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The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters

  • Genetic Transformation and Hybridization
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Abstract

Here we report the effect of the 35S promoter sequence on activities of the tissue- and organ-specific gene promoters in tobacco plants. In the absence of the 35S promoter sequence the AAP2 promoter is active only in vascular tissues as indicated by expression of the AAP2:GUS gene. With the 35S promoter sequence in the same T-plasmid, transgenic plants exhibit twofold to fivefold increase in AAP2 promoter activity and the promoter becomes active in all tissue types. Transgenic plants hosting the ovary-specific AGL5:iaaM gene (iaaM coding an auxin biosynthetic gene) showed a wild-type phenotype except production of seedless fruits, whereas plants hosting the AGL5:iaaM gene along with the 35S promoter sequence showed drastic morphological alterations. RT-PCR analysis confirms that the phenotype was caused by activation of the AGL5:iaaM gene in non-ovary organs including roots, stems and flowers. When the pollen-, ovule- and early embryo-specific PAB5:barnase gene (barnase coding a RNase gene) was transformed, the presence of 35S promoter sequence drastically reduced transformation efficiencies. However, the transformation efficiencies were restored in the absence of 35S promoter, indicating that the 35S promoter might activate the expression of PAB5:barnase in non-reproductive organs such as calli and shoot primordia. Furthermore, if the 35S promoter sequence was replaced with the NOS promoter sequence, no alteration in AAP2, AGL5 or PAB5 promoter activities was observed. Our results demonstrate that the 35S promoter sequence can convert an adjacent tissue- and organ-specific gene promoter into a globally active promoter.

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Abbreviations

AAP2 :

H+/amino acid permease gene 2

AGL5 :

AGAMOUS-like MADS box protein 5

PAB5 :

Poly (A) binding protein 5

iaaM :

IAA monooxygenase

CaMV 35S :

Cauliflower mosaic virus 35S promoter

GUS :

β-Glucuronidase

NOS :

Nopaline synthase

nptII :

Neomycin phosphotransferase

BA :

Benzylaminopurine

NAA :

Naphthaleneacetic acid

X-Gluc :

5-Bromo-4-chloro-3-indolyl-b-d-glucuronic acid

MU :

4-Methyl umbelliferone

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Acknowledgments

We would like to thank Dr. Robert W Hartley (Laboratory of Cellular and Developmental Biology, NIDDK, National Institutes of Health) for generously providing plasmids pMT316 and pMT1002. We also thank Mr. Fengtao Luo for his work in plant genetic transformation and GUS staining analysis. This work was supported by USDA, CPBR/DOE and UConn Research Foundation to Yi Li, and National Natural Science Foundation of China (NSFC: 30530490) and National Basic Research and Development Program (2004CB117300) to Yan Pei.

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Authors and Affiliations

  1. Department of Plant Science, University of Connecticut, Unit 5082, Storrs, CT, 06269, USA

    Xuelian Zheng, Wei Deng, Hui Duan, Yongqin Chen, Richard McAvoy, Yan Pei & Yi Li

  2. Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of Agriculture, Biotechnology Research Center, Southwest University, Chongqing, 400716, China

    Xuelian Zheng, Keming Luo, Shuiqing Song & Yan Pei

  3. Faculty of Life Sciences, Hubei University, Wuhan, 430062, China

    Yongqin Chen

  4. Department of Cell and Structural Biology, University of Illinois, Urbana, IL, 61801, USA

    Hui Duan

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  1. Xuelian Zheng
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  2. Wei Deng
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Correspondence to Yan Pei or Yi Li.

Additional information

Communicated by P. Lakshmanan.

Xuelian Zheng and Wei Deng contributed equally to this work and are considered co-first authors.

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Zheng, X., Deng, W., Luo, K. et al. The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters. Plant Cell Rep 26, 1195–1203 (2007). https://doi.org/10.1007/s00299-007-0307-x

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