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Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa

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Abstract

As perennial plants, Populus species are constantly exposed to environmental stresses, such as wounding and pathogen attack, which lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. In this study, a cDNA clone encoding CHS was isolated from Populus trichocarpa by reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA, named PtrCHS4, was 1,314 bp with a 1,173 bp open reading frame that corresponded to a deduced protein of 391 amino acid residues. Multiple sequence alignments showed that PtrCHS4 shared high homology with CHS proteins from other plants. Phylogenetic analysis revealed that PtrCHS4 was most closely related to PhCHS from Petunia hybrida and NaCHS from Nicotiana attenuata. Semi-quantitative RT-PCR analysis identified that the PtrCHS4 gene was abundantly expressed in the leaves and stems, while its expression was drastically reduced in the roots. Transcript abundance of PtrCHS4 was stimulated by 2.5-fold within 24 h of wounding treatment. Promoter analysis confirmed that the PtrCHS4 promoter was capable of directing expression of the GUS reporter in both wounded and unwounded leaves of transgenic Chinese white poplar (P. tomentosa Carr.), indicating that the PtrCHS4 promoter is systemically responsive to wounding stimuli. Furthermore, promoter deletion analysis showed that the proximal 1,592 bp from the transcription start site were required for promoter activation by jasmonic acid and the −1,096 to −148 region was proved to be necessary for establishing wound-induced pattern of expression.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (3081576), the National Key Project for Research on Transgenic Plant (2011ZX08010-003), the Natural Science Foundation Project of CQ CSTC (CSTC, 2009BA1004, 2009BB0004) and the Fundamental Research Funds for the Central Universities (XDJK2009B018).

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Authors and Affiliations

  1. Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, State Key Laboratory of Eco-environment and Bio-resource of Three Gorges Reservoir Region, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing, 400715, China

    Yiming Sun, Qiaoyan Tian, Li Yuan, Yuanzhong Jiang, Yan Huang, Min Sun, Shaohu Tang & Keming Luo

  2. Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Southwest University, No. 1, Tiansheng Road, Beibei, Chongqing, 400715, China

    Yiming Sun, Qiaoyan Tian, Li Yuan, Yuanzhong Jiang, Yan Huang, Min Sun, Shaohu Tang & Keming Luo

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  1. Yiming Sun
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  2. Qiaoyan Tian
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  3. Li Yuan
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  5. Yan Huang
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Correspondence to Keming Luo.

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Communicated by P. Lakshmanan.

Y. Sun, Q. Tian contributed equally to this work.

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Sun, Y., Tian, Q., Yuan, L. et al. Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa . Plant Cell Rep 30, 1661–1671 (2011). https://doi.org/10.1007/s00299-011-1075-1

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