![]() Expression of PIN proteins will be used as an indicator of auxin transport polarity in root tissues, as cell-specific PIN expression and plasma membrane localization shift in unison with the direction of active auxin transport. ![]() Auxin polarities relevant to lateral root initiation will be compared in wild type and dgt mutant roots in order to assess which are disrupted due to LeCYP1 deficiency. Objective 3 investigates LeCYP1 interactions with auxin-transport and response during lateral root morphogenesis. Lateral root initiation is in specific growth conditions and whether it is preserved among different plants. Analysis of LeCYP1 expression in Arabidopsis, and during genetically or chemically modified lateral root morphogenesis will help to further test how tight the linkage between LeCYP1 expression and Analysis of the timed LeCYP1 expression with the cellular and sub-cellular resolution in the context of lateral root organogenesis and in relation with other auxin-mediated signals and positional cues will help understand whether expression of LeCYP1 is the trigger for lateral root initiation. Objective 2 analyzes the developmental context of LeCYP1 expression and regulation and how important this regulation is for lateral root morphogenesis. An answer to this question will help understand the mechanisms of LeCYP1 action. Objective 1 investigates whether LeCYP1 regulates lateral root morphogenesis by acting only in the cells where this process takes place, or also by contributing long-range signals generated in other tissues, for example the shoot or the primary root meristem. The goal of the proposed research is to understand how LeCYP1 helps integrate auxin signals into developmental changes of root cells leading to morphogenesis of lateral root Thus, while unessential for the primary root growth, cyclophilin LeCYP1 is essential for the auxin-regulated establishment of new root meristems post germination. Instead, pericycle cells in dgt respond to auxin with proliferation that does not lead to morphogenesis. Deficiency in LeCYP1 in three independent tomato mutant alleles, diageotropica (dgt), disrupts the capacity of root pericycle and endodermis cells to respond to auxin with pattern-forming cell division that generates lateral root primordia. A series of preliminary observations have demonstrated that cyclophilin LeCYP1, a peptidyl-prolyl cis-trans isomerase from tomato, functions as an auxin intermediate during the earliest stages of lateral root formation. Goals / Objectives The project addresses the question of how the root system of tomato develops.
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