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LEAVES – The Bundle Ativador Download [Keygen]

The 5-dag seedlings were also morphologically abnormal, in particular, the SAM and leaf primordia showed abnormal differentiation ( Fig 4 ). SAMs were partly fused, and the number of cells in the SAM region was increased in the DEX-treated plants ( Fig 4A ). In addition, the size of the rosette leaves varied among seedlings in the same line, although it was approximately constant among plants in a given line (Fig 4A and 4B ). The 5-dag mock-treated plants were similar to wild-type plants in terms of organ size and shape (Fig 4B ). By contrast, the SAMs of DEX-treated plants were greatly reduced in size. On average, the SAM was reduced to about one quarter of that of wild-type plants (Fig 4B, red arrow). At 5 dag, the 5-dag mock-treated plants were similar to wild-type plants (Fig 4B ). By contrast, the SAM of the DEX-treated plants at 5 dag was severely reduced (Fig 4B, red arrow). The SAMs of both mock-treated and DEX-treated plants at 5 dag were reduced to less than one fifth of those at 10 dag, but the leaf width did not differ significantly between mock- and DEX-treated plants (Fig 4C ). Thus, shorter OsWOX4 knockdown strongly affected not only the size of the SAMs but also the number of leaves in 5-dag plants.

To identify novel rice genes controlling leaf morphogenesis in rice, we used HAs from rice leaf development as baits (Microprobe AS-PR01) on a microarray cDNA library. We hybridized the microarray to compare the gene expression in leaves of 3.5-day-old plants after various treatments. The result revealed one set of genes that were upregulated by ethylene or cytokinin treatment during rice leaf development. We also identified several genes that were downregulated by ethylene or cytokinin treatments.

Although OsWOX4 played a positive role in promoting cell proliferation in the leaf primordia, it also suppressed the expression of OsPNH1 [ 24 ]. Therefore, we further examined whether the cell division was reduced in the vascular bundles of the transgenic plants. After P4 was observed, we focused on the area where the expression of OsPNH1 was silenced, because OsPNH1 expression is relatively strong in this region. The number of longitudinal cell files in the area was not different between mock- and DEX-treated plants ( Fig 11 ). However, in the OsWOX4-silenced plants, the number of cell files was reduced, and the cells were thinner than those in mock-treated plants. The number of cell files in the outer cells of the midrib area was also reduced in the OsWOX4-knockdown plants. These observations indicate that OsWOX4 promotes cell division, and OsWOX4 knockdown strongly suppresses the later vascular differentiation and vascular bundle development.
OsWOX4 is highly expressed in various rice leaf primordia, particularly in the central region of the leaf primordia, similar to DL expression ( S3B Fig ). We confirmed the accumulation of OsWOX4 in the central region of leaf primordia by examining the transiently overexpressing lines. To exclude any developmental defects associated with the overexpression of OsWOX4 in primordia, we generated transgenic lines expressing the OsWOX4 coding region under the control of the maize ubiquitin promoter. We then examined the leaf phenotypes of the newly developed OsWOX4 overexpressing plants. Transgenic lines overexpressing OsWOX4 under the control of the ubiquitin promoter grew normally and produced leaves that exhibited wild-type leaf shapes and architectures ( S4A Fig ). Because the overexpression of OsWOX4 under the maize ubiquitin promoter resulted in normal leaf phenotypes, these results support our finding that OsWOX4 is important in early leaf development. Our findings suggest that OsWOX4 plays a central role in cell proliferation in the central region of leaf primordia for the formation of the midrib.


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