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Direct proembryogenic masses initiation and plant regeneration from immature Torreya grandis embryos

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Abstract

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This paper provides an important direct proembryogenic mass initiation and plant regeneration method that might be supportive and stable for gene transformation and commercial production of Torreya grandis.

Abstract

Torreya grandis is an important and commercially valuable tree that is utilized as a source of essential oil and nuts. Propagation researches of T. grandis have been focused on micropropagation and somatic embryogenesis which derived from callus but limited data regarding direct somatic embryogenesis and plant regeneration are available in the literature. This study investigated the effects of plant growth regulators combinations on direct proembryogenic masses initiation and plant regeneration. Proembryogenic masses were initiated from cotyledons and hypocotyls of immature zygotic embryos directly. The frequency of direct proembryogenic masses initiation (52.6 %) was significantly greater than other treatments when the explants were cultured on Schenk and Hildebrandt medium which supplemented with 0.1 mg l−1 N6-benzylaminopurine, 0.1 mg l−1 Kinetin and 0.1 mg l−1 2,4-dichlorophenoxyacetic acid. Combined application of Abscisic acid and Polyethylene glycol 6000 stimulated cotyledons formation and resulted in morphologically superior somatic embryos. Schenk and Hildebrandt medium supplemented with 10 mg l−1 Abscisic acid and 20 mg l−1 Polyethylene glycol 6000 was optimum (88 %) for somatic embryos maturation. Highest germinated somatic embryos percentage (35.6 %) was obtained on Schenk and Hildebrandt medium supplemented with 2000 mg l−1 activated charcoal, with root and shoot lengths of 2.3 and 1.9 cm, respectively. As proembryogenic masses were initiated without the formation of callus, this study will aid to establish stable genetic transformation and will also be useful in improving understanding of the somatic embryogenesis of T. grandis.

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Abbreviations

BA:

N6-benzylaminopurine

KT:

Kinetin

2,4-D :

2,4-dichlorophenoxyacetic acid

ABA:

Abscisic acid

PEG6000:

Polyethylene glycol 6000

SH medium:

Schenk and Hildebrandt medium

PGRs:

Plant growth regulator(s)

EDTA:

Ethylene diamine tetraacetic acid

SEs:

Somatic embryos

PEMs:

Proembryogenic masses

FAA:

Formalin–acetic acid–alcohol

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Acknowledgments

The authors acknowledge the Major Agricultural Science and Technology Project of Science Technology Department of Zhejiang Province (2012C12002), National Natural Science Foundation of China (31470682, 31100461), National High Technology Research and Development program of China (863 Program, 2013AA102605), Key Agricultural Science and Technology Project of New Varieties Breeding of Zhejiang Province (2012C12904-12), and Spark Key Program of Ministry of Science and Technology of China (2012GA700001) for financial support.

Conflict of interest

The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. School of Forestry and Biotechnology, Zhejiang Agriculture and Forestry University, Lin’an, 311300, Zhejiang, China

      Qixiang Zhang, Weibo Xiang, Jin Yao, Hengkang Hu, Weiwu Yu, Youjun Huang, Jiasheng Wu & Jianqin Huang

    2. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Lin’an, 311300, Zhejiang, China

      Qixiang Zhang, Weibo Xiang, Jin Yao, Hengkang Hu, Weiwu Yu, Youjun Huang, Jiasheng Wu & Jianqin Huang

    3. Lishui Forestry Science Research Institute, Lishui, 323000, Zhejiang, China

      Lianhai Wu

    4. Lanxi Forestry Bureau, Lanxi, 321100, Zhejiang, China

      Jin Yao

    5. Department of Plant Sciences, University of California Davis, One Shields Avenue, Davis, CA, 95616, USA

      Charles A. Leslie

    6. Research Institute of Forestry, Chinese Academy of Forestry, 100091, Beijing, China

      Weibo Xiang

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    1. Qixiang Zhang
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    Correspondence to Jianqin Huang.

    Additional information

    Communicated by J. Lin.

    Q. Zhang and W. Xiang contributed equally.

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    Zhang, Q., Xiang, W., Yao, J. et al. Direct proembryogenic masses initiation and plant regeneration from immature Torreya grandis embryos. Trees 29, 1605–1612 (2015). https://doi.org/10.1007/s00468-015-1243-y

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    • DOI: https://doi.org/10.1007/s00468-015-1243-y

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