Identification of a diverse mini‐core panel of Indian rice germplasm based on genotyping using microsatellite markers

Identification of a small core germplasm set representing the available genetic diversity is essential for its proper evaluation and subsequent utilization in rice improvement programmes. For constituting a small diverse mini‐core panel of Indian rice germplasm, a representative set of 6912 accessions drawn based on their geographic origin from the whole rice germplasm collection available in the National Gene Bank was genotyped using 36 microsatellite markers. Automated fragment analysis of amplicons yielded a total of 435 alleles, with an average 12.4 and range of 3–29 alleles per locus. Polymorphism information content (PIC) ranged from 0.08 (RGNMS190) to 0.86 (RM552) with an average of 0.528. Based on genotyping data, a mini‐core consisting of 98 genotypes was identified. Ninety‐four per cent of the alleles present in the core set were present in the mini‐core. The identified small but diverse panel will be useful for further intensive trait‐specific evaluation and utilization in allele mining.     

An efficient method for the purification and characterization of nematicidal azadirachtins A,B, and H,using MPLC and ESIMS

Azadirachtin A enriched concentrate containing 60% active ingredient (a.i.) was prepared from the methanolic extract of the de-fatted neem (Azadirachta indica A. Juss) seed kernels. Azadirachtins A, B, and H, the three major bioactive constituents of neem seed kernel, were purified from this methanolic concentrate by employing reverse phase medium-pressure liquid chromatography (MPLC), using methanol-water solvent system as an eluant. The three pure azadirachtin congeners thus obtained were characterized by their unique mass spectral fragmentation, using electrospray probe in positive ion mode (ESI). All three azadirachtins exhibited nematicidal and antifungal activities. Azadirachtin B was the most effective against the reniform nematode Rotylenchulus reniformis (EC(50) 96.6 ppm), followed by Azadirachtin A (119.1 ppm) and H (141.2 ppm). At 200-ppm concentration, the test compounds caused 50-65% mortality of Caenorhabditis elegans nematode. Azadirachtin H showed the highest activity against the phytophagous fungi Rhizoctonia solani (EC(50) 63.7 ppm) and Sclerotium rolfsii (EC(50) 43.9 ppm), followed by B and A. The isolation of pure azadirachtins A, B, and H directly by MPLC purification from its concentrate and their characterization by ESIMS are unique and less time-consuming.     

Surgical Correction of Uterine Torsion and Mare–Foal Survival in Advance Pregnant Equine Patients

This article describes the surgical management of uterine torsion by midline celiotomy and cesarean section on 12 mares presented with signs of colic to a teaching veterinary hospital. The mares were either in full term of gestation (n = 7) or in advanced stage of pregnancy (n = 5). Six mares were in first parity. Uterine torsion was diagnosed by per rectal and per vaginal examinations. For surgical intervention, mares were anesthetized using a combination of xylazine (1.1 mg/kg) and ketamine (2.2 mg/kg), intravenously. After intubation, the animals were maintained on halothane (n = 4) or isoflurane (n = 8) inhalation anesthesia. Midline celiotomy was performed, and foals were delivered by cesarean section. In 11 mares, before closing the abdominal wound, the uterus was detorted manually and confirmed for its normal position. Both anesthetic protocols using halothane and isoflurane were found satisfactory for surgical correction of uterine torsion. After long-term follow-up, the study reported 75.0% (9/12) survival rate for mares. One mare was euthanized because of devitalized, necrosed, and adhered uterus to the abdominal wall. Of the nine surviving mares, seven were successfully bred. Three foals were born alive, and only one could survive on long-term basis. Of the nine dead foals, two had umbilical cord torsion.     

Physiological Changes in Sesbania Species to Reducing Light Intensities

Sesbania sesban and S. grandiflora were studied for their physiological characters under different light intensities (100, 75, 50 and 25 %) in a natural environment for 4 months. Rate of photosynthesis (PN) and stomatal conductance (CS) significantly decreased with decreasing light intensities and with intensity reduced to half the full sun light under high shading i.e. at 25 % light intensity, whereas the intercellular CO₂ concentration increased with decreasing light intensities in both the species. The reduction in PN was more in S. grandiflora than S. sesban. S. sesban maintained higher carboxylation efficiency under shade condition as compared to S. grandiflora. Rate of transpiration and water use efficiency (PN/TR) decreased while leaf diffusion resistance increased significantly with decreasing light intensities. Total chlorophyll content was maximal under 50 % light intensity in both plant species. Accumulation of chl b increased but chl a decreases under low light intensities. The higher accumulation of chl b in S. sesban under shade predicted its shade adaptability. The total protein content and NRA reduced under low light intensities. The overall rate of photosynthesis, stomatal conductance, carboxylation efficiency and water-use efficiency were higher in S. sesban as compared to S. grandiflora , indicating its better adaptability under shade condition and that it may be suitable to grow under stress in agroforestry systems in semi-arid tropics.     

Influence of physicochemical parameters of neem (Azadirachta indica A Juss) oils on nitrification inhibition in soil

The technology for the production of neem oil coated urea (NOCU) developed by the Indian Agricultural Research Institute is in the pipeline for adaption by several Indian fertilizer industries. Use of nitrification inhibitors is one of the methods of improving the nitrogen use efficiency (NUE) of nitrogenous fertilizers in agriculture. However, standard specifications for the neem oil as a raw material of NOCU are desired. Accordingly, the present study was undertaken to evaluate 25 samples of neem oils comprising 11 samples of expeller grade (EG) oils, 8 samples of cold-pressed (CP) oils, 3 samples of solvent-extracted oils, and 2 commercial formulations. NOCU was prepared using these oils (5000 ppm of urea-N). The soils fertilized with NOCUs (200 ppm of urea-N) were incubated at 27 degrees C and 50% water-holding capacity for a period of 15 days. Nitrapyrin (0.5% of N) coated urea served as the reference and prilled urea as control. Samples were analyzed for NH4+-N, NO2--N, and NO3--N using standard methods. The percent nitrification inhibition (NI) was calculated, and the results revealed that all of the neem oils caused NI ranging from 4.0 to 30.9%. Two samples of EG oils and two commercial formulations were found to be the best, causing 27.0-30.9% NI. Iodine, acid, and saponification values and meliacin content of all of the oils were analyzed and correlated with NI. The results revealed the direct influence of meliacin content of the neem oils on NI, which, however, was found to be negatively correlated with saponification and iodine values. There is, therefore, a need to introduce new Bureau of Indian Standards (BIS) specifications for neem oils as raw materials of NOCU.     

Azadirachtin-A and tetrahydroazadirachtin-A concentrates: preparation, LC-MS characterization and insect antifeedant/IGR activity against Helicoverpa armigera (Hübner)

A 60% azadirachtin-A concentrate has been obtained through repeated precipitation with hexane from a methanolic solution of a 20% concentrate. Azadirachtin-A (90%) has been obtained by medium-pressure liquid chromatography of the 60% concentrate with an RP-18 column and a methanol + water (1 + 1 by volume) solvent system. Catalytic hydrogenation of the 60 and 90% azadirachtin concentrates yielded the corresponding tetrahydroazadirachtin concentrates. Dihydroazadirachtin and tetrahydroazadirachtin formed during the first 5 h of hydrogenation were identified by LC-ESI-MS on the basis of their unique mass fragmentation pattern. The efficacy of tetrahydroazadirachtin concentrates in inhibiting the feeding and growth of Helicoverpa armigera (Hübner) larvae has been compared with that of azadirachtin concentrates. They were in general more active and deterred larvae from feeding at all concentrations. Tetrahydroazadirachtin-A (90%) and azadirachtin-A (90%) with respective IC(50) values of 280 and 390 mg L(-1) were effective as insect growth regulators, while tetrahydroazadirachtin-A (90%) displayed higher antifeedant activity (AI(50) 14 mg L(-1)) against the test insect.     

Reactivity of epicatechin in aqueous glycine and glucose maillard reaction models: quenching of C2, C3, and C4 sugar fragments

Mechanisms of how epicatechin alters the pathways of the Maillard reaction were investigated. Carbon-13 and nitrogen-15 labeling studies were utilized to define the reactivity of epicatechin with glucose, glycine, and/or reaction products in an aqueous model (pH 7, 125 degrees C for 30 min) via GC, GC/MS and HPLC/MS analysis. Quantification of the volatile reaction product isotopomers by GC/MS from a 1:1 labeled to unlabeled glucose (carbohydrate module labeling technique) plus glycine model system indicated the formation of 2,3-butanedione and acetol were primarily formed via intact C4 and C3 sugar fragments, whereas pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine, and cyclotene were primarily formed via intact C2/C2, C2/C3, C3/C3, C3/C3, and C3/C3 sugar fragment pairs, respectively. The formation of these seven compounds was also reported by GC analysis to be dramatically inhibited when epicatechin was added to the glucose/glycine model system (observed 9-113-fold reduction). HPLC/MS analysis of both the glucose-labeled and glycine-labeled model systems with and without epicatechin indicated that epicatechin reacted directly with C2, C3, and C4 sugar fragments, while epicatechin did not report any direct reactivity with glycine. In conclusion, the quenching of sugar fragmentation products via epicatechin was correlated with the observed inhibition on volatile compound formation when epicatechin was added to a glucose/glycine aqueous reaction model system.     

Bio-availability of iron from spinach (Spanicia oleracea) cultivated in soil fortified with graded levels of iron

In vitro availability of iron along with ascorbic acid, oxalic acid and phosphorus content of two varieties of spinach (Pusa Jyoti and Allgreen) cultivated in soil with different levels of added iron was determined. Addition of graded levels of iron to soil markedly increased the total iron and phosphorus contents and significantly decreased the bio-availability of iron, ascorbic acid and oxalic acid contents of spinach. Ascorbic acid and oxalic acid contents markedly exerted a positive influence while phosphorus exerted a negative influence on the bio-availability of iron.     

Reactions of hydrated electrons with triazine derivatives in aqueous medium

A study is made of the kinetics and mechanism of the reaction of radiolytically produced hydrated electron (e-(aq)) with some triazine derivatives [1,3,5-triazine (T), 2,4,6-trimethoxy-1,3,5-triazine (TMT), 2,4-dioxohexahydro-1,3,5-triazine (DHT), 6-chloro N-ethyl N-(1-methylethyl)-1,3,5-triazine 2,4-diamine (atrazine, AT), and cyanuric acid (CA)] in aqueous medium using pulse and steady-state radiolysis techniques. The second-order rate constants were determined from the pseudo first-order decay of e(-)(aq) in the presence of triazines at 720 nm, and the values obtained with T, TMT, AT, and CA are in the order of 10(9) dm(3) mol(-1) s(-1) and that of DHT was 10(8) dm(3) mol(-1) s(-1) at pH 6. The transient absorption spectra from the reaction of e(-)(aq) with T and TMT are characterized by their lambda(max) at 310 nm, and those of DHT and CA are around 280 and 290 nm, respectively. However, a very weak and featureless absorption spectrum is obtained from AT. On the basis of the spectral evidence and on the quantitative electron transfer from the transient intermediates to the oxidant, methyl viologen (MV(2+)), the intermediate radicals are assigned to N-protonated electron adducts (with the unpaired spin density at carbon) of triazines. The degradation profiles, monitored as the disappearance of parent triazine concentrations as a function of dose, obtained with AT, TMT, CA, and DHT, highlight the potential use of e-(aq) in the degradation of triazines.     

Antifungal activity of some Himalayan medicinal plants and cultivated ornamental species

Extracts of roots of Rumex nepalensis, Berberis aristata, Arnebia benthamii, bark of Taxus wallichiana, Juglans regia and petals of Jacquinia ruscifolia were tested for their antifungal activity against twelve different fungal pathogens. Ethanolic extracts of R. nepalensis and J. ruscifolia extracts showed a broad spectrum of activity.