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Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

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Abstract

This study investigated the delivery of a SV40 promoter driving lacZ gene into cells of Kappaphycus alvarezii using particle bombardment. Thallus pieces 0.5–0.8 mm in diameter and 1 cm in length were prepared as gene recipients. Bombardment parameters of 450 psi (rupture pressures) × 6 cm (particle travel distances), 650 psi × 6 cm, 1,100 psi × 6 cm and 1,100 psi × 9 cm were used. A significant increase in transformation efficiency from about 33% under the rupture pressure of 450 psi to 87% at 650 psi was observed in transformed thalli. Most of the positive cells appeared in epidermal cells bombarded at 450 psi, whereas positive signals were seen in both epidermal and medullary cells at 650 psi. No positive transient expression was detected at a bombardment of 1,100 psi, or in negative or blank controls. For the conditions tested, the best parameter was obtained at 650 psi at a distance of 6 cm. Thus, the strategy of taking vegetative thalli as recipients, using particle bombardment, and combining this with micro-propagation, together with developing an in vivo selectable marker, is a viable way to produce stable transformants, to eliminate chimeric expression, and to achieve transgenic breeding in K. alvarezii.

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Abbreviations

GUS:

β-glucuronidase gene

lacZ :

β-galactosidase gene

CaMV:

Cauliflower mosaic virus

SV40:

Simian virus 40

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Acknowledgments

This work was funded by Special Project for Marine Public Welfare Industry (200705010), National High Technology Research and Development Program of China (2009AA10Z106, 2006AA100311, 20060110A402) and the Innovative Key Project of the Chinese Academy of Sciences (KZCX2-YW-209, KSCX2-YW-G-002).

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Authors and Affiliations

  1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Nanhai Road 7, 266071, Qingdao, China

    Jinfeng Wang, Peng Jiang, Yulin Cui, Xiangyuan Deng, Fuchao Li, Jianguo Liu & Song Qin

  2. Graduate University of Chinese Academy of Sciences, 100049, Beijing, China

    Jinfeng Wang, Yulin Cui & Xiangyuan Deng

Authors
  1. Jinfeng Wang
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  2. Peng Jiang
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  3. Yulin Cui
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  4. Xiangyuan Deng
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  5. Fuchao Li
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  6. Jianguo Liu
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  7. Song Qin
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Correspondence to Peng Jiang or Song Qin.

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Wang, J., Jiang, P., Cui, Y. et al. Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement. Aquacult Int 18, 1027–1034 (2010). https://doi.org/10.1007/s10499-010-9320-0

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