Determination of the structures of cutin monomers by a novel depolymerization procedure and combined gas chromatography and mass spectrometry

Walton, Terence J.; Kolattukudy, Pappachan E.

doi:10.1021/bi00760a025

Cited by 203 publications

(121 citation statements)

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“…First, ABA, which induces coating formation, is known to induce suberization (4,24). The concentration of ABA required to stimulate the coating response was 10-6 M, whereas previous work had indicated that a concentration of 10-4 M was necessary to induce suberization in potato callus cultures (4 (26). Because the alkane-a,w-diols did not originate from the cuticle, they most likely were generated by the reduction of the c-hydroxy fatty acids and dicarboxylic acids of suberin from the vascular coating material.…”

Section: Discussionmentioning

confidence: 99%

“…The diol fraction was then silylated with N,Obis(trimethylsilyl)acetamide and analyzed by capillary GC-MS on a 15-m SPB-5 (Supelco) column in a Hewlett-Packard 5890 gas chromatograph attached to a Hewlett-Packard 59888A mass spectrometer equipped with a Chemstation computer data system. Mass spectra, recorded at 70 eV, were used for identification of the diols (26 (Fig. 1, a-f), were observed with all suberin stains but were not observed in controls.…”

Section: Analysis Of Suberinmentioning

confidence: 99%

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Chemical Characterization of Stress-Induced Vascular Coating in Tomato

Robb¹,

Lee²,

Mohan³

et al. 1991

Plant Physiol.

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Indirect evidence suggests that vascular coatings formed by plants in response to stress consist of suberin-like substances containing lipid and phenolic compounds. To provide more direct chemical evidence that coatings are suberin, we used a natural pathogen, Verticillium albo-atrum, or a stress-responsive hormone, abscisic acid, to induce coating in two isolines of tomato (Lycopersicon esculentum L. cultivar Craigella) that are resistant or susceptible to the pathogen. Using treated petioles that had been monitored cytologically, chemical depolymerization followed by combined gas-liquid chromatography-mass spectrometry analysis of alkane-a,w-diol levels confirmed the presence of suberin after induction of coating and showed quantitative differences between the isolines that correlated with cytological measurements of the coating response. Northem analysis of suberization-associated anionic peroxidase mRNA showed corresponding increases, and tissue blot analysis further indicated that induction of the mRNA was localized in the responding vascular bundles, as determined by suberin histochemistry. Taken together, these results provide chemical evidence that the coatings are mainly suberin. Stress, whether biotic or abiotic, frequently induces the vascular coating response in plants (for review see ref. 19). A recent study (25) ofearly interactions between the wilt fungus, Verticillium albo-atrum, and its tomato host indicated that the coating of xylem vessel walls involves the secretion and deposition of normal or modified cell wall components. The nature of the polymeric materials deposited on the cell wall remains unknown. Although pathogen-induced wall coatings have been variously designated as lignin, induced lignin, lignin-like, and suberin, and although most studies have not satisfactorily distinguished between lignification and suberization, in most cases the deposits probably result from suberization (9, 15). The same process also is known to be injuryinduced in all plant organs (5).According to the current model, suberin consists of an aromatic domain and an aliphatic domain that is probably attached to the aromatic domain (10). The phenolic monomers derived from the phenylpropanoid pathway are polym-

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Section: Discussionmentioning

confidence: 99%

Section: Analysis Of Suberinmentioning

confidence: 99%

Chemical Characterization of Stress-Induced Vascular Coating in Tomato

Robb¹,

Lee²,

Mohan³

et al. 1991

Plant Physiol.

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“…Powdered cutin from apple leaf and fruits of apple, tomato, peach, and pear were obtained as previously described (11). A 500 mg sample of each cutin was degraded in refluxing tetrahydrofuran (25 ml) with an excess of LiAIH4 for a period of 24 to 48 hr.…”

Section: Isolation Of Phenolic Compounds From Cutin and Suberin Aftermentioning

confidence: 99%

Evidence for Covalently Attached p-Coumaric Acid and Ferulic Acid in Cutins and Suberins

Riley¹,

Kolattukudy²

1975

Plant Physiol.

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144

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p-Couimaric acid (4-hydroxycinnamic acid) and ferulic acid (4-hydroxy-3-methoxycinnamic acid) have been identified as conistituen-ts of cutin. Their reduction products were isolated fromi a phenolic fraction released from the cutin of the fruits of apple, peach, pear, and two varieties of tomato and apple leaf by treatment with LiAlH4 or LiAlD4. They were identified by comlibined gas chromatography and mass spectrormetry. p-Cotimaric acid was present in all samples of cutill (0.07-O.5i3% by weight), whereas only peach and pear cutin con-tainied measuirable aniotunits of ferulic acid (0.007% and 0.035.%, respectively). Both p-coutnaric acid and ferulic acid were idenitified to be conistituients of the insoluble material recovered after partial hydrolysis (12-42% loss) of cutin in 1 M NaOH at 80 C. A significanit part (48%) of the p-coumlaric acid contain-ed inl tomato cutin was contained in the insoluible material recovered after partial degradation (7.4%) of this cutin with 0.01 m NaOH. These data indicate that these phenolic components are tightly (possibly covalently) bountid to ctutin. Similar analysis of the phenolic fractions froni the stuberins of potato, sweet potato, turnip, rutabaga, carrot, anid red beet revealed that they contained onily fertilic acid (0.0.5-0.22%). Ferulic acid was identified as a constituen-t of the insoluible m-laterial recovered after partial hv-drolvsis of potato and beet suberins (34% and 32% loss, respectively) in 1 m NaOH at 80 C. A major part (65%) of the fertulic acid conitain-ed in potato suberin was contained in the inisoluble inaterial recovered after partial (26.8% loss) degradation of this suberin with 0.01 M NaOH. Fertulic acid appears to be tightly (probably covalently) bound to suiberin.Cutin, the structural component of plant cuticle, is a polymer of hydroxy fatty acids (8). The common major components of this polymer are palmitic acid, 16-hydroxypalmitic acid, 10,16-dihydroxypalmitic acid, or its positional isomers, oleic acid, 18-hydroxyoleic acid, 18-hydroxy-9, 10-epoxystearic acid, and 9, 10, 18-trihydroxystearic acid. The composition of the aliphatic components of suberin from the underground parts of plants is somewhat similar to that of cutin; however, there are significant quantitative differences between the composition of these two types of protective polymers (5-8). One major difference appears to be that suberin preparations contain quite large proportions of phenolic materials, while cutin preparations contain very little phenolic compounds, although the phenolics which might be present in cutin preparations have not been examined. In this paper we report experimental evidence which strongly suggests that p-coumaric acid and 'or ferulic acid are covalently attached to cutin and suberin. MATERIALS AND METHODS Isolation of Phenolic Compounds from Cutin and Suberin afterLiAlH1 and LiAID4 Degradation. Powdered cutin from apple leaf and fruits of apple, tomato, peach, and pear were obtained as previously described (11). A 500 mg sample of each cutin...

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“…Cutin attaches to the epidermis with a pectinaceous glue layer. Disruption of the pectinaceous glue that attaches the cuticle to the epidermal layer by enzymes or chemicals is required to release the cuticular layer (21). ␣,-Dicarboxylic fatty acid and in-chain hydroxy fatty acids have been reported to be the characteristic monomers of cutin.…”

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confidence: 99%

Defective in Cuticular Ridges (DCR) of Arabidopsis thaliana, a Gene Associated with Surface Cutin Formation, Encodes a Soluble Diacylglycerol Acyltransferase

Sapa

Krishna

Saha

et al. 2010

Journal of Biological Chemistry

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A key step in the triacylglycerol (TAG) biosynthetic pathway is the final acylation of diacylglycerol (DAG) by DAG acyltransferase. In silico analysis has revealed that the DCR (defective in cuticular ridges) (At5g23940) gene has a typical HX 4 D acyltransferase motif at the N-terminal end and a lipid binding motif VX 2 GF at the middle of the sequence. To understand the biochemical function, the gene was overexpressed in Escherichia coli, and the purified recombinant protein was found to acylate DAG specifically in an acyl-CoA-dependent manner. Overexpression of At5g23940 in a Saccharomyces cerevisiae quadruple mutant deficient in DAG acyltransferases resulted in TAG accumulation. At5g23940 rescued the growth of this quadruple mutant in the oleate-containing medium, whereas empty vector control did not. Lipid particles were localized in the cytosol of At5g23940-transformed quadruple mutant cells, as observed by oil red O staining. There was an incorporation of 16-hydroxyhexadecanoic acid into TAG in At5g23940-transformed cells of quadruple mutant. Here we report a soluble acyl-CoA-dependent DAG acyltransferase from Arabidopsis thaliana. Taken together, these data suggest that a broad specific DAG acyltransferase may be involved in the cutin as well as in the TAG biosynthesis by supplying hydroxy fatty acid. Triacylglycerol (TAG),3 an acyl ester glycerol, is the most efficient storage form of energy in eukaryotic cells (1, 2). In plants, TAG is the major component of the seed oils and is mainly deposited during seed and fruit development. Plant oils have a tremendous socioeconomic value in the food industry as well as in the production of lubricants and biofuels. They are also becoming increasingly important raw materials in the petrochemical industry as a substitute for non-renewable crude oil reserves (3). Thus, numerous conventional and molecular genetic strategies have been explored in attempts to increase the TAG content and modify the fatty acid composition of the plant seed oils. Isolation of the genes involved in oil biosynthesis offer exciting opportunities for overexpressing the desired gene of interest so as to redesign the plant oil metabolism for greater yields (3).TAG is usually synthesized by the sequential incorporation of acyl groups to glycerol 3-phosphate by substrate-specific acyltransferases to give lysophosphatidic acid and phosphatidic acid, respectively (4, 5). The synthesized phosphatidic acid is dephosphorylated to diacylglycerol (DAG). The last step in this pathway is catalyzed by acyl-CoA:DAG acyltransferase (DGAT; EC 2.3.1.20) that transfers an acyl group to DAG to form the TAG. DAG lies at the branch point between phospholipid and storage lipid biosynthesis. Therefore, much research has been focused on the acyl-CoA-dependent reaction catalyzed by the DAG acyltransferase, which is an integral endoplasmic reticulum protein (6) and has also been shown to be present in the oil bodies (7) and plastids (8). Several lines of evidence suggest that the TAG biosynthesis and DGAT activity are ...

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Determination of the structures of cutin monomers by a novel depolymerization procedure and combined gas chromatography and mass spectrometry

Cited by 203 publications

References 21 publications

Chemical Characterization of Stress-Induced Vascular Coating in Tomato

Chemical Characterization of Stress-Induced Vascular Coating in Tomato

Evidence for Covalently Attached p-Coumaric Acid and Ferulic Acid in Cutins and Suberins

Defective in Cuticular Ridges (DCR) of Arabidopsis thaliana, a Gene Associated with Surface Cutin Formation, Encodes a Soluble Diacylglycerol Acyltransferase

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