Cloning and functional analysis of two gibberellin 3β-hydroxylase genes that are differently expressed during the growth of rice

被引:245
作者
Itoh, H
Ueguchi-Tanaka, M
Sentoku, N
Kitano, H
Matsuoka, M
Kobayashi, M
机构
[1] RIKEN, Tsukuba Inst, Tsukuba, Ibaraki 3050074, Japan
[2] Nagoya Univ, Biosci Ctr, Nagoya, Aichi 4648501, Japan
[3] Nagoya Univ, Grad Sch Bioagr Sci, Nagoya, Aichi 4648501, Japan
关键词
D O I
10.1073/pnas.141239398
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have cloned two gibberellin (CA) 3 beta -hydroxylase genes, OsCA3ox1 and OsGA3ox2, from rice by screening a genomic library with a DNA fragment obtained by PCR using degenerate primers. We have used full-scan CC-MS and Kovats retention indices to show function for the two encoded recombinant fusion proteins. Both proteins show 3 beta -hydroxylase activity for the steps GA(20) to GA(1), GA(5) to GA(3), GA(44) to GA(38) and GA(9) to GA(4). In addition, indirect evidence suggests that the OsGA3ox1 protein also has 2,3-desaturase activity, which catalyzes the steps GA(9) to 2,3-dehydro-GA(9) and GA(20) to GA(5) (2,3-dehydro GA(20)), and 2 beta -hydroxylase activity, which catalyzes the steps GA(1) to GA(8) and GA(4) to GA3(4) Molecular and linkage analysis maps the OsGA-ox1 gene to the distal end of the short arm of chromosome 5; the OsGA3ox2 gene maps to the distal end of the short arm of chromosome 1 that corresponds to the D18 locus. The association of the OsGA3ox2 gene with the d18 locus is confirmed by sequence and complementation analysis of three d18 alleles. Complementation of the d18-AD allele with the OxGA3ox2 gene results in transgenic plants with a normal phenotype. Although both genes show transient expression, the highest level for OsGA3ox1 is from unopened flower. The highest level for OsGA3ox2 is from elongating leaves.
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页码:8909 / 8914
页数:6
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