Characterisation of the maize leaf patterning mutants Wavy auricle in blade1-R and milkweed pod1-R : a thesis presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Plant Biology at Massey University, Palmerston North, New Zealand

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Massey University
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The maize leaf has three main axes of growth, with an asymmetric distribution of tissue types along each axis. This study focuses on three mutants, Wavy auricle in blade1-R (Wab 1-R), liguleless1-R (lg1-R) and milkweed pod1-R (mwp1-R) that disrupt axial patterning of maize leaves. Dominant Wab1 mutations disrupt both medial-lateral and proximal-distal patterning. Wab1 leaf blades are narrow and ectopic auricle and sheath-like tissues extend into the leaf blade. Previous analyses have shown that Lg1 acts cell-autonomously to specify ligule and auricle tissues. The current study reveals additional roles in defining leaf shape. The recessive lg1-R mutation exacerbates the Wab1-R phenotype; in the double mutants, most of the proximal blade is deleted and sheath tissue extends along the residual blade. A mosaic analysis of Wab1-R was conducted in Lg1 and lg1-R backgrounds to determine if Wab1-R affects leaf development in a cell-autonomous manner. Normal tissue identity was restored in all wab1/- sectors in a lg1-R mutant background, and in three quarters of sectors in a Lg1 background. These results suggest that Lg1 can influence the autonomy of Wab1-R. In both genotypes, leaf-halves with wab1/- sectors were significantly wider than non-sectored leaf-halves, suggesting that Wab1-R acts cell-autonomously to affect lateral growth. mwp1-R is a recessive mutation that specifically affects patterning of sheath tissue. Characterisation of the mwp1-R phenotype revealed that mwp1-R husk leaves and the sheaths of vegetative leaves develop pairs of outgrowths on the abaxial surface associated with regions of adaxialised tissue. In situ hybridisation confirmed that disruptions to adaxial-abaxial patterning are correlated with misexpression of leaf polarity genes. Leaf margins and fused organs such as the prophyll are most severely affected by mwp1-R. The first two husk leaves normally fuse along adjacent margins to form the bi-keeled prophyll. In the most severe cases the mwp1-R prophyll is reduced to an unfused, two-pronged structure and keel outgrowth is significantly reduced. We speculate that the adaxial-abaxial patterning system has been co-opted during evolution to promote outgrowth of the keels in normal prophyll development. The results of this study place Mwp1, wab1 and Lg1 in a network of genes that regulate leaf polarity and axial patterning.
Corn genetics, Maize leaf growth