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Published on October 11, 2007; 10.1104/pp.107.107391

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Received August 14, 2007
Accepted September 25, 2007

A ligation-independent cloning TRV vector for high-throughput virus induced gene silencing identifies roles for NbMADS4-1 and -2 in floral development

Yiyu Dong , Tessa M. Burch-Smith , Yule Liu , Padmavathi Mamillapalli , and Savithramma P. Dinesh-Kumar *

Peking-Yale Joint Center of Plant Molecular Genetics and Agrobiotechnology, College of Life Sciences, Peking University, Beijing 100871, China; Department of Molecular, Cellular and Developmental Biology, Yale University, 219 Prospect Street, New Haven, CT06520-8103

* Corresponding author; email: savithramma.dinesh-kumar{at}yale.edu.

Virus induced gene silencing (VIGS) is a widely used, powerful technique for reverse genetics. VIGS vectors derived from the Tobacco rattle virus (TRV) are among the most popular for VIGS. We have developed a TRV RNA2 vector that allows the insertion of gene silencing fragments by ligation-independent cloning (LIC). This new vector has several advantages over previous vectors, particularly for applications involving the analysis of large numbers of sequences, since TRV-LIC vectors containing the desired insert are obtained with 100% efficiency. Importantly, this vector allows the high-throughput cloning of silencing fragments without the use of costly enzymes required for recombination, as is the case with GATEWAY®-based vectors. We generated a collection of silencing vectors based on 400 tomato ESTs in this TRV-LIC background. We have used this vector to identify roles for SlMADS1 and its N. benthamiana homologs, NbMADS4-1 and -2 in flowering. We find that NbMADS4-1 and -2 act non-redundantly in floral development and silencing of either gene results in loss of organ identity. This TRV-LIC vector should be a valuable resource to the plant community.




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J. R. Tuttle, A.M. Idris, J. K. Brown, C. H. Haigler, and D. Robertson
Geminivirus-Mediated Gene Silencing from Cotton Leaf Crumple Virus Is Enhanced by Low Temperature in Cotton
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Compiled by, F. Tooke, T. Chiurugwi, and N. Battey
Flowering Newsletter bibliography for 2007
J. Exp. Bot., July 18, 2008; (2008) ern109v1.
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