Author
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Matthews, Benjamin - Ben |
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YOUSSEF, REHAM - El-Fayoum University |
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Submitted to: Bio-protocol
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 12/1/2016 Publication Date: 1/20/2016 Citation: Matthews, B.F., Youssef, R.M. 2016. Agrobacterium rhizogenes - based transformation of soybean roots to form composite plants. Bio-protocol. 6:e1708. Interpretive Summary: Transgenic soybean roots are used to overexpress genes, silence genes, test promoters, and they have a variety of other useful applications. However, production of transgenic soybean roots using tissue culture and sterile procedures is tedious, time-consuming, and can be frustrating if the cultures become contaminated. Here, we describe a technique to produce composite soybean plants with genetically transformed roots and untransformed shoots. This technique is relatively easy and does not use sterile tissue culture. This technique is of use to scientists who use genetic transformation to study gene function in roots, because it will save them time and money. Technical Abstract: Composite plants are a powerful tool to rapidly analyze the effects of gene overexpression, gene silencing, and examine test promoter expression in transgenic roots. No sterile tissue culture is needed. This avoids loss of valuable material due to contamination of sterile cultures. This method uses simple techniques that require less training and care than techniques using sterile tissue culture. Furthermore, it takes less time to produce transgenic roots than techniques using sterile tissue culture. If the transgenic roots are to be challenged with a pathogen, there is no need to produce axenic pathogen for root inoculation with this technique, because sterile medium is not used. Therefore, there is no agar medium for contaminants to grow on and obscure results. Here, we describe the production of composite soybean plants with transgenic roots using Agrobacterium rhizogenes. The composite plants can be grown in the greenhouse. |
