Tricalcium phosphate solubilization and nitrogen fixation by newly isolated Aneurinibacillus aneurinilyticus CKMV1 from rhizosphere of Valeriana jatamansi and its growth promotional effect

Abstract Aneurinibacillus aneurinilyticus strain CKMV1 was isolated from rhizosphere of Valeriana jatamansi and possessed multiple plant growth promoting traits like production of phosphate solubilization (260 mg/L), nitrogen fixation (202.91 nmol ethylene mL-1 h-1), indole-3-acetic acid (IAA) (8.1 µg/mL), siderophores (61.60%), HCN (hydrogen cyanide) production and antifungal activity. We investigated the ability of isolate CKMV1 to solubilize insoluble P via mechanism of organic acid production. High-performance liquid chromatography (HPLC) study showed that isolate CKMV1 produced mainly gluconic (1.34%) and oxalic acids. However, genetic evidences for nitrogen fixation and phosphate solubilization by organic acid production have been reported first time for A. aneurinilyticus strain CKMV1. A unique combination of glucose dehydrogenase (gdh) gene and pyrroloquinoline quinone synthase (pqq) gene, a cofactor of gdh involved in phosphate solubilization has been elucidated. Nitrogenase (nif H) gene for nitrogen fixation was reported from A. aneurinilyticus. It was notable that isolate CKMV1 exhibited highest antifungal against Sclerotium rolfsii (93.58%) followed by Fusarium oxysporum (64.3%), Dematophora necatrix (52.71%), Rhizoctonia solani (91.58%), Alternaria sp. (71.08%) and Phytophthora sp. (71.37%). Remarkable increase was observed in seed germination (27.07%), shoot length (42.33%), root length (52.6%), shoot dry weight (62.01%) and root dry weight (45.7%) along with NPK (0.74, 0.36, 1.82%) content of tomato under net house condition. Isolate CKMV1 possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent and this is the first study to include the Aneurinibacillus as PGPR.

Domain Analysis and Isolation of Coniferyl Alcohol Dehydrogenase Gene from Pseudomonas nitroreducens Jin1.

Aim: The Aim of present study is to analyse conserved functional Short Dehydrogenase Reductase (SDR) domain from bacteria. Based on the domain analysis selection of coniferyl alcohol dehydrogenase gene for isolation from Pseudomonas nitroreducens Jin1. Place and Duration of Study: Department of Biotechnology, Punjabi University, Patiala. From July, 2012 to November, 2012. Methodology: Bioinformatics tools were used to analyse various calA genes from bacteria based on the presence of conserved domain in members of SDR family. Based on insilico analysis, Pseudomonas nitroreducens Jin1 calA was selected. PCR was used for amplification of the gene from the genome of Pseudomonas nitroreducens Jin1. Result: Multiple sequence alignment results for conserved domains amongst members of SDR family identified presence of all domains of Short Dehydrogenase Reductase members in Pseudomonas nitroreducens Jin1 calA gene. Amongst the various sequences compared the P. nitroreducens Jin1, calA was found to be the smallest in size. The locus of calA in the genome resides at 103513-104280 bases. It was amplified from the genome of Pseudomonas nitroreducens Jin1. The calA gene that was amplified is of size 768bp. Conclusion: calA gene isolated from Pseudomonas nitroreducens Jin1 is a small gene with all the functional domains and can be used for biotransformation of coniferyl alcohol to coniferyl aldehyde.

Standardization of PCR-RFLP analysis of nsSNP rs1468384 of NPC1L1 gene.

Niemann-Pick C1-like 1 (NPC1L1) protein, a newly identified sterol influx transporter, located at the apical membrane of the enterocyte, which may actively facilitate the uptake of cholesterol by promoting the passage of sterols across the brush border membrane of the enterocyte. It effects intestinal cholesterol absorption and intracellular transport and as such is an integral part of complex process of cholesterol homeostasis. The study of population data for the distribution of these single nucleotide polymorphisms (SNP) of NPC1L1 has lead to the identification of six non-synonymous single nucleotide polymorphisms (nsSNP). The in vitro analysis using the software MuPro and StructureSNP shows that nsSNP M510I (rs1468384), which involves A→G base pair change leads to decrease in the stability of the protein. A reproducible and a cost-effective PCR-RFLP based assay was developed to screen for the SNP among population data. This SNP has been studied in Caucasian, Asian, and African American populations. Till date, no data is available on Indian population. The distribution of M510I NPC1L1 genotype was estimated in the North Western Indian Population as a test case. The allele distribution in Indian Population differs significantly from that of other populations. The methodology thus proved to be robust enough to bring out these differences.

Apolipoprotein E isotype distribution amongst Ramgharias of Punjab.

Apolipoprotein E is a major constituent of chylomicrons and HDL fractions of the normal blood plasma involved in lipid transfer systems. The three common alleles apoE2, apoE3 and apoE4 with six possible phenotypes have been identified at this locus in all the populations of the world studied so far, with apoE3 being the most common. The present study employed PCR restriction isotyping techniques to estimate allele frequency distribution amongst Ramgharias an artisan caste group of Punjab in North India. The methodology was developed to visualise the isotypes on PAGE using silver staining. The major genotypes observed were E2/3, E3/3 and E3/4 with allele frequencies APO E2=0.107, APOE3=0.714 AND APO E4=0.179.