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An endochitinase gene expressed at high levels in the stylar transmitting tissue of tomatoes

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Abstract

A gene (pMON9617; Chi2;1) identified by screening a tomato pistil cDNA library has been found to encode a protein similar in sequence to class II chitinases. Using a baculovirus expression system we show that the Chi2;1 protein is an active endochitinase. The Chi2;1 protein is most similar in sequence to a previously described stylar chitinase (SK2) isolated from potato. Both proteins lack the diagnostic N-terminal cysteine-rich regions and the C-terminal vacuolar targeting signals of class I chitinases and appear to define a novel type of class II endochitinases associated with flowers. Chi2;1 is expressed predominantly in floral organs and its expression within these organs is temporally regulated. The highest level of expression is found in the transmitting tissue of the style where Chi2;1 mRNA begins to accumulate just prior to anthesis. In vegetative tissue, low levels of Chi2;1 mRNA were detected, and these levels did not increase in response to wounding or treatment with ethephon. mRNA from Chi2;1 orthologs was not detected in most other angiosperms tested, even including some members of the Solanaceae, and it is therefore unlikely that Chi2;1 is essential for stylar function. A fragment containing 1.4 kilobase pairs of 5′-flanking DNA from the Chi2;1 gene was shown to drive high-level expression of an attached reporter gene in the styles of transgenic tomatoes. This fragment has utility for engineering expression of other coding regions in styles.

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  1. K. Harikrishna

    Present address: Department of Biotechnology, Faculty of Biotechnology and Food Science, University Pertanian Malaysia, UPM-Serdang, 43400, Selangor Darul Ehsan, Malaysia

Authors and Affiliations

  1. Section of Molecular and Cellular Biology, Division of Biological Sciences, University of California, 95616, Davis, CA, USA

    K. Harikrishna, Rachanee Jampates-Beale, Stephen B. Milligan & Charles S. Gasser

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  1. K. Harikrishna
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  2. Rachanee Jampates-Beale
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Harikrishna, K., Jampates-Beale, R., Milligan, S.B. et al. An endochitinase gene expressed at high levels in the stylar transmitting tissue of tomatoes. Plant Mol Biol 30, 899–911 (1996). https://doi.org/10.1007/BF00020802

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