Knockdown of dorsal/relish increases larvae susceptibility to microbes in Spodoptera litura

The tobacco cutworm, Spodoptera litura has developed high levels of resistance to frequently-used insecticides, and failures of chemical control has frequently been reported in China. Moreover, the control efficiency of biological control agents was not satisfactory by farmers, and thus the development of novel or combined techniques for such pest control becomes an imperative requirement. In this study the potential application of innate immune-related genes in insect control was investigated. Sldorsal and Slrelish were identified from S. litura. Sldorsals were mainly expressed in epidermal and fatbody of larvae, and Slrelish highly expressed in fatbody and haemolymph of larvae. Their expression characteristic suggests potential roles in defense against fungi or bacteria. The expression of Sldorsal was up-regulated by the challenge of entomogenous fungi Isaria javanica and Beauveria bassiana, however, the expression of Slrelish could only be induced by Escherichia coli challenge. Injection of Sldorsal or Slrelish dsRNA down-regulated the expression level of the corresponding gene and broke specific signaling transduction pathway of innate immunity in S. litura. Knockdown of Sldorsal by RNAi significantly increased the mortality of S. litura larvae caused by I. javanica and B. bassiana, and Slrelish RNAi highly increased the mortality caused by E. coli. Sldorsal and Slrelish are essential genes for innate immunity, their knockdown by RNAi increases S. litura susceptibility to insect pathogens, therefore enhances the mortalities caused by I. javanica, B. bassiana and E. coli respectively.     

Intraspecific mitochondrial genome comparison identified CYTB as a high-resolution population marker in a new pest Athetis lepigone

A new outbreak pest, Athetis lepigone (Möschler) (Lepidoptera: Noctuidae), has caused severe economic loss in maize crops in China. In order to conduct population genetics study with a more polymorphic and scientific mitochondrial marker, we sequenced the complete mitochondrial genomes of 13 different A. lepigone individuals. Intraspecific comparison of all PCGs showed that the NADH dehydrogenase and cytochrome b genes had the highest nucleotide diversity. We also found evidence of episodic positive selection on two amino acids, which are encoded by NADH dehydrogenase genes (ND3 and ND4L), against a background of widespread neutral selection of all other mitochondrial PCGs. The genetic divergence observed in this study indicated that the cytochrome b gene (CYTB) is better than COI at recovering population structure. The preliminary population genetic analysis illustrated strong gene flow among A. lepigone populations in China. Our study provides basic information for further research on population genetics of A. lepigone.     

Heat exposure affected the reproductive performance of pregnant mice: Enhancement of autophagy and alteration of subcellular structure in the corpus luteum

To investigate whether autophagy and subcellular changes are involved in the corpus luteum after heat exposure, a total of 30 early pregnant mice were divided equally into heat stress (HS) and non-HS (NHS) groups (n = 15). Mice in the HS group were exposed to 40.5 ± 0.2 ℃ for 7 consecutive days. Ovaries were collected for immunohistochemistry (IHC), western blot (WB) analysis and transmission electron microscopy (TEM). Serum was collected to determine progesterone by RIA and uteri were collected to count the implantation sites. Results showed that heat exposure increased rectal temperature, decreased body weight and number of implantation sites. WB analysis revealed that ovarian expression of LC3B and Atg7 was up-regulated, while p62 was down-regulated in the HS group. IHC results demonstrated that ovarian staining intensity of LC3B was more intense in the HS group than that of the NHS group. LC3B was mainly localized in the granulosa cells, oocytes and luteal steroidogenic cells of the HS group. TEM results revealed double-layered separated membranes indicative of autophagosomes in the luteal steroidogenic cells of the HS group. Moreover, TEM showed that the mitochondrial cristae became dearth, structure-less, swollen after HS. Additionally, the nucleus expanded and accumulation of lipid droplets increased after HS. Results also showed that heat exposure decreased serum progesterone level and ovarian P450scc expression. These results indicate that HS enhanced autophagy and altered the subcellular structure of luteal steroidogenic cells, which may contribute to interfering with the maintenance of luteal function in early pregnant mice.     

Diverse toxic effectors are harbored by vgrG islands for interbacterial antagonism in type VI secretion system

The type VI secretion system (T6SS) is considered as one of the key competition strategies by injecting toxic effectors for intestinal pathogens to acquire optimal colonization in host gut, a microenviroment with high-density polymicrobial community where bacteria compete for niches and resources. Enterotoxigenic Escherichia coli (ETEC), a major cause of infectious diarrhea in human and animals, widely encode T6SS clusters in their genomes. In this report, we first identified VT1, a novel amidase effector in ETEC, significantly hydrolyzed D-lactyl-L-Ala crosslinks between N-acetylmuramoyl and L-Ala in peptidoglycan. Further study showed that the VT1/VTI1 effector/immunity pair is encoded within a typical vgrG island, and plays a critical role for the successful establishment of ETEC in host gut. Numerous putative effectors with diverse toxin domains were found by retrieving vgrG islands in pathogenic E. coli, and designated as VT modules. Therein, VT5, a lysozyme-like effector widely encoded in ETEC, was confirmed to effectively kill adjacent cells, suggesting that VT toxin modules may be critical for pathogenic E. coli to seize a significantly competitive advantage for optimal intestinal colonization. To expand our analyses for large-scale search of VT antibacterial effectors based on vgrG island, >200 predicted effectors from 20 bacterial species were found and classified into 11 predicted toxins. This work reports a new retrieval strategy for screening T6SS effectors, and provides an example how pathogenic bacteria antagonize and displace commensal microbiome to successfully colonize in the host niches through a T6SS-dependent manner.     

Design,synthesis and biological evaluation of novel desloratadine derivatives with anti-inflammatory and H1 antagonize activities

To improve the anti-inflammatory activity of desloratadine, we designed and synthesized a series of novel desloratadine derivatives. All compounds were evaluated for their anti-inflammatory and H₁ antagonistic activities. Among them, compound 2c showed the strongest H₁ antagonistic and anti-inflammatory activity. It also exhibited promising pharmacokinetic profiles and low toxicity. All these results suggest that compound 2c as a novel anti-allergic agent is worthy of further investigation.     

Correlation between population size of pear Psylla (Cacopsylla chinensis) and leaf structure features in different pear cultivars

The feeding preference of pear psylla (Cacopsylla chinensis) varies among pear cultivars. To clarify whether leaf structural factors are related to C. chinensis resistance, eleven pear cultivars of three pear species and their hybrids in China were studied. The population size of C. chinensis in plots of different cultivars was investigated, and leaf paraffin sections of each pear cultivar were observed. Correlation analysis results showed that the thickness of palisade tissue (PT) and the thickness ratio (TR) of PT to leaf (L) were the main factors influencing resistance to C. chinensis. The leaves of cultivar Wujiuxiang and those species from France and the USA exhibited the thickest PT and the highest TR; these three cultivars demonstrated much higher resistance to C. chinensis than did the other cultivars. The cultivar Gold Nijisseiki, with a large C. chinensis population size, showed low PT and TR. The findings of this study may provide valuable information to pear growers, experts and scientists regarding plant protection, cultivation and breeding when selecting suitable varieties to culture from the perspective of pest control.     

Genomic evidence for the population genetic differentiation of Misgurnus anguillicaudatus in the Yangtze River basin of China

Misgurnus anguillicaudatus, an important aquatic species, is mainly distributed in the Yangtze River basin. To reveal the population genetic structure of M. anguillicaudatus distributed in the Yangtze River basin, genotyping by sequencing (GBS) technique was employed to detect the genome wide genetic variations of M. anguillicaudatus. A total of 30.03 Gb raw data were yielded from 70 samples collected from 15 geographic sites located in the Yangtze River basin. Subsequently, 2092 high quality SNPs were genotyped across these samples and used for a series of genetic analysis. The results of genetic analysis showed that high levels of genetic diversity were observed and the populations from upper reaches (UR) were significantly differentiated from the middle and lower reaches (MLR) of Yangtze River basin. Meanwhile, no significant isolation by distance was detected among the populations. Ecological factors (e.g. complicated topography and climatic environment) and anthropogenic factors (e.g. aquaculture and agriculture cultivation) might account for the genetic disconnectivity between UR and MLR populations. This study provided valuable genetic data for the future breeding program and also for the conversation and scientific utilization of those abundant genetic resources stored in the Yangtze River basin.     

Residual contact vial method for the rapid on-site detection of insecticide resistance in Thrips palmi

A residual contact vial plus water (RCVpW) bioassay method, in which water was supplemented to minimize control mortality, was established to monitor insecticide resistance in field populations of the melon thrips, Thrips palmi. In the RCVpW, median lethal doses (LD₅₀) of six insecticides commonly used in T. palmi control, were determined at 8 h post-treatment, using a susceptible RDA strain according to the RCVpW protocol. Diagnostic doses for on-site resistance monitoring of the six insecticides, which were determined as doses two-fold higher than required to achieve LD₉₀ in the RDA strain, were in the range of 0.299 to 164.3 μg^?1 cm². Insecticide resistance levels in five field populations of T. palmi were evaluated to test the applicability of RCVpW in monitoring the pest. Although the RDA strain exhibited 100% mortality to diagnostic doses, field populations showed a reduced mortality in response to all test insecticides, indicating different degrees of resistance. In particular, all test field populations exhibited a significantly low mortality in response to spinosad, suggesting a wide distribution of spinosad resistance. Synergistic bioassay revealed that cytochrome P450-mediated metabolic factor is involved in spinosad resistance in the Korean population. Interestingly, an apparently reduced mortality to emamectin benzoate and chlofenapyr was observed in some field populations, perhaps suggesting uneven distribution of resistance to these insecticides in field populations. Our study showed that the RCVpW protocol can be employed both as an on-site resistance monitoring method for major thrip species, and in the selection of appropriate insecticides for their control.     

Therapeutic potential of the methanolic extract of Lepidium sativum seeds on mice infected with Trypanosoma evansi

The present study aimed to investigate the therapeutic potential of the methanolic extract of Lepidium sativum seeds in mice experimentally infected with Trypanosoma evansi. A total of thirty-two male Swiss albino mice were randomly divided into four groups: the first group was the normal control, while the second, third and fourth groups were infected intraperitoneally with 1 × 10⁴ trypanosomes. The third and fourth groups were treated with 100 μl of Lepidium sativum seed extract (LSSE) at a dose of 200 mg/kg body weight intraperitoneally (infected + LSSEI) and orally (infected + LSSEO) respectively, once a day, for a period of four days.Parasitaemia was found to be significantly raised in the untreated infected group, reaching 2 × 10⁷ at day 4 post-infection, but was significantly reduced by 65.5% and 88% in the mice treated orally and intraperitoneally with LSSE, respectively. The erythrocyte count, HCT, haemoglobin content, leucocyte count and the percentage of lymphocytes was significantly reduced in the untreated infected group, while the treatment with LSSE returned these parameters to their pre-infection values. In addition, our study proved that LSSE provided protection against liver tissue damage and decreased the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The present study also established that intraperitoneal injection of LSSE is more effective than oral administration in the treatment of trypanosome infection in mice. In conclusion, the infection caused haematological, biochemical and histological changes that were ameliorated following treatment with LSSE.     

Prevalence of Escherichia coli strains resistance to antibiotics in wound infections and raw milk

Antibiotic-resistant Escherichia coli strains including extended-spectrum β-lactamase (ESBL) isolates are globally widespread in medical, food, and environmental sources. Some of these strains are considered the most pathogenic bacteria in humans. The present work examined the predominance of antibiotic resistance in E. coli strains in wound infections comparing with E. coli strains isolated from a raw milk as a potential source of those strains. The wound infections included abdomen, anus, arm, back, buttock, chest, foot, hand, head, leg, lung, mouth, neck, penis, thigh, toe, and vagina infections. In total, 161 and 153 isolates identified as E. coli were obtained from wound infections and raw milk, respectively. A Vitek 2 system innovated by bioMérieux, France was applied to perform the identification and susceptibility tests. The E. coli isolates that have ability to produce ESBL were detected by an ESBL panel and NO45 card (bioMérieux). Over half of the E. coli were from abdomen, back, and buttock wound infections. More than 50%of the E. coli isolates obtained from wound infections were resistant to cefazolin, ampicillin, cefuroxime, ciprofloxacin, mezlocillin, moxifloxacin, piperacillin, and tetracycline; 70% of the isolates from wound infections and 0% of the isolates from raw milk were E. coli isolates produced ESBL. The data showed that the strains resistance to multi-antibiotic and produced ESBL are more widespread among wound infections than in raw milk.     

Common variants of ROCKs and the risk of hypertension,and stroke: Two case-control studies and a follow-up study in Chinese Han population

The Rho kinases (ROCKs) are recognized as a critical regulator of vascular functions in cardiovascular disorders. It is crucial to illustrate the association of ROCKs genetic variation and hypertension and/or stroke events. Herein we aimed at investigating the association of ROCK1 and ROCK2 with hypertension and stroke in Chinese Han population. Seven tagSNPs at ROCK1 and ROCK2 were genotyped in a community-based case-control study consisting of 2012 hypertension cases and 2210 normotensive controls and 4128 subjects were further followed up. In stroke case-control study, 1471 ischemic stroke (IS) inpatients and 607 hemorrhagic stroke (HS) inpatients were collected, and 2443 age-matched controls were selected from the follow-up population. Risks were estimated as odds ratio (OR) and hazard ratio (HR) by logistic and Cox regression. The community-based case-control study didn't identify any significant tagSNPs associated with hypertension even after adjustment for covariates. The follow-up analysis showed that rs1481280 of ROCK1 significantly associated with incident hypertension (HR = 1.130, P = 0.048) after adjusting for covariates. rs7589629 and rs978906 of ROCK2 were significantly associated with incident IS (HR = 1.373, P = 0.004; HR = 1.284, P = 0.026) respectively. In stroke case-control study, rs288980, rs1481280 and rs7237677 were significantly associated with IS and the adjusted ORs (P values) of additive model were 0.879 (0.010), 0.895 (0.036) and 0.857 (0.002) respectively. Furthermore, rs288980, rs7237677 and rs978906 were significantly associated with HS and the adjusted ORs (P values) of additive model were 0.857 (0.025), 0.848 (0.018) and 0.856 (0.027) respectively. Our findings suggest that ROCK1 and ROCK2 contribute to the genetic susceptibility of hypertension and stroke.     

Biophysical properties and finger print of Boswellia Sp. Burseraceae

For the first time a finger print analysis via high-performance thin layer chromatography (HPTLC) of Boswellia Sp. Burseraceae was accomplished. A preliminary investigation of the Boswellia Sp. Burseraceae displayed the presence of chemical constituents that could be involved in the production of innovative pharmaceuticals for an array of antiviral, anticancer, and antibacterial uses. Moreover, the finger print analysis would deem useful for establishing HPTLC standardization for natural and herbal photochemical constituents.     

Advances in the understanding of same-sex and opposite-sex sexual harassment

Sexual harassment has traditionally been studied as men's harassment of women. This has led to a lack of knowledge about same sex harassment, and women harassing peers. This has also downplayed the inherent sexual nature of sexual harassment acts. While keeping in mind that sexual harassment is undesirable and causes distress, one needs to consider that many acts that are perceived as unwanted may not primarily be motivated by a wish to derogate but rather by an interest in soliciting short-term sex. In the current study we examined both perpetrators as well as victims of harassment, and specified both sex of perpetrator and target (a total of eight sex constellations). We reproduced the previously found association between unrestricted sociosexuality and sexual harassment in a representative sample of 1326 high school students (57% women). In all regression models sociosexuality outcompeted traditional measures such as porn exposure, rape stereotypes and hostile sexism. Based on the original work we divided the harassment acts into two groups of tactics: sexual solicitation and competitor derogation. Men were particularly subject to derogatory tactics from other men, while women were particularly subject to solicitation from opposite sex peers. Sexual harassment may be understood better from a human sexual strategies perspective, including competitor derogation and mate solicitation. As such, sociosexual orientation predicts both same sex derogation and opposite sex solicitation. The current results highlight the importance of considering the sex of both perpetrator and target. This advanced understanding of the inherently sexual nature of sexual harassment needs to inform future prevention studies. Unrestricted sociosexuality predicts sexual harassment in all constellations better than traditional social science models.     

Extracellular vesicles derived from Echinococcus granulosus hydatid cyst fluid from patients: isolation,characterization and evaluation of immunomodulatory functions on T cells

Cystic echinococcosis is a chronic and complex zoonotic disease. The mechanisms underlying the parasite’s establishment, growth and persistence are not completely understood, and are thought be modulated by a crosstalk through extracellular vesicles. Here, EVs were isolated from the hydatid cyst fluid of patients with cystic echinococcosis and protoscolex culture supernatant. Proteomic analysis of these EVs revealed several parasite- and human-derived proteins. Very few studies have performed proteomic analysis of EVs isolated from HCF and PCS. Our proteomic analysis of the EVs derived from HCF and PCS facilitated identification of 1175 proteins, wherein 1026 and 38 proteins were exclusively identified in the EVs derived from HCF (HCF-EVs) and PCS (PCS-EVs), respectively, and 111 proteins were shared in both. The results of co-culture of PCS-EVs with murine peripheral blood mononuclear cells showed that PCS-EVs significantly regulated T lymphocyte functions in a dose-dependent manner. Collectively, our results provide valuable information on parasite survival strategies and new insights into the role of these EVs in the establishment and persistence of hydatid cysts.     

Olfactory responses of adult Potato Tuber Moth,Phthorimaea operculella (Zeller) measured by attraction relative to the tomato leaf volatiles

The potato tuber moth (PTM) is an oligophagous herbivore and a severe pest of solanaceous crops in many countries of the world. Previously, we reported host expansion and damage potential of PTM on tomato, a congeneric crop of potato. Here we tested adult olfactory behaviour of PTM to leaves of five different cultivated tomato varieties including Moneymaker, Campari, Ailsa craig, LA3475 and E6203, and one wild species, Solanum pimpinellifolium. Tomato leaf hydro-distilled oils of Moneymaker, Campari, Ailsa craig, S. pimpinellifolium and E6203 showed strong attractiveness and LA3475 exhibited repulsiveness for adult PTM of both sexes in two-armed bioassays. Volatiles of Moneymaker, Campari, Ailsa Craig, S. pimpinellifolium and E6203 showed attractiveness for mated adult PTM of both sexes (> 70%). The extracted oil of all tomato leaves contains monoterpenes, sesquiterpenes, diterpenes and C₆ alcohols as well as other components. Phytol is one of the common and major compounds in all tomato varieties. Phytol showed weak attractiveness: 60.8–63.6% and 57.6–60.6% for male and female PTM, respectively when tested at concentrations of 0.1–10 mg/mL. Therefore, the attractiveness of tomato leaf volatiles might be due to synergistic effects of plant volatile mixtures. Identification PTM attractant volatile in tomato leaf is important in its control in environmentally friendly manner.     

Expression of Leptospira membrane proteins Signal Peptidase (SP) and Leptospira Endostatin like A (Len A) in BL-21(DE3) is toxic to the host cells

Heterologous expression of Integral Membrane Proteins (IMPs) is reported to be toxic to the host system in many studies. Even though there are reports on various concerns like transformation efficiency, growth properties, protein toxicity, inefficient expression and protein degradation in IMP overexpression, no studies so far addressed these issues in a comprehensive way. In the present study, two transmembrane proteins of the pathogen Leptospira interrogans, namely Signal peptidase (SP), and Leptospira Endostatin like A (Len-A) were taken along with a cytosolic protein Hydrolase (HYD) to assess the differences in transformation efficiency, protein toxicity, and protein stability when over expressed in Escherichia coli (E. coli). Bioinformatics analysis to predict the transmembrane localization indicated that both SP and Len are targeted to the membrane. The three proteins were expressed in full length in the E. coli expression strain, BL 21 (DE3). Significant changes were observed for the strains transformed with IMP genes under the parameters analysed such as, the transformation efficiency, survival of colonies on IPTG-plate, culture growth kinetics and protein expression compared to the strain harbouring the cytosolic protein gene.     

Reproductive biology and morphology of Apis mellifera jemenitica (Apidae) queens and drones

The current study aimed to investigate the important reproductive biology and morphology of A.m. jemenitica queens and drones through measuring the weight of virgin and mated queens, size and weight of spermathecae, weight of ovaries, number of ovarioles, quantity and viability of semen in queen and drones. Accordingly, the average weights of 0.139 ± 0.01 g and 0.143 ± 0.013 g recorded for virgin and mated queens respectively. The sizes of spermathecae were 1.248 ± 0.103 mm and 1.25 ± 0.022 mm for virgin and mated queens respectively. The mean weight of ovaries was 0.013 ± 0.003 g and the numbers of ovarioles varied from 124 to 163 with the mean of 142.9 ± 9.47 and with no significant difference between virgin and mated queens. The average number of stored sperm per spermathecae of mated queen was estimated to be 4.202 ± 0.613 million with the viability of 80.39%. The average number of sperm per drone recorded was 8,763,950 ± 1,633,203.15 with viability of 79.54 ± 6.70%. In general, the current study revealed that the values recorded for reproductive biology and morphological characters of A. m. jemenitica queens and drones were relatively lower than values recorded for other Apis mellifera races. This mainly could be associated with the body size of the race which is known to be the smallest race among A. mellifera races. Moreover, the harsh environmental conditions of the regions, high temperature, low humidity and limited resources may have contributed for the smaller biological and morphological values. The information will serve as a base in future selection and breeding of program of the race.     

Hybrid mesoporous silicates: A distinct aspect to synthesis and application for decontamination of phenols

Water pollution due to organic compounds is of great concern and efforts are being made to develop efficient adsorbents for remediation of toxic pollutants. The development of new functionalized materials with increased performance is growing to meet the regulatory standards in response to public concerns for environment. In this study, an attempt has been made to investigate the influence of synthesis parameters like the reaction temperature, the surfactant-to-silica ratio and reaction time on the structural and textural properties of novel ordered mesoporous silica hybrids. In order to understand the effect of different synthesis parameters, all the prepared materials were systematically characterized by various analytical, spectroscopic and imaging techniques such as XRD, BET, TG etc. It was deduced from these studies that the synthesis temperature influence greatly the structural order whereas both the P104/Na₂SiO₃ molar ratio and reaction time found to influence textural properties significantly. However, under optimized experimental condition, we could achieve the functionalized silica hybrids that offers successful incorporation of -Amino, -Glucidoxy, -Methacrylate, -Vinyl and -Phenyl moieties indicated by FTIR peaks at 793 cm^?1, 2870 cm^?1, 796 cm^?1, 1630 cm^?1 and 954 cm^?1. XRD studies reveal orthorhombic and tetragonal symmetry for the hybrids and these materials were found to be thermally stable due to incorporation of organic moiety in silica matrix. Functionalized silica hybrids then applied as adsorbents demonstrated efficient and comparable removal of 4-aminophenol and p-nitrophenol in 20 min facilitated through organic moiety. Detailed modeling of the sorption using equilibrium and kinetic isotherms has been carried out to get an insight into the transport process. The adsorption isotherms of phenol derivatives are well-fitted with the Langmuir, Freundlich and Temkin Isotherms and the adsorption kinetics follows the pseudo second order model. The modeling confirms that the uptake is a chemisorption process.     

Susceptibility of mice strains to oxidative stress and neurotransmitter activity induced by Plasmodium berghei

This study investigated the susceptibility of female C57Bl/6 and Swiss Albino mice to oxidative stress and neurotransmitters activity induced by Plasmodium berghei. On day 9 p.i. with P. berghei infected erythrocytes, the mice reduced in weight. This weight loss was markedly higher in SW mice and reached about ?14%. Also, the infection was able to cause oxidative damage to the brain tissue. Catalase activity as well as glutathione, malondialdehyde and nitric oxide levels were different in the two mice strains. Moreover, the brain content of neurotransmitters, epinephrine, norepinephrine, dopamine and serotonin in mice brain was higher in SW mice than B6 mice. We concluded that, the strain of mice is one factor that could alter the response of mice to P. berghei infection.     

A comparative study of bacterial diversity based on culturable and culture-independent techniques in the rhizosphere of maize (Zea mays L.)

ObjectiveMaize is an important crop for fodder, food and feed industry. The present study explores the plant-microbe interactions as alternative eco-friendly sustainable strategies to enhance the crop yield.MethodologyBacterial diversity was studied in the rhizosphere of maize by culture-dependent and culture-independent techniques by soil sampling, extraction of DNA, amplification of gene of interest, cloning of desired fragment and library construction.ResultsCulturable bacteria were identified as Achromobacter, Agrobacterium, Azospirillum, Bacillus, Brevibacillus, Bosea, Enterobacter, Microbacterium, Pseudomonas, Rhodococcus, Stenotrophomonas and Xanthomonas genera. For culture-independent approach, clone library of 16S ribosomal RNA gene was assembled and 100 randomly selected clones were sequenced. Majority of the sequences were related to Firmicutes (17%), Acidobacteria (16%), Actinobacteria (17%), Alpha-Proteobacteria (7%), Delta-proteobacteria (4.2%) and Gemmatimonadetes (4.2%) However, some of the sequences (30%) were novel that showed no homologies to phyla of cultured bacteria in the database. Diversity of diazotrophic bacteria in the rhizosphere investigated by analysis of PCR-amplified nifH gene sequence that revealed abundance of sequences belonging to genera Azoarcus (25%), Aeromonas (10%), Pseudomonas (10%). The diazotrophic genera Azotobacter, Agrobacterium and Zoogloea related nifH sequences were also detected but no sequence related to Azospirillum was found showing biasness of the growth medium rather than relative abundance of diazotrophs in the rhizosphere.ConclusionThe study provides a foundation for future research on focussed isolation of the Azoarcus and other diazotrophs found in higher abundance in the rhizosphere.