The extent of genetic diversity among Vanilla species: Comparative results for RAPD and ISSR

Vanilla is a large genus of about 110 species in the orchid family (Orchidaceae), including the species Vanilla planifolia from which commercial vanilla flavoring is derived. Since most species of vanilla are considered rare and endangered there is an urgent need to conserve them through genetic analysis and propagation/conservation studies on this crop.The present study investigated the genetic diversity among nine leafy- and leaf-less Vanilla species employing 30 decamer RAPD primers and 10 ISSR primers. The species under study were diverse and displayed a range of variability (0–66% and 0–81% for RAPD and ISSR, respectively). A total of 154 RAPD polymorphic markers (83.24%, h = 0.378) and 93 ISSR polymorphic markers (86.11%, h = 0.363) were used to generate a genetic similarity matrix followed by the cluster analysis. Specific groupings were revealed by each cluster analysis with slight variation between two different markers. Among the nine species studied, V. planifolia, Vanilla aphylla and Vanilla tahitensis revealed very low level of variation within their collections, thus indicating a narrow genetic base. The large genetic distance of Vanilla andamanica from other species suggests its different origin. A close genetic affinity was observed between the pairs V. planifolia, V. tahitensis and Vanilla albida, V. aphylla. These are the first comparative results for RAPD and ISSR reporting inter-relationship among nine cultivated, wild and hybrid Vanilla species.     

Geostatistical and fuzzy clustering approach for delineation of site-specific management zones and yield-limiting factors in irrigated hot arid environment of India

Precision Agriculture - Delineation of management zones (MZs) are needed to manage fields in order to maximize economic return, minimize environmental impact, and improve soil and crop management....     

Impact of Aeration on the Growth Performance of Silver Barb,Puntius gonionotus,during Fingerling Rearing

Impact of aeration on growth of silver barb, Puntius gonionotus during fingerling rearing was studied through a 100‐d rearing experiment conducted in 18 concrete tanks of 50 m² (10 × 5 × 1.2 m) size. Fry (0.74 ± 0.27 g, 35 ± 6 mm) were stocked in the experimental tanks at three stocking densities (25, 50, and 75 fry/m²) and were evaluated with and without provision of 6 h (2400–0600 h) of night time aeration. Aeration resulted in higher pH and dissolved oxygen regime and increased fingerling length and weight. The results suggest a rearing density of 75/m² to be ideal for rearing fry to fingerling of this species when aeration is provided, whereas, under non‐aerated condition, rearing the fry to fingerling stage at 50/m² was found advantageous over those at 25 and 75/m².     

Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA

The taxonomy and phylogeny of Indian Citrus is revisited using PCR-RFLP of the trnD-trnT and rbcL-ORF 106 regions as well as sequence data analysis of the trnL-trnF intergenic spacer region of cpDNA. The study was based on 50 accessions of Citrus genotypes, collected from wild, semi-wild and domesticated stocks. Of the 13 restriction enzymes (RE) used for restriction digestion of the polymerase chain reaction (PCR) amplicons, four (Hinf I, Msp I, Alu I, Hae III) generated 47 restriction fragments, of which 24 (51%) were polymorphic. PCR-RFLP data showed a genetic distance ranging from 0 to 0.79 among 50 accessions of Citrus, and a cluster analysis, based on Neighbor-Joining (NJ) method, placed all the accessions in eight major clusters. Analysis of trnL-trnF sequences from 23 representative accessions of Citrus showed a pair-wise sequence divergence rate in the range of 0–0.064. NJ, minimum evolution (ME) and maximum parsimony (MP) analyses of trnL-trnF sequences produced phylogenetic trees, which placed all the 23 accessions in five clusters. PCR-RFLP analysis resulted in a well resolved phylogenetic tree with branches supported by moderate to high bootstrap values, while the trnL-trnF sequence-based trees showed only moderate to low bootstrap support for the internal tree branches, indicating uncertain origin of some Citrus genotypes. This study shows that the trnL-trnF spacer sequence data can detect genetic variation in Indian Citrus genotypes, but the utility of the data in inferring phylogeny at intra and inter-specific levels is limited probably by factors such as hybridization, bud mutations, apomixis and polyploidy. However, PCR-RFLP and trnL-trnF data supported the recognition of C. maxima, C. medica, and C. reticulata as the basal species of edible Citrus.     

Nursery rearing of kalbasu, Labeo calbasu (Hamilton), at different stocking densities in outdoor concrete tanks

The growth and survival of kalbasu, Labeo calbasu, was evaluated at stocking densities of 5, 10 and 15 million spawn ha⁻¹ in nursery rearing in concrete tanks of 50 m², each for a period of 25 days. Survival of fry was density dependent and significantly higher (54.5%) at 5 million ha⁻¹ than those at 10 million ha⁻¹ (50.1%) and 15 million ha⁻¹ (46.9%). Similarly, growth and specific growth rate were inversely related to the stocking density and varied significantly among the three densities.     

Effect of sub-lethal nitrite on selected haematological parameters in fingerling Catla catla (Hamilton)

A sub‐lethal nitrite toxicity trial was conducted using static conditions for a period of 96 h with fingerlings of Catla catla (21.4±3.6 g). Fingerlings were exposed to five concentrations of nitrite, that is, 1, 2, 4, 8 and 10.4 mg L^?1 and a nitrite‐free control to study changes in haematological parameters. Nitrite caused an increase in immature erythrocyte population (7–24%) in lower concentrations (0–4 mg L^?1) at 6 h while they were absent in higher concentrations. The total erythrocyte count was reduced at 6 h followed by an increase at 12 h with further reduction up to 96 h in all concentrations of nitrite. The 96‐h exposure resulted in 21.2–31.8% reduction in erythrocyte population in 1–10.4 mg L^?1 nitrite. The haemoglobin content decreased progressively with increasing nitrite concentrations as well as exposure periods. Total leukocyte count decreased initially at 6 h in all treatments followed by an increase after 12 h, signifying development of a protective response of the body to nitrite stress. Blood glucose decreased initially up to 24 h followed by an increase through 96 h. Serum protein level decreased continuously with increasing exposure period. The study revealed that exposure to nitrite caused changes in almost all the haematological parameters in the fingerlings depending on the concentration as well as exposure period. Nitrite being one of the important inorganic nutrients often recorded at higher levels in intensively cultured ponds, the present study highlights its adverse impact on fish and stressed the need for the management of this nutrient in culture ponds.     

Comparative changes in water quality and role of pond soil after application of different levels of organic and inorganic inputs

Changes in water parameters were studied in a yard experiment for 7 weeks after application of cow dung at 20, 50 and 100 t ha^?1, poultry manure at 4, 10 and 20 t ha^?1, feed mixture (groundnut oil cake and rice bran at 1:1) at 10, 20 and 30 t ha^?1 and inorganic fertilizers at 100 kg urea+50 kg single super phosphate (SSP) ha^?1, twice this dose (2x ha^?1) and thrice this dose (3x ha^?1). To study the role of soil in the mineralization process, each treatment was divided into two groups – one with and the other without soil substrate. Higher degree of changes in water parameters was observed at higher input levels. Both organic amendment and inorganic fertilization caused significant reduction (P<0.05) in dissolved oxygen and increase in free CO₂, dissolved organic matter, total ammonia, nitrite, nitrate and phosphorus contents of water. Organic inputs significantly decreased (P<0.05) water pH and increased total alkalinity and hardness. In contrast, inorganic fertilization caused a significant increase in pH; alkalinity and hardness increased significantly in the presence of soil, but reduced in its absence. In organic input, presence of soil substrate caused significantly lower value of pH, dissolved oxygen, dissolved organic matter and phosphate‐phosphorus and significantly higher free CO₂, alkalinity, hardness, ammonia, nitrite and nitrate contents, compared with those in the absence of soil, revealing enhanced microbial mineralization in the presence of soil.     

Characterization of Ca-ATPase activity in gill microsomes of freshwater mussel, Lamellidens marginalis (Lamarck) and heavy metal modulations

Freshwater mussel Lamellidens marginalis showed a Ca²⁺-stimulated ATPase activity in the microsomal fraction of gill. Mg²⁺ was also found to be an activating cation for this ATPase enzyme. Ca²⁺ ATPase showed maximal activity at 40 °C and between pH 7.5–8.0. Substrate specificity of Ca²⁺ ATPase was highest with ATP, followed by GTP and other trinucleotides such as UTP, CTP and ADP also showed some amount of hydrolysis. Ca²⁺ ATPase showed slight inhibition with ruthenium red and sodium vanadate and is insensitive to sodium azide, ouabain, oligomycin B and phenylalanine. Heavy metals like Hg²⁺, Cu²⁺ and Zn²⁺ showed 50% inhibition of Ca²⁺ ATPase activity at concentrations of 0.25 mM, 0.5 mM and 1 mM, respectively whereas Cd²⁺ and Ni²⁺ showed 39% and 28% inhibition of enzyme activity at 1 mM concentrations.     

Spin conservation accounts for aluminum cluster anion reactivity pattern with O2

The reactivity pattern of small (approximately 10 to 20 atoms) anionic aluminum clusters with oxygen has posed a long-standing puzzle. Those clusters with an odd number of atoms tend to react much more slowly than their even-numbered counterparts. We used Fourier transform ion cyclotron resonance mass spectrometry to show that spin conservation straightforwardly accounts for this trend. The reaction rate of odd-numbered clusters increased appreciably when singlet oxygen was used in place of ground-state (triplet) oxygen. Conversely, monohydride clusters AlnH-, in which addition of the hydrogen atom shifts the spin state by converting formerly open-shell structures to closed-shell ones (and vice versa), exhibited an opposing trend: The odd-n hydride clusters reacted more rapidly with triplet oxygen. These findings are supported by theoretical simulations and highlight the general importance of spin selection rules in mediating cluster reactivity.     

Cell secretion and membrane fusion

Secretion occurs in all cells of multicellular organisms and involves the delivery of secretory products packaged in membrane-bound vesicles to the cell exterior. Specialized cells for neurotransmission, enzyme secretion or hormone release utilize a highly regulated secretory process. Secretory vesicles are transported to specific sites at the plasma membrane, where they dock and fuse to release their contents. Similar to other cellular processes, cell secretion is found to be highly regulated and a precisely orchestrated event. It has been demonstrated that membrane-bound secretory vesicles dock and fuse at porosomes, which are specialized supramolecular structures at the cell plasma membrane. Swelling of secretory vesicles results in a build-up of pressure, allowing expulsion of intravesicular contents. The extent of secretory vesicle swelling dictates the amount of intravesicular contents expelled during secretion. The discovery of the porosome, its isolation, its structure and dynamics at nm resolution and in real time, its biochemical composition and functional reconstitution into artificial lipid membrane, have been determined. The molecular mechanism of secretory vesicle fusion at the base of porosomes, and vesicle swelling, has also been resolved. These findings reveal the molecular mechanism of cell secretion.