Genetic diversity of bitter gourd (Momordica charantia L.) genotypes revealed by RAPD markers and agronomic traits

Bitter gourd or bitter melon (Momordica charantia L.) is considered as minor cucurbitaceous vegetable in spite of having considerable nutritional and medicinal properties. Although some reports on genetic diversity based on morphological characterization are available, no work has been conducted to estimate genetic diversity using molecular markers in this crop. In the present study, 38 genotypes of M. charantia including few commercially cultivars collected from different parts of India based on agro-ecological zones were analysed for diversity study both at morphological and molecular levels. Genomic DNA was extracted from young healthy leaves following the procedure of Doyle and Doyle [Doyle, J.J., Doyle, J.L., 1990. A rapid DNA isolation procedure from small quantity of fresh leaf material. Phytochem. Bull. 119, 11–15]. Pair-wise comparison of genotypes was calculated as per the procedure of Jaccard [Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223–270]. Dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA) and the computation for multivariate analysis was done using the computer programme NTSYS-pc Version 2.0 [Rohlf, F.J., 1998. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.01. Exeter Software, Setauket, NY, USA]. Diversity based on yield related traits and molecular analysis was not in consonance with ecological distribution. Among 116 random decamer primers screened 29 were polymorphic and informative enough to analyse these genotypes. A total of 208 markers generated of which 76 (36.50%) were polymorphic and the number of bands per primer was 7.17 out of them 2.62 were polymorphic. Pair-wise genetic distance (GD) based on molecular analysis ranged from 0.07 to 0.50 suggesting a wide genetic base for the genotypes. The clustering pattern based on yield related traits and molecular variation was different.     

Temperature shifts induce adaptive changes in the physical state of carp (Cyprinus carpio L.) erythrocyte plasma membranes in vitro

Blood, freshly collected from warm- and cold-acclimated carp, Cyprinus carpio L., was cooled to 5°C for 4h or warmed to 25°C for 4h, respectively, and the fluorescence anisotropy of washed red blood cells was recorded using the fluorescent dye 3-(p-(6-phenyl-1,3,5-hexatrienyl) phenyl propionic acid [DPH-PA] (which is restricted to the outer leaflet of the plasma membrane) before and after the temperature shift. Despite individual variation, the plasma membrane of cold-exposed erythrocytes became more fluid while that of warm-exposed cells became more rigid following the temperature shift. This response was rapid and reversible. Cold-exposed cells from warm-acclimated fish became more fluid within 40–60 minutes and reverted to their original fluidity within the same time on warming, at a rate of 1°C/min; erythrocytes, from cold-adapted carp displayed an opposite change in fluidity over a similar time period. Cells from warm-acclimated, temperature down-shifted carp hyperfluidized their plasma membranes in the cold, whereas cells from cold-acclimated fish up-shifted in temperature showed no similar effect. These cells showed a complete adjustment of membrane physical state to the temperature. Total phospholipids obtained from warm-acclimated temperature down-shifted cells became more rigid than they were, when assayed at the acclimation temperature. In contrast, phospholipids obtained from cold-acclimated cells became more rigid when exposed to increasing temperatures. No significant changes occurred to the polar head groups, or to the fatty acid composition of the total phospholipids. It was concluded that the lipids play only a secondary role in the control of the physical state of plasma membrane in carp erythrocytes, and that some non-lipid components of these structures might be involved in these regulatory processes.     

Effect of high inclusion of rendered animal by‐product ingredients on growth,digestibility and economic performances in climbing perch Anabas testudineus

A 70‐day growth trail was conducted to investigate the effects of inclusion of high levels of meat and bone meal (MBM) and protein concentrate (PC) on growth, digestibility and economic performances of climbing perch, Anabas testudineus. Four isonitrogenous diets were formulated by lowering the level of dietary fishmeal protein at 0 (D₁, control), 70 (D₂), 85 (D₃) and 100% (D₄) with a mixture of MBM and PC protein (1:1). Triplicate groups of 300 fish (mean weight of 0.80 g) stocked in each 40 m² pond and fed the respective test diets. A digestibility trial was conducted after the growth trial in indoor glass aquarium. The result showed that growth parameters were significantly decreased (P < 0.05) with fishmeal replacement levels. However, significant differences were not found in feed conversion ratio and survival of fish. No difference was also found in protein efficiency ratio among D1, D2 and D3. Similar to growth parameters, total fish production was highest in D1, intermediate in D2 and D3; and lowest in D4. Apparent digestibility coefficients of dry matter, protein and lipid were highest (P < 0.05) in D1 and lowest in D4. The economic analysis revealed that the benefit cost ratio was ranked by D₃ (1.81), D₂ (1.71), D₁ (1.66) and D₄ (1.46) respectively. Upon considering the overall performances and unavailability of finite protein sources, it can be concluded that 70–85% fishmeal could be replaced with a mixture of MBM and PC (1:1) in practical diets for climbing perch.     

Soil boron status: impact of lime and fertilizers in an Indian long-term field experiment on a Typic Paleustalf

In Indian agriculture, nitrogen (N) and phosphorus (P) fertilizers are predominantly used by the farmers, often ignoring secondary and micronutrients. Significance of boron (B) in nutrient management studies has been increasingly underlined under intensive cropping systems particularly in acid soils. In order to understand the distribution of soil native B in different fractions and their contribution to plant B uptake as influenced by nutrient management, soil samples collected after wheat (2009–2010) from a long-term experiment (LTE) continuing since 1972–1973 on Typic Paleustalf of Ranchi were subjected to sequential fractionation of soil B. Treatments included N alone, NP, NPK, 150% of recommended NPK, NPK + farmyard manure (FYM), NPK + lime, and an unfertilized-control. Five soil B fractions were determined along with hot CaCl₂-extractable (available) B. Averaged across the treatments, the soil had low organic carbon (C), pH and cation exchange capacity (CEC), and high free sesquioxides. Total B content was 21.7 mg kg^?1. Among different B fractions, residual B was the major contributor to total B and other fractions collectively shared 7% of total B only. Application of N alone depleted readily soluble, specifically adsorbed and organically bound B bringing the contents even below unfertilized-control. Conjoint use of lime or FYM with NPK increased significantly these fractions, whereas a decrease in oxide bound B was noticed under these treatments. Available B was positively correlated with these fractions indicating their significance in controlling B availability in the soil. The study revealed that use of lime or FYM helped modifying the distribution of soil B in different fractions by way of changing soil pH and organic C content, resulting in enrichment of plant available pool. A drastically low available B content in different treatments receiving fertilizers alone, however, suggested the necessity of B fertilization at prescribed rates for maintaining soil B fertility as also high crop yields.     

Nitrogen,Phosphorus, and Carbon Budgets in Polyculture Ponds of Indian Major Carps and Giant Freshwater Prawn in Orissa State,India

A nutrient budget was quantified for six polyculture ponds of giant freshwater prawn (Macrobrachium rosenbergii) and Indian major carps (Catla catla, Labeo rohita, and Cirrhinus mrigala). The initial stocking size of prawn postlarva was 16.50 ± 0.54 mm with a body weight of 0.05 ± 0.001 g. Body weight of major carps was 9.0 ± 0.89 g. The duration of culture was 280 days. Feed accounted for 82% of total nitrogen (N), 93% of total phosphorus (P), and 95% of organic carbon (OC) inputs. Harvest of prawn and carps recovered 44% N, 1% P, and 19% OC. N, P, and OC accumulated in sediment were 47%, 73%, and 69%, respectively. Nutrient load in the harvest water was 0.67 ± 0.21 kg inorganic N, 0.15 ± 0.01 kg P, and 7.72 ± 0.62 kg OC per ton of Indian major carps and prawn.     

Deciphering the cross-talk of implantation: advances and challenges

Implantation involves a series of steps leading to an effective reciprocal signaling between the blastocyst and the uterus. Except for a restricted period when ovarian hormones induce a uterine receptive phase, the uterus is an unfavorable environment for blastocyst implantation. Because species-specific variations in implantation strategies exist, these differences preclude the formulation of a unifying theme for the molecular basis of this event. However, an increased understanding of mammalian implantation has been gained through the use of the mouse model. This review summarizes recognized signaling cascades and new research in mammalian implantation, based primarily on available genetic and molecular evidence from implantation studies in the mouse. Although the identification of new molecules associated with implantation in various species provides valuable insight, important questions remain regarding the common molecular mechanisms that govern this process. Understanding the mechanisms of implantation promises to help alleviate infertility, enhance fetal health, and improve contraceptive design. The success of any species depends on its reproductive efficiency. For sexual reproduction, an egg and sperm must overcome many obstacles to fuse and co-mingle their genetic material at fertilization. The zygote develops into a blastocyst with two cell lineages (the inner cell mass and the trophectoderm), migrates within the reproductive tract, and ultimately implants into a transiently permissive host tissue, the uterus. However, the molecular basis of the road map connecting the blastocyst with the endometrium across species is diverse (1) and not fully understood. Recent advances have identified numerous molecules involved in implantation (1-4), yet new discoveries have not yielded a unifying scheme for the mechanisms of implantation.     

Solvent tuning of electrochemical potentials in the active sites of HiPIP versus ferredoxin

A persistent puzzle in the field of biological electron transfer is the conserved iron-sulfur cluster motif in both high potential iron-sulfur protein (HiPIP) and ferredoxin (Fd) active sites. Despite this structural similarity, HiPIPs react oxidatively at physiological potentials, whereas Fds are reduced. Sulfur K-edge x-ray absorption spectroscopy uncovers the substantial influence of hydration on this variation in reactivity. Fe-S covalency is much lower in natively hydrated Fd active sites than in HiPIPs but increases upon water removal; similarly, HiPIP covalency decreases when unfolding exposes an otherwise hydrophobically shielded active site to water. Studies on model compounds and accompanying density functional theory calculations support a correlation of Fe-S covalency with ease of oxidation and therefore suggest that hydration accounts for most of the difference between Fd and HiPIP reduction potentials.     

Hydrous Ferric Oxide (HFO)—a Scavenger for Fluoride from Contaminated Water

Groundwaters contaminated with excess fluoride (above 1.5 mg/L) posed some risks to the public health in India. Methods available for fluoride contaminated water treatment are cogaulation–filtration using alum + lime + bleaching powder (Nalgonda technique) and adsorption using activated alumina. Use of aluminium compounds for water treatment purpose needs replacement on neurological health problem. Objective to this, the synthesis and fluoride adsorption behavior of hydrous ferric oxide (HFO) are reported here. It is seen that fluoride adsorption density varies as a function of pH, contact time, aging time, drying temperature and particle size of HFO. Highest adsorption density for fluoride is found to be at pH 4.0. Effects of competing anions in removing fluoride from solution were tested. Arsenite, arsenate, phosphate and sulfate show strong interfering effect at high anions to fluoride molar ratio in solution. Adsorption of fluoride on HFO follows the Freundlich isotherm and the Lagergren first-order kinetic model. It was also determined that HFO is a better adsorbent in removing fluoride from high fluoride groundwater than some other adsorbents. Regeneration of fluoride-rich HFO results showed that 1.0 M NaOH solution could be used up to a maximum of 75% regeneration.     

In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method

Macrophomina phaseolina (Tassi) Goid. is the most important pathogen of jute and primarily causes seedling blight, leaf spot and stem rot. The pathogen was detected from field samples by a simple method of direct PCR (dPCR) which obviates the steps of DNA extraction. The leaf bits were treated with a lysis buffer at 65°C for 25 min, whereas the stem pieces were initially incubated at 65°C for 5 min followed by incubation at 60°C for 25 min and the lysate was used as PCR template. Based on the type of tissue, the composition and concentration of lysis buffer systems were optimized. For leaf samples the optimized buffer system composed of 20 mmol l ^-1 tris (hydroxymethyl aminomethane (Tris)-Cl (pH 8.0), 1.5 mmol l ^-1 ethylene diamine tetra acetate (EDTA) (pH 8.0), 1.4 mol l ^-1 sodium acetate and 200 μg/ml proteinase K. Further, 3% PVP (w/v) and β-mercaptoethanol (1% w/v) were added into the buffer. In case of stem samples, PVP was not applied and higher concentrations were used for other components. M. phaseolina could be detected from both leaf and stem samples generating amplicon of 350 bp. This is the first report of detecting M. phaseolina by a direct PCR method without DNA extraction.     

<Emphasis Type="Italic">Fusarium</Emphasis> elicitor-dependent calcium influx and associated ros generation in tomato is independent of cell death

Fusarium oxysporum f. sp. lycopersici elicitor (EFOL-2) treatment induces cytosolic influx of calcium in Fusarium-resistant tomato suspension culture. The calcium signature was found to be biphasic, which is characteristic of recognition of oligosaccharides in the elicitor preparation. Further, several lines of evidence such as, (i) attainment of saturation level of the [Ca²⁺]_cyt at a definite extra-cellular calcium concentration (ii) prominent reduction in EFOL-2-induced influx in [Ca²⁺]_cyt on treatment with the calcium channel blockers verapamil and diltiazem and (iii) establishment of a refractory stage of [Ca²⁺]_cyt level upon repeated stimulation by EFOL-2, is indicative of receptor-mediated activation of the calcium channel for cytosolic elevation. In addition, inhibition of EFOL-2-induced [Ca²⁺]_cyt increase by protein kinase inhibitor staurosporine and wortmannin indicate phosphorylation is a regulatory event of calcium influx. Additionally, monitoring of cell death on EFOL-2 treatment indicated that the degree of ROS generation is not capable of inducing cell death. Inhibition of ROS generation on two separate occasions such as, calcium-free media and on treatment with inhibitors causing calcium channel occlusion revealed ROS generation as a successive event of calcium influx.