Root and shoot suberization in transgenic lines of grey poplar (Populus × canescens), an economically important tree species

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This project aims to thoroughly characterize suberin CRISPR-Cas9 knock-out and 35S-promoter over-expression lines of grey poplar and compare them to the corresponding wild-type. Besides primary roots, this project will for the first time extend the investigations on the formation, structure, and function of suberized periderms in secondary roots and shoots of poplar forming in 8-12 week-old plants. This will include the following work packages: (i) Generation of transgenic (CRISPR-Cas9 knock-out and over-expression) poplar lines of suberin key genes. (ii) Optimization of growth protocols to attain further developed roots in their secondary developmental state and shoots, both undergoing secondary thickening and periderm formation. (iii) Detailed anatomical characterization of primary roots, secondarily thickened roots, and shoot periderms to identify tissue-specific deposition or absence of suberin in comparison to the wild-type and alterations in response to abiotic stress (salt and drought). (iv) Chemical analysis of suberin and associated wax amount and composition in primary roots, secondarily thickened roots, and shoot periderms to identify modifications in comparison to the wild-type and in response to abiotic stress. (v) Characterization of water and solute transport in roots and shoot periderms in comparison to the wild-type and in response to abiotic stress. (vi) Analysis of modified gene expression (RNA-seq) in suberin mutant lines in comparison to their wild-type reference and in response to abiotic stress. Based on these experiments described above, we intend to further elucidate the function of suberin in primary and secondary roots and shoot periderm in adaptation to the abiotic environmental stress factors drought and salt.