Ribozymes targeted to stearoyl-ACP delta9 desaturase mRNA produce heritable increases of stearic acid in transgenic maize leaves.
Ribozymes are RNAs that can be designed to catalyze the specific cleavage or ligation of target RNAs. We have explored the possibility of using ribozymes in maize to downregulate the expression of the stearoyl-acyl carrier protein (Delta9) desaturase gene. Based on site accessibility and catalytic activity, several ribozyme constructs were designed and transformed into regenerable maize lines. One of these constructs, a multimer hammerhead ribozyme linked to a selectable marker gene, was shown to increase leaf stearate in two of 13 maize lines. There were concomitant decreases in Delta9 desaturase mRNA and protein. The plants with the altered stearate phenotype were shown to express ribozyme RNA. The ribozyme-mediated trait was heritable, as evidenced by stearate increases in the leaves of the R1 plants derived from a high-stearate line. The increase in stearate correlated with the presence of the ribozyme gene. A catalytically inactive version of this ribozyme did not produce any significant effect in transgenic maize. This is evidence that ribozymes can be used to modulate the expression of endogenous genes in maize.
Merlo
A O
AO
Biotechnology and Plant Genetics Department, Dow AgroSciences, 9330 Zionsville Road, Indianapolis, Indiana 46268, USA.
Cowen
N
N
Delate
T
T
Edington
B
B
Folkerts
O
O
Hopkins
N
N
Lemeiux
C
C
Skokut
T
T
Smith
K
K
Woosley
A
A
Yang
Y
Y
Young
S
S
Zwick
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D005227:Fatty Acids / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics*; D008969:Molecular Sequence Data; D010641:Phenotype; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics* / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism; D013229:Stearic Acids / Q000378:metabolism; D003313:Zea mays / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D018744:DNA, Plant; D005227:Fatty Acids; D016337:RNA, Catalytic; D012333:RNA, Messenger; D018749:RNA, Plant; D013229:Stearic Acids; C031183:stearic acid; D006899:Mixed Function Oxygenases; C018747:acyl-(acyl-carrier-protein)desaturase