Sequence analysis of the non-structural 3A and 3C protein-coding regions of foot-and-mouth disease virus serotype Asia1 field isolates from an endemic country

A total of 18 foot-and-mouth disease virus (FMDV) serotype Asia1 field isolates belonging to two different lineages (including the divergent group) as delineated earlier in VP1-based phylogeny were sequenced in the non-structural 3A and 3C protein-coding regions. The phylogenetic trees representing the regions coding for the non-structural proteins were very similar to that of the structural VP1 protein-coding region. Phylogenetic comparison at 3C region revealed clustering of Asia1 viruses with the isolates of serotypes O, A and C in the previously identified clade. Comparison of amino acid sequences identified lineage-specific signature residues in both the non-structural proteins. Overall analysis of the amino acid substitutions revealed that the 3A coding region was more prone to amino acid alterations than 3C region.     

Graded regulation of the Kv2.1 potassium channel by variable phosphorylation

Dynamic modulation of ion channels by phosphorylation underlies neuronal plasticity. The Kv2.1 potassium channel is highly phosphorylated in resting mammalian neurons. Activity-dependent Kv2.1 dephosphorylation by calcineurin induces graded hyperpolarizing shifts in voltage-dependent activation, causing suppression of neuronal excitability. Mass spectrometry-SILAC (stable isotope labeling with amino acids in cell culture) identified 16 Kv2.1 phosphorylation sites, of which 7 were dephosphorylated by calcineurin. Mutation of individual calcineurin-regulated sites to alanine produced incremental shifts mimicking dephosphorylation, whereas mutation to aspartate yielded equivalent resistance to calcineurin. Mutations at multiple sites were additive, showing that variable phosphorylation of Kv2.1 at a large number of sites allows graded activity-dependent regulation of channel gating and neuronal firing properties.     

Emergence of antigenic variants with in serotype A foot and mouth disease virus in India and evaluation of a new vaccine candidate panel

Emergence of genetically and antigenically divergent lineages/genotypes and poor intergenotypic antigenic coverage is a major concern in serotype A foot-and-mouth-disease virus (FMDV) in India. In 2009, to cover antigenic diversity emerged in serotype A virus field isolates, IND40/2000 was selected as the new vaccine strain for incorporation in the trivalent FMD vaccine formulation used in India. Although current vaccine strain (IND40/2000) covers most isolates antigenically, a few VP3(59)-deletion group isolates showed low r-value in routine vaccine matching exercise. The VP3(59)-deletion group within genotype 18 emerged first in late part of 2002 and in 2007 causing outbreaks along with non-deletion isolates of the same genotype. In case of emergence or re-emergence of more antigenically divergent isolates in future, a need for a new vaccine candidate to cover maximum isolates of both deletion and non-deletion group may arise. Four alternate candidate vaccine strains (IND281/2003, IND195/2007, IND360/2007 and IND123/2008) were selected based on set criteria and antigenic relationships with field isolates sampled between 2002 and 2009 were analyzed using a micro-neutralization test. Phylogenetic analysis based on capsid region of serotype A isolates revealed existence of two broad distinct clusters (VP3(59)-deletion and non-deletion group) within genotype 18. The VP3(59)-deletion group has diversified genetically with time giving rise to three different sub-lineages (clade18a, 18b and 18c). The present study indicates that the virus candidates IND281/2003 (VP3(59)-deletion group) and IND195/2007 (non-deletion group) can be used as an adjunct or alternative strain to currently used vaccine strain IND40/2000 in case of emergence of more antigenically divergent isolates in future.     

Effect of micronutrient supplementation around calving on the plasma cortisol levels of Murrah buffaloes and Sahiwal and Karan Fries cows

Micronutrients when fed around peripartum may reduce the stress induced by cortisol. With this objective, 24 Sahiwal (SW) and 24 Karan Fries (KF) cows and 24 Murrah (Mu) buffaloes were taken and divided into four groups of six each. Vitamin E (VE), zinc (Zn) and copper were supplemented from 30 days pre- to 30 days postcalving in groups 1, 2 and 3. Animals without supplementation served as control. Blood sampling was done on days 30, 15, 7 and 3 precalving; at calving; and on days 3, 7, 15 and 30 postcalving. Plasma cortisol levels were measured by ELISA. Supplementation of VE and Zn significantly (P?<?0.05) reduced plasma cortisol levels at calving and at 30 days postcalving. KF cows exhibited best results with VE, whereas SW cows and Mu buffaloes responded best to Zn. Copper increased the peripartum cortisol levels. Reduction in cortisol levels may help in keeping the animal healthy and stronger to fight immuno-suppression generally observed around the period of peripartum.     

AFLP Analysis of the Phenetic Organization and Genetic Diversity in the Sugarcane Complex, Saccharum and Erianthus

Amplified fragment length polymorphism (AFLP) markers were evaluated for determining the phylogenetic relationships, and the diversity in the Saccharum complex using 30 clones belonging to S. officinarum, S. robustum, S. spontaneum, S. barberi, S. sinense and the related genus Erianthus. The phenetic tree of the species clones based on AFLP data was consistent with the known taxonomical relationships. AFLP gave higher resolution of closely related species into discrete groups than that by RAPD and RFLP markers, reported earlier. The levels of diversity within the various Saccharum species were also found to be higher than those obtained previously with the same set of clones using RAPD markers. The intraspecies similarity in S. barberi and S. sinense was much higher than interspecies similarity suggesting a clear separation of the two, which are considered ‘horticultural species’. The genetic similarity matrix derived from a single primer combination highly correlated (r = 0.980) with that obtained from all the 12 primer combination used in the study, thus highlighting the efficiency of a single primer combination in delineating species relationships. All the primer combinations could identify markers that are specific to each of the species and the genus Erianthus. Among the species, specific markers were highest in S. spontaneum followed by S. robustum, S. barberi, S. officinarum and S. sinense. Erianthus had a distinct profile with 30% of the total amplified fragments being specific to it. This offers great scope for identifying intergeneric hybrids, which has been very difficult using morphological traits and RAPD markers. High degree of correspondence between the results from the cluster analysis based on Jaccard's similarity index, Neighbour Joining tree based on Sokal and Michener distance matrix and AFTD (Analyses Factorielle on Table of Distances) analysis clearly demonstrated that AFLP markers would be an appropriate tool in providing better information about the relationships among the species, estimation of diversity, and in revealing species and genus specific markers that could be directly applied in sugarcane breeding programmes.     

Identification and classification of aromatic rices based on DNA fingerprinting

Aromatic rices are preferred by the consumers all over the world due to its flavour and palatability. Although a large number of these collections are available, little systematic analysis of genetic diversity has been carried out. With the objective of identification and classification of aromatic rice genotypes, RAPD profiling was employed using 58 random decamer primers. Most of these primers (96.5%) detected polymorphism among the genotypes. Of the 465 amplified bands, 314 were polymorphic. Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped all the traditional tall, photosensitive, low yielding, long grained ‘basmati’ aromatics together. The short grained aromatic cultivars, formed a different cluster with high level of average similarity among themselves. The dendrogram based on 58 primers was highly similar to that based on 10 and 15 primers with matrix correlation (r) of 0.88 and 0.91, respectively. This suggested that a set of 10 primers can be employed for an initial assessment of genetic diversity in a large number of collections. All the rice genotypes included in the study could be distinguished from each other at the level of 19 to 186 polymorphic bands between individuals in pair wise comparison over all the 58 primers. Probability of identical profiles by chance suggested that about 10⁴¹ genotypes can be unambiguously differentiated by RAPD fingerprints obtained by 58 primers. A diagrammatic mode of presentation of DNA fingerprints of the aromatic rices based on 10 of the informative primers was developed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Trace elemental characterization of some food crustacean tissue samples by EDXRF technique

The trace elemental analysis of two species of mud crab and three species of prawn samples from Chilika lagoon, Orissa, India has been carried out by using Energy Dispersive X-ray Fluorescence (EDXRF) technique available at Institute of Physics, Bhubaneswar. Elements namely K, Ca, Mn, Fe, Cu, Zn, Se, Br, Sr and Pb have been measured in the present investigation. The study indicates the effectiveness of the technique in analyzing biological materials like tissue samples and opens a door for easy analysis of seafood items with a easy, fast, sensitive, simultaneous multi-elemental technique with a simple sample preparation procedure without any chemical treatment. Though all the specimens were collected from the same environment of the lagoon the elemental variation might be due to the differential migration pattern and metabolism or other biological factors.     

Root Growth of a Salt Susceptible and a Salt Resistant Rice (Oryza sativa L.) During Seedling Establishment Under NaCl Salinity

The salt susceptible rice ( Oryza sativa L.) cv. Jaya and salt resistant cv. Damodar seed germination and seedling growth rates were studied in laboratory and field conditions. The effect of salinity on seed germination, shoot and root length, seedling vigour index (SVI) and increase in the root: shoot length ratio in the laboratory was relatively more in cv. Jaya than in cv. Damodar. The relative susceptibility to salinity was more in cv. Jaya than in cv. Damodar in the field also. However, their responses varied with growth period. The root: shoot length and fresh and dry weight ratios increased with salinity at 15 days in cv. Jaya. The root: shoot fresh and dry weight ratio decreased with salinity at 15 days in cv. Damodar. However, the root: shoot fresh weight ratio decreased with salinity at 25 days in the susceptible cv. Jaya. The root and shoot length fresh and dry weight of cv. Damodar was enhanced at 0.5% (w/v) NaCl treatment compared to the control seedlings at 25 days. SVI in cv. Jaya decreased with salinity in the laboratory and field conditions. SVI in cv. Damodar, SVI showed little change at 15 days but decreased with salinity of 1–3% NaCl with an enhancement at 0.5% NaCl level in the laboratory and at 25 days in field conditions.     

Mapping quantitative trait loci (QTL) for grain size in rice using a RIL population from Basmati × indica cross showing high segregation distortion

Grain size is one of the three productivity related traits in rice and hence a major target for genetic improvement. Since understanding genetic variation in grain size between Basmati and indica genotypes is important for rice improvement, a recombinant inbred population was developed from a traditional aromatic cultivar ‘Basmati 370’ and a non-aromatic indica genotype ‘IRBB60’. This population was phenotyped in two locations for grain length (GL), grain breadth (GB), GL/GB ratio (LBR) and grain weight (GW). Though the RIL population reported in the current study exhibited segregation distortion (SD) for 54 % of the markers, they were utilized in analysis using an appropriate statistical package, PROC QTL in the SAS environment. Interval mapping revealed a total of 15 QTLs for GL, seven for GB, 15 for LBR and two for GW. Among them 13 were not reported earlier and thus novel. For a known major QTL identified in the study, GW8 for GB, a PCR based functional marker was designed and validated. This is the first report wherein a very high proportion of markers (>50 %) exhibiting SD have been successfully used for QTL mapping.