Plant Physiology Preview
Published on January 27, 2006; 10.1104/pp.105.071845
Received September 26, 2005
Returned for revision October 11, 2005
Accepted January 24, 2006
Function and Characterization of Starch Synthase I Using Mutants in Rice (Oryza sativa L.)
Naoko Fujita *, Mayumi Yoshida , Noriko Asakura , Takashi Ohdan , Akio Miyao , Hirohiko Hirochika , and Yasunori Nakamura
Department of Biological Production, Akita Prefectural University, Akita City, Akita, 010-0195 Japan; CREST, Japan Science and Technology, Kawaguchi, Saitama, 332-0012 Japan
Department of Biological Production, Akita Prefectural University, Akita City, Akita, 010-0195 Japan
NIAS, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan
* Corresponding author; email: naokof{at}akita-pu.ac.jp.
Four starch synthase I (SSI)-deficient rice (Oryza sativa) mutant lines were generated using retrotransposon Tos17 insertion. The mutants exhibited different levels of SSI activities, and produced significantly lower amount of SSI protein ranging from 0 to 20% of the wild type. The mutant endosperm amylopectin showed decrease in chains with degree of polymerization (DP) 8-12 and increase in chains with DP 6-7 and DP 16-19. The degree of change in amylopectin chain length distribution was positively correlated with the extent of decrease in SSI activity in the mutants. The structural changes in the amylopectin increased the gelatinization temperature of endosperm starch. Chain length analysis of amylopectin in the SSI band excised from native-PAGE/SS activity staining gel showed that SSI preferentially synthesized DP 7-11 chains by elongating DP 4-7 short chains of glycogen or amylopectin. These results show that SSI distinctly generates DP 8-12 chains from short DP 6-7 chain emerging from the branch point in the A or B1 chain of amylopectin. SSI seemingly functions from the very early through the late stage of endosperm development. Yet, the complete absence of SSI, despite being a major SS isozyme in the developing endosperm, had no effect on the size and shape of seeds and starch granules, and the crystallinity of endosperm starch, suggesting that other SS enzymes are probably capable of partly compensating SSI function. In summary the present study strongly suggested that amylopectin chains are synthesized by the coordinated actions of SSI, SSIIa and SSIIIa isoforms.
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