EAGER Collaborative Research: Developing Transformation Technologies for Porphyra

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The genome of the marine red alga Porphyra umbilicalis is being sequenced by the Joint Genome Institute. The sequence information will help scientists address many fundamental questions, because Porphyra spp. belong to an ancient eukaryotic lineage, are important human foods ("nori"), have complex life histories, and---even compared to other intertidal organisms--- possess an unusually stress-tolerant metabolism. Computer-based analyses of the new genomic data will be sufficient to address some research questions, but most studies (e.g., the basis of Porphyra's tolerance to extreme drying or high light) will require experimental approaches based upon bioinformatics analyses. This project will develop the essential technology of stable genetic transformation in Porphyra to make such experimental work possible. The investigators will focus on transforming neutral spores of P. umbilicalis because these abundantly-produced spores lack a cell wall. This should make it possible to transform the cell by electroporation, among other approaches. Native Porphyra promoters of Porphyra genes will be used, based on information provided by the JGI sequencing project, and codon-optimized reporter genes will be synthesized. Spores will be treated with selective antibiotics to recover transformed sporelings, and the stability of the transformation will be assessed as sporelings mature to adults. The PIs and a postdoctoral associate will work across both participating laboratories to develop transformation technologies. The postdoctoral associate will investigate reproductive pathways or stress physiology during the development of transformation techniques. S/he will be well-prepared to make individual and collaborative advances with the Porphyra model system due to the comprehensive postdoctoral training and participation in the Porphyra NSF Research Coordination Network. This project will allow the scientific community to use the data from the whole genome sequencing project in experimental research on Porphyra, leading to fundamental advances in the areas of metabolism, evolution, and developmental biology.