牡丹组间杂交子代真实性的SSR鉴定

doi:10.16178/j.issn.0528-9017.20240351

浙江农业科学 ›› 2025, Vol. 66 ›› Issue (7): 1672-1675.DOI: 10.16178/j.issn.0528-9017.20240351

牡丹组间杂交子代真实性的SSR鉴定

赵海军¹(), 王建民¹, 闫闪闪¹, 张佩¹, 盖伟玲²^,^*()

1.菏泽市牡丹区牡丹研究院,山东菏泽 274000
2.青岛农业大学,山东青岛 266109

收稿日期:2024-03-27 出版日期:2025-07-11 发布日期:2025-07-28
通讯作者: 盖伟玲(1971—),女,山东青岛人,实验师,硕士,主要从事花卉育种栽培研究工作,E-mail:yanpingyu211@outlook.com。
作者简介:赵海军(1970—),男,汉族,山东菏泽人,高级农艺师,本科,主要从事牡丹研究工作,E-mail:15065066999@163.com。
基金资助:
山东省农业良种工程项目(2020LZGC011)

Authenticity identification of inter-subgroup hybridization offspring of Paeonia Sect. Mouton based on SSR markers

ZHAO Haijun¹(), WANG Jianmin¹, YAN Shanshan¹, ZHANG Pei¹, GAI Weiling²^,^*()

1. Peony Research Institute of Mudan District Heze City, Heze 274000, Shandong
2. Qingdao Agricultural University, Qingdao 266109, Shandong

Received:2024-03-27 Online:2025-07-11 Published:2025-07-28

摘要/Abstract

摘要：

以滇牡丹(Paeonia delavayi)和日本牡丹栽培品种为亲本进行亚组间杂交,是培育牡丹优异品种的重要育种途径,对其杂交子代真实性进行早期的分子鉴定,可以缩短育种周期,提高育种效率,降低育种成本。本研究以滇牡丹为母本,以日本牡丹栽培品种岛锦为父本,通过人工授粉杂交获得了2株实生幼苗,利用简单重复序列标记(SSR 分子标记)技术确认了种苗的真实性。结果表明,2株杂交苗在2对 SSR 引物中具有亲本特异性互补型条带,证实了它们是真实杂交种。该研究所应用的 SSR 分子标记技术具有准确、高效的特性,可用于早期鉴定牡丹杂交子代,为培育优良牡丹品种提供技术支持。

关键词: 牡丹亚组间杂交, 实生子代苗, SSR分子标记

Abstract:

Inter-subgroup hybridization between Paeonia delavayi and Japanese peony cultivars as parents is an important breeding pathway to cultivate excellent peony varieties, and early molecular characterization of the authenticity of their hybrid offspring can shorten the breeding cycle, improve the breeding efficiency, and reduce the breeding cost. In this study, two offspring seedlings were obtained by hand-pollination hybridization between Paeonia delavayi as the female parent and Japanese peony cultivar Daojin as the male parent. The authenticity of the hybrid offspring was identified using SSR molecular marker technology. The results revealed that two hybrid seedlings had parental specific complementary bands in two pairs of SSR primers, confirming that they were true hybrid species. The SSR molecular marker technology applied by this research institute has accurate and efficient characteristics, which can be used for early identification of peony hybrid offspring and provide technical support for cultivating excellent peony varieties.

Key words: peony inter-subgroup hybridization, offspring seedling, SSR molecular marker

中图分类号:

S685.11

赵海军, 王建民, 闫闪闪, 张佩, 盖伟玲. 牡丹组间杂交子代真实性的SSR鉴定[J]. 浙江农业科学, 2025, 66(7): 1672-1675.

ZHAO Haijun, WANG Jianmin, YAN Shanshan, ZHANG Pei, GAI Weiling. Authenticity identification of inter-subgroup hybridization offspring of Paeonia Sect. Mouton based on SSR markers[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(7): 1672-1675.

图/表 5

图1 杂交实生子代苗

Fig.1 Hybrid offspring seedlings

表1 PCR反应程序

Table 1 PCR reaction program

步骤	反应温度/℃	时间
1	94	3 min	$35 个循环$
2	94	30 s
3	Tm	30 s
4	72	15 s
5	72	7 min
6	16	Pulse

表1 PCR反应程序

Table 1 PCR reaction program

步骤	反应温度/℃	时间
1	94	3 min	$35 个循环$
2	94	30 s
3	Tm	30 s
4	72	15 s
5	72	7 min
6	16	Pulse

表2 引物信息

Table 2 Primer information

引物	引物序列5'-3'	5'端修饰	重复碱基	退火温度/℃	条带大小/bp
P1	F TGGCGCTCCACACGGTATCTT	Tail C	(CTC)4	58.24	228
	R ATCGTGTTCGGTTCGCCGGT			59.08
P2	F TCGATGCCCGCAGGGTTTCT	Tail C	(GCA)4	58.42	381
	R ACAGGTCTTGTCCGGCCTTGT			58.01
P3	F TCCCAAAAGAGGGCGGTGCT	Tail C	(AAG)4	58.25	306
	R TCCCCTCCGAAACGGCATGT			58.05
P4	F ATGGCGAGATTGCCCCGTCT	Tail C	(TTAA)3	58.20	228
	R TGCAGTCACCACCATCCGTCA			58.10
P5	F CGTCATACTGAACGCCGCCGA	Tail C	(TCTCGC)3	59.85	208
	R TCACGTTGGATGTTCACTGCCCA			58.76
P6	F CGGCGTGCCTTTGAGACGTG	Tail C	(AGA)5	58.91	157
	R TGCCTCGAAAAACTCGCCTTCTCC			59.48
P7	F AGCGTGAAGCAACAAGCCGTG	Tail C	(CATTT)3	58.82	167
	R ACTGCGTTTCACGGCGAGGA			59.01
P8	F AGCAGAGAAGGTAGGCGGCG	Tail C	(ACTGA)	58.59	171
	R AGCGCTGAAGGCCAGTCATGG			59.45
P9	F GGGGACTCAAATCCTTGCGAAAACCA	Tail C	(CAC)4	59.68	189
	R AGGCCTAGTTTTGGTCTGGGCG			58.83
P10	F CAGCTGGCTTCCCACGCAGT	Tail C	(GGT)4	59.63	177
	R TAGCCATGCCGCCTCCACCT			59.97
Tail C	CAGGACCAGGCTACCGTG	FAM		59.00

表3 样品DNA浓度检测结果

Table 3 Results of DNA concentration of samples

样品	类型	浓度值/ (μg·mL^-1)	吸光度					消光因子
样品	类型	浓度值/ (μg·mL^-1)	D₂₆₀	D₂₈₀	D₂₆₀/D₂₈₀	D₂₆₀/D₂₃₀		消光因子
滇牡丹	DNA	583.481	11.670	6.282	1.858	1.970	50.00
岛锦	DNA	382.362	7.647	3.932	1.945	2.074	50.00
子代1	DNA	573.446	11.469	5.959	1.925	1.993	50.00
子代2	DNA	192.732	3.855	1.973	1.954	2.183	50.00

图2 引物P5和P10毛细管电泳结果

Fig.2 Results of capillary electrophoresis of primers P5 and P10

参考文献 7

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牡丹组间杂交子代真实性的SSR鉴定

Authenticity identification of inter-subgroup hybridization offspring of Paeonia Sect. Mouton based on SSR markers

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