control plants; similarly, specific activity of nitrite reductase in 0.5 mM treated plants was also high compared to other low nitrogen treatments followed by control. The results of the present investigation therefore indicate that nitrogen uptake efficiency as revealed by the activities of nitrate reductase and nitrite reductase was high in 0.5 mM nitrate treatment. It also shows that 0.5 mM nitrate is optimum nitrogen concentration for the growth of finger millet in hydroponic condition.     

Organic acids from leaves,fruits, and rinds of Garcinia cowa

Organic acids in fresh leaves, fruits, and dried rinds of Garcinia cowa (G. cowa) were determined by high-performance liquid chromatography. Fresh leaves, fruits, and dried rinds were extracted with water at 120 degrees C for 20-30 min under 15 lbs/in(2) pressure. Also, dried rinds were extracted with solvents (acetone and methanol) using a Soxhlet extractor at 60 degrees C for 8 h each. The samples were injected to HPLC under gradient elution with 0.01 M phosphoric acid and methanol with a flow rate of 0.7 mL/min using UV detection at 210 nm. The major organic acid was found to be (-)-hydroxycitric acid present in leaves, fruits, and rinds to the extent of 1.7, 2.3, and 12.7%, respectively. (-)-Hydroxycitric acid lactone, and oxalic and citric acids are present in leaves, fruits, and rinds in minor quantities. This is the first report on the composition of organic acids from G. cowa.     

INLAND AQUACULTURE IN INDIA: PAST TREND,PRESENT STATUS AND FUTURE PROSPECTS

ABSTRACT

In India, inland aquaculture has emerged as a fast-growing enterprise and a viable alternative to the declining capture fisheries. The present paper is an attempt to assess Indian inland aquaculture with respect to its resource base, output trends, systems and activities, yield gaps, adoption and impact on aquaculturists, economics, returns to inputs, investment needs, and future prospects. The paper is largely based on existing literature and observations made as part of an ICAR-WorldFish demand supply project. Indian aquaculture is primarily limited to inland sector and carp-oriented; for that reason, this activity received special attention. Freshwater aquaculture observed tremendous growth in the past 15 years, but immense scope still exists for horizontal expansion and increases in productivity (vertical expansion). This is evidenced by the fact that the average farm fish yield is only one-third of that achieved in farm trials. The difference was mainly due to much higher input use in on-farm trials. Most of the aquaculturists were practicing extensive aquaculture, but aquaculturists with semi-intensive operations benefited most from adoption of technology. The benefit:cost ratios for different systems of aquaculture varied between 1.22 to 1.86. The return to capital was much higher than the return to labor, due to the low labor input. The semi-intensive aquaculture system would receive the greatest return from projected macrolevel investments, followed by extensive and intensive systems. Dedicated efforts are needed to meet the demand for quality fish seed and feed in order to achieve the desired 45% increase in area and greater than 50% increase in productivity. Based on the observations, activities designed to foster inland aquacultural development in India are recommended.     

Study of the efficiency of chemically assisted enucleation method for handmade cloning in goat (Capra hircus)

The present investigation was carried out to find an efficient chemically assisted procedure for enucleation of goat oocytes related to handmade cloning (HMC) technique. After 22-h in vitro maturation, oocytes were incubated with 0.5 μg/ml demecolcine for 2 h. Cumulus cells were removed by pipetting and vortexing in 0.5 mg/ml hyaluronidase, and zona pellucida were digested with pronase. Oocytes with extrusion cones were subjected to oriented bisection. One-third of the cytoplasm with the extrusion cone was removed with a micro blade. The remaining cytoplasts were used as recipients in HMC. Goat foetal fibroblasts were used as nuclear donors. The overall efficiency measured as the number of cytoplasts obtained per total number of oocytes used was significantly (p < 0.05) higher in chemically assisted handmade enucleation (CAHE) than oriented handmade enucleation without demecolcine (OHE) (80.02 ± 1.292% vs. 72.9 ± 1.00%, respectively, mean ± SEM). The reconstructed and activated embryos were cultured in embryo development medium (EDM) for 7 days. Fusion, cleavage and blastocyst development rate were 71.63 ± 1.95%, 92.94 ± 0.91% and 23.78 ± 3.33% (mean ± SEM), respectively which did not differ significantly from those achieved with random handmade enucleation and OHE. In conclusion, chemically assisted enucleation is a highly efficient and reliable enucleation method for goat HMC which eliminates the need of expensive equipment (inverted fluorescence microscope) and potentially harmful chromatin staining and ultraviolet (UV) irradiation for cytoplast selection.     

Molecular differentiation in Indian Citrus L. (Rutaceae) inferred from nrDNA ITS sequence analysis

Molecular differentiation in 24 accessions representing 19 taxa of Indian Citrus has been examined through sequence analysis of Internal Transcribed Spacer (ITS) region of nrDNA. Sequence length in the 24 accessions of Citrus taxa ranged from 512 to 665 bp (ITS1 & ITS2 partial and 5.8S complete sequence). The ITS sequences were very rich in G+C content ranging from 61.40 to 66.60% with an average of 64.2%. Genetic distance within Citrus group ranged from 0 to 13.4% with an average of 4.6%, showing moderate rate of nucleotide divergence. The phylogeny was inferred using the Maximum parsimony (MP) and Neighbor-Joining (NJ) methods. Both MP and NJ trees separated all the 24 accessions of Citrus into six distinct clusters. The disposition of all the accessions of Citrus in separate clusters in ITS-derived dendrograms was partly in accordance with the morpho-taxonomic affinities of the target taxa. This study supports the concept of Citrus medica (citron), C. reticulata (mandarin), and C. maxima (pummelo) as the basic species of the genus. However, ITS marker could not find any clear cut differentiation between subgenera Citrus and Papeda as proposed in Swingle’s Citrus classification system. The present study also supports the distinctiveness of C. indica (Indian wild orange), C. latipes (Khasi papeda) and C. hystrix (Melanesian papeda) as true species, besides elucidating the probable hybrid origin and relationships among the cultivated species/biotypes, such as Citrus ×aurantiifolia (sour lime) C. ×limon (lemon), C. ×taitensis (Indian rough lemon), C. limettioides (sweet lime), C. ×aurantium (including sour and sweet oranges and grapefruit), and other indigenous varieties of Indian origin: C. megaloxycarpa (sour pummelo), C. karna (karna orange), C. pseudolimon (Hill lemon), ‘Memang athur’, ‘Pummelo-lemon’ and ‘Kathairi nimbu’.     

Effect of feeding frequency on growth, survival and feed utilization in fingerlings of Catla catla (Hamilton), Labeo rohita (Hamilton) and Cirrhinus mrigala (Hamilton) in outdoor rearing systems

Fingerling rearing of Indian major carps, Catla catla, Labeo rohita and Cirrhinus mrigala, were conducted in concrete tanks under a polyculture system at a density of 0.3 million fry ha^?1 to evaluate the effect of one, two or three feedings per day on growth, survival and feed utilization. Fry were fed with formulated supplementary diet at 10% of biomass per day during the first 15 days, followed by 8% in the next 15 days and 6% thereafter, for 60 days. Feeding of fish three times daily led to higher growth (103.9±8.5 mm/10.3±2.4 g) than those feeding twice (100.2±4.8 mm/9.9±1.5 g) or once (97.2±9.8 mm/9.4±2.1 g) daily, although the values were not significantly different (P>0.05). Survival rate ranging between 72.3% and 75.1% also did not vary significantly (P>0.05) among the treatments. However, higher feeding frequencies resulted in better feed utilization as evident from decreasing feed conversion ratio values. The present study suggested requirement of a higher feeding frequency for rohu compared with catla and mrigal.     

Bacteriocins and their applications for the treatment of bacterial diseases in aquaculture: a review

Aquaculture is one of the fastest growing food producing sectors in the world. The increase in the world population requires faster growth in aquaculture for better food availability and to overcome malnutrition. But this worldwide growth of aquaculture is overwhelmed by catastrophic fish diseases and spoilage during cultivation and preservation problems caused by pathogenic bacteria. Various remedies are available for food preservation and also from the bacterial diseases in the fish production. Due to the emergence of antibiotic resistance and adverse effects, an alternative to antibiotic is the need of the hour. The proteins such as bacteriocins, i.e. ribosomally synthesized antimicrobial peptides (AMPs) and possess antagonistic against closely related and other bacteria. These proteins are produced by most lineages of bacteria which are playing key roles in recognition and possess a cognate immunity system for self‐protection as well as host protection from infections. These proteins are potent immunomodulators with broad spectrum inhibition properties which are further used as novel therapeutic agents. In this review, we have tried to summarize the bacteriocins on the basis of their classifications, structural and functional attributes, mode of actions, bacteriocins isolated from fish and gut microbiota and presence of beneficiary bacteria in the fish gut. Further, this study highlighted where further research is a prerequisite to increase our basic understanding and search for novel bacteriocins to elucidate the proteins/peptides having antimicrobial properties for disease control in aquaculture as an alternative to antibiotics.