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Optimization of the induction, germination, and plant regeneration system for somatic embryos in apomictic walnut (Juglans regia L.)

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Abstract

The low efficiency of somatic embryo induction, proliferation, germination, and conversion to plantlets is a major problem in walnut. In this study, we used the cotyledons of immature embryos of apomictic seeds in walnut plus trees ‘Y17’ as explants. The optimum medium and culture conditions of somatic embryo induction, proliferation, and germination were tested through comparative experiments. The results showed that the optimum formula for somatic embryo induction, proliferation, and germination medium was DKW medium with 1.0 mg L−1 6-BA, 2.0 mg L−1 KT, and 0.01 mg L−1 IBA, DKW medium with 30–40 g L−1 sucrose and 7–8 g L−1 agar, and DKW medium with 2.0 mg L−1 concentrations of GA3, respectively. Meanwhile, the optimum time of seedling training was 36 h, and the substrate ratio was perlite: peat soil: vermiculite (1: 2: 1) after plant transplanting. The inter simple sequence repeat marker banding patterns of the primary embryo, secondary embryo and regenerated plants were all identical to that of the mother plant and proved the genetic stability of regenerated plants. An excellent walnut tissue culture system from walnut somatic embryo to the whole plant was formed, which can accelerate the speed of walnut genetic improvement.

Key message

An excellent walnut tissue culture system from walnut somatic embryo to the whole plant was formed, which can accelerate the speed of walnut genetic improvement.

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Abbreviations

DKW:

Driver and kuniyuki walnut

6-BA:

6-Benzylaminopruine

KT:

Kinetin

IBA:

Indole-3-butyric acid

GA3 :

Gibberellic acid (3)

WPM:

Wood plant medium

MS:

Murashige and skoog medium

ISSR:

Inter simple sequence repeats

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Funding

This research was supported by the National Natural Science Foundation of China (32001340), the Improved Variety Program of Shandong Province of China (2020LZGC090102), and the Natural Science Foundation of Shandong Province (ZR2020QC169).

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Author notes

  1. Hongcheng Fang, Yuhui Dong have contributed equally to this work.

Authors and Affiliations

  1. College of Forestry, Shandong Agricultural University, Tai’an, Shandong Province, China

    Hongcheng Fang, Yuhui Dong, Rui Zhou, Qingpeng Wang, Qicheng Duan, Changxi Wang, Yan Bao, Shengyi Xu, Xinya Lang, Shasha Gai & Ke Qiang Yang

  2. State Forestry and Grassland Administration Key Laboratory of Silviculture in the Downstream Areas of the Yellow River, Shandong Agricultural University, Tai’an, Shandong Province, China

    Hongcheng Fang, Yuhui Dong & Ke Qiang Yang

  3. Taian Culaishan Forest Farm, Tai’an, Shandong Province, China

    Rong Chen

Authors
  1. Hongcheng Fang
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  2. Yuhui Dong
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  3. Rui Zhou
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  4. Qingpeng Wang
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  5. Qicheng Duan
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  7. Yan Bao
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  9. Xinya Lang
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  10. Shasha Gai
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  11. Rong Chen
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  12. Ke Qiang Yang
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Contributions

RC and KY contributed the conception, design, and final revision. HF and YD contributed to design and drafting the manuscript. YB, SX, XL, SG, QW and QD analyzed and interpreted the data. RZ and CW contributed to supervise and revise the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Rong Chen or Ke Qiang Yang.

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The authors declare that there is no conflict of interest.

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Communicated by Francisco de Assis Alves Mourão Filho.

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Fang, H., Dong, Y., Zhou, R. et al. Optimization of the induction, germination, and plant regeneration system for somatic embryos in apomictic walnut (Juglans regia L.). Plant Cell Tiss Organ Cult 150, 289–297 (2022). https://doi.org/10.1007/s11240-022-02266-9

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  • DOI: https://doi.org/10.1007/s11240-022-02266-9

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