Strong expression of the rice catalase gene CatB promoter in protoplasts and roots of both a monocot and dicots.
The rice (Oryza sativa L.) catalase (EC 1.11.1.6) gene CatB is expressed in roots and cultured cells. We examined the promoter activity of its 5'-flanking region in a monocot and in two dicots. Transient expression assays in rice Oc and tobacco BY-2 suspension cell protoplasts showed that CatB's 5'-flanking DNA fragments (nucleotides -1066 to +298) had about 20 and 3-4 times as much promoter activity, respectively, as the CaMV 35S promoter. Serial deletion analyses of the CatB promoter region revealed that the shortest fragment (-56 to +298) still had about 10 times as much promoter activity as the CaMV 35S promoter in rice protoplasts. In tobacco protoplasts, the activity of the fragment (-56 to +298) was about half of the CaMV 35S promoter. Transgenic rice and Arabidopsis plants carrying GUS genes driven by the 5'-truncated CatB promoters were generated and their GUS activity was examined. The region ranging from -329 to +298 showed preferential expression in the roots of rice and Arabidopsis, and in the shoot apical meristems of Arabidopsis. In situ hybridization revealed that CatB was highly expressed in branch root primordia and root apices of rice. Fusion of the GUS gene to the region (-329 to +298) conferred strong expression in these same areas, indicating that the presence of this region was sufficient to express CatB specifically in the roots. There may be new regulatory element(s) in this region, because it contained no previously known cis-regulatory elements specific for gene expression in roots.
Iwamoto
Masao
M
Genome and Biodiversity Research Center, National Institute of Agrobiological Sciences, 2-1-2, Kannondai, Tsukuba, Ibaraki 305-8602, Japan. iwamas@nias.affrc.go.jp
Higo
Hiromi
H
Higo
Kenichi
K
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002374:Catalase / Q000235:genetics*; D017343:Genes, Plant; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D018517:Plant Roots / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011523:Protoplasts / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics
D010940:Plant Proteins; D002374:Catalase; D005966:Glucuronidase