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In vitro propagation of four threatened Paphiopedilum species (Orchidaceae)

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Abstract

The effects of seed maturity, media type, carbon source, and organic nutrient additives on seed germination, protocorm development, and plant growth of Paphiopedilum villosum var. densissimum Z. J. Liu et S. C. Chen were investigated. Micropropagation frequency was enhanced through the use of 200-day-old seed, Knudson C (KC) medium, and the presence of both glucose and coconut milk in the medium. The effects of various plant growth regulators on the frequency of shoot organogenesis in four Paphiopedilum species were also investigated. Explants of P. villosum var. densissimum and P. insigne (Lindl.) Stein incubated in the presence of 5 mg l−1 6-benzyladenine (BA) with 0.5 mg l−1 α-naphthalene acetic acid (NAA) and 0.2 mg l−1 BA with 0.1 mg l−1 NAA, respectively, showed a twofold increase in the frequency of shoot organogenesis. For explants of P. bellatulum (Rchb. f.) Stein and P. armeniacum S. C. Chen et F. Y. Liu, the combination of 5.5 mg l−1 BA with 0.5 mg l−1 NAA and 4 mg l−1 BA with 0.1 mg l−1 NAA, respectively, resulted in the highest frequencies of shoot organogenesis.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

AC:

Activated charcoal

BA:

6-Benzyladenine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

KT:

Kinetin

LH:

Lactalbumin hydrolysate

NAA:

α-Naphthalene acetic acid

TDZ:

1-Phenyl-3-(1, 2, 3-thiadiazol-5-yl)-urea

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Acknowledgments

The authors thank Chunyan Han, Guangwan Hu, and Yuan Liu for their kind help. This work was supported by the Natural Science Foundation of Yunnan (2005C0053 M), the Ministry of Science and Technology of China (2005DK21006) and the Ministry of Education of China (B08044 & MUC-985-3-3).

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Authors and Affiliations

  1. Kunming Institute of Botany, Chinese Academy of Sciences, 650204, Kunming, China

    Bo Long, Zhi-ying Cheng & Chun-lin Long

  2. Graduate School, Chinese Academy of Sciences, 100039, Beijing, China

    Bo Long

  3. Department of Horticulture, Virginia Polytechnic and State University, Blacksburg, VA, 24061, USA

    Alex X. Niemiera

  4. College of Life and Environmental Sciences, Minzu University of China, 100081, Beijing, China

    Chun-lin Long

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  1. Bo Long
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  2. Alex X. Niemiera
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  4. Chun-lin Long
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Correspondence to Chun-lin Long.

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Long, B., Niemiera, A.X., Cheng, Zy. et al. In vitro propagation of four threatened Paphiopedilum species (Orchidaceae). Plant Cell Tiss Organ Cult 101, 151–162 (2010). https://doi.org/10.1007/s11240-010-9672-1

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  • DOI: https://doi.org/10.1007/s11240-010-9672-1

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