UNIVERSITÄT ZU KÖLN

Z01: Resource development for plant-fungal interactions

A fundamental understanding of the interactions between plants and fungi in barley, one of the most important European crops, should be investigated to facilitate future agricultural applications. To do this efficiently, genome engineering methods can be used to analyze specific interactions at the molecular and physiological levels. Prerequisites for the efficient investigation of plant-fungal interactions are knowledge of the genomic data and efficient transformation methods, tissue- and cell-type-specific expression systems, and non-invasive reporters. Since the interaction with fungi is restricted to certain parts of the plant (e.g. leaves, roots and ears), constitutive overexpression or suppression of resistance/susceptibility genes often leads to undesirable side effects. Therefore, specific expression systems are needed to overexpress fungal effectors. In addition, improvements have been made to the reporter systems required to visualize transformation success. While earlier reporter genes rely on sophisticated instrumentation (e.g. fluorescent proteins), others require substrates and do not allow further material cultivation. The subproject aims to develop appropriate expression systems and to test them transiently and in stably transgenic barley plants. The RUBY fusion protein, which catalyzes the conversion of tyrosine into the betalain pigment known from beetroot, is used. This pigment is also visible to the naked eye and easily detected. Another aim is to test different barley genotypes for suitability for tissue culture and transformability, as these technologies cannot yet be used independently of genotype. In summary, resources are to be created in the subproject that can be used in the other subprojects to test identified effectors, etc.

For more information visit Hensel lab

Publications

• Hensel G, Kastner C, Oleszczuk S, et al.  Agrobacterium-mediated gene transfer to cereal crop plants: current protocols for barley, wheat, triticale, and maize. Int J Plant Genomics. 2009;2009:835608. doi: 10.1155/2009/835608.
• Hensel G, Himmelbach A, Chen W, et al. Transgene expression systems in the Triticeae cereals. J Plant Physiol. 2011 Jan 1;168(1):30-44. doi: 10.1016/j.jplph.2010.07.007.
• Becker M and G Hensel: Ribonucleoprotein (RNP)-mediated targeted mutagenesis in barley (Hordeum vulgare L.). In: Yang B, W Harwood and Q Que (Eds.): Plant Genome Engineering. Methods in Molecular Biology, vol 2653. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3131-7_12
• Pouramini P and G Hensel: Precise gene editing of cereals using CRISPR/Cas technology. Ricroch A, Miladinović D, Sweet J, Eriksson D, Van Laere K and Woźniak-Gientka E (Eds.). A Roadmap for Plant Genome Editing. Springer,  https://doi.org/10.1007/978-3-031-46150-7_9
• Yeo FK, Hensel G, Vozábová T, et al. Golden SusPtrit: a genetically well-transformable barley line for studies on the resistance to rust fungi. Theor Appl Genet. 2014 Feb;127(2):325-37. doi: 10.1007/s00122-013-2221-7.