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First published online 14 July 2004
doi: 10.1242/dev.01262


Development 131, 3921-3929 (2004)
Published by The Company of Biologists 2004


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The establishment of axial patterning in the maize leaf

Toshi Foster1,*,{dagger}, Angela Hay2,*, Robyn Johnston3 and Sarah Hake4

1 The Horticulture and Food Research Institute of New Zealand, Private Bag 11 030, Palmerston North, New Zealand
2 Department of Plant Sciences, Oxford University, South Parks Road, Oxford, OX1 3RB, UK
3 Institute of Molecular BioSciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand
4 Plant Gene Expression Center, 800 Buchanan Street, Albany, CA 94710, USA



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Fig. 1. Leaf and whole plant phenotypes 8 weeks after planting. (A) Wild-type, (B) Wab1, (C) lg1-R;Wab1 and (D) lg1-R plants. (E-H) Adaxial view of blade-sheath boundary of (E) wild-type, (F) Wab1, (G) lg1-R;Wab1, and (H) lg1-R leaves. a, auricle; b, blade; ae, auricle extension; ea, ectopic auricle; es, ectopic sheath; lg, ligule; s, sheath. Arrowhead in G indicate presumptive blade-sheath boundary in lg1-R;Wab1 leaf.

 


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Fig. 2. Scanning electron and light micrographs illustrating epidermal and histological features of wild-type and mutant leaves. (A-C) SEM of adaxial surface of wild-type (A) blade, (B) auricle and (C) sheath. (D-G) Transverse section through wild-type (D) blade, (E) auricle, (F) internal sheath and (G) marginal sheath tissue. (H) Cartoon depicting regions where tissue was sampled. (I-K) Transverse sections through (I) ectopic auricle in Wab1 blade, (J) ectopic sheath in Wab1 blade and (K) ectopic sheath in lg1-R;Wab1 blade. (L-M) SEM of adaxial surface of (L) Wab1 ectopic auricle and (M) lg1-R;Wab1 ectopic sheath. All sections are oriented with the adaxial surface upwards. Arrows in A and D indicate multicellular base of macrohair. Normal bundle-sheath anatomy indicated by arrowhead in D, abnormal bundle-sheath anatomy indicated by arrowheads in I and J. Scale bars: 100 µm.

 


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Fig. 3. lg1 and lg2 expression in Wab1 leaves. RT-PCR gel blot analysis of lg1 and lg2 expression in 4-week-old seedlings of wild-type and Wab1/+ at the following stages of leaf development: SAM including P1-5 (S), P6-8 pre-ligule region (PL), P6-8 blade (B), P9-10 ligule region (L). Lanes 9-16 are as for lanes 1-8 using a four-fold dilution of cDNA. Control with no cDNA included in PCR (C). Control amplification of ubiquitin (ub) indicates equal amounts of cDNA present in each sample.

 


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Fig. 4. Sector phenotypes in Wab1 mutants. (A-E) Abaxial view of Wab1 leaves with auricle extension and/or ectopic auricle phenotypes with albino sectors exhibiting normal blade characteristics. (F) Scanning electron micrograph of adaxial surface of boxed region in C, illustrating unexpanded, dovetailed auricle-like cells in Wab1/wab1+ ectopic auricle tissue (right of arrowhead), and normal blade epidermal characteristics such as macrohairs in the wab1+/– sector (left of arrowhead). (G) Fluorescence micrograph of a transverse section through a sector adjacent to auricle extension, illustrating a sharp boundary between green, Wab1/wab1+ auricle-like tissue (which fluoresces red), and albino, wab1+/– tissue (which appears blue-green). (H) SEM of adaxial surface of sector boundary near that shown in G, with hairy, fully expanded auricle-like cells in the Wab1/wab1+ tissue (right of arrowhead) and normal blade cells including prickle hairs and macrohairs in the wab1+/– tissue (left of arrowhead). (I) Fluorescence micrograph of a transverse section through the sector shown in J. Green, Wab1/wab1+ tissue is very thin with widely spaced veins, a hairless adaxial surface and abaxial hairs characteristic of marginal sheath tissue. Albino wab1+/– tissue has the histological organization of normal blade tissue. (J) Abaxial view of sector adjacent to ectopic sheath tissue, arrowhead marks sector boundary. (K) Cartoon depicting an albino non-mutant (w3 wab1+/–) sector in a green Wab1/wab1+ leaf. Scale bar: 500 µm (F) and 100 µm (G-I).

 


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Fig. 5. Sector phenotypes in lg1-R;Wab1 mutants. (A) Abaxial view of yellow v4, wab1+/– sector, and (B) adaxial and (C) abaxial view of white, w3 wab1+/– sectors adjacent to ectopic sheath tissue. Arrowheads in A and B mark sector borders. (E) Fluorescence micrograph of a transverse section through the inner sector boundary of leaf shown in C. Green lg1-R;Wab1 tissue has the histological organisation of sheath, and albino wab1+/– tissue that of the blade. (D) SEM of adaxial surface of sector boundary shown in C and D, illustrating the sharp boundary between epidermal cell types in lg1-R;Wab1/wab1+ tissue (right of arrowhead) and lg1-R;wab1+/– tissue (left of arrowhead). (F) Fluorescence micrograph of a transverse section through another sector adjacent to marginal sheath-like tissue. The adaxial surface of green sheath-like tissue in D and F is hairless, whereas the albino tissue has hairs specific to blade. Scale bar: 100 µm (D-F).

 


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Fig. 6. lg1 affects cell autonomy of the Wab1 phenotype. (A) Cartoon illustrating spread of the Wab1 phenotype (wavy lines) into a wab1+/– sector and spread of normal phenotype (no wavy lines) from a sector into Wab1/wab1+ tissue. We propose that lg1 may transmit both correct and incorrect positional signals towards the margins of Wab1 leaves (red arrows). (B) In the absence of lg1, the Wab1 phenotype is strictly cell-autonomous.

 


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Fig. 7. Model for lg1 function in leaf morphogenesis. (A) Cartoon of wild-type leaf illustrating distal blade (green) and proximal sheath (blue) separated by ligule and auricle (red). (B) In the absence of lg1, ligule and auricle are deleted, the blade-sheath boundary is shifted distally, and the base of the blade is narrower than in wild-type leaves. (C) Wab1 disrupts proximodistal patterning and restricts lateral growth of the blade. Misexpression of lg1 in Wab1 leaves results in ectopic ligule and auricle, and partially compensates for the narrow leaf phenotype at the base of the blade. The lateral signalling function of lg1 permits some recovery of proximodistal patterning at the margins of Wab1 leaves. (D) In the absence of lg1, Wab1 leaves never establish blade at the margins and are extremely narrow.

 

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© The Company of Biologists Ltd 2004