分子核型分析揭示芍药属组间杂种新倍性及其产生机制

doi:10.3864/j.issn.0578-1752.2025.23.015

摘要/Abstract

摘要：

【目的】 牡丹与芍药杂交产生的芍药属组间杂种（Paeonia Itoh Group）是重要的新兴观赏植物。之前的研究均未在组间杂种中发现三倍体（2n=3x=15，ABC）以外的倍性，以及A、B基因组间的染色体易位。通过扩大研究材料范围、改进核型分析技术，旨在检测组间杂种中是否存在其他倍性和染色体异源易位现象，探究组间杂种性状与染色体组成的关系，为芍药属组间杂种产生机制及性状遗传规律奠定理论基础。【方法】 利用双色基因组原位杂交（genomic in situ hybridization，GISH）和rDNA荧光原位杂交（fluorescence in situ hybridization，FISH）技术，对‘粉云飞荷’芍药（2n=2x=10，CC）×‘金帝’牡丹（2n=2x=10，AB）组合产生的11个组间杂交后代进行核型分析；利用流式细胞术检测组间杂种‘京华幻彩’叶片形态与倍性的关系。【结果】 9个材料为三倍体（2n=3x=15，ABC），其中，在2个三倍体中发现A、B基因组间的染色体易位，包括2A与2B、3A与3B染色体间的相互易位，以及3A与3B染色体间的非相互易位。另外2个组间杂种中，‘京蕊紫’为二倍体（2n=2x=10，AC），而‘京华幻彩’具有二倍体（2n=2x=10，AC）和近三倍体（2n=3x-1=14，AB^-1C）等不同核型，在不同分株苗中，不同核型独立存在或呈嵌合状态，其中，不裂叶片均为二倍体，而开裂叶片为近三倍体、近二倍体或包含近三倍体的嵌合体。不同倍性的组间杂种具有高度一致的性状，包括生活型均为草本、花盘均为革质、雌雄蕊均高度不育等；但叶型、花色、花期等性状在不同倍性间存在明显差异。【结论】 证实了三倍化是组间杂种形成的主要途径，新发现的少数二倍体可能是由三倍体或近三倍体在体细胞有丝分裂过程中发生B基因组染色体丢失形成。倍性差异对组间杂种性状有重要影响，来自亲本的3个亚基因组均与组间杂种性状的形成有密切关系。揭示了芍药属组间杂种染色体核型的多样性及其形成机制。

关键词: 芍药, 牡丹, 核型, 二倍体, 三倍体, 非整倍体, 易位, 性状

Abstract:

【Objective】 Inter-sectional hybrids (Paeonia Itoh Group) produced by crossing between tree peonies and herbaceous peonies are important emerging ornamental plants. No previous studies have found ploidy other than triploid (2n=3x=15, ABC), or translocation between A and B genomes in intersectional hybrids. By employing the wider range of materials and the improved karyotyping technology, this research aimed to test whether there is other ploidy or inter-genome translocation in intersectional hybrids, and to explore how the intersectional hybrid traits are related to chromosome compositions, which could lay a theoretical foundation for the generation mechanism of inter-sectional hybrids in genus Paeonia and the genetic rules of their traits. 【Method】 In this study, we did molecular karyotype analysis on 11 intersectional hybrids from P. lactiflora Fen Yun Fei He (2n=2x=10, CC) × P. × lemoinei ‘L’ Esperance’ (2n=2x=10, AB) by bicolor genomic in situ hybridization (GISH) and rDNA fluorescence in situ hybridization (FISH). The relationship between morphology and ploidy of leaves from the inter-sectional hybrid Jing Hua Huan Cai was examined by flow cytometry. 【Result】 The results showed that 9 of them were triploids (2n=3x=15, ABC), in 2 of which chromosomal translocations between A and B genomes were found, including reciprocal translocations between 2A and 2B, or 3A and 3B chromosomes, and non-reciprocal translocations between 3A and 3B chromosomes. In the rest two intersectional hybrids, Jing Rui Zi was diploid (2n=2x=10, AC), but Jing Hua Huan Cai were diploid (2n=2x=10, AC), or near triploid (2n=3x-1=14, AB^-1C) or in more other ploidy, which were found to exist independently or in chimeric state in different divided seedlings. Therein, the unlobed leaves of Jing Hua Huan Cai were all diploids, while the lobed leaves were near triploid, near diploid, or chimera involving near triploid. The intersectional hybrids with different ploidy had highly consistent traits, including herbaceous life form, leathery flower disc and high sterility of both pistils and stamens. However, leaf morphology, flower color and flowering time were significantly different among different ploidy. 【Conclusion】 The results above confirm that triploidization is the main pathway for the formation of intersectional hybrids, and the newly discovered few diploids might be formed by the loss of B genome chromosomes during mitosis of somatic cells in triploids or near triploids. The traits of intersectional hybrids are strongly influenced by their ploidy level. The three subgenomes from the parents were all closely related to the trait formation of intersectional hybrids. These findings reveal the diversity of chromosome karyotypes and the formation mechanism of intersectional hybrids in Paeonia.

Key words: herbaceous peony, tree peony, karyotype, diploid, triploid, aneuploid, translocation, trait

关紫恒, 季润泽, 荣琪, 徐雨杰, 钟原, 成仿云. 分子核型分析揭示芍药属组间杂种新倍性及其产生机制[J]. 中国农业科学, 2025, 58(23): 4998-5012.

GUAN ZiHeng, JI RunZe, RONG Qi, XU YuJie, ZHONG Yuan, CHENG FangYun. The New Ploidies of Intersectional Hybrids in Paeonia and Their Generation Mechanisms Revealed by Molecular Karyotype Analysis[J]. Scientia Agricultura Sinica, 2025, 58(23): 4998-5012.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2025.23.015

https://www.chinaagrisci.com/CN/Y2025/V58/I23/4998

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材料名称 Name of materials	基因组构成 Genome structure	核型公式 Karyotype formula	sm染色体编号 Number of sm chromosomes	异源易位染色体 Chromosomes with inter-genome translocations
N2-1	ABC	2n=3x=15=10m+2sm+3st	4A, 4C	-
N2-10	ABC	2n=3x=15=11m+1sm+3st	4C	-
N3-1	ABC	2n=3x=15=10m+2sm+3st	4A, 4C	-
N5-6	ABC	2n=3x=15=12m+3st	-	-
S2-2	ABC	2n=3x=15=11m+1sm+3st	4C	-
京蕊黄Jing Rui Huang	ABC	2n=3x=15=9m+3sm+3st	4A, 4B, 4C	-
京桂美Jing Gui Mei	ABC	2n=3x=15=10m+2sm+3st	4A, 4B	-
京华旭日Jing Hua Xu Ri	ABC	2n=3x=15=10m+2sm+3st	4A, 4B	2A, 2B, 3A, 3B
京华朝霞Jing Hua Zhao Xia	ABC	2n=3x=15=11m+1sm+3st	4C	3A, 3B
京蕊紫Jing Rui Zi	AC	2n=2x=10=6m+2sm+2st	4A, 4C	-
京华幻彩Jing Hua Huan Cai	AC	2n=2x=10=7m+1sm+2st	4C	-
京华幻彩Jing Hua Huan Cai	AB^-1C	2n=3x-1=14=10m+1sm+3st	4A	-

材料名称 Name of materials	样品数 No. of samples	DNA含量（峰值荧光强度） DNA content (Fluorescence intensity at peak)	估测基因组大小 Estimated genome size (Gb)	倍性计算 Ploidy calculation		估测倍性 Estimated ploidy
凤丹P. ostii Feng Dan（2n=2x=10）*	2	53.16±0.21	12.28	2.00	2.06	二倍体Diploid
京蕊黄Jing Rui Huang（2n=3x=15）*	2	77.57±1.60a	17.91±0.37	2.92	3.00	三倍体Diploid
京蕊紫Jing Rui Zi（2n=2x=10）*	3	39.39±0.53d	9.10±0.12	1.48	1.52	二倍体Diploid
京华幻彩不裂叶片 Jing Hua Huan Cai unlobed leaves	9	43.36±4.02cd	10.01±0.92	1.63	1.68	二倍体Diploid
京华幻彩开裂叶片Ⅰ Jing Hua Huan Cai lobed leaves Ⅰ	3	50.18±2.75c	11.59±0.64	1.89	1.94	近二倍体 Near diploid
京华幻彩开裂叶片Ⅱ Jing Hua Huan Cai lobed leaves Ⅱ	4	69.58±3.83b	16.07±0.88	2.62	2.69	近三倍体 Near triploid
京华幻彩开裂叶片Ⅲ Jing Hua Huan Cai lobed leaves Ⅲ	4	47.31±1.69c	10.93±0.39	1.78	1.83	近二倍体 Near diploid
京华幻彩开裂叶片Ⅲ Jing Hua Huan Cai lobed leaves Ⅲ	4	69.49±2.67b	16.05±0.62	2.61	2.69	近三倍体 Near triploid
京华幻彩开裂叶片Ⅳ Jing Hua Huan Cai lobed leaves Ⅳ	1	67.36	15.56	2.53	2.61	近三倍体 Near triploid
京华幻彩开裂叶片Ⅳ Jing Hua Huan Cai lobed leaves Ⅳ	1	79.05	18.26	2.97	3.06	三倍体Triploid