The aim of the study was to establish the frequencies of CYP2C9*1, *2, *3 and CYP2C19*1, *2 and *3 in the south Indian population and to compare them with the inter-racial distribution of the CYP2C9 and CYP2C19 genetic polymorphisms. Genotyping analyses of CYP2C9 and CYP2C19 were conducted in unrelated, healthy volunteers from the three south Indian states of Andhra Pradesh, Karnataka and Kerala, by the polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). The allele frequencies of the populations of these three states were then pooled with our previous genotyping data of Tamilians (also in south India), to arrive at the distribution of CYP2C9 and CYP2C19 alleles in the south Indian population. Frequencies of CYP2C9 and CYP2C19 alleles and genotypes among various populations were compared using the two-tailed Fisher's exact test. The frequencies of CYP2C9*1, *2 and *3 in the south Indian population were 0.88 (95% CI 0.85-0.91), 0.04 (95% CI 0.02-0.06) and 0.08 (95% CI 0.06-0.11), respectively. The frequencies of CYP2C9 genotypes *1/*1, *1/*2, *1/*3, *2/*2, *2/*3 and *3/*3 were 0.78 (95% CI 0.74-0.82), 0.05 (95% CI 0.03-0.07), 0.15 (95% CI 0.12-0.18), 0.01 (95% CI 0.0-0.02), 0.01 (95% CI 0.0-0.02) and 0.0, respectively. CYP2C19*1, *2 and *3 frequencies were 0.64 (95% CI 0.60-0.68), 0.35 (95% CI 0.31-0.39) and 0.01 (95% CI 0.0-0.03), respectively. As a result of a significant heterogeneity, the data on CYP2C19 genotype frequencies were not pooled. The frequency of CYP2C9*2 mutant alleles in south Indians was higher than in Chinese and Caucasians, while CYP2C9*3 was similar to Caucasians. CYP2C19*2 was higher than in other major populations reported so far. The relatively high CYP2C19 poor-metabolizer genotype frequency of 12.6% indicates that over 28 million south Indians are poor metabolizers of CYP2C19 substrates.
Positional cloning recently identified the mutation causing copper toxicosis (CT) in Bedlington terriers. Isolation of the MURR1 gene will be of great value in developing a reliable diagnostic test for the breeding of a copper toxicosis-free stock. It will replace the current diagnostic test using the CT-linked marker, C04107, which is located in intron 1 of the MURR1 gene with a distance of approximately 8 kb from the exon 2 deletion. Despite the short distance between C04107 and the CT mutation, possible recombinant dogs have been reported with C04107. Although these dogs have a normal phenotype, they carry the C04107 allele 2, which is associated with CT. To study the origin of this possible recombination event we collected a pedigree consisting of two unaffected American Bedlington terriers and their litter of four pups, which were all homozygous for the C04107 2,2 genotype. Mutation analysis showed that two dogs were heterozygous for the CT exon 2 deletion mutation, whereas four dogs were homozygous for the wild-type (WT) allele. Haplotype analysis was performed using two DNA markers in the MURR1 gene and four DNA markers flanking the gene and spanning a region of approximately 600 kb. Surprisingly, we identified a new haplotype (haplotype C) that contains allele 2 of marker C04107 in combination with the WT MURR1 allele. Analysis of the flanking markers suggests there are different genetic backgrounds in the Bedlington terrier population.
The gram pod borer, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), is infected by nucleopolyhedrosis virus (HearNPV), which is the most promising microbial biocontrol agent of the pest. A genetic diversity analysis of geographically distinct isolates of HearNPV was done, using the polyhedrin (polh) gene of the viruses that encodes a major structural protein of the occlusion bodies. The gene was amplified and isolated from eight Indian isolates, using the polymerase chain reaction (PCR). These sequences were compared with the polh genes of other HearNPV from different geographical regions of the world. A phylogenetic tree was constructed, using polh nucleotide/deduced amino acid sequences to know their genetic relatedness. The polh gene of isolates originating from nearby locations clustered together with the gene of isolates in the present study; however, some showed relatedness with gene isolate from other geographically distinct isolates, with respect to the genetic distances among them. The Indian isolate (Ban-PDBC-In) shared lower genetic distance of 0.0020 to 0.0040 substitutions per site with the Spanish isolates SP1A-Sp and SP1B-Sp and clustered in one group based on nucleotide sequences. The isolates showed different a clustering pattern in phylogenetic tree based on deduced amino acid sequences than that of the nucleotide sequences. The overall genetic distances between polh nucleotides ranged from 0.0000 to 0.0203 substitutions per site, while it was 0.0000 to 0.0121 between deduced amino acid sequences. Among different geographical groups of isolates, the Indian group showed the highest genetic diversity based on both polh nucleotide (0.0070 ± 0.0002 substitutions per site) and deduced amino acid (0.0057 ± 0.0003 substitutions per site) sequences among different groups of geographical isolates. A diversity analysis of virus isolates can aid in the selection and identification of virulent virus isolates for the development of a virus-based bio-pesticide formulation.
Understanding the genetic diversity of BmNPV isolates causing grasserie viral disease of mulberry silkworm Bombyx mori L. is essential for adoption of management strategies including biotechnological tools. The present study was aimed at the use of restriction fragment length polymorphism (RFLP) profiling for studying the molecular diversity analysis of six BmNPV isolates collected from Devanahalli, Kolar, Shidlaghatta, Hosakote, Tumakuru, and Ramanagara areas in Karnataka, India (BmNPV-Ko, BmNPV-Ho, BmNPV-SG, BmNPV-DH, BmNPV-TM, BmNPV-Ram respectively). DNA was extracted from each of these isolates and subjected to digestion with different restriction enzymes EcoR1, BamH1, Sma1, Nco1, and a combination of BamH1+Nco1 and electrophoresed in 0.8% w/v agarose gel to visualize restriction enzyme profile. The analysis revealed that all the six BmNPV isolates had similar Nco1 and Sma1 restriction patterns, although there was variation in low molecular weight fragments. The EcoR1 and BamH1 restriction patterns were nearly the same for all the isolates except for the presence of an approximately 4kb and an additional 1.5kb polymorphic band only in BmNPV-TM and BmNPV-Ram isolates. BamH1+Nco1 digestion of the DNA from each isolate yielded numerous fragments, which was different in BmNPV-Ram isolate. Molecular diversity analysis can helps in understanding the evolution and phylogeny of the virus, enhance the knowledge on its pathogenicity and can help to develop and adopt suitable measures to combat and diagnose the disease to reduce crop loss and increase income generating ability of the farmer.
Fall Armyworm, Spodoptera frugiperda (J E Smith) (Lepidoptera, Noctuidae) is native to the tropical and subtropical region of America. Being a polyphagous pest known to cause major damage to economically important cultivated grasses. This insect is one of the most important pests of maize, being firstly registered in the India in 2018 and it causes a 33% yield loss in the plant production. The use of chemical pesticides as a prophylactic method causes some problems such as ecological instability, pollution, high costs and death of natural enemies. In the current study, screening of native B. thuringiensis isolates and their insecticidal activities were tested against the 2 nd instar larvae of maize fall armyworm, Spodoptera frugiperda under laboratory condition. Preliminary bioassay was conducted using diet incorporation method and treated with spore crystal lysates prepared from the native Bacillus thuringiensis isolates. Upon screening 50.0 per cent (of the B. thuringiensis isolates exhibited mortality in the range of 26-50 per cent while 10.0 per cent of the B. thuringiensis isolates showed more than 75 per cent mortality after 7 days of exposure. Among the seventy isolates, Bt-Oa1 and Bt-257 recorded maximum of 86.84% mortality after 7 days after treatment.
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