Deoxyribonucleic Acid Analysis Research Articles

Genetic imprint of the Mongol: signal from phylogeographic analysis of mitochondrial DNA

Mitochondrial deoxyribonucleic acid (DNA) from 201 unrelated Mongolian individuals in the three different regions was analyzed. The Mongolians took the dominant East Asian-specific haplogroups, and some European-prevalent haplogroups were detected. The East Asians-specific haplogroups distributed from east to west in decreasing frequencies, and the European-specific haplogroups distributed conversely. These genetic data suggest that the Mongolian empire played an important role in the maternal genetic admixture across Mongolians and even Central Asian populations, whereas the Silk Road might have contributed little in the admixture between the East Asians and the Europeans.

Cutaneous Granulomas with Predominantly CD8+ Lymphocytic Infiltrate in a Child with Severe Combined Immunodeficiency

Combined immunodeficiency disorders comprise a heterogeneous group of diseases characterized by both humoral and cell-mediated immunodeficiency. Cutaneous granulomas manifestations in children with combined immunodeficiency are rare. We report the case of a 6-year-old boy who presented with disseminated cutaneous granulomas and a history of multiple infections. Laboratory evaluation revealed severe combined immunodeficiency, and deoxyribonucleic acid (DNA) analysis confirmed mutations on a gene of chromosome 19 that encodes an enzyme called Janus kinase 3 (Jak-3). Immunohistochemistry revealed expression of CD8(+) in the perivascular lymphocytic infiltrate Disseminated granulomatous lesions in children with a history of frequent infections should prompt the clinician to initiate detailed immunocompetence evaluation as they might prove to be the first manifestation of immunologic impairment.

Insect collections and DNA analyses: how to manage collections?

Numerous papers have been published recently regarding deoxyribonucleic acid (DNA) studies of museum insect specimens. Despite the great enthusiasm for such analyses, there continues to be a potential conflict between specimen maintenance and their use for experimental procedures, since DNA extraction methods are sample destructive. This has created a strong limitation for studies on museum specimens, making it impossible to work on rare or irreplaceable species. However, new methods for improving insect preservation have been recently published, together with new procedures for performing less destructive DNA extractions that combine specimen maintenance and DNA extraction.

Characterization of a methanogenic consortium enriched from a coalbed methane well in the Powder River Basin, U.S.A.

Abstract Well-bore water samples from the Fort Union Formation in the Powder River Basin of Wyoming tested positive for the presence of living microbial communities capable of generating methane from Wyodak coal under laboratory conditions. The methanogens in this consortium produced methane from acetate and methanol, but did not produce methane from a H2–CO2 headspace. This was consistent with a phylogenetic analysis of archaeal 16S ribosomal deoxyribonucleic acid (rDNA) sequences from the enrichment culture, which revealed just two phylotypes, both closely related to Methanosarcina mazei. Phylogenetic analysis of bacterial 16S ribosomal ribonucleic acid (rRNA) genes revealed phylotypes similar to the acetogens Acidaminobacter hydrogenoformans and Syntrophomonas sp., and to known fermentative species. This methane-producing consortium was maintained on a defined microbial medium supplemented with Wyodak coal plus 50 mg/L yeast extract as the sole carbon substrates. At 22 °C, the maximum methane production rate was 0.084 m3/t coal/day (2.7 scf/ton/day); in comparison, total methane reserves in the Powder River Basin are approximately 1.6 to 2.2 m3/t (50 to 70 scf/ton). When the incubation temperature was increased from 22 °C to 38 °C, the rate of methane production increased by 300%; similarly, when the culture medium pH was lowered from 7.4 to 6.4, the methanogenesis rate increased by 680%. Increasing the coal particle surface area by 890% via smaller particle size increased methane production rates by over 200%. Microbial methane production in coal slurries was also enhanced by the addition of the solvent N,N-dimethylformamide (DMF). These results suggest an opportunity to enhance coalbed methane reserves by stimulating the activity of existing methanogenic consortia in-situ; in particular, reservoir treatments that enhance coal solubility and dissolution rates may be beneficial.

Spores of the mycorrhizal fungus Glomus mosseae host yeasts that solubilize phosphate and accumulate polyphosphates

The present paper reports the presence of bacteria and yeasts tightly associated with spores of an isolate of Glomus mosseae. Healthy spores were surface disinfected by combining chloramine-T 5%, Tween-40, and cephalexin 2.5 g L(-1) (CTCf). Macerates of these spores were incubated on agar media, microorganisms were isolated, and two yeasts were characterized (EndoGm1, EndoGm11). Both yeasts were able to solubilize low-soluble P sources (Ca and Fe phosphates) and accumulate polyphosphates (polyPs). Sequence analysis of 18S ribosomal deoxyribonucleic acid showed that the yeasts belong to the genera Rhodotorula or Rhodosporidium (EndoGm1) and Cryptococcus (EndoGm11). Results from inoculation experiments showed an effect of the spore-associated yeasts on the root growth of rice, suggesting potential tripartite interactions with mycorrhizal fungi and plants.

Feasibility of 2,4,6-Trichlorophenol and Malonic Acid as Metabolic Uncoupler for Sludge Reduction in the Sequence Batch Reactor for Treating Organic Wastewater

The activated sludge process generates a large amount of excess sludge as a byproduct, which is one of the most serious challenges in biological wastewater treatment. In the present study, the feasibility of 2,4,6-trichlorophenol (TCP) and malonic acid (MA) as metabolic uncouplers to reduce sludge generation in the sequence batch reactor (SBR) for treating organic wastewater for a long period was studied. The results showed that 2 mg/L TCP could reduce sludge generation by about 47%, while chemical oxygen demand (COD) removal efficiency and sludge settlability were not obviously influenced. Although 10 mg/L MA could also reduce excess sludge production by about 30% while slightly affecting COD removal, it seriously deteriorated sludge settlability. Accordingly, TCP is a better uncoupler for sludge reduction for a longer period in the SBR for treating organic wastewater, and MA can only be used as a short-term or transitional uncoupler. Microscopic and 16S ribosomal deoxyribonucleic acid analyses showed that the microbial population of sludge varied when uncouplers were fed to the activated sludge system. Occurrence of large amounts of filament and the disappearance of protozoa may be the main reason for the aggravation of sludge settlability under uncoupled metabolic conditions caused by MA.

High abundance and dark CO2 fixation of chemolithoautotrophic prokaryotes in anoxic waters of the Baltic Sea

We determined the abundance and distribution of chemolithoautotrophic prokaryotes in the redoxcline in two basins (Gotland Deep, Landsort Deep) of the central Baltic Sea by combining dark CO2 fixation measurements with flow cytometric cell sorting. Maximum CO2 fixation rates were recorded in sulfidic waters about 20 m below the chemocline. Flow cytometric analyses of deoxyribonucleic acid (DNA)‐stained bacterioplankton revealed the existence of at least five different prokaryotic clusters in water samples collected below the chemocline. Dark CO2 fixation in these clusters was determined by flow cytometric sorting after anoxic incubations with NaH14CO3 tracer. Two clusters, representing about 30% of total prokaryotes, were responsible for 65% to 100% of the total dark fixation. Calculated cell‐specific CO2 fixation rates in the two basins ranged from 3.5 to 24.7 fg C cell−1 d−1 and suggested that these clusters are dominated by chemolithoautotrophic prokaryotes. Mean cell‐specific fixation rates reached more than 10 fg C cell−1 d−1 in most cases, indicating relatively high growth rates (doubling times 1‐2 d) of chemolithoautotrophic prokaryotes. Our results provide the first evidence of such high cell‐specific CO2 uptake and abundance of chemolithoautotrophic prokaryotes in a pelagic marine environment. However, the identity of the organisms as well as the mechanisms fueling CO2 dark fixation in the anoxic zone remain unknown.

Molecular and cytogenetic evidence of three sibling species of the Anopheles barbirostris Form A (Diptera:Culicidae) in Thailand

Nine isoline colonies of Anopheles barbirostris Form A, derived from individual isofemale lines from Chiang Mai, Phetchaburi, and Kanchanaburi, were established in our insectary at Chiang Mai University. All isolines shared the same mitotic karyotype (X(1), X(2), Y(1)). Molecular analysis of deoxyribonucleic acid (DNA) sequences and polymerase chain reaction (PCR) products of ITS2, COI, and COII regions revealed three distinct groups: A1 (Chiang Mai), A2 (Phetchaburi), and A3 (Kanchanaburi). Crossing experiments among the three groups exhibited strong reproductive isolation, producing low and/or non-hatched eggs, and inviable and/or abnormal development of the reproductive system of F(1)-progenies. Asynaptic regions along the five polytene chromosome arms of F(1)-hybrid larvae clearly supported the existence of three sibling species within A. barbirostris Form A, provisionally named species A1, A2, and A3.

A non-invasive genetic method to identify the sympatric mustelids pine marten (Martes martes) and stone marten (Martes foina): preliminary distribution survey on the northern Iberian Peninsula

The closely related mustelids European pine marten (Martes martes) and stone marten (Martes foina) sympatrically inhabit a large area of Europe. However, given our limited knowledge of their bioecological relationships, their extremely elusive behaviour and the fact that their faeces cannot be distinguished on the basis of morphology alone, it is very difficult to monitor their populations. In this study, we describe a reliable non-invasive polymerase chain reaction (PCR)–restriction fragment length polymorphism (PCR-RFLP) method for distinguishing between M. martes and M. foina based on the analysis of deoxyribonucleic acid extracted from faeces samples. The method was specifically designed to avoid possible interference from potential prey mammals and other sympatric carnivores. The procedure consists of PCR amplifying a mitochondrial D-loop region followed by digesting the resulting 276-bp-long amplicons with the restriction enzymes HaeIII and RsaI. To assess the efficiency of this technique, we conducted a preliminary field study across the potential sympatric distribution areas of both marten species in the northern Iberian Peninsula. Out of 359 faeces samples collected, we identified 80 as specimens from the stone marten and 235 from the pine marten. Unequivocal species identification was thus possible in 88% of the faeces samples collected. These findings reveal the combined use of non-invasive genetic sampling and GIS technology to be a reliable and cost-effective procedure for improving our knowledge of the spatial distributions of sympatric marten species. This protocol could also be used to identify and improve information gaps, to develop effective research and management programmes and in population and landscape genetics studies of marten species.

Surface fine topography and PCR-based determination of metacercaria of Paragonimus sp. from edible crabs in Arunachal Pradesh, Northeast India

In several mountainous regions of Northeastern India, foci of Paragonimus infection reportedly involving species that are known to prevail in China have been identified. The present study was undertaken to demonstrate the surface fine topography and sequence analysis of the ribosomal deoxyribonucleic acid (rDNA; second internal transcribed spacer, ITS2) of the metacercarial stages of the lung fluke collected from a mountain stream of the area (Miao, Changlang District in Arunachal Pradesh). The encysted metacercariae were oval in shape and had a smooth surface. The newly excysted metacercaria had a ventral sucker larger than the oral; the body surface was covered with numerous single-pointed and thorn-like tegumentary spines, of which those on the anterior part of the body were bigger in size and showed a gradual reduction in length and number towards the posterior end; dome-shaped papillae in variable numbers were seen around the rim of the oral sucker and were sparsely distributed all over the body surface. The polymerase chain reaction-amplified rDNA ITS2 sequences of the metacercariae were aligned with known sequences for the various species of Paragonimus, and the expectation value was found to be most significant with P. westermani, revealing an absolute match. The surface topography including the number and distribution of papillae and spination patterns and the ITS2 sequences of the metacercariae strongly suggest that the Paragonimus species, prevalent in the region of India, is in fact P. westermani.

A taxonomic review of the genus Phaeocystis

Phaeocystis is recognized both as a nuisance and as an ecologically important phytoplankton species. Its polymorphic life cycle with both colonial and flagellated cells causes many taxonomic problems. Sequence variation among 22 isolates representing a global distribution of the genus has been compared using three molecular markers. The ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO) spacer is too conserved to resolve species. The most conserved 18S ribosomal deoxyribonucleic acid (rDNA) analysis suggests that an undescribed unicellular Phaeocystis sp. (isolate PLY559) is a sister taxon to the Mediterranean unicellular Phaeocystis jahnii; this clade branched prior to the divergence of all other Phaeocystis species, including the colonial ones. The internal transcribed spacer (ITS) region shows sufficient variation that some spatial population structure can be recovered, at least in P. antarctica. P. globosa and P. pouchetii have multiple different ITS copies, suggestive of cryptic species that are still able to hybridize. A molecular clock has been constructed that estimates the divergence of the cold water colonial forms from the warm-water colonial forms to be about 30 Ma and the divergence of P. antarctica and P. pouchetii to be about 15 Ma. A short description of the colonial stage and the flagellated stage for each formally recognized species is provided. Morphological information is also provided on a number of undescribed species. These include the strain Ply 559, consisting of non-colonial cells with peculiar tubular extrusomes, a second non-colonial species from the north western Mediterranean Sea producing a lot of mucus, and a colonial species with scale-less flagellates found in Italian waters. In addition, three flagellated morphotypes with scales different from those of P. antarctica were reported in the literature from Antarctic waters. The picture emerging from both molecular and morphological data is that the number of species in the genus is still underestimated and that cryptic or pseudocryptic diversity requires a sound assessment in future research of this genus. Based on all published observations, an emended description of the genus is provided.

Analysis of Mitochondrial DNA for Authentication of Meats from Chamois (Rupicapra rupicapra), Pyrenean Ibex (Capra pyrenaica), and Mouflon (Ovis ammon) by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

The prevention of fraudulent labeling of game meat constitutes an important part of food regulatory control and quality assurance systems. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis based on mitochondrial deoxyribonucleic acid (DNA) was developed for authentication of meats from chamois (Rupicapra rupicapra), pyrenean ibex (Capra pyrenaica), and mouflon (Ovis ammon). Amplification and restriction site analysis of a DNA fragment about 720 base pairs (bp) from the mitochondrial 12S rRNA gene of all analyzed species permitted the selection of Msel and Apol endonucleases for meat speciation. The 12S rRNA restriction profiles obtained allowed the unequivocal identification of chamois, pyrenean ibex, and mouflon/sheep and their differentiation from meats of domestic species such as cattle, goat, and swine. The highly variable mitochondrial D-loop gene was also targeted to attempt discrimination between mouflon and sheep meats. A D-loop region (700-1000 bp) was amplified and sequenced in all game and domestic species analyzed, and a primer set was designed for the selective amplification of a 370 bp DNA fragment from mouflon and sheep. PCR-RFLP analysis with the selected Maell enzyme generated a single electrophoretic profile characteristic for sheep, whereas 3 different fragment patterns were obtained for mouflon meats. Consequently, the PCR-RFLP technique developed can be routinely applied in inspection programs in order to verify the correct labeling of game species.

Scanning tunneling spectroscopy of single-strand deoxyribonucleic acid for sequencing

The authors performed scanning tunneling spectroscopy (STS) analysis of single-strand deoxyribonucleic acid (DNA) with fluorescein isothiocyanate (FITC) dye at the 5′ terminal (FITC-5′-agctgtac-3′). They obtained high-resolution topographic scanning tunneling microscope images of the DNA that showed eight nucleotide bases and one FITC molecule. The current versus voltage (I-V) data of FITC had local maxima or plateaus, whereas those of DNA had simple growth curves. They also found that in the STS analysis, the I-V data of the DNA were fitted well by an exponential curve in the positive and negative bias regions. They also discuss the possibility of whether four nucleotides (adenine, guanine, cytosine, and thymine) can be differentiated through STS experiments.

Genotyping of samples lacking expected antibodies in ABO blood group

We report nine donations with ABO inconsistency in reverse typing caused by partly or entirely missing antibodies. A and B antigens and antibodies were examined by serological blood typing, and ABO deoxyribonucleic acid (DNA) analyses were performed by sequence-specific priming and sequencing. A B101 allele was demonstrable in a case with O phenotype. The molecular mechanisms in deficiency of natural ABO antibody could be partly clarified. The ABO genotyping technique is an accurate method for determining the blood samples involved in ABO grouping discrepancies and is a valuable complement to serology for correct determination of donor blood status. The mechanisms involved in the absence of potent natural antibodies directed against A and B antigen lacking on an individual's own red cell membranes remain to be further investigated.

The Cytocompatibility and Early Osteogenic Characteristics of an Injectable Calcium Phosphate Cement

In this study, the cytocompatibility and early osteogenic characteristics of rat bone marrow cells (RBMCs) on injectable calcium phosphate (CaP) cement (Calcibon) were investigated. In addition to unmodified CaP cement discs, 2 other treatments were given to the discs: preincubation in MilliQ and sintering at different temperatures. After primary culture, RBMCs were dropwise seeded on the discs and cultured for 12 days. The samples were evaluated in terms of cell viability, morphology (live and dead assays and scanning electron microscopy (SEM)), cell proliferation (deoxyribonucleic acid (DNA) analyses), early cell differentiation (alkaline phosphatase (ALP) activity), and physicochemical analyses (x-ray diffraction (XRD)). The live and dead, DNA, and SEM results showed that Calcibon discs without any additional treatment were not supporting osteoblast-like cells in vitro. There were fewer cells, and cell layers were detached from the disc surface. Therefore, different preincubation periods and sintering temperatures were evaluated to improve the cytocompatibility of the CaP cement. Preincubating discs in MilliQ for periods of 1, 4, 8, and 12 weeks resulted in the hydrolysis of alpha-tri calcium phosphate (TCP) into an apatite-like structure with some beta-TCP, as shown with XRD, but the material was not cytocompatible. Sintering the discs between 800 degrees C and 1100 degrees C resulted in conversion of alpha-TCP to beta-TCP with some hydroxyapatite and an increase in crystallinity. Eventually, the discs sintered at 1100 degrees C achieved better cell attachment, more-abundant cell proliferation, and earlier differentiation than other sintered (600 degrees C, 800 degrees C, and 1000 degrees C), preincubated, and unmodified specimens. On basis of our results, we conclude that in vivo results with CaP-based cements do not guarantee in vitro applicability. Furthermore, unmodified Calcibon is not cytocompatible in vitro, although preincubation of the material results in a more-favorable cell response, sintering of the material at 1100 degrees C results in the best osteogenic properties. In contrast to in vivo studies, the Calcibon CaP cement is not suitable as a scaffold for cell-based tissue-engineering strategies.

MELANOTIC NONPSAMMOMATOUS TRIGEMINAL SCHWANNOMA AS THE FIRST MANIFESTATION OF CARNEY COMPLEX

Melanotic schwannoma is a rare neoplasm, classifiable as a peripheral nerve sheath tumor, and differentiated from a typical schwannoma by heavy pigmentation. Psammoma bodies can be visualized in more than 50% of melanotic schwannomas. Half of patients with such "psammomatous melanotic schwannomas" have Carney complex, a dominantly transmitted autosomal disorder. Most recently, the tumor suppressor gene, PRKAR1A, coding for the Type 1alpha regulatory subunit of protein kinase A was found to be mutated in approximately half of the known Carney complex families. Although cranial schwannomas have been described in patients with Carney complex, their numbers are too small to be considered a definite part of the syndrome. Furthermore, only melanotic schwannomas with psammoma bodies are included as diagnostic criteria for Carney complex. The objective of this report is to communicate a case of trigeminal nonpsammomatous melanotic schwannoma as the first manifestation of Carney complex. A 34-year-old woman presented with odontalgia, right V3 hypoesthesia, V2 paresthesia, and diplopia. Magnetic resonance imaging scans of the brain revealed a small tumor with homogenous contrast in the right trigeminal pathway. We performed an extradural approach to the right cavernous sinus by a middle fossa approach. The lateral wall was opened between the cranial nerves, and a soft and black tumor was resected in a piecemeal fashion. Histology and immunohistochemical analysis of the tumor were compatible with melanotic schwannoma, but no psammomatous bodies were identified. Endocrine evaluation showed that this patient's symptoms fulfilled the diagnostic criteria of Carney complex, with lentiginosis, multiple breast ductal adenomas, multiple hypoechoic nodules on thyroid ultrasonography, and a 4 x 5-cm asymptomatic atrial cardiac myxoma, which was removed 15 days after the neurosurgery. Three months later, a recurrence of melanotic schwannoma was identified. Molecular analyses of genomic and somatic deoxyribonucleic acid from the patient found a 578 to 579delTG mutation of PRKAR1A. We present the unusual case of a nonpsammomatous trigeminal melanotic schwannoma associated with Carney complex, with confirmed PRKAR1A gene mutation. Our case highlights that neurosurgeons, in the presence of a melanotic schwannoma, should be aware of the features of the Carney complex because, in such cases, pre- and postoperative management is significantly affected. We also postulate that the absence of psammoma bodies or cranial localization do not exclude this diagnosis.

Evolutionary computation in bioinformatics: a review

This paper provides an overview of the application of evolutionary algorithms in certain bioinformatics tasks. Different tasks such as gene sequence analysis, gene mapping, deoxyribonucleic acid (DNA) fragment assembly, gene finding, microarray analysis, gene regulatory network analysis, phylogenetic trees, structure prediction and analysis of DNA, ribonucleic acid and protein, and molecular docking with ligand design are, first of all, described along with their basic features. The relevance of using evolutionary algorithms to these problems is then mentioned. These are followed by different approaches, along with their merits, for addressing some of the aforesaid tasks. Finally, some limitations of the current research activity are provided. An extensive bibliography is included

A Second Decorin Frame Shift Mutation in a Family With Congenital Stromal Corneal Dystrophy

To identify the genetic defect in a Belgian family with congenital stromal corneal dystrophy. Case report and result of deoxyribonucleic acid (DNA) analyses. DNA sequencing of polymerase chain reaction (PCR) products generated from amplification of exons and adjacent introns of the decorin gene. The family consisted of a mother and her son, both suffering from congenital stromal corneal dystrophy. In both individuals, a single base pair deletion (c.941delC) in the coding sequence of the decorin gene was demonstrated, predicting a C-terminal truncation of the decorin protein (p.Pro314fsX14). This is the second family with congenital stromal corneal dystrophy of the cornea in which a frame shift mutation in the decorin gene has been detected. Both in this family and in a previously reported Norwegian family, a decorin protein missing the 33 C-terminal amino acids is predicted. This observation strongly supports a role for decorin in the pathogenesis of this disorder.

Microfluidic systems for extracting nucleic acids for DNA and RNA analysis

This paper is to review the differences in the developments of microfluidic chips for extracting genomic deoxyribonucleic acid (DNA) and viral ribonucleic acid (RNA) from blood by the Biosensor Focus Interest Group (BFIG) in Singapore. DNA was extracted in a multi-step process by isolating and lysing white blood cells (WBC), typically ∼10 μm in diameter. Viral RNA was extracted directly from the submicron viruses in the blood. In terms of basic microfluidic components required, both DNA and RNA extractions used similar mixers for mixing reagents, filters for capturing or separating the blood cells, and a binder for capturing and purifying the DNA/RNA molecules. The designs of the filters were adapted to either capture WBC for DNA isolation or capture all virus particles for RNA isolation. The designs of these two kinds of filters had to be different. Besides the differences in the sizes of WBC and viruses, the concentration of the virus particles is usually much lower than WBC. Thus, a much higher volume of blood for filtering would be required for extracting viral RNA, especially for the intention to detect the viruses at early onset of infection. With proper modifications of the protocols, it has been demonstrated that both genomics DNA and viral RNA could be extracted successfully in these microfluidic chips. The quality of the extracted samples was verified by polymerase chain reaction (PCR) and gel-electrophoresis after the extractions.

Plant growth-promoting bacteria as biofertilizer

Seventeen rhizobacteria isolated from different ecological regions, i.e. Brazil, Indonesia, Mongolia and Pakistan were studied to develop inoculants for wheat, maize and rice. Almost all the bacterial isolates were Gram-negative, fast-growing motile rods and utilized a wide range of carbon sources. These isolates produced indole-3-acetic acid at concentrations ranging from 0.8-42.1 μg/ml, irrespective of the region. Fifteen isolates fixed N at rates ranging from 20.3-556.8 nmole C 2 H 2 reduced/h/vial. Isolate 8N-4 from Mongolia produced the highest amount of indole-3-acetic acid (42.1 μg/mL), produced siderophores (0.3 mg/mL) and was the only isolate that solubilized phosphate (188.7 tag P/mL). Inoculation of the wheat variety Orkhon with 8N-4 isolate resulted in the maximum increase in plant biomass, root length, and total N and P contents in plants. Random amplified polymorphic deoxyribonucleic acid (RAPD) analysis, conducted with 60 decamer primers, revealed a high level of polymorphism among the bacterial isolates from different geographic regions and a low level of polymorphism among isolates from the same region. The complete 16S rRNA gene sequence analysis demonstrated that 8N-4 is a Bacillus pumilus strain (Accession number AY548949). It was concluded that Bacillus pumilus 8N-4 can be used as a bio-inoculant for biofertilizer production to increase the crop yield of wheat variety Orkhon in Mongolia.