Table 2. Identification of 16S rRNA gene terminal restriction fragments of class Deltaproteobacteria from Baltic Sea sediments.
T-RF size (bp) a with | |||||||||||||
HaeIII | HhaI | MspI | RsaI | ||||||||||
clone | expectedb | observedc | observedd,e | expectedb | observedc | observedd,e | expectedb | observedc | observedd,e | expectedb | observedc | observedd,e | lowest rankf |
JML-43 | 38 | nd | 29 | 92 | 88/90 | 88 | 504 | 503/505/506 | 504 | 487 | 487/488/490 | 488 | Bacteriovorax (g) |
JML-48 | 67 | 62/61 | 61 | 96 | 90/92 | 91 | 165 | 161/162 | 161 | 494 | 495 | 494 | Desulfobacterium (g) |
JML-70 | 71 | 71 | 65 | 83 | 76 | 78 | 200 | 201 | 199 | 441 | 440 | 440 | Deltaproteobacteria (c) |
Paila10-32 | 145 | 141 | 143 | 95 | 88/90/92 | 90 | 127 | 122 | 123 | nd | 490/492 | 491 | Desulfomonile (g) |
JML-67 | 178 | nd | 176 | 92 | 88/90 | 88 | 140 | 140 | 139 | 83 | 78/91 | 79 | Deltaproteobacteria (c) |
JML-37 | 183 | nd | 180 | 96 | 92 | 92 | 134 | 129/130 | 131/133g | 57 | 52 | 50 | Desulfobacteraceae (f) |
JML-2 | 192 | 191/193 | 191/192h | 94 | 88/90/92 | 90 | 163 | 162 | 163 | 213 | 211/213 | 212 | Desulfobacterales (o) |
JML-55 | 200 | 201/203 | 202 | 92 | 90/92 | 91 | 504 | 506/508 | 508 | 487 | 490/492 | 491 | Desulfovibrio (g) |
Paila10-52 | 204 | 203/205 | 203 | 94 | 88/90/92 | 90 | 143 | nd | 143 | nd | 490/492 | 492 | Deltaproteobacteria (c) |
JML-22 | 204 | 205/206 | 205 | 92 | 88/90/92 | 90 | nd | 510/511/512 | 512 | 225 | 226/227 | 226 | Desulfobulbaceae (f) |
JML-64 | 207 | 205/206 | 204 | 231 | 227 | 227 | 138 | 136/137 | 137 | 81 | 77/78 | 77 | Desulfobacterales (o) |
JML-5 | 208 | 205/206 | 205 | 96 | 88/90/92 | 90 | 69 | 62 | 62/64g | 247 | 242/243 | 243 | Desulfobacteraceae (f) |
JML-29 | 208 | 205/206 | 206 | 379 | 378 | 378 | 165 | 165 | 164 | 57 | 52 | 51 | Desulfobacteraceae (f) |
Paila10-17 | 215 | 214/216 | 214 | 92 | 88/90 | 89 | 130 | 126/127/128 | 127 | 57 | 52 | 51 | Desulfuromonadaceae(f) |
Paila10-65 | 217 | 216/217 | 216 | 94 | 90/92 | 91 | 132 | 128/129/130 | 129 | 57 | 52/54 | 52 | Desulfuromonadales(o) |
JML-7 | 239 | 237/238 | 238 | 95 | 90/92 | 91 | 164 | 160/161/162 | 161 | 246 | 245/246 | 245 | Desulfobacteraceae (f) |
GF1-6 | 239 | 237/238 | 238 | 96 | 90/92 | 92 | 164 | 160/161/ | 161 | 246 | 245/246 | 245 | Desulfobacterium (g) |
Paila10-20 | 240 | 235/237 | 236 | 94 | 88/90/92 | 90 | 209 | 206/209 | 208 | nd | 290 | 289 | Deltaproteobacteria (c) |
GF1-20 | 240 | 237/238 | 238 | 96 | 90/92 | 92 | 165 | 165/167 | 164/166g | 247 | 246/245 | 245 | Desulfobacterium (g) |
GF1-41 | 254 | 254/255 | 254 | 92 | 88/90 | 88 | 130 | 126/127/128 | 127 | 463 | 462 | 463 | Desulfuromusa (g) |
Paila10-80 | 270 | 270/271 | 269 | 94 | 88/90/92 | 90 | 163 | 160/161/162 | 161 | 243 | 242/244 | 242 | Desulfobulbaceae (f) |
GF1-21 | 272 | 270/271 | 271 | 96 | 90/92 | 92 | 165 | 162/165 | 164 | 332 | 332 | 332 | Desulfobacula (g) |
GF1-39 | 272 | 270/271 | 271 | 96 | 90/92 | 92 | 165 | 162/165 | 162/163/164g | nd | 468/470/471 | 469/471g | Desulfobacula (g) |
GF1-34 | 423 | 423 | 423 | 96 | 90/92 | 92 | 165 | 165 | 164 | 332 | 332 | 332 | Desulfobacula (g) |
T-RF = terminal restriction fragment, nd = not detected.
Only those T-RFs that were identified with at least three restriction endonucleases are shown.
Expected T-RFs derived from virtual digestion of partial (approximately 400–500 bp) 16S rRNA gene clone sequences.
Observed T-RFs (between 50–700 bp) of 16S rRNA genes derived from terminal restriction fragment length polymorphism analysis of sediment samples.
Observed T-RFs (between 30–700 bp) of 16S rRNA genes derived from terminal restriction fragment length polymorphism analysis of 16S rRNA gene clones.
Shift of 0–2 bp between observed T-RFs from sediment samples and from 16S rRNA gene clones was allowed since repeats of restriction enzyme digestions of one 16S rRNA gene clone resulted 0–2 bp difference in lengths of observed T-RFs.
16S rRNA gene clone sequences used in virtual digestion were assigned to class level and the lowest rank (c = class, o = order, s = suborder, f = family, g = genus) using taxonomic Classifier (version 2.2, RDP training set 6) of Ribosomal Database Project (RDP) with 80% confidence threshold [39].
T-RFs of different size derived from one restriction enzyme digestion of one 16S rRNA gene clone.
T-RFs of different size derived from repeats of restriction enzyme digestions of one 16S rRNA gene clone.