Partial male sterility in transgenic tobacco carrying an antisense gene for alternative oxidase under the control of a tapetum-specific promoter |
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Authors: | Kitashiba Hiroyasu Kitazawa Erina Kishitani Sachie Toriyama Kinya |
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Institution: | (1) Laboratory of Plant Breeding, Faculty of Agriculture, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan |
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Abstract: | The alternative oxidase of plant mitochondria is the terminal oxidase of the cyanide-insensitive respiratory pathway and is
encoded by a nuclear gene. A 1 kb genomic fragment including exon 3 of the alternative oxidase was amplified by PCR from the
genome of Arabidopsis thaliana. This fragment was connected to a tapetum-specific promoter in the antisense orientation and
then introduced into tobacco. The pollen viability in three transgenic plants ranged from 2% to 60%. The reduced pollen viability
cosegregated with the transgene in a selfed progeny. Immunolocalization of alternative oxidase protein in the immature flower
bud section indicated that expression of alternative oxidase protein in tapetum of the transgenic plant was much lower than
that of the non-transformant. The histological observation and protein gel-blot analysis showed that the development of pollen
grains in the transgenic plant did not progress after the degradation of the tapetum, and the amount of alternative oxidase
in pollen grains of the transgenic plant became lower than that of the non-transformant. These results suggested that the
alternative oxidase activity in the tapetum has a significant effect on the pollen development.
This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | alternative oxidase antisense male-sterility tapetum-specific promoter tobacco transformation |
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