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Plant Physiol, November 2001, Vol. 127, pp. 1136-1146
Significant Accumulation of C4-Specific Pyruvate,
Orthophosphate Dikinase in a C3 Plant,
Rice1
Hiroshi
Fukayama,
Hiroko
Tsuchida,
Sakae
Agarie,2
Mika
Nomura,
Haruko
Onodera,
Kazuko
Ono,
Byung-Hyun
Lee,3
Sakiko
Hirose,
Seiichi
Toki,
Maurice S.B.
Ku,
Amane
Makino,
Makoto
Matsuoka, and
Mitsue
Miyao*
National Institute of Agrobiological Sciences, Tsukuba 305-8602,
Japan (H.F., H.T., S.A., H.O., K.O., B.-H.L., S.H., S.T., Mi.M.);
Faculty of Agriculture, Kagawa University, Kagawa 761-0701, Japan
(M.N.); Botany Department, Washington State University, Pullman,
Washington 99164-4238 (M.S.B.K.); Graduate School of Agricultural
Sciences, Tohoku University, Sendai 981-8555, Japan (A.M.); and
BioScience Center, Nagoya University, Chikusa, Nagoya 461-8601, Japan
(Ma.M.)
The C4-Pdk gene encoding the
C4 enzyme pyruvate, orthophosphate dikinase (PPDK) of maize
(Zea mays cv Golden Cross Bantam) was introduced into
the C3 plant, rice (Oryza sativa cv
Kitaake). When the intact maize C4-Pdk gene,
containing its own promoter and terminator sequences and exon/intron
structure, was introduced, the PPDK activity in the leaves of some
transgenic lines was greatly increased, in one line reaching 40-fold
over that of wild-type plants. In a homozygous line, the PPDK protein
accounted for 35% of total leaf-soluble protein or 16% of total leaf
nitrogen. In contrast, introduction of a chimeric gene containing the
full-length cDNA of the maize PPDK fused to the maize
C4-Pdk promoter or the rice
Cab promoter only increased PPDK activity and protein
level slightly. These observations suggest that the intron(s) or the terminator sequence of the maize gene, or a combination of both, is
necessary for high-level expression. In maize and transgenic rice
plants carrying the intact maize gene, the level of transcript in the
leaves per copy of the maize C4-Pdk gene was
comparable, and the maize gene was expressed in a similar
organ-specific manner. These results suggest that the maize
C4-Pdk gene behaves in a quantitatively and
qualitatively similar way in maize and transgenic rice plants. The
activity of the maize PPDK protein expressed in rice leaves was
light/dark regulated as it is in maize. This is the first reported
evidence for the presence of an endogenous PPDK regulatory protein in a
C3 plant.
1
This study was supported in part by the
Bio-Oriented Technology Research Advancement Institution of Japan
(PROBRAIN grant to Mi.M. and Ma.M.).
2
Present address: Faculty of Agriculture, Saga
University, Saga 840-8502, Japan.
3
Present address: College of Agriculture, Gyeongsang
National University, Chinju 660-701, Korea.
*
Corresponding author; e-mail mmiyao{at}affrc.go.jp; fax
81-298-38-7073.
© 2001 American Society of Plant Physiologists
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