Integrative approaches to improve litchi (Litchi chinensis Sonn.) plant health using bio-transformations and entomopathogenic fungi.

Bio-transformations refer to the chemical modifications made by an organism on a chemical compound that often involves the interaction of plants with microbes to alter the chemical composition of soil or plant. Integrating bio-transformations and entomopathogenic fungi into litchi cultivation can enhance symbiotic relationships, microbial enzymatic activity in rhizosphere, disease suppression and promote overall plant health. The integration of biological formulations and entomopathogenic fungi can significantly influence growth, nutrient dynamics, physiology, and rhizosphere microbiome of air-layered litchi (Litchi chinensis Sonn.) saplings. Biological modifications included, K-mobilizers, AM fungi, Pseudomonas florescence and Azotobacter chroococcum along with Metarhizium, entomopathogenic fungi have been used. The treatments included, T1-Litchi orchard soil + sand (1:1); T2-Sand + AM fungi + Azotobacter chroococcum (1:2:1); T3-Sand + Pseudomonas florecence + K-mobilizer (1:1:1); T4- AM fungi + K-mobilizers (1:1); T5, P. Florecence + A. chroococcum + K-mobilizer (1:1:1); T6-Sand + P. florecence (1:2) and T7-Uninoculated control for field performance. Treatments T4-T6 were further uniformly amended with drenching of Metarrhizium in rhizosphere. T2 application significantly increased resident microbe survival, total chlorophyll content and root soil ratio in seedlings. A. chroococcum, Pseudomonas, K-mobilizers and AM fungi increased in microbial biomass of 2.59, 3.39, 2.42 and 2.77 times, respectively. Acidic phosphatases, dehydrogenases and alkaline phosphatases were increased in rhizosphere. Leaf nutrients reflected through DOP were considerably altered by T2 treatment. Based on Eigen value, PCA-induced changes at biological modifications showed maximum total variance. The study inferred that the bio-transformations through microbial inoculants and entomopathogenic fungi could be an encouraging strategy to enhance the growth of plants, health and productivity. Such practices align well with the goals of sustainable agriculture through biological means by reducing dependency on chemical inputs. By delving into these aspects, the research gaps including microbial processes, competitive and symbiotic relationships, resistance in microbes and how complex interactions among bio-transformations, entomopathogenic fungi and microbes can significantly impact the health and productivity of litchi. Understanding and harnessing these interactions can lead to more effective and sustainable farming practices.

Grain yield, adaptation and progress in breeding for early-maturing and heat-tolerant wheat lines in South Asia.

Maintaining wheat productivity under the increasing temperatures in South Asia is a challenge. We focused on developing early maturing wheat lines as an adaptive mechanism in regions suffering from terminal heat stress and those areas that require wheat adapted to shorter cycles under continual high temperature stress. We evaluated the grain yield performance of early-maturing heat-tolerant germplasm developed by CIMMYT, Mexico at diverse locations in South Asia from 2009 to 2014 and estimated the breeding progress for high-yielding and early-maturing heat-tolerant germplasm in South Asia. Each year the trial comprised of 28 new entries, one CIMMYT check (Baj) and a local check variety. Locations were classified by mega environment (ME); ME1 being the temperate irrigated locations with terminal high temperature stress, and ME5 as hot, sub-tropical, irrigated locations. Grain yield (GY), days to heading (DTH) and plant height (PH) were recorded at each location. Effect of temperature on GY was observed in both ME1 and ME5. Across years, mean minimum temperatures in ME1 and mean maximum temperatures in ME5 during grain filling had significant negative association with GY. The ME1 locations were cooler that those in ME5 in the 5 years of evaluations and had a 1-2 t/ha higher GY. A mean reduction of 20 days for DTH and 20 cm in PH was observed in ME5. Negative genetic correlations of -0.43 to -0.79 were observed between GY and DTH in South Asia during 2009-2014. Each year, we identified early-maturing germplasm with higher grain yield than the local checks. A positive trend was observed while estimating the breeding progress across five years for high-yielding early-maturing heat tolerant wheat compared to the local checks in South Asia. The results suggests the potential of the high-yielding early-maturing wheat lines developed at CIMMYT in improving wheat production and maintaining genetic gains in South Asia.

Mean performances, character associations and multi-environmental evaluation of chilli landraces in north western Himalayas.

Even though many varieties have been recommended across agro-climate zones of Himachal Pradesh, yet the information on stability is lacking in this State. Hence, the present investigation was carried out to identify high yielding stable genotypes among various pre-adapted landraces. The material consists of 20 chilli landraces including check i.e. DKC-8. The experiment was laid out in a RCBD. The data were recorded and analyzed to work out mean performances and the inferences were drawn for parameters of variability, correlation coefficients, path coefficients and stability analysis. As per mean performances, CS7 and CS9 were earliest in flowering, CS13 is earliest in days to ripe maturity, CS10 had highest plant height and CS9 had highest average fruit weight and ripe fruit yield plant-1. High PCV and GCV were recorded for ripe fruit yield plant-1. Heritability and genetic advance were recorded maximum for plant height in summer seasons and were recorded maximum for number of ripe fruits plant-1 in winter season. Correlation coefficients showed that number of ripe fruits plant-1 and average ripe fruit weight were positively and significantly correlated with ripe fruit yield plant-1. Path coefficient analysis in summer and winter seasons showed that average ripe fruit weight had the highest positive direct effect on ripe fruit yield plant-1. The pooled data over environments were analyzed to estimate the interaction effects between genotypes × environment. The mean sum of squares due to genotypes, environments and genotypes × environment interaction were significant for all the characteristics. CS1, CS3, CS6, CS10, CS13, CS15 were adapted to all environments, CS7 and CS9 were specifically adapted to favourable environment and CS2 was specifically adapted to unfavorable environment for 50% flowering, landraces CS1, CS2 and CS3were well adapted to all environments for ripe maturity whereas landraces CS6, CS10 and CS19 were well adapted to all environment for number of ripe fruit and ripe fruit yield.

Antiarthritic activity of a standardized, multiherbal, Ayurvedic formulation containing Boswellia serrata: in vitro studies on knee cartilage from osteoarthritis patients.

A validated in vitro model of cartilage damage and published data were used showing that this model measures the chondroprotective and antiinflammatory effects of different antiarthritic drugs. In this report, this model was used to evaluate the effects of a new antiarthritic Ayurvedic formulation containing Zingiber officinale root, Tinospora cordifolia stem, Phyllanthus emblica fruit and oleoresin of Boswellia serrata. Glucosamine sulphate was used as a positive control in the study. Aqueous extracts of each drug were tested on explant cultures of knee cartilage obtained from osteoarthritis patients undergoing knee replacement surgery. The new formulation caused a sustained and statistically significant inhibition in the release of glycosaminoglycans and aggrecan by cartilage explants from these patients. This formulation also induced a transient antiinflammatory effect as measured by a reduction in the levels of nitric oxide released by explants. Furthermore, the data strongly suggest that oleoresin of B. serrata plays a crucial role in the chondroprotective and antiinflammatory activity of this formulation. In summary, this report provides the first, direct, in vitro biochemical evidence of anti-arthritic activity a new Ayurvedic formulation. This formulation significantly reduced damage of articular knee cartilage from chronic osteoarthritis patients.

Natural language processing.

Natural language processing (NLP) is the study of mathematical and computational modeling of various aspects of language and the development of a wide range of systems. These include spoken language systems that integrate speech and natural language; cooperative interfaces to databases and knowledge bases that model aspects of human-human interaction; multilingual interfaces; machine translation; and message-understanding systems, among others. Research in NLP is highly interdisciplinary, involving concepts in computer science, linguistics, logic, and psychology. NLP has a special role in computer science because many aspects of the field deal with linguistic features of computation and NLP seeks to model language computationally.

Screening for disease resistance in barley cultivars against Bipolaris sorokiniana using callus culture method.

Screening for resistant barley genotypes in response to fungal toxin of Bipolaris sorokiniana was assessed on standing barley plants as well as in selected callus lines of the same. For the standing lines tested, those manifesting chlorosis in response to toxin infiltration showed a significantly slower disease progress as compared to the necrotic lines. Also, necrosis in the callus tissues of the susceptible cultivar in MS medium supplemented with different concentrations of the crude toxin was significantly higher than in the callus tissues of the chlorotic lines studied. Similar host response to the toxin in in vitro and field situations open up the possibility of screening barley cultivars for resistance to spot blotch using callus culture as against classical methods of screening in order to increase accuracy and save time and space.

Calculation of antibody and antigen concentrations from ELISA data using a graphical method.

A graphical method for determining the concentration of either the antibody or the antigen from ELISA data is presented in the form of a GWBASIC program. In the program, ELISAEQ, optical densities (OD) obtained from a 96-well ELISA plate can be input either directly by interfacing a microplate reader to the computer or manually. The program uses standard sample data, and selects the semilogarithmic linear range. Over this range, a least-squares method is used to determine the concentrations of interest. In addition, a hyperbolic interpolation formula is derived over the entire range for estimating the antibody or antigen concentration of the unknown samples whose OD is beyond the linear range.

Rapid polymerase chain reaction amplification using intact bacterial cells.

We have demonstrated that efficient polymerase chain reaction amplifications from chromosomal DNA can be carried out using whole bacterial cells as the starting material. Cells from the liquid or solid cultures can be used directly, without any pre-treatment, thus eliminating the need for DNA isolation.

Production of L-phenylacetylcarbinol by free and immobilized yeast cells.

Production of L-phenylacetylcarbinol (L-PAC) through biotransformation of benzaldehyde by free and immobilized cells of the yeast Saccharomyces cerevisiae has been attempted. L-PAC production was found to be maximum (0.4 microliter/ml) when anaerobically grown free cells were used as biocatalyst during aerobic biotransformation for two hours with magnetically stirred bioreactor. Growth under oxygen limited conditions led to accumulation of higher amount of pyruvate decarboxylase enzyme and co-substrate, pyruvate, resulting in higher L-PAC formation. L-PAC yield was low when biotransformations were carried out anaerobically either for aerobically or anaerobically grown free cells. Free cells were found to be more efficient biocatalyst for L-PAC production, as compared with the immobilized cells, with the investigated benzaldehyde concentration (0.3% v/v) and cell density (17.5% w/v). The study has explored and indicated the possibility of optimizing the yield of L-PAC by growing the yeast cells under oxygen limited condition for suitable aerobic mode of benzaldehyde biotransformation.

Prevention of perinatal hepatitis B virus transmission in an obstetric/infant population.

STUDY DESIGN: This retrospective study reviewed the screening practice and seroprevalence of hepatitis B surface antigen (HBsAg) among all mothers with live births at a teaching hospital in Montreal between November 1, 1990 and April 30, 1991. RESULTS: Most women (94%) were screened prenatally and 5.2% postnatally. Screening status could not be determined for 0.8% of women. One-quarter of all postnatal screening results were available only at 48 h or more postdelivery. No infants born to women with postnatal screening or to women with unknown screening status were immunized expectantly. The maternal seroprevalence was 1.08% (95% confidence interval from 0.6, 1.4). All 22 infants born to HBsAg-positive mothers had received hepatitis B immune globulin within 12 h of birth and the first dose of hepatitis B vaccine within 24 h. Follow-up of infants revealed that only 50% had received the second and third doses according to the recommended protocol, with 83% completing the immunization series. CONCLUSION: These results indicate that a program of prenatal HBsAg screening and neonatal prophylaxis against hepatitis B can be successfully instituted in a high volume obstetric hospital, and that better monitoring of infants is required to ensure completion of vaccination.

Clonal variability and its relevance in generation of new pathotypes in the spot blotch pathogen, Bipolaris sorokiniana.

Spot blotch pathogen Bipolaris sorokiniana of wheat was investigated with threefold objectives: to establish a relationship between morphological and pathological variability of isolates, identify clonal genotype(s) acting as a source for the generation of new variability, and to determine the mechanism of generation of such variability in the pathogen. Isolates were collected from the leaves and seeds of field-grown wheat crop at four different sites in eastern Gangetic plains of India. Eighty-six clonal isolates derived from a single isolate (gray with white patches, Group III), which segregated in an equal proportion of parental and nonparental types, were studied. Morphological characters-i.e., colony morphology, growth rate, and sporulation-were studied along with disease-causing ability of the isolate clones. Clonal isolates were grouped into three categories. Microscopic analysis of nuclei was done to determine the causes of such variability. Morphological variability appeared to be related to the pathological variability. The isolate having epidemic potential appeared different than that acting as the reservoir for variability. The cause of such variability could be attributed either to hyphal fusion and heterokaryosis, nuclear migration and occurrence of multinucleate state, or a combination of these factors. Random Amplified Polymorphic DNA (RAPD) assay suggested that the unique fragments for different groups could be utilized as molecular markers to identify the isolates of specific groups.

Identification of molecular marker and aggressiveness for different groups of Bipolaris sorokiniana isolates causing spot blotch disease in wheat (Triticum aestivum L.).

One hundred fifty-five isolates of Bipolaris sorokiniana of wheat were studied for their morphopathological characterization. These isolates were grouped in five categories--black, brown/dull black, gray cottony growth, dull white/greenish black, and white--on the basis of their growth pattern. The frequency of the black suppressed type was maximum (45.63%), whereas the white isolate displayed lowest frequency (6.96%) in the natural population. Twenty RAPD (random amplified polymorphic DNA) primers were used to observe the variability among the identified groups of B. sorokininana. From each group, eight random isolates were investigated. A total of 143 bands were amplified, out of which 107 (74.83%) were polymorphic and 36 (25.17%) were monomorphic. On an average, the total numbers of bands generated per primer were 7.15, of which 5.35 and 1.80 were polymorphic and monomorphic, respectively. Dendrograms based on molecular polymorphism unveiled a considerable amount of diversity among the isolates. Specific DNA bands were identified for selected isolates. The distinct markers appeared to be potential enough to be employed as genetic fingerprints for future strain identification and classification. The study indicated that the RAPD primers provide an easy, rapid, and simple technique for the preliminary assessment of genetic diversity among the fungal isolates.

Conservative management of a severely alloimmunized pregnancy: case report.

A pregnancy complicated by severe alloimmunization because of anti-E antibodies was evaluated with fetal movement counts, nonstress tests, and serial ultrasonography-amniocentesis. The successful outcome of this case suggests that such treatment may be an acceptable alternative to intrauterine transfusions for very select cases of severe alloimmunization.

Construction, expression, and characterization of a mutated animal fatty acid synthase deficient in the dehydrase function.

The location of the dehydrase domain in the multifunctional animal fatty acid synthase has been determined by engineering a fatty acid synthase mutant deficient in dehydrase activity. A full-length fatty acid synthase cDNA encoding a mutated histidine residue (His878-->Ala) was constructed and expressed in insect Sf9 cells using a baculoviral vector. The mutated recombinant fatty acid synthase retained all partial activities of the multifunctional complex except the dehydrase and was unable to synthesize fatty acids. beta-Hydroxybutyryl moieties were formed by the mutant fatty acid synthase from acetyl-CoA, malonyl-CoA, and NADPH and slowly released as the CoA thioester, confirming that this protein cannot perform the dehydration reaction. This finding points to an important catalytic role for His878 in the dehydration reaction and establishes that the dehydrase domain is located immediately adjacent to the carboxyl terminus of the transferase domain. Examination of the completed domain map for the animal fatty acid synthase indicates that the catalytic domains are clustered in two groups separated by a central structural core: the ketoacyl synthase, malonyl/acetyltransferase, and dehydrase in the amino-terminal half and the enoyl reductase, ketoreductase, acyl carrier protein, and thioesterase in the carboxyl-terminal half. A model is proposed in which the two centers for acyl chain initiation, elongation and termination, are formed by the cooperation of the three amino-terminal domains of one subunit with the four carboxyl-terminal domains of the other subunit.

Construction of a cDNA encoding the multifunctional animal fatty acid synthase and expression in Spodoptera frugiperda cells using baculoviral vectors.

A cDNA encoding the 2505-residue multifunctional rat fatty acid synthase has been constructed and expressed as a catalytically active protein in Spodoptera frugiperda (Sf9) cells using Autographa californica nuclear polyhedrosis virus (baculovirus). The 7.5 kb cDNA was engineered by the amplification and sequential splicing together of seven fragments contained in overlapping cDNAs that collectively spanned the entire rat fatty acid synthase coding sequence. The full-length cDNA was cloned into a baculoviral transfer vector and used together with linearized baculoviral DNA to co-transfect Sf9 cells. Recombinant viral clones were purified and identified by Western blotting. The recombinant fatty acid synthase was expressed maximally 2 days after infection of the Sf9 cells, constituting up to 20% of the soluble cytoplasm, and could be conveniently separated from the insect host fatty acid synthase by high-performance anion-exchange chromatography. The catalytic properties of the purified recombinant fatty acid synthase are indistinguishable from those of the best preparations of the natural protein obtained from rat liver. These results indicate that, in the insect cell host, all seven catalytic components of the 2505-residue recombinant fatty acid synthase fold correctly, the acyl-carrier-protein domain is appropriately phosphopantetheinylated post-translationally, and the multifunctional polypeptide forms catalytically competent dimers. Thus the baculoviral system appears to be well suited for the expression of specific fatty acid synthase mutants that can be used to explore the mechanism by which the seven domains of this multifunctional homodimer co-operate in the biosynthesis of fatty acids.

Characterization of the interthiol acyltransferase reaction catalyzed by the beta-ketoacyl synthase domain of the animal fatty acid synthase.

The enzyme activity responsible for translocation of saturated acyl chains from the 4'-phosphopantetheine of the acyl carrier protein to the active site cysteine of the beta-ketoacyl synthase in the animal fatty acid synthase has been identified. An enzyme assay was devised that allows uncoupling of the interthiol transfer step from the condensation reaction. Experiments with various fatty acid synthase mutants indicate clearly that catalysis of the transfer of saturated acyl moieties from the 4'-phosphopantetheine thiol to the active site cysteine thiol, Cys-161, is an inherent property of the beta-ketoacyl synthase domain. Catalytic efficiency of the interthiol transferase increases from C2 to C12 and decreases with increasing chain-lengths beyond C12. Malonyl, beta-hydroxybutyryl, and crotonyl thioesters are not substrates for the transferase, whereas the beta-ketobutyryl moiety is a poor substrate. These features of the substrate specificity are exactly as predicted for a transferase that fulfills the proposed role in the fatty acid synthase reaction sequence and indicate that this activity plays an important role in determining the overall specificity of the beta-ketoacyl synthase reaction.