Mechanism of Vein Pattern Formation in Arabidopsis Thaliana Leaves: testing the Canalization Hypothesis

Abstract

Several mechanisms have been proposed to explain the process of vein pattern formation in plant tissues. The most widely accepted amongst biologists is the canalization hypothesis, derived from pea root and stem experiments. According to this hypothesis, a signal, thought to be the phytohormone auxin, is transported polarly from cell to cell from the shoot to the root and is canalized progressively into narrow channels of high auxin fluxes that later differentiate to become vascular tissue. In this project, we set out to test whether auxin canalization drives vein pattern formation, using Arabidopsis thaliana mutants with increased auxin transport (max4-1, max3-9, max2-1 and max1-1). We predicted that the mutants would have distinct vein patterns and especially different angles between the primary and secondary veins, compared to the wild type. First rosette leaves of 15 plants per genotype were harvested for analysis each day from 7 to 17 days after sowing, giving a total of eight hundred twenty-five leaf samples to analyze. Venation patterns were extracted and analyzed using custom-made software written with Matlab. Overall, compared with the wild type, mutants with the highest auxin transport (max4-1 and max3-9) had different vein patterns at early developmental stages, confirming a role for auxin transport in vein patterning. However, veins of mutants and wild type connected at similar angles, which is not consistent with the auxin canalization hypothesis, as originally formulated.