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Plant Physiol, October 1999, Vol. 121, pp. 461-470
Auxin and Cytokinin Have Opposite Effects on Amyloplast
Development and the Expression of Starch Synthesis Genes in Cultured
Bright Yellow-2 Tobacco Cells1
Yutaka
Miyazawa,*
Atsushi
Sakai,2
Shin-ya
Miyagishima,
Hiroyoshi
Takano,3
Shigeyuki
Kawano,4 and
Tsuneyoshi
Kuroiwa
Department of Biological Sciences, Graduate School of Science,
University of Tokyo, Hongo, Tokyo, 113-0033 Japan
In
cultured Bright Yellow-2 (BY-2) tobacco (Nicotiana
tabacum) cells, the depletion of auxin
(2,4-dichlorophenoxyacetic acid) in the culture medium induces the
accumulation of starch. This is accelerated by the addition of
cytokinin (benzyladenine). Light and electron microscopic observations
revealed that this amyloplast formation involves drastic changes in
plastid morphology. The effects of auxin and cytokinin on amyloplast
development were investigated by adding auxin or cytokinin to cells
grown in a hormone-free culture. Auxin repressed amyloplast
development, whereas cytokinin accelerated starch accumulation
regardless of the timing of hormone addition. RNA gel-blot analysis
revealed that the accumulation of the ADP-glucose pyrophosphorylase
small subunit gene (AgpS), granule-bound starch
synthase, and starch branching enzyme transcripts were also affected by
hormonal conditions. High levels of AgpS, granule-bound
starch synthase, and starch branching enzyme transcripts accumulated in
amyloplast-developing cells grown in auxin-depleted conditions.
Furthermore, the addition of auxin to the cells cultured in
hormone-free medium reduced the level of AgpS
transcripts, whereas the addition of cytokinin increased it,
irrespective of the timing of hormone addition. These results suggest
that auxin and cytokinin exert opposite effects on amyloplast
development by regulating the expression of the genes required for
starch biosynthesis.
1
This work was supported by a research fellowship
(no. 5122 to Y.M.) from the Japan Society for the Promotion of Science
for Young Scientists, by a Grant-in-Aid for Scientific Research in Priority Areas (no. 10170208 to A.S.), by a grant-in-aid (no. 11163206 to T.K.) from the Ministry of Education, Science, Sports and Culture of
Japan and the Ministry of Agriculture, Forestry and Fisheries of Japan.
2
Present address: Department of Biological
Science, Faculty of Science, Nara Women's University, Nara, 630-8506 Japan.
3
Department of Biological Science, Faculty of
Science, Kumamoto University, Kumamoto, 860-8555 Japan.
4
Department of Integrated Biosciences, Graduate
School of Frontier Sciences, University of Tokyo, Hongo, Tokyo,
113-0033 Japan.
*
Corresponding author; e-mail miyazawa{at}biol.s.u-tokyo.ac.jp; fax
81-3-3814-1408.
© 1999 American Society of Plant Physiologists
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