PMID- 33158141 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20201128 IS - 2304-8158 (Print) IS - 2304-8158 (Electronic) IS - 2304-8158 (Linking) VI - 9 IP - 11 DP - 2020 Nov 4 TI - Microbial Ecology of Greek Wheat Sourdoughs, Identified by a Culture-Dependent and a Culture-Independent Approach. LID - 10.3390/foods9111603 [doi] LID - 1603 AB - The aim of the present study was to assess the microecosystem of 13 homemade spontaneously fermented wheat sourdoughs from different regions of Greece, through the combined use of culture-dependent (classical approach; clustering by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) and identification by PCR species-specific for Lactiplantibacillus plantarum, and sequencing of the 16S-rRNA and 26S-rRNA gene, for Lactic Acid Bacteria (LAB) and yeasts, respectively) and independent approaches [DNA- and RNA-based PCR-Denaturing Gradient Gel Electrophoresis (DGGE)]. The pH and Total Titratable Acidity (TTA) values ranged from 3.64-5.05 and from 0.50-1.59% lactic acid, respectively. Yeast and lactic acid bacteria populations ranged within 4.60-6.32 and 6.28-9.20 log CFU/g, respectively. The yeast: LAB ratio varied from 1:23-1:10,000. A total of 207 bacterial and 195 yeast isolates were obtained and a culture-dependent assessment of their taxonomic affiliation revealed dominance of Lb. plantarum in three sourdoughs, Levilactobacillus brevis in four sourdoughs and co-dominance of these species in two sourdoughs. In addition, Companilactobacillusparalimentarius dominated in two sourdoughs and Fructilactobacillussanfranciscensis and Latilactobacillus sakei in one sourdough each. Lactococcus lactis, Lb. curvatus, Leuconostoc citreum, Ln. mesenteroides and Lb. zymae were also recovered from some samples. Regarding the yeast microbiota, it was dominated by Saccharomyces cerevisiae in 11 sourdoughs and Pichia membranifaciens and P. fermentans in one sourdough each. Wickerhamomyces anomalus and Kazachstania humilis were also recovered from one sample. RNA-based PCR-DGGE provided with nearly identical results with DNA-based one; in only one sample the latter provided an additional band. In general, the limitations of this approach, namely co-migration of amplicons from different species to the same electrophoretic position and multiband profile of specific isolates, greatly reduced resolution capacity, which resulted in only partial verification of the microbial ecology detected by culture-dependent approach in the majority of sourdough samples. Our knowledge regarding the microecosystem of spontaneously fermented Greek wheat-based sourdoughs was expanded, through the study of sourdoughs originating from regions of Greece that were not previously assessed. FAU - Syrokou, Maria K AU - Syrokou MK AD - Laboratory of Food Quality Control and Hygiene, Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos St., 11855 Athens, Greece. FAU - Themeli, Christina AU - Themeli C AD - Laboratory of Food Quality Control and Hygiene, Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos St., 11855 Athens, Greece. FAU - Paramithiotis, Spiros AU - Paramithiotis S AUID- ORCID: 0000-0002-5112-974X AD - Laboratory of Food Quality Control and Hygiene, Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos St., 11855 Athens, Greece. FAU - Mataragas, Marios AU - Mataragas M AUID- ORCID: 0000-0003-0782-8954 AD - Department of Dairy Research, Institute of Technology of Agricultural Products, Hellenic Agricultural Organization "DEMETER", 3 Ethnikis Antistaseos St., 45221 Ioannina, Greece. FAU - Bosnea, Loulouda AU - Bosnea L AD - Department of Dairy Research, Institute of Technology of Agricultural Products, Hellenic Agricultural Organization "DEMETER", 3 Ethnikis Antistaseos St., 45221 Ioannina, Greece. FAU - Argyri, Anthoula A AU - Argyri AA AUID- ORCID: 0000-0003-3539-8699 AD - Institute of Technology of Agricultural Products, Hellenic Agricultural Organization "DEMETER", 1 Sof. Venizelou St., 14123 Lycovrissi, Greece. FAU - Chorianopoulos, Nikos G AU - Chorianopoulos NG AUID- ORCID: 0000-0002-7695-8261 AD - Institute of Technology of Agricultural Products, Hellenic Agricultural Organization "DEMETER", 1 Sof. Venizelou St., 14123 Lycovrissi, Greece. FAU - Skandamis, Panagiotis N AU - Skandamis PN AD - Laboratory of Food Quality Control and Hygiene, Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos St., 11855 Athens, Greece. FAU - Drosinos, Eleftherios H AU - Drosinos EH AD - Laboratory of Food Quality Control and Hygiene, Department of Food Science and Human Nutrition, Agricultural University of Athens, 75 Iera Odos St., 11855 Athens, Greece. LA - eng GR - T1EDK-05339/European Union and Greek national funds/ PT - Journal Article DEP - 20201104 PL - Switzerland TA - Foods JT - Foods (Basel, Switzerland) JID - 101670569 PMC - PMC7694216 OTO - NOTNLM OT - Greek wheat sourdough OT - PCR-DGGE OT - RAPD-PCR OT - identification OT - lactic acid bacteria OT - yeasts COIS- The authors declare no conflict of interest. EDAT- 2020/11/08 06:00 MHDA- 2020/11/08 06:01 PMCR- 2020/11/04 CRDT- 2020/11/07 01:03 PHST- 2020/10/07 00:00 [received] PHST- 2020/10/29 00:00 [revised] PHST- 2020/11/02 00:00 [accepted] PHST- 2020/11/07 01:03 [entrez] PHST- 2020/11/08 06:00 [pubmed] PHST- 2020/11/08 06:01 [medline] PHST- 2020/11/04 00:00 [pmc-release] AID - foods9111603 [pii] AID - foods-09-01603 [pii] AID - 10.3390/foods9111603 [doi] PST - epublish SO - Foods. 2020 Nov 4;9(11):1603. doi: 10.3390/foods9111603.