Plant Physiology Preview
Published on July 25, 2002; 10.1104/pp.006353
Received April 3, 2002
Returned for revision April 30, 2002
Accepted May 1, 2002
hpRNA-Mediated Targeting of the Arabidopsis FAD2 Gene Gives Highly Efficient and Stable Silencing
Peter A. Stoutjesdijk , Surinder P. Singh *, Qing Liu , Clive J. Hurlstone , Peter A. Waterhouse , and Allan G. Green
Commonwealth Scientific and Industrial Research Organization Plant Industry, P.O. Box 1600, Canberra, Australian Capital Territory 2601, Australia
* Corresponding author; email: surinder.singh{at}csiro.au.
The endogenous  12-desaturase gene (FAD2) in Arabidopsis was targeted for silencing using seed-specific cosuppression (CS), hairpin (HP) RNA (hpRNA), and intron-spliced HP (iHP) constructs. The iHP construct, incorporating the 120-bp 3'-untranslated region of the FAD2 gene, gave the highest degree of silencing. In some iHP lines 12-desaturase activity was reduced to levels as low as those in the null fad2-1 mutant, and every primary transformant showed a pronounced reduction in FAD2 activity. One highly silenced iHP line was propagated for five generations and showed no reversion or diminution in its degree of silencing. About 75% of plants transformed with the HP construct, targeting the FAD2 coding region, gave dramatically reduced 12-desaturase activity, whereas approximately 50% of plants transformed with the CS construct, containing the same coding region sequence, showed silencing at a much less profound level. In all three types of constructs, the degree of silencing was increased when the transgenes were homozygous, but this was much more pronounced for the CS constructs. All three types of construct could give a single locus that was capable of effective silencing, but in the one such CS line where this was the case, the locus had a complex insertion pattern. This is consistent with the concept that posttranscriptional gene silencing is induced by double-stranded, or self-complementary, RNA that is formed in cases of CS by complex insertion patterns at a single locus and that the most effective way of generating profoundly silenced plants is by the use of constructs that encode hpRNAs. Furthermore, these results demonstrate for the first time, to our knowledge, that iHP constructs targeted against an endogenous seed-expressed gene are clearly able to generate phenotypic changes that are inherited stably over several generations, making this approach a reliable technique for genetic modification of seed quality and possibly other traits in agricultural plants.
This article has been cited by other articles:
|
|
|
|
 
J. McNaughton, M. Roberts, B. Smith, D. Rice, M. Hinds, C. Sanders, R. Layton, I. Lamb, and B. Delaney
Comparison of Broiler Performance When Fed Diets Containing Event DP-3O5423-1, Nontransgenic Near-Isoline Control, or Commercial Reference Soybean Meal, Hulls, and Oil
Poult. Sci.,
December 1, 2008;
87(12):
2549 - 2561.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. de la Luz Gutierrez-Nava, M. J. Aukerman, H. Sakai, S. V. Tingey, and R. W. Williams
Artificial trans-Acting siRNAs Confer Consistent and Effective Gene Silencing
Plant Physiology,
June 1, 2008;
147(2):
543 - 551.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Avisar, A. I. Prokhnevsky, K. S. Makarova, E. V. Koonin, and V. V. Dolja
Myosin XI-K Is Required for Rapid Trafficking of Golgi Stacks, Peroxisomes, and Mitochondria in Leaf Cells of Nicotiana benthamiana
Plant Physiology,
March 1, 2008;
146(3):
1098 - 1108.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. J. Wassom, V. Mikkelineni, M. O. Bohn, and T. R. Rocheford
QTL for Fatty Acid Composition of Maize Kernel Oil in Illinois High Oil x B73 Backcross-Derived Lines
Crop Sci.,
January 16, 2008;
48(1):
69 - 78.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Dafny-Yelin, S.-M. Chung, E. L. Frankman, and T. Tzfira
pSAT RNA Interference Vectors: A Modular Series for Multiple Gene Down-Regulation in Plants
Plant Physiology,
December 1, 2007;
145(4):
1272 - 1281.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Travella, T. E. Klimm, and B. Keller
RNA Interference-Based Gene Silencing as an Efficient Tool for Functional Genomics in Hexaploid Bread Wheat
Plant Physiology,
September 1, 2006;
142(1):
6 - 20.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Van Houdt, A. Bleys, and A. Depicker
RNA Target Sequences Promote Spreading of RNA Silencing
Plant Physiology,
January 1, 2003;
131(1):
245 - 253.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Liu, S. P. Singh, and A. G. Green
High-Stearic and High-Oleic Cottonseed Oils Produced by Hairpin RNA-Mediated Post-Transcriptional Gene Silencing
Plant Physiology,
August 1, 2002;
129(4):
1732 - 1743.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
