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Changes in freezing tolerance in hybrid poplar caused by up- and down-regulation of PtFAD2 gene expression

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Abstract

In plant species, the level of polyunsaturated fatty acids (PUFAs) is essential for cold acclimation. To test whether changes in PUFA levels can lead to the alteration of freezing tolerance in poplar trees, we up- and down-regulated a Populus tomentosa Δ-12 fatty acid desaturase gene (PtFAD2) in the hybrid poplar (P. alba × P. glandulosa) clone 84 K. Real-time PCR results demonstrated that compared to untransformed control lines, the transcriptional level of PtFAD2 increased by up to 90% in over-expressing poplar lines (line OE-1) and decreased in down-regulated RNAi lines by up to 64% (line DR-1). As a result, the content of linoleic (C18:2) and linolenic (C18:3) unsaturated fatty acids (FAs) in total FAs increased by 7.5 and 3.9%, respectively, in the OE-1 line and decreased by 14.4 and 5.4% in the DR-2 line when compared to non-transgenic lines. After freezing treatment at −4°C for 3 h without pre-cold acclimation, the survival rates of the PtFAD2-over-expressing cuttings were significantly higher (60% for OE-1) than those of non-transgenic plants (36.7%) and down-regulated lines (10% for DR-2). These results clearly demonstrate that the expression level of PUFAs substantially affected the freezing tolerance of hybrid poplar cuttings and could thus be utilized as an effective strategy to improve poplar anti-freezing traits through genetic engineering biotechnology.

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Acknowledgments

We thank Dr. Liu Qing of the CSIRO Plant Industry in Australia for critical review of this manuscript and for providing the pHurricane vector. This study was supported by grants from the High Tech Development Program (2006AA100109) and Introduction of Foreign Advanced Agricultural Science and Technology into China (No. 2006-4-C01).

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Authors and Affiliations

  1. Lab of Biotechnology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing, 100091, People’s Republic of China

    Zhou Zhou, Min-Jie Wang, Shu-Tang Zhao, Jian-Jun Hu & Meng-Zhu Lu

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  1. Zhou Zhou
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  2. Min-Jie Wang
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  3. Shu-Tang Zhao
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  4. Jian-Jun Hu
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  5. Meng-Zhu Lu
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Correspondence to Meng-Zhu Lu.

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Z. Zhou and M.-J. Wang have the same contribution to this paper.

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11248_2009_9349_MOESM1_ESM.tif

Supplementary Figure 1 Diagrammatic representation of the PtFAD2 up-regulated (A) and down-regulated (B) constructs. NPTII: neomycin phosphotransferase II; CaMV35S: Cauliflower mosaic virus 35S promoters; At FAD2 intron 1.2 kb: the 1.2-kb fragment of the second intron of the Arabidopsis thaliana AtFAD2 served as the spacer for the hairpin structure formation in pHurricane; NOS Pro and NOS Ter: Nopaline synthase promoter and terminator, respectively. (TIFF 1886 kb)

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Zhou, Z., Wang, MJ., Zhao, ST. et al. Changes in freezing tolerance in hybrid poplar caused by up- and down-regulation of PtFAD2 gene expression. Transgenic Res 19, 647–654 (2010). https://doi.org/10.1007/s11248-009-9349-x

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  • DOI: https://doi.org/10.1007/s11248-009-9349-x

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