Molecular cloning and characterization of a novel gene encoding limonoid UDP-glucosyltransferase in Citrus.
We isolated a cDNA clone encoding limonoid UDP-glucosyltransferase (limonoid GTase) from the albedo of Satsuma mandarin (Citrus unshiu Marc.) and investigated the contribution to limonoid glucoside accumulation in fruit. The isolated cDNA clone (CitLGT) was 1732 bp in length encoding 511 deduced amino acids with a predicted molecular mass of 57.5 kDa. The products of in vitro translation from an expression vector had the limonoid GTase activity. Southern blot analysis of genomic DNA indicated that CitLGT was present as a single copy gene in the Citrus genome. The amount of transcript corresponding to CitLGT mRNA changed the same way as the fluctuation of limonin glucoside content during fruit development of navel orange (Citrus sinensis Osb.). This indicates that the transcription of CitLGT regulates the conversion of limonoid aglycones to glucosides in citrus fruit.
Kita
M
M
United Graduate School, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan. mkita@okt.affrc.go.jp
Hirata
Y
Y
Moriguchi
T
T
Endo-Inagaki
T
T
Matsumoto
R
R
Hasegawa
S
S
Suhayda
C G
CG
Omura
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002957:Citrus / Q000737:chemistry* / Q000201:enzymology / Q000378:metabolism; D003001:Cloning, Molecular; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D036701:Limonins; D008969:Molecular Sequence Data; D018749:RNA, Plant / Q000032:analysis; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D014315:Triterpenes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
D036701:Limonins; D018749:RNA, Plant; D014315:Triterpenes; D005964:Glucosyltransferases; C406404:limonoid GTase; C001546:limonin