A distinct member of the basic (class I) chitinase gene family in potato is specifically expressed in epidermal cells View Full Text


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Article Info

DATE

1999-04

AUTHORS

Gema Ancillo, Bärbel Witte, Elmon Schmelzer, Erich Kombrink

ABSTRACT

We have isolated cDNA clones encoding class I chitinase (ChtC) from potato leaves which share a high degree of nucleotide and amino acid sequence similarity to other, previously described basic (class I) chitinases (ChtB) from potato. Despite this similarity, characteristic features distinguish ChtC from ChtB, including an extended proline-rich linker region between the hevein and catalytic domains and presence of a potential glycosylation site (NDT) in the deduced protein. These differences are in accordance with the properties of purified chitinase C which is glycosylated and hence has a higher molecular mass in comparison to chitinase B. In contrast to the coding sequences, the 3′-untranslated regions of ChtC and ChtB exhibited a low degree of similarity, which allowed us to generate gene-specific probes to study the genomic organization and expression of both types of gene. Genomic DNA blots suggest that ChtC and ChtB are each encoded by one or two genes per haploid genome. RNA blot analysis showed that in healthy potato plants ChtC mRNA is most abundant in young leaves, the organs which also contain high levels of chitinase C. By contrast, ChtB mRNA abundance is highest in old leaves, which accumulate chitinase B. By in situ RNA hybridization with gene-specific probes we could demonstrate that ChtC mRNA in leaves is restricted to epidermal cells, whereas ChtB mRNA showed no distinct pattern of cell-type-specific localization. Infection of potato leaves with Phytophthora infestans, or treatment with fungal elicitor, ethylene, or wounding resulted in accumulation of both ChtC and ChtB mRNAs; however, for ChtC, in contrast to ChtB, no corresponding accumulation of the encoded protein could be detected, suggesting a post-transcriptional mechanism of regulation. Salicylic acid treatment did not induce accumulation of either mRNA. The possible functional implications of these findings for pathogen defence and developmental processes are discussed. More... »

PAGES

1137-1151

References to SciGraph publications

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  • 1983-09. A plant DNA minipreparation: Version II in PLANT MOLECULAR BIOLOGY REPORTER
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1006178425803

    DOI

    http://dx.doi.org/10.1023/a:1006178425803

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/10380801


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    42 schema:description We have isolated cDNA clones encoding class I chitinase (ChtC) from potato leaves which share a high degree of nucleotide and amino acid sequence similarity to other, previously described basic (class I) chitinases (ChtB) from potato. Despite this similarity, characteristic features distinguish ChtC from ChtB, including an extended proline-rich linker region between the hevein and catalytic domains and presence of a potential glycosylation site (NDT) in the deduced protein. These differences are in accordance with the properties of purified chitinase C which is glycosylated and hence has a higher molecular mass in comparison to chitinase B. In contrast to the coding sequences, the 3′-untranslated regions of ChtC and ChtB exhibited a low degree of similarity, which allowed us to generate gene-specific probes to study the genomic organization and expression of both types of gene. Genomic DNA blots suggest that ChtC and ChtB are each encoded by one or two genes per haploid genome. RNA blot analysis showed that in healthy potato plants ChtC mRNA is most abundant in young leaves, the organs which also contain high levels of chitinase C. By contrast, ChtB mRNA abundance is highest in old leaves, which accumulate chitinase B. By in situ RNA hybridization with gene-specific probes we could demonstrate that ChtC mRNA in leaves is restricted to epidermal cells, whereas ChtB mRNA showed no distinct pattern of cell-type-specific localization. Infection of potato leaves with Phytophthora infestans, or treatment with fungal elicitor, ethylene, or wounding resulted in accumulation of both ChtC and ChtB mRNAs; however, for ChtC, in contrast to ChtB, no corresponding accumulation of the encoded protein could be detected, suggesting a post-transcriptional mechanism of regulation. Salicylic acid treatment did not induce accumulation of either mRNA. The possible functional implications of these findings for pathogen defence and developmental processes are discussed.
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