Karyological and molecular studies between six species of Plantago in the Northern border region at Saudi Arabia

Figures & data

Table 1. List of investigated taxa and their codes.

			GPS
No.	Taxa	Code	Longitudes (E)	Latitudes (N)
1	Plantago albicans L.	P1	41.04°	30.98°
2	Plantago ciliata Desf.	P2
3	Plantago amplexicaulis Cav. subsp. bauphula (Edgew.)	P3
4	Plantago lanceolata L.	P4
5	Plantago psammophilia Agnew.	P5
6	Plantago ovata Forssk.	P6

Table 2. List of RAPD, ISSR and SCoT primers and the sequences used in this study, number of total bands, polymorphic bands and percentage of polymorphism of each primer generated.

Marker	Primer name	Band size range (bp)	Sequence	Total bands	Polymorphic bands	% Polymorphism
RAPD	OP-A7	430–1260	5′ GAA AGG GGT G 3′	7	4	57.14
	OP-A12	320–1360	5′ GTG ATC GCA G3′	11	9	81.82
	OP- B2	330–1270	5′ TCG GGG ATA G 3′	6	6	100
	OP-B11	360–1460	5′ GTA GAC CCG T 3′	10	5	50
	OP-C4	330–780	5′ CCG CAT CTA C 3′	7	3	42.86
	OP-C9	300–1340	5′ CTC ACC GTC C 3′	10	9	90
	OP-D1	280–1365	5 ACC GCG AAG G 3′	12	12	100
ISSR	44A	270–405	5 CTC TCT CTC TCT CTC TTG 3′	3	2	66.67
	44B	415–1340	5′ CTC TCT CTC TCT CTC TGC 3′	4	2	50
	HB-9	340–1335	5′ CAC CAC CAC GC 3′	6	4	66.67
	HB-11	360–670	5′ GTG TGT GTG TGT GTG CC 3′	5	1	20
	HB-13	400–810	5′ GAG GAG GAG C 3′	5	4	80
SCoT	SCoT 1	375–1000	ACG ACA TGG CGA CCA CGC	7	4	57.14
	SCoT 2	380–1170	ACC ATG GCT ACC ACC GGC	6	2	33.33
	SCoT 3	265–615	ACG ACA TGG CGA CCC ACA	4	4	100
	SCoT 4	460–570	ACC ATG GCT ACC ACC GCA	3	3	100
	SCoT 6	400–620	CAA TGG CTA CCA CTA CAG	3	2	66.67

Table 3. The morphological characters of the studied Plantago species.

			Leaves									Corolla
Species	V. t.	Pl. Len.	Position	L. Blade	L. margin	L. apex	L. surface	Scapes	Spike	Bracts	Sepals	tube	lobes	Capsule	Seeds
P. albicans	P.	Up to c. 30 cm tall	Radical	Linear-lanceolate	Entire	Acute	Lanate with whitish hairs	Longer than leaves	Cylindrical	Ovate	Ovate, glabrous	Glabrou	Glabrou	Ellipsoid, 2-seeded	Ellipsoid, brown
P. amplexicaulis	A.	Up to 30 cm tall	Cauline	Narrow lanceolate	Entire or sparingly toothed	Acuminate	Sparsely pilose to nearly glabrous	Longer than leaves	Ovoid-oblong	Glabrous, ovate-orbicular	Ovate, glabrous	Glabrou	Glabrou	Ellipsoid, 2-seeded	Ellipsoid, concave, brown to blackish
P. ciliata	A./P.	Up to 10 cm tall	Radical	Lanceolate-spathulate to obovate-spathulate	Entire to minutely denticulate	Acuminate	Covered densely with long white to greyish hairs	As long as leaves	Oblong-cylindrical	Ovate	Obovate-spathulate	Glabrou	Long ciliate on back	Ellipsoid, 2-seeded	Elliptic, brown
P. lanceolata	P.	Small to 30 (−60) cm tall	Radical	Narrow lanceolate to narrow elliptic	Entire	Acute	Sulcate, sparsely covered with white hairs	Longer than leaves	Conic-cylindrical	Ovate	Obovate	Glabrou	Glabrou	Ellipsoid, 2-seeded	Ellipsoid to oblong, brown to dark brown
P. ovata	A.	Up to 5 (−10) cm tall	Radical	Narrow linear	Entire or distantly denticulate	Obtuse to acute	Densely covered with appressed wooly hairs	As long as leaves	Ovoid-oblong	Obovate-orbicular, concave	Ovate-elliptic	Glabrous	Glabrous	Ellipsoid	Elliptic-ovate, brown
P. psammophilia	A.	Up to 25 cm tall	Cauline	Linear-oblanceolate	Entire	Acute	Densely covered with greyish-white, long appressed hairs	Shorter than leaves	Cylindrical	Ovate	Obovate	Glabrous	Densely ciliate on back	Ellipsoid	Elliptic, brown

Table 4. Karyotype parameters of somatic chromosomes of genus Plantago in KS.

Taxa code	Chr. No.	DCL	MCL	Mean arm ratio (MAR)	Mean centromeric index (MCI)	Rec	Syi index	TF%	A1	A2	A	K F
P. albicans	2n = 10	448.753	89.751	2.101 ± 0.328+	0.330 ± .029	88.6	54.219	35.157	0.424	0.087	0.297	2nsm(+) +4nsm(–) +4 nm
P. ciliata	2n = 10	363.963	72.793	1.603 ± 0.238	0.390 ± 0.027	80.2	64.625	42.795	0.353	0.144	0.223	6 nsm(–) +4 nm
P. amplexicaulis	2n = 10	459.944	91.989	1.927 ± 0.218	0.362 ± 0.02	82.5	59.057	37.129	0.421	0.150	0.282	2 nsm(+)+ 2nsm(–) + 6 nm
P. lanceolata	2n = 12	433.072	72.179	2.189 ± 0.263	0.335 ± 0.022	80.1	51.274	39.256	0.486	0.183	0.343	2 nsm(+)+ 6nsm(–) + 4 nm
P. psammophilia	2n = 10	369.068	73.814	1.810 ± 0.166	0.353 ± 0.018	90.1	57.393	36.465	0.433	0.096	0.281	4nsm(–) + 6 nm
P. ovata	2n = 8	308.242	77.061	2.071 ± 0.215	0.346 ± 0.017	91.1	53.427	34.823	0.480	0.060	0.330	2nsm(+) + 4 nsm(–) +2 nm

Notes: DCL: diploid chromosome number; A1: intrachromosomal asymmetry index; MCL: mean chromosome length; A2: interchromosomal asymmetry index; Rec: resemblance between chromosomes; A: degree of asymmetry of karyotype; Syi: the systematic indices. KF: karyotype formula; TF%: total form percentage.

Figure 1. Chromosomes of six species of Plantago in KSA(X = 1000). A: P. albicans. B: P. ciliata. C: P. amplexicaulis. D: P. lanceolata. E: P. psammophilia. F: P. ovata.

Figure 2. Karyogram of the selected species of Plantago (Bar = 25 µm). A: P. albicans. B: P. ciliata. C: P. amplexicaulis. D: P. lanceolata. E: P. psammophilia. F: P. ovata.

Figure 3. Ideogram representation of the haploid chromosome length of six species of Plantago. A: P. albicans. B: P. ciliata. C: P. amplexicaulis. D: P. lanceolata. E: P. psammophilia. F: P. ovata.

Table 5. Bands characteristics produced by molecular markers (RAPD, ISSR & SCoT) in different Plantago species.

No	Parameters	RAPD	ISSR	SCoT
1	Studied species	6	6	6
2	No. of primers	7	5	5
3	Markers range	280–1460	270–1340	265–1170
4	Total bands	63	23	23
5	Polymorphic bands	48	13	15
6	Unique bands	11	3	2
7	Monomorphic bands	15	10	8
8	% polymorphism	74.56%	56.67%	71.43%

Figure 4. The amplification profile of seven RAPD primers for six studied taxa of Plantago species in KSA. 1: P. albicans. 2: P. ciliata. 3: P. amplexicaulis. 4: P. lanceolata. 5: P. psammophilia. 6: P. ovata.

Figure 5. The amplification profile of five ISSR primers for six studied taxa of Plantago species in KSA. 1: P. albicans. 2: P. ciliata. 3: P. amplexicaulis. 4: P. lanceolata. 5: P. psammophilia. 6: P. ovata.

Figure 6. The amplification profile of five SCoT primers for six studied taxa of Plantago species in KSA. 1: P. albicans. 2: P. ciliata. 3: P. amplexicaulis. 4: P. lanceolata. 5: P. psammophilia. 6: P. ovata.

Figure 7. Percptual mapping (Biplot) of the studied species of Plantago L. based on cytological parameters and molecular attributes. P 1: P. albicans. P2: P. ciliata. P 3: P. amplexicaulis. P4: P. lanceolata. P5: P. psammophilia. P6: P. ovata.

Figure 8. Cluster phenogram showing the relationships between the studied taxa of Plantago using Euclidean coefficient and single linkage method. P 1: P. albicans. P2: P. ciliata. P 3: P. amplexicaulis. P4: P. lanceolata. P5: P. psammophilia. P6: P. ovata.

Table 6. Similarity indicies between the six studied species of Plantago L.

Taxa	P1	P2	P3	P4	P5	P6
P1	1.0
P2	0.876	1.0
P3	0.703	0.625	1.0
P4	0.723	0.666	0.784	1.0
P5	0.465	0.439	0.762	0.763	1.0
P6	0.648	0.675	0.662	0.500	0.689	1.0

Note: P1: P. albicans; P2: P. ciliata; P 3: P. amplexicaulis; P4: P. lanceolata; P 5: P. psammophilia; P6: P. ovata.

Figure 9. UPGAMA phenogram showing the relationships between the studied taxa of Plantago based on morphological, cytological parameters and molecular attributes.

Figure 10. Scatter diagram of 6 species in the genus Plantago.