Assessment of Genetic Variability in the Coding Sequence of Melatonin Receptor Gene (MTNR1A) in Tropical Arid Sheep Breeds of India

Seasonal behaviour in sheep, which varies in tropical and temperate environmental conditions, is a matter of study, because it can provide a clue to address the problem of seasonality in sheep. Melatonin receptor is the membrane‐bound G‐coupled receptor, sensing the message of photoperiodic cues thorough melatonin. Restriction fragment length polymorphism (RFLP) studies were carried out to assess the variability of gene at G612A and C606T SNPs in MTNR1A gene, which have been studied to be markers for out‐of‐season breeding. Allelic frequency distribution corresponded to higher frequency of GG and CC genotype, in tropical arid sheep breed in comparison with temperate region sheep breed. PCR amplification of MTNR1A gene of 30 animals was performed and single nucleotide polymorphisms (SNPs) identification was carried out using Lasergene software. Seven SNPs/mutations were identified, but most of them were synonymous, except the one G706A, leading to substitution of valine by isoleucine. Polyphen‐2 analysis of G706A mutation revealed that it is a benign mutation. Two important SNPs C426T and G555A, which were identified in temperate sheep breeds, could not be traced in Magra and Marwari breeds of sheep. Thus, the Magra and Marwari breeds of tropical, arid region demonstrated the presence of both polymorphic SNPs markers G612A and C606T, associated with out‐of‐season breeding. GG and CC genotypes were having a higher prevalence in the studied population.     

Protocol: a medium-throughput method for determination of cellulose content from single stem pieces of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Background

Lignocellulosic biomass is an important renewable resource for biofuels and materials. How plants synthesise cellulose is not completely understood. It is known that cellulose synthase complex (CSCs) moving in the plasma membrane synthesise the cellulose. CESA proteins are the core components of CSC. In Arabidopsis, in vitro mutagenesis of proteins followed by complementation analysis of mutants lacking the gene represents an important tool for studying any biological process, including cellulose biosynthesis. Analysis of a large number of plants is crucial for these types of studies.

Results

By using aspiration rather than centrifugation to remove liquids during various stages of protocol, we were able to increase the throughput of the method as well as minimise the sample loss. As a test case, we determined cellulose content of wild type and secondary wall cesa mutants across the length of primary shoot which was fond to be rather uniform in 7-week-old plants. Additionally, we found that the cellulose content of single mutants was comparable to the higher order mutants.

Conclusions

Here we describe a medium-throughput adaptation of Updegraff’s method that allowed us to determine cellulose content of 200 samples each week.

     

Sialylation of Helicobacter bizzozeronii lipopolysaccharides modulates Toll-like receptor (TLR) 2 mediated response

Sialic acid in lipopolysaccharides (LPS) of mucosal pathogens is known to be an important virulence factor. Few strains of Helicobacter pylori express sialyl-Lewis-X and we have reported that human and canine Helicobacter bizzozeronii strains express sialyl-lactoseamine in their LPS. However, the role of sialyation of Helicobacter LPS in the interaction with the host cells is still unknown. In this study H. bizzozeronii LPS is shown to activate the TLR2 in a dose and strain dependent manner in the in vitro HEK-293 cells model expressing TLR2, but not the cells expressing TLR4. These results indicate that TLR2 is the specific receptor for H. bizzozzeronii LPS, as previously described for H. pylori. To further explore the role of sialylation of H. bizzozeronii LPS on TLR2 response, H. bizzozeronii Δhbs2 mutant strains deficient in sialyltransferase activity were constructed by homologous recombination. LPS from H. bizzozeronii Δhbs2 strains enhanced the NF-ĸB induction via TLR2 compared to the respective wild types, leading to the conclusion that the sialylation of H. bizzozeronii LPS in wild-type strains may modulate host immune response.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0133-4) contains supplementary material, which is available to authorized users.     

Investigation of an outbreak of sheeppox among native sheep breeds in the Western Himalayas of India

Veterinary Research Communications - An outbreak of sheeppox was investigated in a cluster of villages situated in Western Himalayan ranges of a Northern Indian state. Non-migratory sheep...     

Effect of selected single nucleotide polymorphisms in SLC11A1, ANKRA2, IFNG and PGLYRP1 genes on host susceptibility to Mycobacterium avium subspecies paratuberculosis infection in Indian cattle

Paratuberculosis (PTB) is a chronic infectious enteritis of ruminants, caused by Mycobacterium avium subspecies paratuberculosis (MAP) that brings huge economic loss to the dairy farmers. The study was conducted to explore the association of selected SNPs in IFNG, SLC11A1, ANKRA2 and PGLYRP1 genes with resistance to PTB disease in Indian cattle population. A case-control resource population was established based on the results of diagnostic tests used for detection of MAP infection status viz. ELISA, Johnin PPD test, faecal microscopy and IS900 blood PCR. The PCR-RFLP method was used for genotyping of SNPs. SNPs rs109453173 in SLC11A1, rs110853455 in IFNG and rs41933863 in ANKRA2 genes were significantly (P<0.05) associated with resistance to MAP infection. For SNP rs109453173, GG genotype and G allele was found to be associated with resistance against MAP infection than CC and CG genotypes and C allele, respectively. For SNP rs110853455, AG genotype was found to be associated with susceptibility to MAP infection than AA and GG genotype. For SNP rs41933863, the AG genotype provided three and six times more resistance against MAP infection than GG and AA genotype. The results of this study are suggestive of SNPs rs109453173, rs110853455 and rs41933863 as potential markers for screening MAP resistant cattle and a breeding programme favouring GG genotype and G allele for rs109453173, AG genotype for rs41933863 and against AG genotype for rs110853455 might confer resistance against MAP infection in Indian cattle. However, investigation of these SNPs in an independent and larger population will warrant the strength of association for resistance against MAP infection in cattle.

     

The use of circular and hybrid external skeletal fixation systems to repair open tibial fractures in large ruminants: a report of six clinical cases

Veterinary Research Communications - A prospective clinical study consisting of six bovines (200–300&nbsp;kg body weight) undergoing open tibial fractures was carried out, to evaluate the...     

The sensitivity of growth and yield of dwarf wheat to water stress at three growth stages

Summary Response of dwarf wheat (Triticum aestivum L. em Thell) to three different levels of water stress at three growth stages — seeding to maximum tillering, maximum tillering to flowering and flowering to maturity, was studied under field conditions for two seasons. At each of these three stages, plants were subject to three ratios of irrigation water to cumulative pan evaporation (IW/CPE) –0.45, 0.60 and 0.75. During the remaining stages the plants were irrigated with an IW/CPE ratio of 0.9. Thus mild, moderate and severe stress treatments were compared with a no-stress control. At all stages moderate and severe water stress decreased plant height, leaf area, ear number, 1000-grain weight, grain yield and water-use efficiency. In stage 3 the effect of water stress on straw yield was not marked. Wheat was most sensitive to water stress during stage 1 when the reduction in grain yield was caused by a reduction in numbers of ears and grains per ear. In stage 2, grain yield reduction was due to fewer grains perear and a lower 1000-grain weight. On rewatering, mild stressed plants showed recovery of plant height, tiller number and in consequence, yield. Results indicate that under the conditions of this study the wheat crop should be irrigated at a IW/CPE ratio of 0.75 when water resources are limited. With an unlimited water supply the ratio may be increased to 1.2 in stage 2 to maximise the yield.     

Rice straw recycling: A sustainable approach for ensuring environmental quality and economic security

One of the major challenges in Asian countries is the effective management of rice straw. To ensure food security for their ever-growing population, Asian countries grow more rice, leading to increasing rice straw generation. Burning of rice straw, a common practice, is detrimental to both environmental and human health. However, if managed effectively, rice straw has the potential to safeguard the sustainability of agricultural ecosystems and to uplift the economic security of the population de...     

‘Neodomesticates’ of the Himalayan allium spices (Allium species) in Uttarakhand,India and studies on eco-geography and morphology

Genetic Resources and Crop Evolution - The study on Himalayan allium spices was undertaken with specific reference to commercially important taxa belonging to the genus Allium viz. A. stracheyi...