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Plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis

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Abstract

A protocol for plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis was developed. Viable protoplasts were isolated from embryogenic cell suspensions at a yield of 1.2 × 107 protoplasts/ml packed cell volume (PCV). Liquid and feeder layer culture systems with medium-A and medium-B were used for protoplast culture. In liquid culture system, medium-B was more efficient for inducing cell division (17.5% at 14 days) and colony formation (6.7% at 28 days) than medium-A. However, all protoplast-derived cell colonies (PDCC) obtained from liquid culture system could not develop further. In feeder layer culture system, there was no significant difference between medium-A and medium-B on cell division and colony formation of the cultured protoplasts, and the cell division frequency at 14 days and colony formation frequency at 28 days were 24.5% and 11.2%, respectively, in medium-B. Comparative study on the effects of BAP (2.2 μM, 4.4 μM, 8.8 μM), zeatin (0.4 μM, 0.8 μM, 1.2 μM) and TDZ (0.2 μM, 0.4 μM, 0.6 μM) on embryo formation of PDCC from feeder-layer culture indicated that TDZ was best. TDZ at 0.4 μM induced 7906 mature embryos per ml PCV PDCC, which was 4-fold the frequency as with BAP at 4.4 μM, 7.5-fold as with zeatin at 0.8 μM and 150-fold as control medium (no mentioned cytokinins) after 45 days on M3 medium. About 44% of the mature embryos were converted into plantlets with poor root system after subculture on M4 medium. Root further development of regenerated plantlets was promoted by addition of activated charcoal (AC) to MS basal medium.

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Abbreviations

AC:

Activated charcoal

BAP:

6-benzylaminopurine

2,4-D:

2,4-dichlorophenoxyacetic acid

ECS:

Embryogenic cell suspensions

FDA:

Fluorescein diacetate

Gln:

Glutamine

IAA:

Indolacetic-3-acid

MES:

2-N-morpholino ethanesulfonic acid

NAA:

1-naphthaleneacetic acid

PCV:

Packed cell volume

PDCC:

Protoplast-derived cell colonies

TDZ:

Thidiazuron

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Acknowledgements

This work was supported by grants from National Natural Science Foundation of China (30400287), Guangdong Natural Science Foundation of China (04300538, 06023159, and 06300470), Guangdong Scientific and Technical Foundation (2006B20101014) and Guangzhou Scientific and Technical Foundation (2006Z3-E0281).

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Authors and Affiliations

  1. The Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Zhongshan (Sun Yat-sen) University, Guangzhou, 510275, Guangdong, China

    Wang Xiao, Xue-Lin Huang, Xia Huang, Ya-Ping Chen, Xue-Mei Dai & Jie-Tang Zhao

  2. Department of Biology, Guangdong Institute of Education, Guangzhou, 510303, Guangdong, China

    Wang Xiao

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  1. Wang Xiao
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  2. Xue-Lin Huang
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  3. Xia Huang
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Correspondence to Xue-Lin Huang or Xia Huang.

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Xiao, W., Huang, XL., Huang, X. et al. Plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis. Plant Cell Tiss Organ Cult 90, 191–200 (2007). https://doi.org/10.1007/s11240-007-9241-4

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