TY - JOUR

T1 - Laser microsampling of soil microbial community

AU - Gorlenko, M. V.

AU - Chutko, E. A.

AU - Churbanova, E. S.

AU - Minaev, N. V.

AU - Kachesov, K. I.

AU - Lysak, L. V.

AU - Evlashin, S. A.

AU - Cheptsov, V. S.

AU - Rybaltovskiy, A. O.

AU - Yusupov, V. I.

AU - Zhigarkov, V. S.

AU - Davydova, G. A.

AU - Chichkov, B. N.

AU - Bagratashvili, V. N.

N1 - Funding information: This work was supported by the Russian Federal Agency of Scientific Organizations (Agreement No 007-GZ/C3363/26) in part of development of new method and systems for new 3d printer technologies of laser transfer; the Russian Foundation for Basic Research (RFBR) Project No. 16–02-00955 in part of microbiological research and Project No. 18–32-00607 in part research of laser transfer processes. S.A. Evlashin was granted by Scholarship of the President of the Russian Federation SP-1493.2016.4.

PY - 2018/11/28

Y1 - 2018/11/28

N2 - Standard microorganism isolating technology applied for complex multiphase environmental samples such as soil or sediment needs pre-treatment steps to remove living cells from their mixed-phase microniche, by creating a liquid-phase sample. This process removes synergetic relationships, which help to maintain viability of yet-to-be-cultured and hard-to-culture bacteria. In this paper we demonstrate a high throughput Laser Micro-Sampling (LMS) technology for direct isolation of pure microbial cultures and microbial consortia from soil. This technology is based on laser printing of soil microparticles by focusing near-infrared laser pulses on specially prepared samples of a soil/gel mixture spread onto a gold-coated glass plate. Microsamples of soil are printed on glucose-peptone-yeast agar plates, to estimate the LMS process influence on functional and taxonomic microbial diversity, and on «Eco-log» sole carbon sources microplates, to investigate functional diversity by "metabolic fingerprinting". The obtained results are compared with traditionally treated soil samples. It was shown that LMS treatment leads to increasing of cultured biodiversity and modifies the functional diversity. The strain of rare genus Nonomuraea was isolated by LMS from complex natural environment without using media selective for this genus.

AB - Standard microorganism isolating technology applied for complex multiphase environmental samples such as soil or sediment needs pre-treatment steps to remove living cells from their mixed-phase microniche, by creating a liquid-phase sample. This process removes synergetic relationships, which help to maintain viability of yet-to-be-cultured and hard-to-culture bacteria. In this paper we demonstrate a high throughput Laser Micro-Sampling (LMS) technology for direct isolation of pure microbial cultures and microbial consortia from soil. This technology is based on laser printing of soil microparticles by focusing near-infrared laser pulses on specially prepared samples of a soil/gel mixture spread onto a gold-coated glass plate. Microsamples of soil are printed on glucose-peptone-yeast agar plates, to estimate the LMS process influence on functional and taxonomic microbial diversity, and on «Eco-log» sole carbon sources microplates, to investigate functional diversity by "metabolic fingerprinting". The obtained results are compared with traditionally treated soil samples. It was shown that LMS treatment leads to increasing of cultured biodiversity and modifies the functional diversity. The strain of rare genus Nonomuraea was isolated by LMS from complex natural environment without using media selective for this genus.

KW - Biodiversity

KW - Laser cell printing

KW - Metabolic fingerprinting

KW - Microbe isolation

KW - Nonomuraea

KW - Unculturable

UR - http://www.scopus.com/inward/record.url?scp=85057506827&partnerID=8YFLogxK

U2 - 10.1186/s13036-018-0117-4

DO - 10.1186/s13036-018-0117-4

M3 - Article

C2 - 30519282

AN - SCOPUS:85057506827

VL - 12

SP - 27

JO - Journal of biological engineering

JF - Journal of biological engineering

SN - 1754-1611

IS - 1

M1 - 27

ER -