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Exploration of nifH gene through soil metagenomes of the western Indian Himalayas.

ABSTRACT: This group has previously highlighted the prevalence of Csp genes from cold Himalayan environments. However, this study has explored the uncultured diazotrophs from metagenomes of western Indian Himalayas. The metagenomic nifH gene clone library was constructed from the Temperate, Subtropical and Tarai soils of Western Himalaya, India followed by polymerase chain reaction (PCR) amplification. After preliminary screening, selected clones were sequenced. In silico analysis of the clones was done, which documented 83.33 % similarities with unculturable sequence database and more than 70 % similarity with culturable bacterial database. Detailed sequence analysis of 24 nifH clones showed similarity to the corresponding genera of diazotrophs belonging to alpha-, beta-, gamma- and delta-proteobacteria. The prominent diazotrophs were Azotobacter spp., Agrobacterium tumefaciens, Methylococcus capsulatus, Geobacter bemidjiensis, Dechloromonas aromatica, Burkholderia xenovorans, Xanthobacter autotrophicus and Sideroxydans lithotrophicus, respectively. Alignment of these clones with culturable bacterial database suggests that most of the sequences belong to ?-proteobacterium group.

SUBMITTER: Soni R

PROVIDER: S-EPMC4711282 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Exploration of nifH gene through soil metagenomes of the western Indian Himalayas.

Soni Ravindra R Suyal Deep Chandra DC Sai Santosh S Goel Reeta R

3 Biotech 20160111 1

This group has previously highlighted the prevalence of Csp genes from cold Himalayan environments. However, this study has explored the uncultured diazotrophs from metagenomes of western Indian Himalayas. The metagenomic nifH gene clone library was constructed from the Temperate, Subtropical and Tarai soils of Western Himalaya, India followed by polymerase chain reaction (PCR) amplification. After preliminary screening, selected clones were sequenced. In silico analysis of the clones was done, ...[more]

PMID: 28330093

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Project description:Escalation of human-wildlife conflict (HWC) is a barrier to the conservation of ecological corridors across the globe. The existing mechanisms to counter HWC are either economically and socially taxing, or ineffective for long-term management. We assessed HWC in the corridor linking the Rajaji and Corbett Tiger Reserves in Uttarakhand, India, and its drivers, along with the benefits derived by local communities from the forest. We designed an innovative incentive-based mitigation mechanism to encourage coexistence of people and wildlife around the corridor. Costs incurred due to conflict and benefits derived from the forest were assessed using semi-structured questionnaire-based personal interviews (n = 757) with representatives from forest dependent households (hh). Fuelwood (1678.7 ± 131 kg hh-1 year-1), fodder (4772 ± 186 kg hh-1 year-1) and green/dry grass (3359 ± 104 kg hh-1 year-1) contributed 3 ± 1%, 6 ± 0.5% and 9 ± 1%, respectively, to the annual income of dependent households. 69% of the households practising agriculture reported crop damage by wild animals, 19% of the households that owned livestock reported livestock loss, and 1.58% reported attack on humans resulting in injuries. The cost incurred due to crop raiding and livestock depredation was US $ 159.83 ± 1.0 hh-1 year-1 and US $ 229.32 ± 34.0 hh-1 year-1, respectively. Crop loss was positively associated with the number of crops grown per season and cultivation of sugarcane, wheat and pulses, and negatively with distance from forest and cultivation of fodder and finger millet. Livestock depredation was negatively associated with distance from forest and positively with number of livestock owned, primarily calves. The accounting profit from cultivating a hectare of land, in the absence of crop depredation by wild animals, was estimated at US $ 3571.84 ha-1 year-1 and US $ 361.44 ha-1 year-1 for the plains and hills, respectively. This value can be used to calculate the payments to be paid to local communities to encourage them to adopt HWC resistant agricultural and pastoralism practices. The net present value of benefits from participating in the payments to encourage coexistence programme for 5 years, discounted at 12%, was US $ 12,875.7 ha-1 for the plains and US $ 1302.9 ha-1 for the hills.

Project description:Substrate-induced gene expression (SIGEX) is a high-throughput promoter-trap method. It is a function-based metagenomic screening tool that relies on transcriptional activation of a reporter gene green fluorescence protein (gfp) by a metagenomic DNA library upon induction with a substrate. However, its use is limited because of the relatively small size of metagenomic DNA libraries and incompatibility with screening metagenomes from anaerobic environments. In this study, these limitations of SIGEX were addressed by fine-tuning metagenome DNA library construction protocol and by using Evoglow, a green fluorescent protein that forms a chromophore even under anaerobic conditions. Two metagenomic libraries were constructed for subseafloor sediments offshore Shimokita Peninsula (Pacific Ocean) and offshore Joetsu (Japan Sea). The library construction protocol was improved by (a) eliminating short DNA fragments, (b) applying topoisomerase-based high-efficiency ligation, (c) optimizing insert DNA concentration, and (d) column-based DNA enrichment. This led to a successful construction of metagenome DNA libraries of approximately 6 Gbp for both samples. SIGEX screening using five aromatic compounds (benzoate, 3-chlorobenzoate, 3-hydroxybenzoate, phenol, and 2,4-dichlorophenol) under aerobic and anaerobic conditions revealed significant differences in the inducible clone ratios under these conditions. 3-Chlorobenzoate and 2,4-dichlorophenol led to a higher induction ratio than that for the other non-chlorinated aromatic compounds under both aerobic and anaerobic conditions. After the further screening of induced clones, a clone induced by 3-chlorobenzoate only under anaerobic conditions was isolated and characterized. The clone harbors a DNA insert that encodes putative open reading frames of unknown function. Previous aerobic SIGEX attempts succeeded in the isolation of gene fragments from anaerobes. This study demonstrated that some gene fragments require a strict in vivo reducing environment to function and may be potentially missed when screened by aerobic induction. The newly developed anaerobic SIGEX scheme will facilitate functional exploration of metagenomes from the anaerobic biosphere.

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Exploration of nifH gene through soil metagenomes of the western Indian Himalayas.

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Exploration of nifH gene through soil metagenomes of the western Indian Himalayas.

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