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United States Department of Agriculture

Agricultural Research Service

Title: Virus vector for gene silencing in wheat

Authors
item Tai, Yin Shan
item Bragg, Jennifer - MICRO BIOL UOFCA BERKELEY
item Edwards, Michael

Submitted to: Biotechniques
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: July 27, 2005
Publication Date: September 1, 2005
Citation: Tai, Y.-S., Bragg, J., Edwards, M.C. 2005. Virus vector for gene silencing in wheat. BioTechniques. 39:310-314.

Interpretive Summary: Wheat is one of the major cereal crops consumed world wide. Under the pressure of global food demand, farmers are eager to have disease resistant crops released from breeding programs. Usually it takes many years to breed a useful resistant plant line. In addition to the traditional breeding processes, a high-throughput screening method to identify candidate genes for resistance is a critical step needed to shorten the time required for breeding programs. “Virus-induced gene silencing” (VIGS) is a powerful tool for plant research to investigate genes in a high-throughput manner. To our knowledge, there is no publication to date about the application of VIGS in wheat, although it is commonly used in dicot plants (for example, tomato, pepper, and pea). In this study, we verify the possibility of using VIGS in wheat. We tested three marker genes to determine if they could be visibly silenced (shut-off) using VIGS. These phenotypes include photo-bleaching, yellowing, or necrosis of leaves. Furthermore, we confirmed gene silencing due to the decreased RNA levels of each gene by a technique called “reverse-transcription polymerase chain reaction” (RT-PCR). Our overall goal is to use VIGS in functional genomics analyses of wheat; we intend to use it to study the molecular mechanisms of wheat resistance against various fungal pathogens.

Technical Abstract: Wheat is one of the major cereal crops consumed world wide. Under the pressure of global food demand, farmers are eager to have disease resistant crops released from breeding programs. Usually it takes many years to breed a useful resistant plant line. In addition to the traditional breeding processes, a high-throughput screening method to identify candidate genes for resistance is a critical step needed to shorten the time required for breeding programs. “Virus-induced gene silencing” (VIGS) is a powerful tool for plant research to investigate genes in a high-throughput manner. To our knowledge, there is no publication to date about the application of VIGS in wheat, although it is commonly used in dicot plants (for example, tomato, pepper, and pea). In this study, we verify the possibility of using VIGS in wheat. We tested three marker genes to determine if they could be visibly silenced (shut-off) using VIGS. These phenotypes include photo-bleaching, yellowing, or necrosis of leaves. Furthermore, we confirmed gene silencing due to the decreased RNA levels of each gene by a technique called “reverse-transcription polymerase chain reaction” (RT-PCR). Our overall goal is to use VIGS in functional genomics analyses of wheat; we intend to use it to study the molecular mechanisms of wheat resistance against various fungal pathogens.

Last Modified: 8/29/2014
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