Karyotype analysis and fluorescence in situ hybridization localization of ribosomal DNA and telomeric repeats in root tip chromosomes of Hyacinthoides non-scripta

doi:10.3969/j.issn.1004-1524.20250246

Acta Agriculturae Zhejiangensis ›› 2026, Vol. 38 ›› Issue (4): 687-695.DOI: 10.3969/j.issn.1004-1524.20250246

• Horticultural Science • Previous Articles Next Articles

Karyotype analysis and fluorescence in situ hybridization localization of ribosomal DNA and telomeric repeats in root tip chromosomes of Hyacinthoides non-scripta

LIU Huanhuan¹^,²(), SU Xiaoqian², DU Wenkai¹, HU Fengrong²^,^*()

¹ School of Landscape Architecture, Jiangsu Vocational College of Agriculture and Forestry, Zhenjiang 212400, Jiangsu, China
² College of Landscape Architecture, Nanjing Forestry University, Nanjing 210037, China

Received:2025-03-27 Online:2026-04-25 Published:2026-05-08
Contact: HU Fengrong

Abstract

Abstract:

To elucidate the genetic relationship and evolutionary variations among different flower colors of bluebell (Hyacinthoides non-scripta), a cytological analysis was conducted to provide reliable evidence for polyploid breeding strategies. Karyotype analysis and fluorescence in situ hybridization (FISH) using 45S rDNA, 5S rDNA, and telomere repeats as probes were performed on white-flowered and blue-flowered bluebells. Karyotype analysis revealed that the white bluebell was diploid, with a chromosome number of 2n=2x=16, with an average arm ratio of 4.07, a karyotype asymmetry coefficient of 75.18%, and a karyotype formula of 2n=2x=16=2m+6sm+4st+4t(sat). The blue bluebell was triploid, with a chromosome number 2n=3x=24, with an average arm ratio of 4.27, a karyotype asymmetry coefficient of 75.20%, and a karyotype formula of 2n=3x=24=3m+9sm+6st+6t(sat). Both varieties exhibited a 3B karyotype, with satellite DNA located at the ends of the short arms of two pairs of telocentric (t) chromosomes. The 45S rDNA signals were localized on the chromosomal secondary constrictions and 5S rDNA signals were localized on the top of the long arm of the metacentric (m) chromosome by FISH results. Telomeric repeat sequences were uniformly distributed at both ends of each chromosome, without interstitial signals. White bluebells displayed four 45S rDNA and two 5S rDNA signal sites, while blue bluebells exhibited six 45S rDNA and three 5S rDNA signal sites. Karyotype analysis and FISH signal localization indicated that blue bluebells likely originated from whole-genome duplication of the white bluebell, providing a cytological theoretical basis for polyploid breeding in bluebells.

Key words: Hyacinthoides non-scripta, chromosome, karyotype analysis, fluorescence in situ hybridization (FISH)

CLC Number:

S682.2

LIU Huanhuan, SU Xiaoqian, DU Wenkai, HU Fengrong. Karyotype analysis and fluorescence in situ hybridization localization of ribosomal DNA and telomeric repeats in root tip chromosomes of Hyacinthoides non-scripta[J]. Acta Agriculturae Zhejiangensis, 2026, 38(4): 687-695.

Figures/Tables 7

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序号 No.	相对长度/% Relative length/%			臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
序号 No.	短臂 Short arm	长臂 Long arm	全长 Full length	臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
1	2.78±0.04	7.13±0.08	9.91±0.11	2.57±0.03	sm	28.02	1.59	L
2	2.70±0.06	6.79±0.26	9.49±0.20	2.52±0.15	sm	28.48	1.52	L
3	1.09±0.07	6.58±0.34	7.67±0.39	6.04±0.27	st	14.25	1.23	M2
4	1.22±0.15	5.90±0.14	7.12±0.19	4.98±0.60	st	17.09	1.14	M2
5	3.12±0.05	3.68±0.11	6.80±0.13	1.18±0.03	m	45.91	1.09	M2
6	2.95±0.02	3.45±0.16	6.40±0.14	1.17±0.06	m	46.07	1.02	M2
7	0.77±0.04	5.50±0.11	(6.27±0.09)+(1.09±0.08)	7.14±0.51	t	12.87	1.00	M2
8	0.74±0.10	5.28±0.06	(6.02±0.15)+(1.02±0.01)	7.13±1.03	t	12.74	0.96	M1
9	0.71±0.09	5.23±0.05	(5.94±0.13)+(0.61±0.10)	7.37±0.86	t	12.46	0.95	M1
10	0.68±0.09	5.01±0.06	(5.69±0.10)+(056±0.12)	7.37±1.01	t	11.90	0.91	M1
11	1.80±0.06	3.79±0.05	5.59±0.081	2.11±0.07	sm	32.22	0.89	M1
12	1.82±0.04	3.66±0.02	5.48±0.03	2.01±0.05	sm	33.19	0.88	M1
13	0.92±0.10	4.19±0.21	5.11±0.24	4.63±0.50	st	18.06	0.82	M1
14	0.79±0.05	4.04±0.20	4.83±0.21	5.19±0.43	st	16.29	0.77	M1
15	1.42±0.21	2.56±0.11	3.98±0.31	1.87±0.23	sm	35.71	0.64	S
16	1.31±0.14	2.38±0.10	3.69±0.22	1.85±0.16	sm	35.47	0.59	S

序号 No.	相对长度/% Relative length/%			臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
序号 No.	短臂 Short arm	长臂 Long arm	全长 Full length	臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
1	2.78±0.04	7.13±0.08	9.91±0.11	2.57±0.03	sm	28.02	1.59	L
2	2.70±0.06	6.79±0.26	9.49±0.20	2.52±0.15	sm	28.48	1.52	L
3	1.09±0.07	6.58±0.34	7.67±0.39	6.04±0.27	st	14.25	1.23	M2
4	1.22±0.15	5.90±0.14	7.12±0.19	4.98±0.60	st	17.09	1.14	M2
5	3.12±0.05	3.68±0.11	6.80±0.13	1.18±0.03	m	45.91	1.09	M2
6	2.95±0.02	3.45±0.16	6.40±0.14	1.17±0.06	m	46.07	1.02	M2
7	0.77±0.04	5.50±0.11	(6.27±0.09)+(1.09±0.08)	7.14±0.51	t	12.87	1.00	M2
8	0.74±0.10	5.28±0.06	(6.02±0.15)+(1.02±0.01)	7.13±1.03	t	12.74	0.96	M1
9	0.71±0.09	5.23±0.05	(5.94±0.13)+(0.61±0.10)	7.37±0.86	t	12.46	0.95	M1
10	0.68±0.09	5.01±0.06	(5.69±0.10)+(056±0.12)	7.37±1.01	t	11.90	0.91	M1
11	1.80±0.06	3.79±0.05	5.59±0.081	2.11±0.07	sm	32.22	0.89	M1
12	1.82±0.04	3.66±0.02	5.48±0.03	2.01±0.05	sm	33.19	0.88	M1
13	0.92±0.10	4.19±0.21	5.11±0.24	4.63±0.50	st	18.06	0.82	M1
14	0.79±0.05	4.04±0.20	4.83±0.21	5.19±0.43	st	16.29	0.77	M1
15	1.42±0.21	2.56±0.11	3.98±0.31	1.87±0.23	sm	35.71	0.64	S
16	1.31±0.14	2.38±0.10	3.69±0.22	1.85±0.16	sm	35.47	0.59	S

序号 No.	相对长度/% Relative length/%			臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
序号 No.	短臂 Short arm	长臂 Long arm	全长 Full length	臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
1	2.00±0.09	4.49±0.07	6.49±0.14	2.26±0.08	sm	30.77	1.56	L
2	1.74±0.04	4.33±0.08	6.07±0.11	2.49±0.05	sm	28.67	1.46	L
3	1.74±0.03	3.93±0.09	5.66±0.09	2.27±0.07	sm	30.67	1.36	L
4	0.99±0.14	4.37±0.25	5.36±0.12	4.63±0.79	st	18.46	1.29	L
5	0.80±0.04	4.27±0.14	5.06±0.12	5.37±0.40	st	15.73	1.21	M2
6	0.83±0.07	4.12±0.17	4.95±0.10	5.07±0.58	st	16.71	1.19	M2
7	2.19±0.06	2.47±0.13	4.66±0.19	1.13±0.04	m	47.00	1.12	M2
8	2.15±0.08	2.37±0.10	4.53±0.18	1.10±0.02	m	47.50	1.09	M2
9	2.11±0.08	2.25±0.06	4.36±0.13	1.07±0.02	m	48.32	1.05	M2
10	0.53±0.02	3.80±0.16	(4.33±0.15)+(0.80±0.03)	7.20±0.47	t	12.24	1.04	M2
11	0.49±0.04	3.73±0.14	(4.23±0.10)+(0.79±0.06)	7.73±0.95	t	11.67	1.01	M2
12	0.44±0.03	3.69±0.13	(4.13±0.10)+(0.81±0.08)	8.48±0.96	t	10.73	0.99	M1
13	0.48±0.04	3.63±0.12	(4.11±0.09)+(0.41±0.03)	7.73±0.94	t	11.76	0.99	M1
14	0.48±0.04	3.57±0.10	(4.06±0.08)+(0.40±0.01)	7.52±0.78	t	11.90	0.97	M1
15	0.47±0.03	3.52±0.09	(3.99±0.06)+(0.39±0.04)	7.55±0.76	t	11.85	0.96	M1
16	1.18±0.02	2.58±0.10	3.76±0.11	2.17±0.08	sm	31.50	0.90	M1
17	1.13±0.08	2.50±0.21	3.63±0.19	2.27±0.31	sm	31.04	0.87	M1
18	1.13±0.11	2.34±0.15	3.47±0.19	2.11±0.27	sm	32.63	0.83	M1
19	0.47±0.08	2.73±0.05	3.21±0.06	6.07±0.94	st	14.76	0.77	M1
20	0.44±0.01	2.61±0.03	3.06±0.02	5.90±0.23	st	14.50	0.73	S
21	0.41±0.03	2.46±0.08	2.87±0.08	6.05±0.46	st	14.30	0.69	S
22	0.88±0.01	1.89±0.13	2.76±0.14	2.15±0.13	sm	31.76	0.66	S
23	0.85±0.01	1.84±0.12	2.68±0.11	2.18±0.16	sm	31.55	0.64	S
24	0.87±0.05	1.70±0.05	2.57±0.10	1.96±0.05	sm	33.81	0.62	S

序号 No.	相对长度/% Relative length/%			臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
序号 No.	短臂 Short arm	长臂 Long arm	全长 Full length	臂比(长臂/ 短臂) Arm ratio (L/S)	着丝粒位置 Position of centromere	着丝粒指数/% Centromeric index/%	相对长度系数 Coefficient of relative length	相对长度类型 Type of relative length
1	2.00±0.09	4.49±0.07	6.49±0.14	2.26±0.08	sm	30.77	1.56	L
2	1.74±0.04	4.33±0.08	6.07±0.11	2.49±0.05	sm	28.67	1.46	L
3	1.74±0.03	3.93±0.09	5.66±0.09	2.27±0.07	sm	30.67	1.36	L
4	0.99±0.14	4.37±0.25	5.36±0.12	4.63±0.79	st	18.46	1.29	L
5	0.80±0.04	4.27±0.14	5.06±0.12	5.37±0.40	st	15.73	1.21	M2
6	0.83±0.07	4.12±0.17	4.95±0.10	5.07±0.58	st	16.71	1.19	M2
7	2.19±0.06	2.47±0.13	4.66±0.19	1.13±0.04	m	47.00	1.12	M2
8	2.15±0.08	2.37±0.10	4.53±0.18	1.10±0.02	m	47.50	1.09	M2
9	2.11±0.08	2.25±0.06	4.36±0.13	1.07±0.02	m	48.32	1.05	M2
10	0.53±0.02	3.80±0.16	(4.33±0.15)+(0.80±0.03)	7.20±0.47	t	12.24	1.04	M2
11	0.49±0.04	3.73±0.14	(4.23±0.10)+(0.79±0.06)	7.73±0.95	t	11.67	1.01	M2
12	0.44±0.03	3.69±0.13	(4.13±0.10)+(0.81±0.08)	8.48±0.96	t	10.73	0.99	M1
13	0.48±0.04	3.63±0.12	(4.11±0.09)+(0.41±0.03)	7.73±0.94	t	11.76	0.99	M1
14	0.48±0.04	3.57±0.10	(4.06±0.08)+(0.40±0.01)	7.52±0.78	t	11.90	0.97	M1
15	0.47±0.03	3.52±0.09	(3.99±0.06)+(0.39±0.04)	7.55±0.76	t	11.85	0.96	M1
16	1.18±0.02	2.58±0.10	3.76±0.11	2.17±0.08	sm	31.50	0.90	M1
17	1.13±0.08	2.50±0.21	3.63±0.19	2.27±0.31	sm	31.04	0.87	M1
18	1.13±0.11	2.34±0.15	3.47±0.19	2.11±0.27	sm	32.63	0.83	M1
19	0.47±0.08	2.73±0.05	3.21±0.06	6.07±0.94	st	14.76	0.77	M1
20	0.44±0.01	2.61±0.03	3.06±0.02	5.90±0.23	st	14.50	0.73	S
21	0.41±0.03	2.46±0.08	2.87±0.08	6.05±0.46	st	14.30	0.69	S
22	0.88±0.01	1.89±0.13	2.76±0.14	2.15±0.13	sm	31.76	0.66	S
23	0.85±0.01	1.84±0.12	2.68±0.11	2.18±0.16	sm	31.55	0.64	S
24	0.87±0.05	1.70±0.05	2.57±0.10	1.96±0.05	sm	33.81	0.62	S

Karyotype analysis and fluorescence in situ hybridization localization of ribosomal DNA and telomeric repeats in root tip chromosomes of Hyacinthoides non-scripta

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