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Accumulation of sweet protein monellin is regulated by thepsbA 5′UTR in tobacco chloroplasts

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Abstract

Post-transcriptional RNA processing and translational regulations are important steps for gene expression. To analyze the 5′UTRof psbA that enhances translation of the sweet protein monellin in chloroplasts, we cloned the monellin gene, with and without thepsbA 5′UTR, into the chloroplast expression vector for chloroplast transformation. Transgenic plants were identified as being transplastomic via PCR and Southern blot analyses. We also observed non-specific recombination during tobacco chloroplast transformation. Analyses of the transcription patterns showed that intercistronic cleavage of the psbA mRNA 5′ untranslated (UTR) region was functional at the mature stage, with the monocistronic mRNA ofmonellin increasing while its dicistronic mRNA decreased. Moreover, monellin accumulation accounted for 2.3% of the total soluble protein at the mature stage, but only 1.3% at the young stage in transplastomic lines that contained the 5′UTRof psbA. These results suggest that activation of the endonucleolytic cleavage of thepsbA 5′UTR element depends on chloroplast developmental conditions, and that it enhances the accumulation of sweet protein monellin in those chloroplasts.

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Authors and Affiliations

  1. National Institute of Agricultural Biotechnology, RDA, 441-707, Suwon, Korea

    Kyung Hee Roh, Kong-Sik Shin, Yeon-Hee Lee, Seok-Cheol Seo & Seung-Bum Lee

  2. Department of Horticultural Science, Seoul National University, 151-742, Seoul, Korea

    Hyo-Guen Park

  3. Department of Molecular Biology and Microbiology, Biomolecular Science, University of Central Florida, 32816-2364, Orlando, FL, USA

    Henry Daniell

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  1. Kyung Hee Roh
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  2. Kong-Sik Shin
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Correspondence to Seung-Bum Lee.

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Roh, K.H., Shin, KS., Lee, YH. et al. Accumulation of sweet protein monellin is regulated by thepsbA 5′UTR in tobacco chloroplasts. J. Plant Biol. 49, 34–43 (2006). https://doi.org/10.1007/BF03030786

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