Journal Article

Transcriptional Expression Characteristics and Subcellular Localization of ADP-Glucose Pyrophosphorylase in the Oil Plant <i>Perilla frutescens</i>

Sang-Bong Choi, Kyung-Hwan Kim, Ibrahim Halil Kavakli, Seong-Kon Lee and Thomas W. Okita

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 2, pages 146-153
Published in print February 2001 | ISSN: 0032-0781
Published online February 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce019
Transcriptional Expression Characteristics and Subcellular Localization of ADP-Glucose Pyrophosphorylase in the Oil Plant Perilla frutescens

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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Three ADP-glucose pyrophosphorylase clones were isolated from the cotyledon cDNA library of the oil plant, Perilla frutescens, and their intracellular localization investigated. Two of three cDNAs (PfagpS1 and PfagpS2) were homologous to the catalytic small subunit of AGPases found in other plants, while the third clone (PfagpL) was highly similar to the large subunit type. Transcripts for PfagpS1 and PfagpS2 were observed in both photosynthetic and non-photosynthetic tissue, showing the highest expression in the stem, while PfagpL transcripts were abundantly expressed in stem and cotyledon. To evaluate the subcellular localization of PfagpS2 and PfagpL as well as the maize BT2, N-terminus-GFP DNA fusion were constructed and transformed into tobacco plants. Immunoblot analysis showed that the expressed PfagpS2- and PfagpL-GFP fusions were targeted to the plastid in the heterologous tobacco system whereas the BT2-GFP remained intact, suggesting a cytoplasmic location. These intracellular assignments were confirmed by direct confocal microscopic examination. GFP signals were localized to the cytoplasm as well as in the nucleus in BT2-GFP plants, and to the plastids in PfagpS2- and PfagpL-GFP plants. Our results indicate that Perilla cotyledons contain multiple AGPase subunits, of which at least two isoforms and very likely the third, are plastidial in nature.

Keywords: Key words: ADP-glucose pyrophosphorylase — BT2 — GFP localization — Perilla frutescens — Plastids.; Abbreviations: AGPase, ADP-glucose pyrophosphorylase; PfagpS1, small subunit 1 of Perilla AGPase; PfagpS2, small subunit 2 of Perilla AGPase; PfagpL, large subunit of Perilla AGPase; SDS-PAGE, SDS-polyacrylamide gel electrophoresis; BT2 and SH2, small and large subunit of maize endosperm AGPase.; The nucleotide sequences reported in this paper have been submitted to GenBank under accession numbers AF249915 (PfagpS1), AF249916 (PfagpS2) and AF249917 (PfagpL).

Journal Article.  5352 words.  Illustrated.

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

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