Functional characterization of Fe-dependent mechanisms promoting the stimulation of adventitious root formation in petunia cuttings

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Adventitious root (AR) formation is prerequisite for vegetative propagation of important crops and common practice to maintain the genetic identity among progenies. With increasing use of soil-free media in high-throughput propagation and growing demand for sustainable horticultural practices, a deep understanding of the role of mineral nutrition in AR formation is essential for the improvement of existing propagation protocols. Our previous work demonstrated that Fe is the most limiting nutrient for AR formation and provided compelling evidence for a unique unexploited role of Fe in stimulation of meristematic cell division, associated with its high accumulation in the nuclei of meristematic cells. The proposed project aims to study the physiological and biochemical mechanisms underlying Fe-stimulated AR formation and to identify genes that can be used to improve AR formation. Initially, a detailed cell biological analysis and ultrastructural localization of Fe will define the major anatomical and physiological processes upon Fe application. A comprehensive transcriptome profiling using RNAseq and MS-based analysis of nucleolar proteins will identify genes and proteins involved in Fe-accelerated AR formation. The function of top candidate genes derived from proteomic and transcriptomic studies will be characterized in transposon-insertion or CRISPR/Cas9-modified transgenic plants using anatomical and physiological approaches. Finally, the Fe-dependent AR formation will be studied in wild petunia accession lines, closely related Solanaceae genera such as tomato gene pool for a better access to additional mutants and several commercial cultivars of horticultural crops. The proposed study will advance the understanding of the unique function of Fe in the plant nucleolus and will pave the ground for a targeted exploitation of Fe nutrition in vegetative propagated crops