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GMO BIOLOGY ---- BIOTECH ---- GEOTECH BIO-GEO IMPORTANCE OF - - PowerPoint PPT Presentation
GMO BIOLOGY ---- BIOTECH ---- GEOTECH BIO-GEO IMPORTANCE OF - - PowerPoint PPT Presentation
GMO BIOLOGY ---- BIOTECH ---- GEOTECH BIO-GEO IMPORTANCE OF INTERDISCIPLINARY RESEARCH DNA 1 Biogeotechnology Fundamental Applicative 2 For any application such as Bio-fertilizer, Bioremediation. Sample collection Biological
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Sample collection
Physico-chemical parameters Biological (Isolation of bacteria)
Taxonomic identification and phylogenetic analysis Screening for specific properties Elucidation of microbe interaction Bioprospecting
For any application such as Bio-fertilizer, Bioremediation….
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Growth of bacteria
Water Food/Nutrients Oxygen Temperature pH
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Serial dilution and viable plate count method
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Optimization of growth conditions C, P, N…. Temperature pH
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Sample collection
Physico-chemical parameters Biological (Isolation of bacteria)
Taxonomic identification and phylogenetic analysis Screening for specific properties Elucidation of microbe interaction Bioprospecting
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Screen for specific propertiesc
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% reduction in cell number on 6h incubation with 120 ppm uranium Phosphate solubilizing bacterial isolates
4.2 43.3 31.7 33.2
Figure : Percent reduction in cell number on incubation in 0.1 N NaCl, pH 4 solution amended with 120 ppm uranium for 6 h. Vertical bars represent the mean ± SD
3.9
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Phylogenetic analysis -- depicts the evolutionary relationships
Phylogenetic tree of seven PSB from sand sample of Someshwara beach based on 16S rRNA gene sequences available from the EMBL database (accession numbers are given in perenthesis) constructed after multiple alignments of the data using CLUSTAL_X. Distances (according to the Kimura two parameter model) and clustering with the neighbour joining method were performed using MEGA version 5. Numbers at the node are the bootstrap percentages. Bacillus safensis FO-036b(T) (AF234854) YU-SS-SB-23 (KF156653) Paenibacillus cineris LMG 18439(T) (AJ575658) YU-SS-SB-26 (KF156656) Curtobacterium citreum DSM 20528(T) (X77436) YU-SS-SB-27 (KF156657) YU-SS-SB-31 (KF156660) Burkholderia latens R-5630(T) (AM747628) YU-SS-SB-25 (KF156655) Acinetobacter pittii LMG 1035(T) (HQ180184) YU-SS-SB-29 (KC433736) Flavobacterium flevense DSM 1076(T) (AM230486) YU-SS-SB-22 (KF156652) 100 100 100 100 100 100 100 99 92 0.02 γ - Proteobacteria β - Proteobacteria Actinobacteria Firmicutes Bacteroidetes
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Are all bacteria culturable??
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2 APPROACHES FOR ANALYSIS CULTURE DEPENDENT Bacteria isolation on selective media CULTURE INDEPENDENT Direct isolation of nucleic acids
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Strain characterization PCR amplification Counting 16S rDNA sequencing & Identification DGGE analysis Sequencing of target gene Bioinformatics Phylogenetic and diversity analysis
Metagenomics
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Microbial interaction
Lab scale studies
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Fourier Transform Infra Red (FTIR) spectra - - - - elucidates the chemical groups
Peak Group Vibrational mode 545 PO4
3-
Bend; ʋ4 663 PO4
3-
Bend; ʋ4 825 UO2
2+
- Sym. Stretch; ʋ1
921 UO2
2+
Antisym. Stretch; ʋ3 970 PO4
3-
Antisym. Stretch; ʋ3 1025 H3O+ Bend; ʋ2 1122 H2O Librational 1654 H2O Bend; ʋ2 1799 H3O+ Bend; ʋ4 3175 H3O+/OH Stretch
Table: Details of the peak Armstrong and Clark, 2010
Matches with UP --- Chernikovite
FTIR spectra of the precipitates formed on addition of (A) 250 ppm and (B) 2000 ppm uranium to the cell-free culture supernatant of Acinetobacter sp. YU-SS-SB-29
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SEM and EDX images of the precipitates formed on addition of 250 ppm uranium to the cell-free culture supernatant of Acinetobacter sp. YU-SS-SB-29
Scanning Electron Microscopy Images - - - - surface information
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XRD spectra of the precipitates formed on addition of A) 250 ppm uranium and B) 2000 ppm uranium to the cell-free culture supernatant of Acinetobacter sp. YU-SS-SB-29
10 20 30 40 50 2000 4000
(111)
Intensity (a u)
(001) (101) (110) (002) (200) (102) (201) (211) (103) (113) (220) (221) (301) (310)
(311)
(302) (204) (303)
2 theta in degrees
A B
X-ray Diffraction (XRD) spectrum
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Field application
E nvir
- nmental
Geotechnologists There is the classic issue encountered when working with bacteria, in that microbial processes are relatively slow and
- ften
unpredictable, especially when environmental factors are involved
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Molecular biology has shown that even the simplest of all living systems on the earth today, bacterial cells, are exceedingly complex objects. Although the tiniest bacterial cells are incredibly small, weighing less than 10-12 gms, each is in effect a veritable micro-miniaturized factory containing thousands of exquisitely designed pieces of intricate molecular machinery, made up altogether of one hundred thousand million atoms, far more complicated than any machine built by man and absolutely without parallel in the nonliving world. Michael Denton
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Due to complexity, the microbial geotech would require an integration of microbiology, ecology, geochemistry and geotechnical engineering knowledge
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