Development of interspecific somatic hybrid cell lines in cultivated jute and their early characterization using jute chloroplast RFLP marker

A strong sexual incompatibility barrier that exists between the two cultivated jute species, Corchorus capsularis and Corchorus olitorius, limits the scope for improvement through genetic introgression. Protoplast fusion was carried out to generate interspecific hybrid cell lines. Cotyledonary cell protoplasts of C. capsularis and anthocyaninpigmented hypocotyl protoplasts of C. olitorius were used in the fusion experiments, which appeared to be visually useful in the early selection of the fused products. A chloroplast DNA (cpDNA) marker was developed in jute, which showed species‐specific hybridization patterns with EcoRI‐digested total genomic DNA of C. capsularis and C. olitorius. This cpDNA marker was used in the characterization of the somatic hybrid cell lines at their early stages of growth. Evidence for the presence of both types of cpDNA in the hybrid cell lines was obtained when the total genomic DNA of 4‐ to 7‐month‐old hybrid cell lines was challenged with the chloroplast DNA marker through Southern analysis. It was shown that the early segregation of the parental chloroplasts did not occur in jute, although this is common in other plant species. The hybrid nature of the fused cell lines could also be identified through peroxidase isozyme analysis. Isozyme banding patterns were complex and varied among the hybrid cell lines.     

Quantification of N2 fixation and annual N benefit from N2 fixation in soybean accrued to the soil under soybean‐wheat continuous rotation

A computational exercise was undertaken to quantify the percent N derived from atmosphere %Ndfa) in soybean and consequent N benefit from biological N₂‐fixation process annually accrued to the soil by the soybean crop using average annual N‐input/‐output balance sheet from a 7 yr old soybean‐wheat continuous rotational experiment on a Typic Haplustert. The experiment was conducted with 16 treatments comprised of combinations of four annual rates of farmyard manure (FYM ? 0, 4, 8, and 16 t ha^–1) and four annual rates of fertilizer N (? 0, 72.5, 145, and 230 kg N ha^–1) applications. The estimated N contributed through residual biomass of soybean (RBN_S) consisting of leaf fall, root, nodules, and rhizodeposition varied in the ranges of 7.02–16.94, 11.65–28.83, 3.31–8.91, and 11.3–23.8 kg N ha^–1 yr^–1, respectively. A linear relationship was observed between RBN_S and harvested biomass N (HBN_S) of soybean in the form of RBN_S = 0.461 × HBN_S – 20.67 (r = 0.989, P < 0.01), indicating that for each 100 kg N assimilated by the harvested biomass of soybean, 25.4 kg N was added to the soil through residual biomass. The Ndfa values ranged between 13% and 81% depending upon the annual rates of application of fertilizer N and FYM. As per the main effects, the %Ndfa declined from 76.4 to 26.0 with the increase in annual fertilizer‐N application from 0 to 230 kg N ha^–1, whereas %Ndfa increased from 40.8 to 65.8 with the increase in FYM rates from 0 to 16 t ha^–1, respectively. The N benefit from biological N₂ fixation accrued to the soil through residual biomass of soybean ranged from 7.6 to 53.7 kg N ha^–1 yr^–1. The treatments having %Ndfa values higher than 78 showed considerable annual contribution of N from N₂ fixation to the soil which were sufficient enough to offset the quantity of N removed from the soil (i.e., native soil N / FYM‐N / fertilizer‐N) with harvested biomass of soybean.     

Effects of inoculation of cyanobacteria on nitrogen status and nutrition of rice (Oryza sativa L.) in an Entisol amended with chemical and organic sources of nitrogen

A field experiment was conducted with wetland rice (Oryza sativa cv. IR-36) in a sandy clay loam soil (Entisol) to study the effect of inoculation with a soil-based mixed culture of four diazotrophic cyanobacteria,Aulosira fertilissima, Nostoc muscorum, N. commune andAnabaena spp., on the N-flux in inorganic NH₄ ⁺+NO₃ ^?+ NO₂ ^?), easily oxidizable, hydrolysable and non-hydrolysable forms of N in soil during vegetative growth periods of the crop. Effects on grain and straw yield and N uptake by the crop were estimated. The effects of applying urea N and N as organic sources, viz.Sesbania aculeata, Neem (Azardirachta indica) cake and FYM, each at the rate of 40 kg N ha^?1, to the soil were also evaluated. Inoculation significantly increased the release of inorganic N, evidenced by its increased concentrations either in soil or in soil solution. However, such increases rarely exceeded even 4% of total N gained in different froms in the soil system by inoculation during the vegetative growth stages of the rice plant, when the nutritional requirement of the plants is at a maximum. Most of the N₂ fixed by cyanobacteria remained in the soil as the hydrolysable form (about 85%) during this period. Inoculation caused an insignificant increase in grain (8%) and straw (11%) yield, which was, however, accompanied by a significant increase in N uptake by the grain (30%) and an increase in total uptake of 15.3 kg N ha 1. Such beneficial effects of inoculation varied in magnitude with the application of organic sources, with farmyard manure (FYM) being the most effective. Application of urea N, on the other hand, markedly reduced such an effect.     

Availability of residual fixed ammonium to crops

Summary To estimate the availability of residual fixed NH ₄ ⁺ -N to crops and its participation in the general nitrogen cycle, a field experiment without N fertilization was conducted with maize as the test crop planted at two densities. The experiments were carried out on the layout of a previous experiment, where the maize was cropped with identical densities but was fertilized with five different N treatments. Irrespective of planting density, the residual fixec NH ₄ ⁺ -N and KOBr-extractable N content in soil decreased with increase in the period of crop growth. N uptake by the plant was significantly higher in plot with the high- than the low-density planting. Availability of residual fixed NH ₄ ^– -N to the crop is discussed.     

Bio-extraction of Potassium from Glauconite Nano-particle in an Alfisols of Southern India

ABSTRACT

Glauconite is a potassium (K) bearing mineral, which generally contains 6–10% K₂O and mostly available in various natural deposits in India. Glauconite mineral was converted to nano-particle form by grinding in a high-energy ball mill. The release of K from glauconite nano-particle in soil was studied in a pot culture experiment. Five different treatments viz., 0 mg K₂O kg^?1, 100 mg K₂O kg^?1, 150 mg K₂O kg^?1, 200 mg K₂O kg^?1 used glauconite nano-particle (GNP) as a K source and 100 mg K₂O kg^?1 from muriate of potash (MOP), were applied in soil to grow maize (Zea mays L.) plant. The highest K⁺ concentration was observed in shoot at the application of 200 mg K₂O kg^?1 through GNP and recorded 2 times more K⁺ concentration in shoots as compared to 0 mg K₂O kg^?1 treatment whereas 1.1 times more than the MOP treated soil at 5^th harvesting stage. Selected plant physiological parameters namely electrical conductivity of plant cell, height, leaf area and nitrate reductase activity were also recorded maximum at 200 mg K₂O kg^?1 GNP treated soil. Overall, glauconite nano-particle could supply K throughout the growth period and enhanced biomass yield of maize plant without showing any K deficiency symptom.     

Toxicokinetics, recovery, and metabolism of metamitron in goat

Toxicokinetic behavior and metabolism studies of metamitron and its effect on the cytochrome P(450) content of liver microsomal pellet were carried out in black Bengal goats after a single oral administration at 278 mg kg(-1) and consecutive oral administration of 30 mg kg(-1) for 7 days. Metamitron was detected in the blood sample at 0.08 h (12.0 +/- 0.87 microg mL(-1)), maximum at 4 h (84.3 +/- 8.60 microg mL(-1)) and minimum (14.6 +/- 1.67 microg mL(-1)) at 36 h blood sample after a single oral administration. The absorption rate constant was 0.69 +/- 0.09 h(-1). The Vd(area) (2.00 +/- 0.08 L kg(-1)) and t(1/2)beta (8.98 +/- 0.70 h) values suggested wide distribution and long persistence of the compound in the body. The values of T approximately B (0.80 +/- 0.04), F(c) (0.55 +/- 0.01), Cl(B) (0.15 +/- 0.00 L kg(-1) h(-1)), and K(21) (0.41 +/- 0.03 h(-1)) suggested that metamitron retained in the blood compared to that in the tissue. Maximum concentration of metamitron residue was found in the adrenal gland followed by bile on day 4 of single oral administration. The higher Cl(R) compared to Cl(H) value indicated the excretion of the major portion (34-40%) through urine compared to feces (20-26%). Maximum concentrations of metamitron and its metabolite, deaminometamitron, were excreted through urine and feces at 48 and 24 h samples, respectively. The recovery of metamitron including its metabolite in terms of parent compound varied from 69.3 to 80.1%, of which contribution of metabolite in terms of parent compound varied from 53.1 to 63.0%. Repeated oral administration of metamitron at 30 mg kg(-1) for 7 days caused induction of the cytochrome P(450) content of liver microsomal pellet of goat, suggesting oxidative deamination of metamitron.     

Observations on the epidemiology of bovine cryptosporidiosis in India

The significance of Cryptosporidium as a causative agent of diarrhea has been assessed in bovine for a period of 2 years. A total of 940 faecal samples (470 samples in each year) both from diarrhoeic and non-diarrhoeic bovine (0-12 months age) were examined during three different seasons (rainy season, summer and winter). Overall Cryptosporidium was detected in 17.46% and 18.04% cases in first and second year, respectively. Out of 50.21% diarrhoeic and 49.79% non-diarrhoeic cases Cryptosporidium was detected in 26.79% and 8.13% in first year and 27.49% and 8.59% in second year. Year did not have any significant effect on the occurrence of cryptosporidiosis in bovine during this study period. The prevalence of cryptosporidiosis, both in diarrhoeic (61.64%) and non-diarrhoeic (47.22%) cases was highest in 0-1-month age group (P<0.01). Such a high percentage of cryptosporidiosis in clinically asymptomatic animals indicated that the particular age group of animals might be reservoir for the parasite. During this study period highest prevalence was recorded in rainy season (27.55%) followed by summer (16.99%) and winter (8.71%) (P<0.01). A total of 166 positive cases were genotyped. Molecular characterization of bovine cryptosporidiosis has been carried out by PCR-RFLP analysis of SSU rRNA gene and results indicated that Cryptosporidium parvum mainly responsible for diarrhea in bovine in India.     

Microsatellite markers and genetic diversity assessment in Lolium temulentum

Lolium and Festuca are two important genera of cool-season forage and turf grasses worldwide. Lolium temulentum L. (darnel ryegrass) has been proposed as a model species for genomics studies of cool-season forage and turf grasses. A study with 41 darnel ryegrass, three tall fescue (Festuca arundinacea Schreb.), two tetraploid fescue (F. glaucescens), and two meadow fescue (F. pratensis) genotypes was initiated to (i) identify a set of microsatellite (simple sequence repeats) markers useful for L. temulentum L., and (ii) to utilize such markers for assessing the genetic variability of L. temulentum accessions collected from different geographical regions of the world. A total of 40 tall fescue (TF) EST-SSRs and 60 Festuca–Lolium (F × L) genomic SSRs were screened on a subset of eight genotypes. The selected 30 tall fescue EST-SSRs and 32 F × L genomic SSRs were used for further analysis of genotypes. The TF-EST- and the F × L genomic-SSRs identified 10.3 and 9.3 alleles per marker, respectively with an average polymorphic information content (PIC) value of 0.66. The phenogram based on 319 EST-SSR and 296 genomic SSR fragments, grouped L. temulentum accessions into three major clusters except for accession ABY-BA 8892.78. Lolium temulentum accession ABY-BA 8892.78 did not cluster with any other accession. The Festuca clusters were distantly related with darnel ryegrass clusters with a similarity coefficient of 0.26. The selected set of tall fescue EST- and F × L genomic SSRs were useful in assessing L. temulentum genetic diversity and could benefit the genetic improvement of members of the Festuca–Lolium complex.