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25 2013 1400 002.224.01 .. : 117312, . , - 60- , .7, . 2.

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XXI . , , (, , 2008). . , , , in situ - ( ., 2011). , 0.22 , , , . .

, (Sherman, Safford, 1928; Kleinberger-Nobel, 1951; , , 1958), . 1981 0.3 , (Torella, Morita, 1981). 0.1 3 (Schut et al., 1997; Velimirov, 2001), (Velimirov, 2001; Panikov, 2005; Duda, 2011; ., 2012). , (Huber et al., 2002; ., 2011), (Audic et al., 2007).

0.22 . , , , (Torella, Morita, 1981; MacDonell, Hood, 1982; Hahn, 2004).

2

. , , Bacteroidetes, Alphaproteobacteria, Betaproteobacteria, Actinobacteria Spirochaetes, (Hahn, 2004; Hahn et al., 2009, 2010, 2012).

, , . , . (Haller et al., 1999), (Miyoshi et al., 2004), (Panikov, 2005; ., 2010). (Wang et al., 2007). , 104 , 0.22 , 0.03 3.6% (Wang et al., 2007). . , , . . - -

, .

: 1. ,

0.22 , , .

2. .

3. 16S , .

3

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. , . , 104 1 . , . , . LDS Euryarchaeota. , -. .

, . 250 16S 300 16S ; GenBank 215 . , . .

: 1. -

, , , 2010. 2. 16- -

XXI , , , 2012. 3. :

, , 2012. 4. 4th Congress of European Microbiologists (FEMS-4), Geneva, Switzerland, 2011.

4

5. 14th International Symposium on Microbial Ecology (ISME-14), Copenhagen, Denmark, 2012.

.

8 : 3 5 . .

, , , , , , - . . .

.. 2009 2013 ... .. .

... .. ... .. ( ). - .. ( ) ... .. ( ). .. ( ).

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. 1. , , , - , , (. 1). 1.

, , ,

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-1 ( )

, , 5810N, 3733E, 06.2009

4.4 3.890.40 106 1.690.53 104 (0.4%)

, , 5810N, 3733E, 06.2009

4.7 1.030.10 106 3.160.43 104 (3.1%)

, ., 5814N, 3812E, 05.2010

4.2 8.060.86108* 3.750.42 106* (0.5%)

, , ., 5818N, 3752E 07.2011

7.0 1.440.08 106 2.280.16 104 (1.6%)

* 1 . 2. , (1:1) Millipore 0.22 1 - (4', 6' -2-). Zeiss Axioplan 2 (. ) Zeiss 02,

6

. 100 , 1 . , 0.22 , 0.09 . 3. . -, , JEM-1400 (JEOL, ) 80 . 4. : 1) R2A (Difco), 100 ; 2) SB2, ( -1): KNO3 50, NH4Cl 50, MgSO4 7H2O 60, aCl2 2H2O 20, 50, Na 300; Difco - 18 /. 2 . ( -1): 1) MgSO4 7H2O 20, NH4NO3 30, 50, Na 100, Na 100, Na 100; 2) MgSO4 7H2O 20, NH4NO3 30, 10, 10, 30; 3) MgSO4 7H2O 20, NH4NO3 30, 10, 10. 5. , (SDS) (Dedysh et al., 1998), - 16S 9f 1492r (Weisburg et al., 1991), . 6. 0.22 Millipore 0.09 . , BagMixer 100 MiniMix (Interscience, ). Millipore 5 , , .

7

FastDNA SPIN kit for soil. () 16S ( 9f 1492r, Weisburg et al., 1991) ( 109f 915r, Grosskopf et al, 1998) PE GeneAmp PCR System 9700 (Perkin Elmer Applied Biosystems, ). pGem-T Easy Vector SystemII (Promega). - . . WizardR Plus Minipreps DNA Purification System (Promega). ABI 377A (Perkin Elmer Applied Biosystems, ). SeqMan (Laser Gene 7.0; DNA Star Package). GenBank BLAST (http://www.ncbi.nlm.nih.gov/BLAST). 16S Bellerophon 3.0 (http://comp_bio.anu.edu.au/bellerophon/bellerophon.pl). ARB (http://www.arb_home.de). Phylip bootstrap- 1000 . 16S GenBank JN825208-JN825301, JQ697092-JQ697150 KC878619- KC878680. 8. MOTHUR (Schloss et al., 2009), SINA Incremental Aligner (Pruesse et al., 2012) SILVA (Quast et al., 2013). 97% () Average neighbor. , , , MOTHUR.

8

1. . 3.890.40 1.030.10106 1, (. 1). , 0.22 1.690.53 3.160.43 104 1, 0.4 3.1% .

1 8.060.86108 . , 0.22 3.750.42 106 -1, 0.5% . ( ., 2010) 1 .

1.440.08106 -1, (, 1971; , 1972). 2.280.16104 -1 1.6% .

, 104 1 106 1 , . 2. - . - (120-250 ), (. 1). , (. 1, 1). , , 0.014 0.06 3 0.14 0.30 (Koch, 1997; Maniloff, 1997; Psenner, Loferer, 1997). , . , , (0.2 ), ( , 1972).

9

. 1. , (, ); , 200 . - (1); , 100 . 3. . , . , R2A (1:100) SB2 0.93102 2.00102 -1, . , 0.5-1.2% .

10 . 16S (97-99%) Mesorhizobium, Bradyrhizobium, Pseudomonas, Sphingomonas Ralstonia ( 2). , 0.3-0.5 (. 2). , , ( 1-2 ), . , (. 4.1).

10

2. , .

16S

, %DSB20 Mesorhysobium amorphae (FG025126) 99 DSB21 Bradyrhizobium elkanii (FG025139) 99 DSB22 Sphingomonas sp. (AJ926494) 98 DSB4 Pseudomonas veronii (DQ525597) 99 DSB11 Pseudomonas veronii (DQ525597) 99 IS252 Sphingomonas rhizogenes (AY962684) 99 IS251 Sphingomonas melonis (AM777422) 98 IS260 Ralstonia picketii (CP001644) 99

ISlds Parvibaculum lavamentivorans

(CP000774) 98

Pln1 Polynucleobacter nessesarius subsp.

asymbioticus (AB470420) 100

PO9 Ralstonia picketii (CP001644) 99 PO4 Microbacterium oxydans (AJ717356) 100

PO15 Ralstonia detusculanse (AF280433) 100

PO65 Raltsonia sp. (GU936705) 100

PO6 Brachybacterium muris (NR024571) 99

RB1 Brevundimonas sp. (JQ396622)

99

IS P3-6 Marmoricola scoriae (FN386750) 98

Pseudomonas, DSB-4 DSB-11, 16S , (. 4.1). , 7 ( 2). , Pln1 (. 2) Polynucleobacter nessesarius subsp. asymbioticus, (Hahn et al., 2009, 2012).

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. 2. , : A DSB4Microbacterium sp. PO4, - Bradyrhizobium sp. DSB21, Ralstonia sp. PO65, Mesorhizobium sp. DSB20, - Brevundimonas sp. RB1, - Pseudomonas sp., - Parvibaculum sp. IS lds , - Polynucleobacter nessesarius subsp. asymbioticus Pln 1. 2 .

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, , , - 16S , . 4. 4.1. . 16S 86 , 25 , 61 . 74 Betaproteobacteria, 4 - Alphaproteobacteria, 2 - Gammaproteobacteria Actinobacteria (. 3).

. 3. , 16S , , Proteobacteria Actinobacteria. , , DB MB . 0.1 .

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, , Oxalobacteraceae, Comamonadaceae and Burkholderiaceae. 16S (94-99%) Herbaspirillum, Herminiimonas, Curvibacter Burkholderia. DB27 DB28, Gammaproteobacteria, 99% 16S pPHK Pseudomonas, (Vardhan Reddy et al., 2009). 16S DSB-4 DSB-11, . , Alphaproteobacteria, (~90% 16S ) . (95-97%) 16S Ellin362, (Sait et al., 2002). , Actinobacteria, (95%) 16S - , Candidatus Planktophila limnetica. 4.2 . 16S 100 , 65 Proteobacteria 35 Bacteroidetes (. 4). Betaproteobacteria (44 ) Gammaproteobacteria (21 ), Bacteroidetes - Flavobacteria (34 ) Sphingobacteria (1 ).

16S , Betaproteobacteria, (98-99%) Janthinobacterium, J. lividum, , , (Shivaji et al., 2011).

Gammaproteobacteria 21 , (99%) 16S Pseudomonas Ps. psychrophila, Ps. antarctica . Acinetobacter (99% ) , .

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16S Flavobacteria. 10 Chryseobacterium 24 - Epilithonimonas. (99%) 16S Sphingobacterium faecium, .

. 4. , 16S , , Proteobacteria Bacteroidetes. , , POB. 0.1 .

4.3 . 16S 61 , 60 Betaproteobacteria (99.0-99.5%) 16S Polynucleobacter necessarius subsp. asymbioticus (. 5). - 0.1 3, (Hahn, 2003; Hahn et al., 2009; Wang et al., 2009).

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Polynucleobacter necessarius subsp. asymbioticus . ( 2). 0.22 (Hahn, 2004). (2.1 . ) , , (Hahn et al., 2012).

16S , RB150, Alphaproteobacteria (. 5). (~90%) 16S , 47, .

. 5. , 16S , , Alpha- Betaproteobacteria. , , RB. 0.1 .

16

. 6. 16S , (), () (). 1 Alphaproteobacteria, 2 - Betaproteobacteria, 3 Gammaproteobacteria, 4 - Actinobacteria, 5 - Bacteroidetes.

6. , Betaproteobacteria. Bacteroidetes. , . 5. . 5.1 . 16S , , 120 Euryarchaeota, 48 72 . (97-99%) 16S Methanobacteriales Methanosarcinales (. 7). 34 Methanobacterium, , (Metje et al., 2005; Cadillo-Quiroz et al., 2008) (Metje et al., 2007). 3- 4- , , 16S Methanobrevibacter Methanosarcina, .

17

. 7. , 16S , ( ), () (). 0.1 .

18

(71-74% 16S ) Euryarchaeota LDS (Lake Dagow Sediment). LDS 2004 , , (Glissman et al., 2003). , , , , , (Galand et al., 2006, 2008; Pouliot et al., 2009). 5.2 . 16S , , 77 , 73 Euryarchaeota 4 - Thaumarchaeota (. 7).

16S Methanobacteriales, Methanomicrobiales Methanosarcinales. (41 ) 16S , Methanomicrobiales 97-98% Methanoregula boonei (Brauer et al., 2011). 0.2-0.3 . , Methanobacteriales, Methanobacterium subterraneum, (0.10 0.15 ) (Kotelnikova et al., 1998). Methanosarcinales , . 16S Euryarchaeota - LDS (22 ) RC-V (Rice Cluster V) (5 ). LDS, , , .

16S Thaumarchaeota. , -. Candidatus Nitrosopumilus martimus, 0.17 0.22

19

(Knneke et al., 2005). , , ( 99%) , . , Thaumarchaeota . 5.3 . 16S 105 , Euryarchaeota (. 7). , 47 16S Methanobacteriales Methanomicrobiales. Methanobacteriales, Methanobacterium subterrenium, , . 18 Methanomicrobiales 16S Methanoregula boonei (Brauer et al., 2011). . 58 - LDS RC-V. , LDS, , , (. 7). 16S , , . , LDS .

8. LDS. RC-V . Methanobacteriales, Methanomicrobiales Methanosarcinales.

20

. 8. 16S , (), () () . 1 - Methanobacteriales, 2 - Methanomicrobiales, 3 - Methanosarcinales, 4 LDS, 5 RC-V, 6 - Thaumarchaeota.

,

, 0.22 , 104 1 . 0.5-1.2% . , , , Mesorhizobium, Bradyrhizobium, Pseudomonas, Sphingomonas, Ralstonia Microbacterium. , ( ., 2012). , , . , . , - 16S , .

16S - 9.

21

. 9. , () 97% 16S () (), , , .

, , (. 9). (10 ) , (3 ). , Polynucleobacter necessarius subsp. asymbioticus, , . , Polynucleobacter necessarius subsp. asymbioticus . , Pseudomonas. , , . .

22

, , , . , , .

(. 9). 10 17 . , LDS, , . Methanoregula boonei , , , Methanobacterium subterrenium, , .

, , , LDS. , GenBank 400 . , , , (Barberan et al., 2011). LDS . , -. .

23

1. , 0.22 , ( ) . , 104 1 106 1 .

2. ,

. 0.5-1.2% .

3. ,

Betaproteobacteria, .

4.

16S (97-100%) , Polynucleobacter, Pseudomonas, Herbaspirillum, Herminiimonas, Curvibacter, Burkholderia, Janthinobacterium, Chryseobacterium Epilithonimonas. , , .

5. ,

LDS Euryarchaeota. 16S 71-74% , , , - .

24

,

1. .., .., .., .. (2012).

. . . 81, 3. . 306-312.

2. .., .., .. (2012). - . . . 81, 5. . 665-671.

3. .., .., .., .. . ( ).

1. .., .. . VI - . , , 26-27 2012. , . . 119-120.

2. .., .. . XXI : 16- - , 16-21 2012, . . 46.

3. .. , . : . 13 2012, . . 13.

4. Fedotova A.V., Belova S.E., Dedysh S.N. Phylogenetic analysis of microorganisms in two acidic boreal lakes that pass through 0.2-mikrometr-pore-size-filters. 4th FEMS Congress of European Microbiologists. June 26 30, 2011. Geneva, Switzerland.

5. Fedotova A.V., Dedysh S.N. Representatives of the uncultured Euryarchaeota clade LDS prevail among filterable archaea in Northern freshwater habitats. 14th

International Symposium on Microbial Ecology. August 19-24, 2012. Copenhagen, Denmark.