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Optimization of in vitro multiple shoot clump induction and plantlet regeneration of Kentucky bluegrass (Poa pratensis)

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Abstract

An efficient protocol for Kentucky bluegrass (Poa  pratensis L.) in vitro culture was established using shoot apices of seedlings as explants. The optimal procedure of this protocol for majority of the genotypes was that meristematic cell clumps and small calluses were firstly induced from the bases of explants on initial culture medium supplemented with 0.9 μM 2,4-d and 8.9 μM 6-BA for 20 d, then were separated and transferred to shoot clumps induction medium containing 8.9 μM 6-BA for the formation of multiple shoot clumps. The percentage of multiple shoot clumps and numbers of shoots per clump were deeply related with the combinations of different plant growth regulators, duration of initial culture, the intensity of illumination and genotypes. Histological observation of the induced explants revealed that the meristematic cell clumps were produced from repeated division of the cortical cells and original meristematic primodium cells of explants, and the multiple shoots were formed via organogenesis pathway in the meristematic cell regions of cultures on shoot clumps induction medium. In this study, plantlets were efficiently regenerated on large scale from seven cultivars of Kentucky bluegrass. Hence the meristematic cell clumps and small calluses in this protocol could be considered good targets for genetic transformation of Kentucky bluegrass.

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Abbreviations

6-BA:

6-benzylaminopurine

2,4-d :

2,4-dichlorophenoxy acetic acid

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Acknowledgement

This research was supported by the National Project for Transgenic Plant Research and Industrialization of China (JY2002-B-006) and Scientific Research Award Fund for Excellent Mid-age and Young Scientists in Shandong Province (2004BS06004).

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Authors and Affiliations

  1. School of life science, Shandong University, Jinan, Jinan 250100, Shandong, China

    X.R. Hu, A.F. Yang, K.W. Zhang, J. Wang & J.R. Zhang

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  1. X.R. Hu
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  2. A.F. Yang
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  3. K.W. Zhang
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  4. J. Wang
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  5. J.R. Zhang
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Correspondence to J.R. Zhang.

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Hu, X., Yang, A., Zhang, K. et al. Optimization of in vitro multiple shoot clump induction and plantlet regeneration of Kentucky bluegrass (Poa pratensis). Plant Cell Tiss Organ Cult 84, 90–99 (2006). https://doi.org/10.1007/s11240-005-9009-7

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