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Enhanced accumulation of fatty acids and triacylglycerols in transgenic tobacco stems for enhanced bioenergy production

Plant Cell Reports volume 33, pages 1041–1052 (2014)Cite this article

Abstract

Key message

We report a novel approach for enhanced accumulation of fatty acids and triacylglycerols for utilization as biodiesel in transgenic tobacco stems through xylem-specific expression of Arabidopsis DGAT1 and LEC2 genes.

Abstract

The use of plant biomass for production of bioethanol and biodiesel has an enormous potential to revolutionize the global bioenergy outlook. Several studies have recently been initiated to genetically engineer oil production in seeds of crop plants to improve biodiesel production. However, the “food versus fuel” issues have also sparked some studies for enhanced accumulation of oils in vegetative tissues like leaves. But in the case of bioenergy crops, use of woody stems is more practical than leaves. Here, we report the enhanced accumulation of fatty acids (FAs) and triacylglycerols (TAGs) in stems of transgenic tobacco plants expressing Arabidopsis diacylglycerol acyltransferase 1 (DGAT1) and LEAFY COTYLEDON2 (LEC2) genes under a developing xylem-specific cellulose synthase promoter from aspen trees. The transgenic tobacco plants accumulated significantly higher amounts of FAs in their stems. On an average, DGAT1 and LEC2 overexpression showed a 63 and 80 % increase in total FA production in mature stems of transgenic plants over that of controls, respectively. In addition, selected DGAT1 and LEC2 overexpression lines showed enhanced levels of TAGs in stems with higher accumulation of 16:0, 18:2 and 18:3 TAGs. In LEC2 lines, the relative mRNA levels of the downstream genes encoding plastidic proteins involved in FA synthesis and accumulation were also elevated. Thus, here, we provide a proof of concept for our approach of enhancing total energy yield per plant through accumulation of higher levels of FAs in transgenic stems for biodiesel production.

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Acknowledgments

This work was supported by the World Class University project of the Ministry of Science and Technology of Korea (R31-2009-000-20025-0). The authors declare that they have no conflict of interest.

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Author notes

  1. Akula Nookaraju

    Present address: Kaveri Seed Company Ltd., S.D.Road, Secunderabad, 500003, India

Authors and Affiliations

  1. Department of Bioenergy Science and Technology, Chonnam National University, Kwangju, 500-757, South Korea

    Akula Nookaraju, Shashank K. Pandey, Takeshi Fujino, Ju Young Kim, Mi Chung Suh & Chandrashekhar P. Joshi

  2. Department of Biological Sciences, Biotechnology Research Center, School of Forest Resources and Environmental Science, Michigan Technological University, Houghton, MI, 49931, USA

    Akula Nookaraju & Chandrashekhar P. Joshi

Authors
  1. Akula Nookaraju
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  2. Shashank K. Pandey
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  3. Takeshi Fujino
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  4. Ju Young Kim
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  5. Mi Chung Suh
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  6. Chandrashekhar P. Joshi
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Corresponding author

Correspondence to Chandrashekhar P. Joshi.

Additional information

Communicated by M. Prasad.

Akula Nookaraju and Shashank K. Pandey have contributed equally to this work.

Electronic supplementary material

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299_2014_1582_MOESM1_ESM.tif

Fig. S1 Schematic representation of T-DNA of binary vectors used for transformation studies. p1A-AtLEC2 contained Arabidopsis LEC2-TF gene driven by PtdCesA8A promoter (a), p1A-AtDGAT1 contained Arabidopsis DGAT1 driven by PtdCesA8A promoter (b) and control vector p1A-GUS (c) is a modified pBI121 vector, where CaMV35S promoter was replaced by PtdCesA8A promoter to drive β-glucuronidase (GUS) gene. The two Arabidopsis genes were cloned between PtdCesA8A promoter and NOS-T replacing GUS. The T-DNA of binary vectors also contained neomycin phosphotransferase (NPTII) gene driven by NOS-P for kanamycin-resistant plant selection. LB, left border and RB, right border (TIFF 69 kb)

299_2014_1582_MOESM2_ESM.tif

Fig. S2 Cellulose contents in stems of control and transgenic lines of tobacco. The error bars represent the SEM of three independent observations (n = 3). Cellulose content in stems of individual lines (a) and average contents for DGAT and LEC2 lines (b) was provided (TIFF 65 kb)

299_2014_1582_MOESM3_ESM.tif

Fig. S3 Glucose release from acetonitrile-washed cell wall residues of control and transgenic stems of tobacco. Glucose release was expressed in terms of mg glucose released per mg tissue. The error bars represent the SEM of three independent observations (n = 3). Glucose release from stems of individual lines (a) and average glucose release from stems of DGAT and LEC2 lines (b) was provided (TIFF 67 kb)

Table S1 Supplementary material 4 (DOCX 12 kb)

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Nookaraju, A., Pandey, S.K., Fujino, T. et al. Enhanced accumulation of fatty acids and triacylglycerols in transgenic tobacco stems for enhanced bioenergy production. Plant Cell Rep 33, 1041–1052 (2014). https://doi.org/10.1007/s00299-014-1582-y

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  • DOI: https://doi.org/10.1007/s00299-014-1582-y

Keywords

  • Biodiesel
  • Renewable energy
  • Tobacco
  • Triacylglycerols
  • Xylem

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