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Plant and Cell Physiology, 1999, Vol. 40, No. 12 1243-1252
© 1999

Localization of the Ca2+-Binding Protein, Bra r 1, in Anthers and Pollen Tubes

Takashi Okada1, Zhaojie Zhang2, Scott D. Russell2 and Kinya Toriyama1,3

1 Laboratory of Plant Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan
2 Department of Botany and Microbiology, University of Oklahoma Norman, Oklahoma, U.S.A.

3 For correspondence

Calcium plays an essential role during pollen development and pollen tube growth, and several Ca2+-binding protein are expressed in anthers. We have previously reported that Brassica pollen allergens encoded by Bra r 1 and Bra r 2 show sequence similarities to Ca2+-binding proteins [Toriyama et al. (1995) Plant Mol. Biol. 29: 1157]. Herein, we report that both genes are expressed in the diploid tapetum and haploid microspores, as detected by in situ RNA hybridization. Immunoblot analysis revealed that Bra r 1 and Bra r 2 were accumulated in anthers during pollen development. When pollen grains were suspended in an aqueous solution, both proteins were mainly detected in the pollen extracellular fraction, indicating that Bra r 1 and Bra r 2 are released from the pollen upon hydration. Localization of Bra r 1 was further investigated in sections of anthers and pollen tubes. Bra r 1 was detected in the tapetum, microspores and pollen grains. In longitudinal sections of cross-pollinated pistils, Bra r 1 was detected throughout pollen tubes elongating in transmitting-tissue. These findings suggest that Bra r 1 may be involved in pollen-pistil interaction and pollen tube growth.

(Received July 16, 1999; Accepted October 8, 1999)
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