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Induction of somatic embryogenesis by anther-derived callus culture and plantlet ploidy determination in poplar (Populus × beijingensis )

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Abstract

In vitro regeneration through somatic embryogenesis using anther-derived callus from poplar (P. × beijingensis) has been reported. Calli induced from anther connective tissue were used to create suspension cell. Embryogenic cells were enriched by sieving and centrifugation, then embryogenic cells were shaking cultured in liquid medium lacking NH4 + to incubate round callus (RC). Thick slices of RC induced somatic embryo (SE) when incubated in solid Murashige and Skoog medium supplemented with either N-(2-chloro-4-pyridyl)-N-phenylurea or 6-benzyladenine, both supplemented with 1-naphthaleneacetic acid. Plantlets germinated from SE in the treatment that had induced SE. Loose calluses (LC) cultured on solid medium also contained few SEs. Histological studies revealed various stages of SE development. Ploidy levels of regenerated plantlets were analyzed using flow cytometry and chromosome counting, the result showed 452 diploids and 102 tetraploids from 554 RC-derived plantlets, while all of the 107 LC-derived plantlets were diploid. No chimera was observed among plantlets. These results suggested that SEs originated from a single cell.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BA:

6-Benzyladenine

KT:

6-Furfurylaminopurine

CPPU:

N-(2-chloro-4-pyridyl)-N-phenylurea

NAA:

1-Naphthaleneacetic acid

IBA:

Indole butyric acid

Gln:

Glutamine

Arg:

Arginine

ME:

Malt extract

CH:

Casein(acid) hydrolysate

DAPI:

4,6-Diamdino-2-phenyloindole

MOPS:

3-(N-morpholino)propanesulfonic acid

PVP:

Polyvinylpyrrolidone

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Acknowledgments

This research was supported by a grant from the Special Fund for Forest Scientific Research in the Public Welfare (201404113), Supported by the 111 Project (B13007) and Supported by Program for Changjiang Scholars and Innovative Research Team in University (IRT13047).

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Authors and Affiliations

  1. National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing, 100083, People’s Republic of China

    Zhen Huang, Congping Xu, Yun Li, Peiqi Wang, Yuan Li & Xiangyang Kang

  2. Key Laboratory for Genetics and Breeding of Forest Trees and Ornamental Plants, Ministry of Education, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, People’s Republic of China

    Zhen Huang, Congping Xu, Yun Li, Peiqi Wang, Yuan Li & Xiangyang Kang

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  1. Zhen Huang
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  2. Congping Xu
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Correspondence to Xiangyang Kang.

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Huang, Z., Xu, C., Li, Y. et al. Induction of somatic embryogenesis by anther-derived callus culture and plantlet ploidy determination in poplar (Populus × beijingensis ) . Plant Cell Tiss Organ Cult 120, 949–959 (2015). https://doi.org/10.1007/s11240-014-0649-3

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