Table 1.
Phylogenetic affinities of the ITS coding sequences detected in the Leonardo Da Vinci's drawing samples
| Screened clones per sample % | Selected clone | Length (bp) | Closest identified phylogenetic relatives [EMBL accession numbers] | E‐values | Similarity (%) | Accession Numbers |
|---|---|---|---|---|---|---|
| Sample CI, swabs collected from the recto of the drawing | ||||||
| 59.% | CI‐K10 | [560] | Caloplaca dichroa [EU563428]. | 0.0 | 99 | KP828156 |
| 2.6% | CI‐K11 | [596] | Caloplaca austrocitrina [EU563420]. | 0.0 | 99 | KP828157 |
| 2.6% | CI‐K34 | [551] | Verrucaria nigrescens [JX848570] from freshwater habitats in Europe. | 0.0 | 92 | KP828158 |
| 2.6% | CI‐K20 | [578] | Uncultured Stephanoascus clone F9‐K2 [KC535141] detected in stuffing material from mummies in Palermo, Italy. | 0.0 | 99 | KP828159 |
| Trichomonascus ciferrii strain CBS 5295 [NR_111160]. | 1e‐118 | 85 | ||||
| 2.6% | CI‐K18 | [583] | Uncultured fungus clone F3‐K10 [KC535113] detected in bones from mummies in Palermo, Italy. | 0.0 | 100 | KP828160 |
| Gymnoascus petalosporus [AB361639]. | 8e‐176 | 86 | ||||
| 12.8% | CI‐K2 | [530] | Uncultured Penidiella clone F4‐K26 [KC535131] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828161 |
| Penidiella venezuelensis strain CBS 106.75 [EU019278]. | 0.0 | 94 | ||||
| 2.6% | CI‐K43 | [575] | Uncultured Phialosimplex clones [KC535136, KC535127] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828162 |
| Phialosimplex salinarum strains [KF274685, KF274686, KF274692] isolated from a hypersaline habitat. | 0.0 | 91 | ||||
| 2.6% | CI‐K30 | [554] | Uncultured Acremonium clone AP3‐F4 [KF983520] detected in the Archimedes Palimpsest parchment book. | 0.0 | 99 | KP828163 |
| Acremonium charticola culture‐collection UOA/HCPF<GRC>:14413 [KC253940] common and emerging mould pathogens in Greece. | 0.0 | 98 | ||||
| 2.6% | CI‐K36 | [570] | Uncultured Acremonium clone F2‐K28 [KC535108] detected in clothes from mummies in Palermo, Italy. | 0.0 | 99 | KP828164 |
| Acremonium sp. CBS 115996 [EF042103]. | 0.0 | 96 | ||||
| 2.6% | CI‐K19 | [527] | Uncultured ascomycete [AM901754] from indoor dust. | 6e‐162 | 98 | KP828165 |
| Capnobotryella spp. MA 4902 [AJ972847] isolated from Antalya/Turkey. | 1e‐158 | 97 | ||||
| 2.6% | CI‐K1 | [636] | Uncultured Trebouxia, photobiont from Caloplaca sp. s222 [JX036246]. | 5e‐178 | 93 | KP828166 |
| Trebouxia sp. clade V LM‐2014 isolate L1401 [KJ754252]. | 3e‐176 | 93 | ||||
| 5.1% | CI‐K13 | [707] | Citrus sp. [JN681150, JN661211]. | 0.0 | 99 | KP828167 |
| Sample CII, swabs collected from the verso of the drawing | ||||||
| 5.7% | CII‐K10 | [601] | Caloplaca austrocitrina [EU563417, EU563419, EU563420]. | 0.0 | 99 | KP828168 |
| 2.8% | CII‐K28 | [597] | Caloplaca dichroa [EU563428]. | 0.0 | 99 | KP828169 |
| 2.8% | CII‐K30 | [541] | Caloplaca aurantia [AY233219]. | 0.0 | 99 | KP828170 |
| 8.6% | CII‐K31 | [588] | Caloplaca dichroa [EU563428]. | 0.0 | 97 | KP828171 |
| 5.7% | CII‐K48 | [598] | Caloplaca dichroa [EU563428]. | 0.0 | 97 | KP828172 |
| 8.6% | CII‐K35 | [578] | Uncultured Stephanoascus clone F9‐K27 [KC535144] detected in stuffing material from mummies in Palermo, Italy. | 0.0 | 99 | KP828173 |
| Trichomonascus ciferrii strain CBS 5295 [NR_111160]. | 1e‐118 | 85 | ||||
| 5.7% | CII‐K3 | [578] | Uncultured Stephanoascus clone F9‐K49 [KC535146] detected in stuffing material from mummies in Palermo, Italy. | 0.0 | 99 | KP828174 |
| Trichomonascus ciferrii strain CBS 5295 [NR_111160]. | 7e‐117 | 85 | ||||
| 11.4% | CII‐K39 | [600] | Uncultured fungal clones [KC535116, KC535114] detected in bones from mummies in Palermo, Italy. | 0.0 | 99 | KP828175 |
| Gymnoascus sp. NFCCI 2672 [JQ517292]. | 5e‐158 | 87 | ||||
| 11.4% | CII‐K47 | [533] | Uncultured Penidiella clone F4‐K50 [KC535138] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828176 |
| Penidiella venezuelensis strain CBS 106.75 [EU019278]. | 0.0 | 99 | ||||
| 8.6% | CII‐K5 | [576] | Uncultured Phialosimplex clones [KC535125, KC535140] detected in muscle and skin from mummies in Palermo, Italy. | 0.0 | 99 | KP828177 |
| Phialosimplex chlamydosporus strain UAMH 10961 [GQ169326] associated with infections in dogs. | 0.0 | 91 | ||||
| 8.6% | CII‐K2 | [576] | Uncultured fungus clone F4‐K2 [KC535126] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828178 |
| Leptospora rubella [HE774478]. | 0.0 | 92 | ||||
| 8.6% | CII‐K38 | [570] | Uncultured fungus clone F4‐K28 [KC535132] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828179 |
| Penicillium sp. strain CBS 190.68 [JX841244]. | 0.0 | 89 | ||||
| 5.7% | CII‐K12 | [520] | Uncultured fungus clone CMH052 [KF800143] fungal diversity in Kansas City indoor environments. | 0.0 | 100 | KP828180 |
| 5.7% | CII‐K40 | [527] | Cladosporium sp. strains [NR_121333, NR_121332]. | 0.0 | 99 | KP828181 |
| Sample NyI, nylon membrane | ||||||
| 34.1% | NyI‐K5 | [554] | Uncultured Acremonium clone AP3‐F4 [KF983520] detected in the Archimedes Palimpsest parchment book. | 0.0 | 99 | KP828182 |
| Acremonium charticola culture‐collection UOA/HCPF<GRC>:14413 [KC253940] common and emerging mould pathogens in Greece. | 0.0 | 98 | ||||
| 2.4% | NyI‐K1 | [554] | Uncultured Acremonium clone AP3‐F4 [KF983520] detected in the Archimedes Palimpsest parchment book. | 0.0 | 99 | KP828183 |
| Acremonium charticola culture‐collection UOA/HCPF<GRC>:14413 [KC253940] common and emerging mould pathogens in Greece. | 0.0 | 97 | ||||
| 2.4% | NyI‐K13 | [554] | Uncultured Acremonium clone AP3‐F4 [KF983520] detected in the Archimedes Palimpsest parchment book. | 0.0 | 99 | KP828184 |
| Acremonium charticola culture‐collection UOA/HCPF<GRC>:14413 [KC253940] common and emerging mould pathogens in Greece. | 0.0 | 97 | ||||
| 4.9% | NyI‐K23 | [540] | Uncultured Acremonium clone F2‐K5 [KC535100] detected in clothes from mummies in Palermo, Italy. | 0.0 | 100 | KP828185 |
| Acremonium nepalense isolate Ppf33 [GU586837]. | 0.0 | 94 | ||||
| 2.4% | NyI‐K7 | [570] | Uncultured Acremonium clone F2‐K28 [KC535108] detected in clothes from mummies in Palermo, Italy. | 0.0 | 99 | KP828186 |
| Acremonium sp. CBS 115996 [EF042103]. | 0.0 | 96 | ||||
| 2.4% | NyI‐K18 | [570] | Uncultured Acremonium clone F2‐K28 [KC535108] detected in clothes from mummies in Palermo, Italy. | 0.0 | 99 | KP828187 |
| Acremonium sp. CBS 115996 [EF042103]. | 0.0 | 96 | ||||
| 14.8% | NyI‐K35 | [574] | Uncultured fungus clone CMH376 [KF800467], fungal diversity in Kansas City indoor environments. | 0.0 | 99 | KP828188 |
| 2.4% | NyI‐K17 | [573] | Uncultured Phialosimplex clones [KC535127, KC535120, KC535119] detected in hairs and muscle from mummies in Palermo, Italy. | 0.0 | 99 | KP828189 |
| Phialosimplex chlamydosporus strain UAMH 10961 [GQ169326] associated with infections in dogs. | 0.0 | 91 | ||||
| 2.4% | NyI‐K43 | [533] | Uncultured Penidiella clone F4‐K50 [KC535138] detected in hairs from mummies in Palermo, Italy. | 0.0 | 100 | KP828190 |
| Penidiella venezuelensis strain CBS 106.75 [EU019278]. | 0.0 | 99 | ||||
| 2.4% | NyI‐K4 | [576] | Uncultured fungus clone F4‐K2 [KC535126] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828191 |
| Leptospora rubella [HE774478]. | 0.0 | 92 | ||||
| 2.4% | NyI‐K30 | [576] | Uncultured fungus clone F4‐K2 [KC535126] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828192 |
| Leptospora rubella [HE774478]. | 0.0 | 92 | ||||
| 2.4% | NyI‐K6 | [570] | Uncultured fungus clone F4‐K28 [KC535132] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828193 |
| Penicillium ornatum strain CBS 190.68 [JX841244]. | 0.0 | 89 | ||||
| 2.4% | NyI‐K16 | [602] | Uncultured Rhizopus clone F4‐K34 [KC535134] detected in hairs from mummies in Palermo, Italy. | 0.0 | 99 | KP828194 |
| Rhizopus oryzae strains [KJ417572, AY803930], human pathogenic species. | 0.0 | 99 | ||||
| 7.4% | NyI‐K29 | [604] | Uncultured Rhizopus clone F4‐K34 [KC535134] detected in hairs from mummies in Palermo, Italy. | 0.0 | 100 | KP828195 |
| Rhizopus oryzae strains [KJ417572, AY803930], human pathogenic species. | 0.0 | 99 | ||||
| 2.4% | NyI‐K2 | [562] | Penicillium chrysogenum strains [KJ413371, KC009826, JQ422603, JN561259]. | 0.0 | 99 | KP828196 |
| 4.9% | NyI‐K25 | [563] | Penicillium chrysogenum strains [KJ413371, KC009826, JQ422603, JN561259]. | 0.0 | 99 | KP828197 |
| 7.4% | NyI‐K45 | [520] | Cryptococcus laurentii strains [JQ247577, JQ247574]. | 0.0 | 99 | KP828198 |
| Sample N2, cellulose nitrate membrane | ||||||
| 91.5% | N2‐K2 | [554] | Uncultured Acremonium clone AP3‐F4 [KF983520] detected in the Archimedes Palimpsest parchment book. | 0.0 | 99 | KP828199 |
| Acremonium charticola culture‐collection UOA/HCPF<GRC>:14413 [KC253940] common and emerging mould pathogens in Greece. | 0.0 | 98 | ||||
| 6.4% | N2‐K4 | [557] | Uncultured Acremonium clone F2‐K5 [KC535100] detected in clothes from mummies in Palermo, Italy. | 0.0 | 99 | KP828200 |
| Acremonium nepalense isolate Ppf33 [GU586837]. | 8e‐171 | 95 | ||||
| 2.1% | N2‐K13 | [562] | Uncultured Acremonium clone F2‐K5 [KC535100] detected in clothes from mummies in Palermo, Italy. | 0.0 | 99 | KP828201 |
| Acremonium nepalense isolate Ppf33 [GU586837]. | 8e‐171 | 95 | ||||
Clone libraries containing the ITS fungal regions (using the primers ITS1/ITS4; White et al., 1990) were performed as described by Piñar and colleagues (2015a). A total of 50 clones from each clone library were screened in a DGGE gel as described by Schabereiter‐Gurtner and colleagues (2001). Selected clones were externally sequenced by Sanger sequencing with a fleet of 16 ABI 3730xl (GATC Biotech, Germany). Comparative sequence analysis was performed by comparing pair‐wise insert sequences with those available in the public online database NCBI, using the blast search program (Altschul et al., 1997). The resulting sequences of the fungal clones have been deposited at the GenBank: Genetic sequence database at the National Center for Biotechnical Information (NCBI) and the corresponding accession numbers are given in this table.