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Paleomicrobiology of Permafrost

#3925


Revival of Non-Culturable Microorganisms from Siberian Permafrost Soil

Tech Area / Field

  • BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
  • BIO-MIB/Microbiology/Biotechnology

Status
8 Project completed

Registration date
17.12.2008

Completion date
10.11.2011

Senior Project Manager
Melnikov V G

Leading Institute
Institute of Biochemistry and Physiology of Microorganisms, Russia, Moscow reg., Puschino

Supporting institutes

  • Institute of Physicochemical and Biological Problems in Soil Science, Russia, Moscow reg., Pushchino

Collaborators

  • Universität Innsbruck / Institut für Mikrobiologie, Austria, Innsbruck

Project summary

Permafrost (PF) is a unique natural ecotope, microbiological study of which is relevant and directed to resolving both fundamental scientific issues and practical ones related to increase in microbial biotechnological potential.

The object of this project is to revive and describe microorganisms of permafrost including bacteria, unculturable under standard conditions but a share of which in microbiota of permafrost hundreds and millions years of age is highly liberal.

Many researchers bring to a focus their study on permafrost (PF) microorganisms but a series of fundamental PF microbiology questions remain poorly clarified. Among these questions the following is noteworthy:

  1. in what form are microbial cells in situ present in PF and what is their ultrastructure?
  2. what are reasons for so low number of spore-forming bacilli against predominance of non-spore-forming bacteria in PF? Although according to canonical representation bacilli are indeed champions among microorganisms as to duration of revival and resistance to impact of unfavorable environmental factors.
  3. in what form are spore-forming microbial cells present in PF – in the form of spores or vegetative cells?
  4. why do microbiologists and molecular biologists provide contradicting conclusions on living cells of microorganisms in PF ~1- 3 million years of age and more ? In compliance with findings of microbiologists (including investigators of this project) a considerable amount of viable cells of microorganisms (~ 0,2-30 x 103 CFU/g of ground) is present in PF. But the results of Willerslev, Cooper, Hansen and other researchers show that no amplifying genes of 16S rDNA are found in the same PF samples and hence, a conclusion has been made on contamination as a reason of CFU observation while inoculating PF sample on nutrient medium. At the same time project authors suppose that negative results of molecular biologists stem first of all from a difficulty of isolation of native DNA from PF and a particular state of microorganisms in situ in PF.

Implementation of the objects and tasks assigned implies realization of wide microbiological, biochemical and cytological research with the use of known state-of–art and original methods developed by project investigators.

Scheduled project study of PF samples with the use of original methods of extraction and concentration of cells by low-temperature fractionation of microorganisms, the use of DNA extraction methods and electron microscopy fill up the gap as to a state of microbial cells in situ in PF samples. The analysis of composition and properties of PF microorganisms may originate concept of ancient microbial communities, their structure and functions. With a description of isolated microorganisms it is possible to study mechanisms that provide their survival under prolonged natural cryoconservation and also to describe previously unknown new species of microorganisms. Microorganisms isolated from PF samples can be used with a view to resolving matters on evolution and biopersity and biotechnology – to obtain novel practically valuable microbial metabolites.

Project realization contributes to cooperation on a number of fundamental scientific matters with subsequent practical use. PF may serve as a model of some Mars ground, the study of this model is of a great importance for space microbiology.

In course of project implementation it is planned to take PF samples in Siberia and unculturable microorganisms which could not be isolated with conventional methods are going to be isolated from them and studied using modern techniques. During a performance of project activity a relationship with collaborator is going to be established via discussion of the results and joint publications.


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