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Haploid plants from anther cultures of poplar (Populus × beijingensis)

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An Erratum to this article was published on 04 December 2013

Abstract

Homozygous genotypes are valuable for genetic and genomic studies in higher plants. However, obtaining homozygous perennial woody plants using conventional breeding techniques is currently a challenge due to a long juvenile period, high heterozygosity, and substantial inbreeding depression. In vitro androgenesis has been used to develop haploid and doubled haploid plants. In the present study, we report the regeneration of haploid lines of poplar (Populus × beijingensis) via anther culture. Anthers at the uninucleate stage were induced to produce callus using three basic media. Two auxins (naphthalene acetic acid [NAA] and 2,4-dichloro-phenoxyacetic acid [2,4-D]), and two cytokinin (kinetin [KT] and 6-benzyladenine [BA]) were tested to explore the influence of plant growth regulators on callus response. H medium (Bourgin and Nitsch 1967) supplemented with 1.0 mg/L NAA and 1.0 mg/L KT induced the highest rate of callus formation. When callus obtained from anthers were subcultured in MS medium containing 1.0 mg/L BA and 0.2 mg/L NAA, followed by transfer to half-strength MS medium supplemented with indole-3-butyric acid (0.2–0.5 mg/L), the formation of regenerated plantlets increased dramatically. Inclusion of gibberellic acid (0.02–0.2 mg/L) in addition to a combination of BA (0.6 mg/L)-NAA (0.2 mg/L) in the culture medium resulted in enhanced frequency of shoot development, as well as greater internode elongation. Ploidy analysis of 580 regenerated plants, using both flow cytometry and chromosome counting, revealed 10.3 % haploid and 1.0 % triploid plantlets. The remaining plantlets were all diploid.

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Abbreviations

MS:

Murashige and Skoog (1962) medium

H:

Bourgin and Nitsch (1967) medium

N6 :

Chu et al. (1975) medium

WPM:

Woody plant medium

PGRs:

Plant growth regulators

2,4-D:

2, 4-dichlorophenoxyacetic acid

BA:

6-benzyladenine

NAA:

Naphthalene acetic acid

KT:

Kinetin

GA3 :

Gibberellic acid

IBA:

Indole-3-butytric acid

DH:

Doubled haploid

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Acknowledgments

This work was supported by the Major State Basic Research Development Program (2012CB114505), Key Projects in the National Science & Technology Pillar Program during the 12th 5-year Plan Period (2012BAD01B0302), and National High-tech R&D Program (2013AA102703, 2011AA100201) of China.

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Authors and Affiliations

  1. National Engineering Laboratory for Tree Breeding (NDRC), Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education (MOE), Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration (SFA), Beijing Forestry University, Beijing, 100083, People’s Republic of China

    Ying Li, Hao Li, Zhong Chen, Le-Xiang Ji, Mei-Xia Ye, Jia Wang, Lina Wang & Xin-Min An

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  1. Ying Li
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Correspondence to Xin-Min An.

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Li, Y., Li, H., Chen, Z. et al. Haploid plants from anther cultures of poplar (Populus × beijingensis). Plant Cell Tiss Organ Cult 114, 39–48 (2013). https://doi.org/10.1007/s11240-013-0303-5

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