May's Cold Spring Harbor Protocols features plant gene expression methods, Xenopus imaging

COLD SPRING HARBOR, N.Y. (Mon., May 3, 2010) -- The generation of transgenic plants can be a lengthy and difficult process. Transient expression assays have been developed as faster and more convenient alternatives for investigating gene function. These assays often take advantage of the ability of Agrobacterium to transfer foreign DNA into plant cells with intact cell walls. Agrobacterium-mediated transformation is, however, inefficient and shows great variability. In the May issue of Cold Spring Harbor Protocols (http://www.cshprotocols.org/TOCs/toc5_10.dtl), Andreas Nebenführ and colleagues from the University of Tennessee (http://www.bio.utk.edu/cellbiol/) present "FAST Technique for Agrobacterium-Mediated Transient Gene Expression in Seedlings of Arabidopsis and Other Plant Species," a quick, efficient and economical assay for gene function in intact plants. The technique involves cocultivation of young plant seedlings and Agrobacterium in the presence of Silwet-77. The Silwet-77 facilitates transformation, thus replacing a wounding or device-dependent vacuum step. As one of May's featured articles, it is freely available on the journal's website (http://cshprotocols.cshlp.org/cgi/content/full/2010/5/pdb.prot5428).

The large size and external development of the frog Xenopus laevis make it an ideal system for in vivo imaging of dynamic cellular activity. Xenopus embryos are amenable to simple genetic manipulation techniques including knockdowns and misexpression, as well as transgenesis. The ease of collecting large numbers of embryos and the larger size of individual cells within an embryo as compared with other vertebrate model systems provides an excellent platform for the observation of cellular behavior and subcellular processes. In the May issue of Cold Spring Harbor Protocols (http://www.cshprotocols.org/TOCs/toc5_10.dtl), John Wallingford and colleagues from the University of Texas (http://www.bio.utexas.edu/faculty/wallingford/) provide a suite of articles detailing live imaging of Xenopus laevis at low magnification, confocal imaging of fixed tissues, and in one of May's featured articles, "High-Magnification In Vivo Imaging of Xenopus Embryos for Cell and Developmental Biology". This protocol describes methods for labeling and high-magnification time-lapse imaging by confocal microscopy and is freely available on the journal's website (http://cshprotocols.cshlp.org/cgi/content/full/2010/5/pdb.prot5427).

Source: Cold Spring Harbor Laboratory