The Wisconsin Crop Innovation Center was selected by the University of Wisconsin as one of the winners of the Cool Science Images 2020 contest. Our submission was an image taken of Cannabis sativa T1 siblings from what we think are the first transgenic hemp plants in the world. The plants are illuminated such that it is possible to view the red light emitted from one of the transgene products, the fluorescent protein tandem Tomato (localized to the endoplasmic reticulum, to enhance brightness) that was included in the T-DNA of the test binary construct I built in support of this project. Null segregants appear green in the image. Contest results can be seen here (our image is #9): https://lnkd.in/dmZDhx9
A picture is worth 1000 words. These micrographs from a variety of plant taxa are simply wonderful.
In my capacity as the Research Manager of the Molecular Technologies Department within the Wisconsin Crop Innovation Center, part of the University of Wisconsin – Madison, I am engaged in several projects in which I utilize Golden Gate cloning. Since first adopting Golden Gate cloning during my post-doctoral work in the Ané lab at the UW, I have worked to develop new TypeIIS mediated cloning tools, to expand the applicability of the MoClo Plant Parts kit, for constructing plant molecular biology focused transcriptional units and binary plasmids. I have noticed during the past year that every time I search for information regarding TypeIIS enzymes, or Golden Gate cloning that that I find exactly the paper I am looking for in the literature, with publication dates are often only months or weeks before I have searched for them. I am very happy to be able to tap into such a vibrant and active area of research. The latest examples are a couple of outstanding papers recently published by the fine folks at New England Biolabs (pub 1) (pub 2). These manuscripts provide a treasure trove of empirical data, related to ligation efficiency and fidelity of 4 nucleotide overhanging sequences, that anyone engaged in non-traditional Golden Gate cloning design would do well to read and use in their future work.
After a long waiting period, the work completed during my post-doc with the USDA in California was finally published in 2018. Our unconventional approach to Agrobacterium-mediated plant transformation was met with skepticism and our manuscript was rejected by several journals. Finally, the sacrifice of nearly all my weekends during 2015 and 2016 was rewarded when our manuscript was accepted. The cherry on top was the selection of our work, including a cover image, as Featured Article in The Plant Journal (volume 95, issue 4, August 2018; https://doi.org/10.1111/tpj.13992).