Journal Article

Expression of <i>Bra r 1</i> Gene in Transgenic Tobacco and <i>Bra r 1</i> Promoter Activity in Pollen of Various Plant Species

Takashi Okada, Yoko Sasaki, Rieko Ohta, Noriyuki Onozuka and Kinya Toriyama

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 41, issue 6, pages 757-766
Published in print June 2000 | ISSN: 0032-0781
Published online June 2000 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/41.6.757
Expression of Bra r 1 Gene in Transgenic Tobacco and Bra r 1 Promoter Activity in Pollen of Various Plant Species

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  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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Bra r 1 encodes a Ca2+-binding protein specifically expressed in anthers of Brassica rapa. In this study, we isolated a genomic clone of Bra r 1 and found sequences similar to Pollen Box core motifs and LAT56/59 box, pollen-specific cis-acting element, in the 5′ upstream region of Bra r 1. Reporter gene fusion revealed that the Bra r 1 promoter directs male gametophytic expression in Nicotiana tabacum, Arabidopsis thaliana and B. napus, showing strong expression in mature pollen grains similar to that of endogenous Bra r 1. Genomic DNA of Bra r 1 was introduced into tobacco plants and the highest accumulation of Bra r 1 protein was observed in mature pollen in the same manner as reporter gene expression. Using in vitrogerminated pollen tubes of transgenic tobacco, we firstly demonstrated the subcellular localization of Bra r 1 in pollen tubes. Bra r 1 protein was distributed throughout the pollen tube of transgenic tobacco and slightly intense signals of Bra r 1 were observed in the tip region. In long-germinated pollen tubes, Bra r 1 was detected only in the cytoplasmic compartments while no signals were observed in the empty part of the pollen tube, indicating that cytoplasmic movement toward the tube tip is accompanied by Bra r 1. Hence, we suggest that Bra r 1 is involved in pollen germination and pollen tube growth.

Keywords: Bra r 1; Ca2+-binding protein; Beta-glucuronidase; Pollen allergen; Pollen tube; Promoter activity

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

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