The promoter of the barley aleurone-specific gene encoding a putative 7 kDa lipid transfer protein confers aleurone cell-specific expression in transgenic rice.
This paper describes the aleurone-specific gene Ltp2 from barley, which encodes a putative 7 kDa non-specific lipid transfer protein. As shown by Northern and in situ hybridization analyses, the Ltp2 transcript is present in barley aleurone cells shortly after the initiation of aleurone cell differentiation. The expression of Ltp2 increases until grain mid-maturity, but the mRNA is absent from mature grains. The Ltp2 transcript is undetectable in the embryo and vegetative tissues, confirming the aleurone specificity of the Ltp2 gene. The ability of the isolated 801 bp Ltp2 promoter to direct aleurone-specific expression in immature barley grains is demonstrated by particle bombardment experiments. In these experiments, the activity of the Ltp2 promoter is 5% of the activity of the strong constitutive Actin1 promoter from rice, as quantified by GUS activity measurements. In stably transformed rice plants containing the Ltp2 promoter-Gus construct, the specificity of the Ltp2 promoter is confirmed in vivo by the presence of GUS activity exclusively in the aleurone layer. This study demonstrates the conserved nature of the regulatory signals involved in aleurone-specific gene transcription in cereal grains.
Department of Biotechnological Sciences, Agricultural University of Norway.
R
R
Kalla
K
K
Shimamoto
R
R
Potter
P S
PS
Nielsen
C
C
Linnestad
O A
OA
Olsen
D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002454:Cell Differentiation; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D015870:Gene Expression; D017343:Genes, Plant; D001467:Hordeum / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012275:Oryza / Q000166:cytology / Q000378:metabolism; D010940:Plant Proteins; D030821:Plants, Genetically Modified; D010957:Plasmids; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014908:Triticum / Q000235:genetics
D052179:Antigens, Plant; D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D010940:Plant Proteins; C511920:lipid transfer proteins, plant