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Paper Information

Title: 

MOLECULAR METHODS APPLIED FOR IDENTIFICATION THE MICROBIAL DIVERSITY IN A HYPERSALINE ENVIRONMENT

Type: PAPER
Author(s): MAKHDOUMI KAKHKI ALI*,AMOOZEGAR MOHAMMAD ALI
 
 *DEPARTMENT OF BIOLOGY, SCHOOL OF SCIENCES, FERDOWSI UNIVERSITY OF MASHHAD, 91775-1436 MASHHAD, IRAN
 
Name of Seminar: CONGRESS OF IRANIAN GENETICS
Type of Seminar:  CONGRESS
Sponsor:  ZAKHAYERE ZHENETICE JAHADE DANESHGAHI
Date:  2014Volume 1
 
 
Abstract: 

MICROBIAL COMMUNITY REPRESENTS MORE THAN HALF THE BIOMASS ON THE EARTH. A GRAM OF SOIL, FOR EXAMPLE, CONTAINS MILLIONS OF MICROORGANISMS OF DIFFERENCE GROUPS (4000 DIFFERENT ONES). HOWEVER MOST OF THE ENVIRONMENTAL MICROORGANISMS ARE UN-CULTURABLE (90-99%) AND COULD NOT BE ISOLATED IN LABORATORY CONDITIONS. THERE ARE HUGE BIOTECHNOLOGICAL CAPABILITIES IN THESE GROUPS OF MICROORGANISMS SINCE IT IS NECESSARY TO CONSIDER THIS KIND OF MICROORGANISMS IN BIODIVERSITY STUDIES. PROKARYOTIC DIVERSITY IN ARAN-BIDGOL SALT LAKE, A THALASOHALINE LAKE IN IRAN, WAS STUDIED BY FLUORESCENCE IN SITU HYBRIDIZATION (FISH), DENATURING GRADIENT GEL ELECTROPHORESIS (DGGE) OF PCR-AMPLIFIED FRAGMENTS OF 16S RRNA GENES AND 16S RRNA GENE CLONE LIBRARY ANALYSIS. TOTAL CELL ABUNDANCES IN THE LAKE DETERMINED BY DAPI DIRECT COUNT WAS 3-4×107 CELLS/ML. THE PROPORTION OF BACTERIA TO ARCHAEA IN THE COMMUNITY DETECTABLE BY FISH, APPLYING ARCHAEAL AND BACTERIAL SPECIFIC PROBES WAS UNEXPECTEDLY HIGH AND RANGED BETWEEN 1:3 AND 1:2. GENOMIC DNA WAS DIRECTLY EXTRACTED FROM ENVIRONMENTS AND 16S RRNA OF BOTH DOMAINS WAS PCR-AMPLIFIED. THE PCR PRODUCTS OF EXPECTED SIZE (1500 BP) WERE GEL PURIFIED (DNA EXTRACTION KIT, ROCHE, GERMANY) LIGATED INTO PGEM-T CLONING VECTOR (PROMEGA, USA) AND USED TO TRANSFORM E.COLI DH5Α CELLS. WE CONSTRUCTED TOTAL OF EIGHT CLONE LIBRARIES. ANALYSIS OF INSERTS OF 100 CLONES FROM THESE LIBRARIES CONSTRUCTED REVEALED A TOTAL OF 37 OTUS. A MAJORITY (63 %) OF THESE SEQUENCES WERE NOT RELATED TO ANY PREVIOUSLY IDENTIFIED TAXA. WITHIN THIS SAMPLING EFFORT WE MOST FREQUENTLY RETRIEVED PHYLOTYPES RELATED TO HALORHABDUS (16 % OF ARCHAEAL SEQUENCES OBTAINED) AND SALINIBACTER (36 % OF BACTERIAL SEQUENCES OBTAINED). OTHER PROKARYOTIC GROUPS THAT WERE ABUNDANT INCLUDED REPRESENTATIVES OF HALOQUADRATUM, THE ANAEROBIC GENERA HALANAEROBIUM AND HALOCELLA, PURPLE SULFUR BACTERIA OF THE GENUS HALORHODOSPIRA AND CYANOBACTERIA.

 
Keyword(s): MOLECULAR METHODS, ARCHAEA, BACTERIA, HALOPHILIC MICROORGANISMS, PROKARYOTIC DIVERSITY
 
 
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