Journal Article

Molecular Cloning and Characterization of a cDNA Encoding a Novel Apoplastic Protein Preferentially Expressed in a Shikonin-Producing Callus Strain of <i>Lithospermum erythrorhizon</i>

Yoshimi Yamamura, F. Pinar Sahin, Akito Nagatsu and Hajime Mizukami

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 44, issue 4, pages 437-446
Published in print April 2003 | ISSN: 0032-0781
Published online April 2003 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcg057
Molecular Cloning and Characterization of a cDNA Encoding a Novel Apoplastic Protein Preferentially Expressed in a Shikonin-Producing Callus Strain of Lithospermum erythrorhizon

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A cDNA (LEPS-2) encoding a novel cell wall protein was cloned from shikonin-producing callus tissues of Lithospermum erythrorhizon by differential display between a shikonin-producing culture strain and a non-producing strain. The LEPS-2 cDNA encoded a polypeptide of 184 amino acids. The deduced amino acid sequence exhibited no significant homology with known proteins. Expression of LEPS-2 gene as well as accumulation of LEPS-2 protein was highly correlated with shikonin production in L. erythrorhizon cells in culture. In the intact plant, expression of LEPS-2 was detected only in the roots where shikonin pigments accumulated. Cell fractionation experiments and immunocytochemical analysis showed that the protein was localized in the apoplast fraction of the cell walls. The shikonin pigments were also stored on the cell walls as oil droplets. These results indicate that expression of the LEPS-2 is closely linked with shikonin biosynthesis and the LEPS-2 protein may be involved in the intra-cell wall trapping of shikonin pigments.

Keywords: Keywords: Apoplastic protein — Differential display — High-producing strain — Lithospermum erythrorhizon — Shikonin biosynthesis.; The nucleotide sequence of LEPS-2 reported in this paper has been deposited in the DDBJ/EMBL/GenBank Nucleotide Sequence Database under the accession number AB047916.

Journal Article.  5745 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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