Selective screening: isolation of fungal strains from contaminated soils in Austria

Figure 1

Figure 2

Figure 3

GC Results: Measured US EPA PAH concentrations and standard deviations of the 12 soil samples (S1-12)_ Values are presented as mg/kg dry weight and standard deviations were calculated (mg/kg dryweight±standard deviation)Tabelle 2_ GC-Ergebnisse: Gemessene US EPA PAK-Konzentrationen und Standardabweichungen der 12 Bodenproben (S1-12) sind in der Tabelle dargestellt_ Die Werte sind also mg/kg Trockengeweicht dargestellt und die Standardabweichung wurde berechnet (mg/kg Trockengewicht±Standardabbweichung)

sample	S1	S2	S3	S4	S5	S6	S7	S8	S9	S10	S11	S12
Naphtalene	5.8±1.2	4.8±1.0	12.8±2.6	9.4±1.9	6.8±1.4	31.5±6.3	0.1±0.02	<0.3(0.0)	<0.3 (0.0)	0.1±0.01	<0.3(0.0)	<0.3(0.0)
Acenaphthylene	5.8±1.2	1.8±0.4	<2.7(0.00)	0.9±0.2	0.9±0.2	5.2±1.0	0.1±0.02	<0.3(0.0)	0.4±0.1	0.2±0.03	0.7±0.1	0.3±0.1
Acenatphene	6.9±1.4	4.3±0.9	11.2±2.2	8.5±1.7	6.8±1.4	24.9±5.0	0.6±0.1	0.4±0.1	0.4±0.1	0.1±0.02	<0.3 (0.0)	0.3±0.1
Flurene	10.1±2.0	6.3±1.3	14.8±3.0	10.3±2.1	8.3±1.7	34.3±6.9	0.5±0.1	0.4±0.1	0.4±0.1	0.1±0.02	0.5±0.1	0.6±0.1
Phenanthrene	55.20±11.0	28.3±5.7	70.2±14.0	48.3±9.7	36.5±7.3	154.0±31.0	5.9±1.2	4.4±0.9	4.1±0.8	1.0±0.2	6.4±1.3	5.7±1.2
Anthracene	22.5±4.5	11.4±2.3	22.2±4.4	18.1±3.6	14.4±2.9	67.7±13.5	1.4±0.3	1.2±0.2	1.6±20.3	0.4±0.01	2.1±0.4	1.4±0.3
Fluoranthene	78.0±15.6	29.1±5.8	48.2±9.6	39.0±7.8	30.8±6.2	141.0±28.0	7.5±1.5	12.6±2.5	10.0±2.0	2.9±0.6	17.5±3.5	9.8±2.0
Pyrene	77.8±15.6	30.4±6.1	47.9±9.6	41.6±8.3	32.2±6.4	108.0±22.0	7.5±1.5	24.4±4.9	10.0±2.0	2.3±0.5	14.5±2.9	7.7±2.0
Benz[a]anthracene	36.5±7.3	13.5±2.7	19.9±4.0	16.6±3.3	13.0±2.6	64.3±12.9	4.7±1.0	23.5±4.7	9.8±2.0	1.9±0.4	9.8±2.0	5.9±1.2
Chrysene	37.4±7.5	13.3±2.7	20.7±4.1	15.8±3.2	12.5±2.5	68.6±13.7	5.9±1.2	27.3±5.5	11.0±2.2	2.2±0.4	10.2±2.0	7.1±1.4
Benzo(b) fluoranthene	33.0±6.6	10.4±2.1	14.4±2.9	10.9±2.2	9.0±1.8	36.1±7.2	6.5±1.3	28.1±5.6	11.9±2.4	1.9±0.4	9.1±1.8	7.1±1.4
Benzo(k) fluoranthene	26.6±5.3	9.5±1.9	12.7±2.5	10.6±2.1	8.5±1.7	45.3±9.1	4.2±0.8	33.0±6.6	10.4±2.1	1.7±0.3	8.0±1.6	5.0±1.0
Benzo(a)pyrene	30.6±6.1	10.9±2.2	14.3±2.9	11.9±2.4	9.76±2.0	44.7±8.9	5.0±1.0	27.1±5.4	9.8±2.0	1.8±0.4	8.5±1.7	5.1±1.0
Indeno(1,2,3-cd)pyrene	21.3±4.3	7.0±1.4	8.5±1.7	8.3±1.7	6.0±1.2	20.1±4.0	3.1±0.6	19.6±3.9	8.2±1.6	1.7±0.3	5.2±1.0	2.8±0.6
Dibenzo(a,h) anthracene	5.9±1.2	1.7±0.4	<2.7(0.0)	1.6±0.3	1.3±0.3	4.8±1.0	1.1±0.2	5.5±1.1	2.0±0.4	0.5±0.1	1.2±0.2	0.7±0.1
Benzo(g,hi,i) perylene	19.6+3.9	6.5±1.3	7.8±1.6	6.4±1.3	5.2±1.1	17.2±3.4	3.4±0.7	22.7±4.5	7.2±1.5	1.8±0.4	4.2±0.8	2.7±0.5
sum	472.9±94.6	189.4±38.0	325.6±65.1	258.1±51.7	202.0±40.4	867.8±173.9	57.4±11.5	230.2±46.0	97.2±39.5	20.6±4.0	97.6±19.4	62.2±12.9

Growth pattern of the chosen fungal isolates for sequencing: + = good growth, ~ = very slow growth and - = no growth_ Hex = hexadecane, Tol = toluene, PCB = PCB 126Tabelle 4_ Wachstumsmuster der zum Sequenzieren ausgewählten Pilzisolate: + = gutes Wachstum, ~ = sehr langsames Wachstum und - = kein Wachstum_ Hex = Hexadekan Tol = Toluol, PCB = PCB 126

No	Hex	Tol	PCB
α 14	+	+	~
O	+	+	-
U	+	+	+
V	+	+	-
X 1	+	+	-
Y	+	+	-
Z	+	+	~
BL 3	+	+	+
BL 4	~	+	~
G	+	-	~
H	+	-	~

Results of the second microtiter plate screening: Number of colonies of having the same growth pattern of all 93 fungal isolates on toluene, hexadecane and PCB 126_ + = good growth, ~ = very slow growth and - = no growth_ Hex = hexadecane, Tol = toluene, PCB = PCB 126Tabelle 3_ Ergebnisse des zweiten Mikrotiterplatten-Screenings: Anzahl der Kolonien mit demselben Wachstumsmuster innerhalb der 93 isolierten Kolonien in Gegenwart von Toluol, Hexadekan und PCB 126_ + = gutes Wachstum, ~ = sehr langsames Wachstum und - = kein Wachstum_ Hex = Hexadekan Tol = Toluol, PCB = PCB 126

colony numbers	Hex	Tol	PCB
21	+	+	-
14	+	+	~
9	+	+	+
8	+	-	-
6	+	~	~
5	+	-	~
4	-	-	-
3	~	+	-
3	+	~	-
2	-	+	-
2	~	-	-
2	~	-	~
2	~	~	~
2	~	+	~
2	+	~	+
2	~	~	+
1	-	~	-
1	-	-	~
1	-	~	~
1	~	~	-
1	-	~	+
1	~	~	~

Phylogenetic classification of the ITS/18S rDNA coding sequences of the fungal isolatesTabelle 1_ Phylogenetische Klassifikation der ITS/18S rDNA kodierenden Sequenzen der Pilzisolate

No	Primer Pair	Closest identified phylogenetic relatives [EMBL accession numbers]	Query cover	Ident	ACBR strain No	accession No
α 14	NL1/NL4	Doratomyces purpureofuscus (Cephalotrichum purpureofuscum) genomic DNA sequence contains 28S rRNA gene, strain UAMH 9209 [LN851018.1]	99%	99%	MA6020	KY454753
O	NS5/NS8	Roussoella intermedia strain CBS 170.96 18S ribosomal RNA gene, partial sequence [KF443390.1]	99%	100%	MA6025	KY454758
Y	NL1/NL4	Purpureocillium sp. JCM 28545 gene for 28S ribosomal RNA, partial sequence [LC134235.1]	100%	99%	MA6015	KY454754
BL 3	NL1/NL4	Ochroconis longiphorum strain CBS 435.76 28S ribosomal RNA gene, partial sequence [KF156135.1]	96%	99%	MA6021	KY454755
BL 4	NL1/NL4	Ochroconis longiphorum strain CBS 435.76 28S ribosomal RNA gene, partial sequence [KF156135.1]	99%	99%	MA6017	KY454756
V	ITS1/ITS4	Penicillium janthinellum [EF634422.1]	100%	99%	MA6016	KY454760
Z	ITS1/ITS4	Pyrenochaeta inflorescentiae [EU552153.1]	97%	98%	MA6019	KY454761
H	ITS1/ITS4	Penicillium canescens [AY373901.1]	100%	99%	MA6018	KY454762
	NL1/NL4	Pyrenochaeta inflorescentiae culture-collection CBS:119222 18S ribosomal RNA gene, partial sequence; internal transcribed spacer 1, 5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence	99%	99%		KY454757
U		[EU552153.1]			MA6024
	NS5/NS8	Pyrenochaeta sp. GMG_PPb7 18S ribosomal RNA gene, partial sequence; internal transcribed spacer 1, 5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence [FJ439593.2]	99%	99%		KY454759
X1	ITS1/ITS4	Purpureocillium lilacinum	99%	100%	MA6022	KY454763
G	ITS1/ITS4	Penicillium canescens [AF033493.1]	99%	99%	MA6023	KY454764