A new method for rapid visualization of defects in leaf cuticle reveals five intrinsic patterns of surface defects in <i>Arabidopsis</i>

Tanaka, Toshihiro; Tanaka, Hirokazu; Machida, Chiyoko; Watanabe, Masaru; Machida, Yasunori

doi:10.1046/j.1365-313x.2003.01946.x

Cited by 256 publications

(252 citation statements)

References 28 publications

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“…Several mutants defective in wax biosynthesis or secretion show a similar petal-folding phenotype, and their petals are sensitive to dye immersion due to the lower repellency of the petal surface ( 2009). We immersed the fop1-1 petals in the dye toluidine blue (Tanaka et al, 2004), but they were not stained (Supplemental Fig. S1, M and N).…”

Section: Discussionmentioning

confidence: 99%

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Physical Interaction of Floral Organs Controls Petal Morphogenesis in Arabidopsis

et al. 2013

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Flowering plants bear beautiful flowers to attract pollinators. Petals are the most variable organs in flowering plants, with their color, fragrance, and shape. In Arabidopsis (Arabidopsis thaliana), petal primordia arise at a similar time to stamen primordia and elongate at later stages through the narrow space between anthers and sepals. Although many of the genes involved in regulating petal identity and primordia growth are known, the molecular mechanism for the later elongation process remains unknown. We found a mutant, folded petals1 (fop1), in which normal petal development is inhibited during their growth through the narrow space between sepals and anthers, resulting in formation of folded petals at maturation. During elongation, the fop1 petals contact the sepal surface at several sites. The conical-shaped petal epidermal cells are flattened in the fop1 mutant, as if they had been pressed from the top. Surgical or genetic removal of sepals in young buds restores the regular growth of petals, suggesting that narrow space within a bud is the cause of petal folding in the fop1 mutant. FOP1 encodes a member of the bifunctional wax ester synthase/diacylglycerol acyltransferase family, WSD11, which is expressed in elongating petals and localized to the plasma membrane. These results suggest that the FOP1/WSD11 products synthesized in the petal epidermis may act as a lubricant, enabling uninhibited growth of the petals as they extend between the sepals and the anthers.

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Section: Discussionmentioning

confidence: 99%

“…The length and width of cleared petals were measured using the Image ProPlus 5.0 software. For whole-mount toluidine blue staining, petals were treated with 0.05% (w/v) toluidine blue for 2 min and washed in distilled water twice (Tanaka et al, 2004). Samples were visualized under an Axiophot 2 microscope (Carl Zeiss).…”

Section: Histology and Microscopesmentioning

confidence: 99%

Physical Interaction of Floral Organs Controls Petal Morphogenesis in Arabidopsis

et al. 2013

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“…Water loss was calculated as a percentage of weight loss. For measurements of cuticle permeability to stain, one MG fruit was collected from each plant of the F2 mapping population and from the parents, and dropped in 1% toluidine blue solution during 6 h as described in Tanaka et al (2004).…”

Section: Water Loss and Cuticle Permeability Measurementsmentioning

confidence: 99%

Analyses of Tomato Fruit Brightness Mutants Uncover Both Cutin-Deficient and Cutin-Abundant Mutants and a New Hypomorphic Allele of GDSL Lipase

et al. 2013

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The cuticle is a protective layer synthesized by epidermal cells of the plants and consisting of cutin covered and filled by waxes. In tomato (Solanum lycopersicum) fruit, the thick cuticle embedding epidermal cells has crucial roles in the control of pathogens, water loss, cracking, postharvest shelf-life, and brightness. To identify tomato mutants with modified cuticle composition and architecture and to further decipher the relationships between fruit brightness and cuticle in tomato, we screened an ethyl methanesulfonate mutant collection in the miniature tomato cultivar Micro-Tom for mutants with altered fruit brightness. Our screen resulted in the isolation of 16 glossy and 8 dull mutants displaying changes in the amount and/or composition of wax and cutin, cuticle thickness, and surface aspect of the fruit as characterized by optical and environmental scanning electron microscopy. The main conclusions on the relationships between fruit brightness and cuticle features were as follows: (1) screening for fruit brightness is an effective way to identify tomato cuticle mutants; (2) fruit brightness is independent from wax load variations; (3) glossy mutants show either reduced or increased cutin load; and (4) dull mutants display alterations in epidermal cell number and shape. Cuticle composition analyses further allowed the identification of groups of mutants displaying remarkable cuticle changes, such as mutants with increased dicarboxylic acids in cutin. Using genetic mapping of a strong cutindeficient mutation, we discovered a novel hypomorphic allele of GDSL lipase carrying a splice junction mutation, thus highlighting the potential of tomato brightness mutants for advancing our understanding of cuticle formation in plants.

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“…By contrast, rosette leaves of the mutants were not permeable to the dye. An increase in permeability to toluidine blue is indicative of cuticular defects (31). The presence in the dye solution of 0.01% Tween 20, a mild detergent, was used to minimize the surface effects of ridges and thus ensure a similar wettability between wild type (WT) and mutants.…”

Section: Identification Of Arabidopsis Mutants Lacking Flower Nanoridmentioning

confidence: 99%

Nanoridges that characterize the surface morphology of flowers require the synthesis of cutin polyester

Li‐Beisson

Pollard

Sauveplane

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

238

252

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Distinctive nanoridges on the surface of flowers have puzzled plant biologists ever since their discovery over 75 years ago. Although postulated to help attract insect pollinators, the function, chemical nature, and ontogeny of these surface nanostructures remain uncertain. Studies have been hampered by the fact that no ridgeless mutants have been identified. Here, we describe two mutants lacking nanoridges and define the biosynthetic pathway for 10,16-dihydroxypalmitate, a major cutin monomer in nature. Using gene expression profiling, two candidates for the formation of floral cutin were identified in the model plant Arabidopsis thaliana: the glycerol-3-phosphate acyltransferase 6 (GPAT6) and a member of a cytochrome P450 family with unknown biological function (CYP77A6). Plants carrying null mutations in either gene produced petals with no nanoridges and no cuticle could be observed by either scanning or transmission electron microscopy. A strong reduction in cutin content was found in flowers of both mutants. In planta overexpression suggested GPAT6 preferentially uses palmitate derivatives in cutin synthesis. Comparison of cutin monomer profiles in knockouts for CYP77A6 and the fatty acid -hydroxylase CYP86A4 provided genetic evidence that CYP77A6 is an in-chain hydroxylase acting subsequently to CYP86A4 in the synthesis of 10,16-dihydroxypalmitate. Biochemical activity of CYP77A6 was demonstrated by production of dihydroxypalmitates from 16-hydroxypalmitate, using CYP77A6-expressing yeast microsomes. These results define the biosynthetic pathway for an abundant and widespread monomer of the cutin polyester, show that the morphology of floral surfaces depends on the synthesis of cutin, and identify target genes to investigate the function of nanoridges in flower biology.10,16-dihydroxypalmitic acid ͉ cuticular ridges ͉ CYP77A6 ͉ CYP86A4 ͉ glycerol-3-phosphate acyltransferase 6

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A new method for rapid visualization of defects in leaf cuticle reveals five intrinsic patterns of surface defects in Arabidopsis

Cited by 256 publications

References 28 publications

Physical Interaction of Floral Organs Controls Petal Morphogenesis in Arabidopsis

Physical Interaction of Floral Organs Controls Petal Morphogenesis in Arabidopsis

Analyses of Tomato Fruit Brightness Mutants Uncover Both Cutin-Deficient and Cutin-Abundant Mutants and a New Hypomorphic Allele of GDSL Lipase

Nanoridges that characterize the surface morphology of flowers require the synthesis of cutin polyester

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