Scientia Agricultura Sinica ›› 2025, Vol. 58 ›› Issue (23): 4998-5012.doi: 10.3864/j.issn.0578-1752.2025.23.015

• HYBRIDIZATION BREEDING AND GERMPLASM INNOVATION IN PAEONIA • Previous Articles     Next Articles

The New Ploidies of Intersectional Hybrids in Paeonia and Their Generation Mechanisms Revealed by Molecular Karyotype Analysis

GUAN ZiHeng(), JI RunZe, RONG Qi, XU YuJie, ZHONG Yuan*(), CHENG FangYun*()   

  1. School of Landscape Architecture, Beijing Forestry University/State Key Laboratory of Efficient Production of Forest Resources/ Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding/National Engineering Research Center for Floriculture/Peony International Institute, Beijing 100083
  • Received:2025-03-18 Accepted:2025-05-06 Online:2025-12-01 Published:2025-12-09
  • Contact: ZHONG Yuan, CHENG FangYun

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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

Fig. 1

The 11 intersectional hybrid progenies produced by crossing the pollen parent, P. × lemoinei ‘L’ Esperance’, to P. lactiflora Fen Yun Fei He a-m: Flowers of the intersectional hybrids. a: N2-1; b: N2-10; c: N3-1; d: N5-6; e: S2-2; f: Jing Rui Huang; g: Jing Gui Mei; h: Jing Hua Xu Ri; i: Jing Hua Zhao Xia. J-l: Jing Hua Huan Cai; m: Jing Rui Zi. n-o: Leaves of the intersectional hybrids, n: Lobed; o: Unlobed. p: Jing Hua Huan Cai has both lobed and unlobed leaves"

Fig. 2

Karyotype of P. × lemoinei ‘L’ Esperance’, the pollen parent of intersectional hybrids a: DAPI stained; b: GISH signals (red) with B-genome as probes; c: rDNA-FISH signals, red: 5S rDNA, green: 45S rDNA; d: Composite picture with GISH and FISH signals; e: Chromosome karyotype; f: Idiogram. Bars=10 μm"

Table 1

Karyotype information of intersectional hybrid progenies"

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 -
AB-1C 2n=3x-1=14=10m+1sm+3st 4A -

Fig. 3

Karyotype of 7 triploid intersectional hybrids without inter-genome translocations a: N2-1; b: N2-10; c: N3-1; d: N5-6; e: S2-2; f: Jing Rui Huang; g: Jing Gui Mei. Small red spots in the middle of the short arms: 5S rDNA signals; The small green spots at the end of the short arms: 45S rDNA signals. Bars=10 μm"

Fig. 4

Karyotype of 2 triploid intersectional hybrids with inter-genome translocations"

Fig. 5

Karyotype of an intersectional hybrid named Jing Rui Zi a: GISH signals (red) with A-genome as probes; b: Absence of GISH signals (red) with B-genome as probes; c: C: GISH signals (red) with C-genome as probes; d: Composited picture with GISH and FISH signals (with A-genome as GISH probes, with 5S and 45S rDNA as FISH probes); e: Chromosome karyotype; f: Idiogram. Bars=10 μm"

Fig. 6

Karyotype of an intersectional hybrid named Jing Hua Huan Cai with 2 different ploidies a-f: Karyotype of the diploid，a: GISH signals (red) with A-genome as probes, b: Absence of GISH signals (red) with B-genome as probes, c: GISH signals (red) with C-genome as probes, d: Composited picture with GISH and FISH signals (with A-genome as GISH probes, with 5S and 45S rDNA as FISH probes), e: Chromosome karyotype, f: Idiogram; g-l: Karyotype of the near triploid aneuploid, g:DAPI stained, h: GISH signals with B-genome (red) and C-genome (green) as probes, i: rDNA-FISH signals (red: 5S rDNA, green: 45S rDNA), j: Composited picture with GISH and FISH signals, k: Chromosome karyotype, l: Idiogram. Bars=10 μm"

Table 2

Genome size and ploidy estimation of leaves with different morphologies in Jing Hua Huan Cai"

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
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
79.05 18.26 2.97 3.06 三倍体Triploid

Fig. 7

Measurement of DNA content of leaves with different morphologies in Jing Hua Huan Cai by flow cytometry a: P. ostii Feng Dan, diploid; b: Jing Rui Huang, triploid; c: Jing Rui Zi, diploid; d: Jing Hua Huan Cai unlobed leaves, diploid; e-h: Different ploidies were detected in Jing Hua Huan Cai lobed leaves, e: Near diploid with a genome slightly larger than diploid; f: Near triploid with a genome slightly smaller than triploid; g: Chimera with near diploid and near triploid; h: Chimera with near triploid and triploid"

Fig. 8

Formation mechanism of different ploidy of intersectional hybrids in Paeonia"

Fig. 9

Relationship between different ploidy and characters of intersectional hybrids"

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