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Genetic manipulation, a feasible tool to enhance unique characteristic of Chlorella vulgaris as a feedstock for biodiesel production

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Abstract

Developing a reliable technique to transform unicellular green algae, Chlorella vulgaris, could boost potentials of using microalgae feedstock in variety of applications such as biodiesel production. Volumetric lipid productivity (VLP) is a suitable variable for evaluating potential of algal species. In the present study, the highest VLP level was recorded for C. vulgaris (79.08 mg l−1 day−1) followed by 3 other strains studied; C. emersonii, C. protothecoides, and C. salina by 54.41, 45 and 18.22 mg l−1day−1, respectively. Having considered the high productivity of C. vulgaris, it was selected for the preliminary transformation experiment through micro-particle bombardment. Plasmid pBI 121, bearing the reporter gene under the control of CaMV 35S promoter and the kanamycin marker gene, was used in cells bombardment. Primary selection was done on a medium supplemented by 50 mg l−1 kanamycin. After several passages, the survived cells were PCR-tested to confirm the stability of transformation and then were found to exhibit β-glucuronidase (GUS) activity in comparison with the control cells. Southern hybridization of npt II probe with genomic DNA revealed stable integration of the cassette in three different positions in the genome. The whole process was successfully implemented as a pre-step to transform the algal cells by genes involved in lipid production pathway which will be carried out in our future studies.

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Acknowledgments

The authors would also like to thank Agricultural Biotechnology Research Institute of Iran (ABRII) and Biofuel Research Team (BRTeam) for financing this study.

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Authors and Affiliations

  1. Biotechnology and Plant Breeding Department, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

    Ahmad Farhad Talebi & Abdolreza Bagheri

  2. Energy Crops Genetic Engineering Group, Biofuel Research Team (BRTeam), Microbial Biotechnology and Biosafety Department, Agricultural Biotechnology Research Institute of Iran (ABRII), Mahdasht Road, 31535-1897, Karaj, Iran

    Ahmad Farhad Talebi, Masoud Tohidfar, Meisam Tabatabaei & Seyed Kaveh Mohtashami

  3. Genetic Engineering and Tissue Culture Department, Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran

    Masoud Tohidfar & Motahhareh Mohsenpor

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  1. Ahmad Farhad Talebi
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  2. Masoud Tohidfar
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  3. Meisam Tabatabaei
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  4. Abdolreza Bagheri
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  5. Motahhareh Mohsenpor
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  6. Seyed Kaveh Mohtashami
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Correspondence to Meisam Tabatabaei.

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Talebi, A.F., Tohidfar, M., Tabatabaei, M. et al. Genetic manipulation, a feasible tool to enhance unique characteristic of Chlorella vulgaris as a feedstock for biodiesel production. Mol Biol Rep 40, 4421–4428 (2013). https://doi.org/10.1007/s11033-013-2532-4

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  • DOI: https://doi.org/10.1007/s11033-013-2532-4

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