SRAP markers for characterization of the genetic diversity and differentiation of Pinus nigra populations in protected forested areas in Bulgaria, Greece, and Cyprus

Figures & data

Figure 1. Locations of the analyzed P. nigra Arn. Populations in Bulgaria, Greece, and Cyprus. The designations and geographic coordinates of the populations are presented in Table 1.

Table 1. Geographic coordinates of Pinus nigra Arn populations used in this study. BG, Bulgaria; GR, Greece; CY, Cyprus.

P. nigra Arn. population	Population designation	Geographic coordinates	Natura 2000 protected area	Number of samples
Lepenitsa (BG)	LE	41.96675 N, 23.97714 E	Rodopi-Zapadni	22
Chepino (BG)	CH	41.99581 N, 23.9856 E	Rodopi-Zapadni	22
Draginovo (BG)	DR	42.07095 N, 24.02333 E	Yadenitsa	22
Konitsa (GR)	KO	40.0591 N, 20.77605 E	n/a	21
Samarina (GR)	SA	40.09765 N, 21.04548 E	n/a	20
Aoos lake (GR)	AO	39.84352 N, 21.07539 E	Valia Kalnta kai Techniti Limni Aoou	20
Troodos mountains-1 (CY)	TR1	34.93566 N, 32.90276 E	Ethniko Dasiko Parko Troodous	25
Troodos mountains-2 (CY)	TR2	34.94624 N, 32.85111 E	Ethniko Dasiko Parko Troodous	25

Table 2. Nucleotide sequences of SRAP primers used in the study.

Forward primer		Reverse primer
EM2	5′-TGAGTCCAAACCGGAGC-3′	ME3	5′-GACTGCGTACGAATTATT-3′
EM7	5′-TGAGTCCAAACCGGTCC-3′	ME1	5′-GACTGCGTACGAATTAAA-3′
EM1	5′-TGAGTCCAAACCGGAAA-3′	ME8	5′-GACTGCGTACGAATTCAC-3′

Table 3. Summary of SRAP genotyping data obtained following SRAP analysis of 175 studied black pine samples.

Primer pair	Total number of SRAP loci	Number of polymorphic loci	Percent of polymorphic loci
EM7 × ME1	609	252	41.4%
EM3 × ME3	526	138	26.2%
EM1 × ME8	669	256	38.3%
Total	1804	646	38.3%
Average per primer pair	601 ± 72	215 ± 67

Figure 2. Genetic structure and Principal Coordinate Analysis (PCoA) based on SRAP marker data: (a) estimation of the most probable number of genetic clusters (K), (b) bar plot representing the genetic structure at K = 2, (c) bar plot representing the genetic structure at K = 4, (d) PCoA (Coord. 1 vs. Coord. 2), (e) PCoA (Coord. 2 vs. Coord. 3). LE, CH, DR, KO, SA, AO, TR1, and TR2—designations of the studied populations according to Table 1.

Table 4. Pairwise comparison of Fst values between populations as a measure of their genetic differentiation. LE, CH, DR, KO, SA, AO, TR1, and TR2—designations of the studied populations according to Table 1.

Country and population		Bulgaria			Greece			Cyprus
		LE	CH	DR	KO	SA	AO	TR1	TR2
Bulgaria	LE		0.01	0.01	0.01	0.11	0.21	0.17	0.17
	CH	0.01		0.00	0.01	0.11	0.20	0.15	0.16
	DR	0.01	0.00		0.00	0.10	0.20	0.15	0.15
Greece	KO	0.01	0.01	0.00		0.11	0.20	0.14	0.15
	SA	0.11	0.11	0.10	0.11		0.06	0.12	0.11
	AO	0.21	0.20	0.20	0.20	0.06		0.18	0.18
Cyprus	TR1	0.17	0.15	0.15	0.14	0.12	0.18		0.00
	TR2	0.17	0.16	0.15	0.15	0.11	0.18	0.00

Data availability statement

All data included in this study are available from the corresponding author [I.A.] upon request.