An overexpression of chalcone reductase of Pueraria montana var. lobata alters biosynthesis of anthocyanin and 5'-deoxyflavonoids in transgenic tobacco.
We isolated the chalcone reductase (pl-chr) gene of Pueraria montana var. lobata by using a PCR strategy from cDNA pools of storage roots. A high level of expression of RNA was found in both stems and roots. The genomic Southern blot result suggests that pl-chr exists as a member of a small gene family. By introducing a pl-chr gene under the control of the 35S CaMV promoter into the pink-flowering Xanthi line of Nicotiana tabacum, the flower color was changed from pink to white-to-pink. The contents of anthocyanin in the flowers of the transgenic lines were dramatically decreased by 40%, but the total UV absorption compounds remained unchanged. The production of liquiritigenin in pl-chr overexpressed transgenic tobacco lines was confirmed by HPLC and MS analysis. The introduction of pl-chr gene provides a method to redirect the flavonoid pathway into 5'-deoxyflavonoid production in non-legume crops, in order to manipulate the phenylpropanoid pathway for isoflavonoid production.
Joung
Jae-youl
JY
Plant Cell Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology, Daejon 305-333, Republic of Korea.
Kasthuri
G Mangai
GM
Park
Ji-young
JY
Kang
Won-jin
WJ
Kim
Hyun-soon
HS
Yoon
Bong-sik
BS
Joung
Hyouk
H
Jeon
Jae-heung
JH
D016960:Agrobacterium tumefaciens / Q000235:genetics; D000429:Alcohol Oxidoreductases / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D000872:Anthocyanins / Q000096:biosynthesis* / Q000235:genetics; D015152:Blotting, Northern; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D044950:Flavanones; D005419:Flavonoids / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D029908:Pueraria / Q000201:enzymology*; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014466:Ultraviolet Rays
D000872:Anthocyanins; D018076:DNA, Complementary; D044950:Flavanones; D005419:Flavonoids; D012333:RNA, Messenger; D000429:Alcohol Oxidoreductases; C093070:chalcone reductase; C083152:liquiritigenin