Molecular Breeding

, 39:48 | Cite as

Genetic variation and development of a SCAR marker of anemone-type flower in chrysanthemum

  • Xiaodong Yang
  • Yangyang Wu
  • Jiangshuo Su
  • Ni Ao
  • Zhiyong Guan
  • Jiafu Jiang
  • Sumei Chen
  • Weimin Fang
  • Fadi Chen
  • Fei ZhangEmail author


An anemone shape featuring elongated and pigmented disk florets is an important target in breeding for flower shape in chrysanthemum. A good understanding of the inheritance of anemone-related traits and specific molecular markers are vital for accelerating the breeding progress. In this study, two segregating intercultivar F1 populations of the anemone-type ‘Nannong Xuefeng’ × the nonanemone-type ‘QX096’ (80 lines) and the same anemone-type ‘Nannong Xuefeng’ × ‘Monalisa’ (144 lines) were employed to investigate the genetic variation of anemone traits, and the anemone-specific markers were developed via the bulked segregant analysis (BSA) method by constructing two extreme bulks from each 10 anemone-type and nonanemone-type F1 lines, respectively. A moderate high level of variation coefficient (~ 24 to 57%) was obtained for most tubular floret traits relevant to the anemone type in both segregating populations. The anemone morphology of the hybrids was intermediate between that of their parents, and some transgressive individuals were observed, indicating great potential for the selection of lines with the desirable anemone type. In the BSA, four sequence-related amplified polymorphism (SRAP) primer combinations were identified as informative between the bulks of the anemone and nonanemone F1 individuals. After cloning and sequencing, only one SRAP marker, i.e., M11E1_272, was successfully converted to the sequence-characterized amplified region (SCAR) marker SCAR168. The SCAR marker was validation in the two F1 populations demonstrated that the marker SCAR168 could truly discriminate anemone and nonanemone types with a high coincidence ratio 87.86~92.5%. In addition, a general linear model (GLM)-based association analysis (P < 0.01) revealed that the SCAR marker exhibited a positive effect on the anemone traits and explained 20.83~33.66% phenotypic variation. Results of the current study show the inheritance pattern of anemone traits, and the developed anemone-specific SCAR marker paves the way for the marker-assisted selection of flower shape in chrysanthemum.


Chrysanthemum Bulked segregant analysis Flower shape Inheritance Molecular markers 



Amplified fragment length polymorphism


Bulked segregant analysis


Inter-simple sequence repeat


General linear model


Marker-assisted selection


Random amplified polymorphic DNA


Sequence-characterized amplified regions


Sequence-related amplified polymorphism


Simple sequence repeats



We would like to thank the anonymous reviewers for their constructive comments and suggestions that significantly improved the presentation of this paper.

Author contribution

FZ conceived and designed the project. FC, WF, and ZG provided the materials. XY and YW conducted experiments. XY and FZ analyzed the data and wrote the manuscript. All authors read and approved the manuscript.

Funding information

This work was financially supported by the National Natural Science Foundation of China (31370699, 31572152) and the Program for Key Research and Development, Jiangsu, China (BE2018426, BE2017412).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11032_2019_958_MOESM1_ESM.docx (20 kb)
Table S1 Sequences of SRAP primers (19 kb)
11032_2019_958_MOESM2_ESM.docx (15 kb)
Table S2 Phenotypic values of six anemone-related floral traits in the segregating F1 population of ‘Nannong Xuefeng’ × ‘QX096’ (DOCX 15 kb)
11032_2019_958_MOESM3_ESM.docx (15 kb)
Table S3 Phenotypic values of six anemone-related floral traits in the segregating F1 population of ‘Nannong Xuefeng’ × ‘Monalisa’ (DOCX 15 kb)
11032_2019_958_MOESM4_ESM.doc (27 kb)
Table S4 DNA sequence of the SRAP amplified fragment M11E1_272 (DOC 27 kb)
11032_2019_958_MOESM5_ESM.docx (19 kb)
Table S5 Segregation of anemone and nonanemone types in segregating F1 populations (DOCX 18 kb)
11032_2019_958_Fig5_ESM.png (350 kb)
Fig. S1

Flower morphology of anemone-type chrysanthemum ‘Nannong Xuefeng’ and nonanemone type ‘QX096’. a, d: Inflorescence morphology; b, e: Disk floret; c: Ray floret. Lo: Lobe; Ov: Ovule; St: Style; An: Anther. (PNG 350 kb)

11032_2019_958_MOESM6_ESM.tif (16.8 mb)
High Resolution Image (TIF 17250 kb)
11032_2019_958_Fig6_ESM.png (12.4 mb)
Fig. S2

Amplification result in segregating F1 individuals of anemone-type ‘Nannong Xuefeng’ and anemone-type ‘Monalisa’ by SCAR168. A: Anemone-type individuals; N: Nonanemone-type individuals. The specific recombination events are indicated by red rectangle, the unclear band patterns are indicated by blue rectangle. (PNG 12742 kb)

11032_2019_958_MOESM7_ESM.tif (20 mb)
High Resolution Image (TIF 20518 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Xiaodong Yang
    • 1
  • Yangyang Wu
    • 1
  • Jiangshuo Su
    • 1
  • Ni Ao
    • 1
  • Zhiyong Guan
    • 1
  • Jiafu Jiang
    • 1
  • Sumei Chen
    • 1
  • Weimin Fang
    • 1
  • Fadi Chen
    • 1
  • Fei Zhang
    • 1
    Email author
  1. 1.State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of HorticultureNanjing Agricultural UniversityNanjingPeople’s Republic of China

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