Dataset Information

Petal abscission in rose is associated with the differential expression of two ethylene-responsive xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2.

ABSTRACT: Abscission is a process that involves shedding of plant organs from the main plant body. In this study it is shown that the process of petal separation in the fragrant rose, Rosa bourboniana, is accompanied by the expression of two xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2. The sequences of the two genes show 52% amino acid identity but are conserved at the catalytic site. The genes are up-regulated soon after the initiation of the abscission process and their transcription is associated with the progression of abscission, being faster in ethylene-treated flowers but slower during field abscission. Transcription is ethylene responsive, with the ethylene response being tissue-specific for RbXTH1 but largely tissue-independent for RbXTH2. Expression is correlated with an increase in xyloglucan endotransglucosylase (XET) action in petal abscission zones of both ethylene-treated and field abscising flowers. Proximal promoters of both the genes drive ?-glucuronidase expression in an ethylene-responsive and abscission-related manner in agrobacteria-infiltrated rose petals, indicating that cis-elements governing ethylene-responsive and abscission-related expression probably lie within the first 700 nucleotides upstream of the translational initiation codon. The results show that cell wall remodelling of the xyloglucan moieties through the XET action of XTHs may be important for cell separation during abscission.

SUBMITTER: Singh AP

PROVIDER: S-EPMC3193013 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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Petal abscission in rose is associated with the differential expression of two ethylene-responsive xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2.

Singh Amar Pal AP Tripathi Siddharth Kaushal SK Nath Pravendra P Sane Aniruddha P AP

Journal of experimental botany 20110715 14

Abscission is a process that involves shedding of plant organs from the main plant body. In this study it is shown that the process of petal separation in the fragrant rose, Rosa bourboniana, is accompanied by the expression of two xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2. The sequences of the two genes show 52% amino acid identity but are conserved at the catalytic site. The genes are up-regulated soon after the initiation of the abscission process and their transcript ...[more]

PMID: 21765161

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Project description:Background and aimsShoot elongation of arrowhead tubers (Sagittaria pygmaea Miq.) is stimulated by anoxia, ethylene and CO2. The aim of this study was to characterize anoxic elongation by comparison with elongation stimulated by ethylene and CO2.MethodsThe effects of the inhibitors aminoethoxyvinylglycine (AVG) as an ethylene biosynthesis inhibitor, 1-methylcyclopropene (1-MCP) as a potent inhibitor of ethylene action, and pyrazol, an inhibitor of alcohol dehydrogenase, on shoot elongation were examined. Moreover, the effects of these gaseous factors on expression of genes possibly involved in modification of cell wall architecture were examined by polymerase chain reaction (PCR) methods.Key results and conclusionsIn air, promotion by 5% CO2 and 5 microL L-1 ethylene of shoot elongation occurred. At 1% O2, ethylene also stimulated shoot elongation but CO2 did not. Pyrazol inhibited shoot elongation in hypoxia but not in normoxia, suggesting that alcohol fermentation contributes to elongation enhanced by hypoxia. AVG and 1-MCP partially prevented shoot elongation both in normoxia and in hypoxia, but they did not have significant effects in anoxia, suggesting that endogenous ethylene acts as a stimulator of shoot elongation in normoxia and in hypoxia but not in anoxia. Ethylene is not involved in anoxia-enhanced elongation. We cloned four cDNAs (SpEXPA1, 2, 3 and 4) encoding alpha-expansin (EXPA) and five cDNAs (SpXTH1, 2, 3, 4 and 5) encoding xyloglucan endotransglucosylase/hydrolase (XTH) from shoots of arrowhead tubers. The transcript levels of SpEXPA1 and 2 were increased by anoxia and those of SpEXPA2 were increased by 5% CO2. Ethylene slightly elevated the level of SpEXPA4 transcripts. Anoxia enhanced the transcript levels of SpXTH1 and 4; neither ethylene nor CO2 had any effect. CO2 enhanced transcript levels of SpXTH3 and depressed those of SpXTH5. Ethylene decreased transcript levels of SpXTH5. These results suggest that four SpEXPA genes and five SpXTH genes are differently responsive to anoxia, CO2 and ethylene. Enhancement of SpEXPA1 and 2, and SpXTH1 and 4 transcript levels suggests that these gene products are involved in anoxic shoot elongation through modification of cell wall architecture.

Project description:BACKGROUND AND AIMS:Xyloglucan endotransglucosylase/hydrolase (XTH) proteins that possess xyloglucan endotransglucosylase (XET) activity contribute to cell-wall assembly and remodelling, orchestrating plant growth and development. Little is known about in-vivo XET regulation, other than at the XTH transcriptional level. Plants contain 'cold-water-extractable, heat-stable polymers' (CHPs) which are XTH-activating factors (XAFs) that desorb and thereby activate wall-bound XTHs. Because XAFs may control cell-wall modification in vivo, we have further explored their nature. METHODS:Material was cold-water-extracted from 25 plant species; proteins were precipitated by heat-denaturation, then CHP was ethanol-precipitated. For XAF assays, CHP (or sub-fractions thereof) was applied to washed Arabidopsis thaliana cell walls, and the enzymes thus solubilized were assayed radiochemically for XET activity. In some experiments, the CHP was pre-treated with trifluoroacetic acid (TFA), alkali (NaOH) or glycanases. KEY RESULTS:CHP specifically desorbed wall-bound XTHs, but not ?-glucosidases, phosphatases or peroxidases. CHP preparations from 25 angiosperms all possessed XAF activity but had no consistent monosaccharide composition. Of 11 individual plant polymers tested, only gum arabic and tamarind xyloglucan were XAF-active, albeit less so than CHP. On gel-permeation chromatography, XAF-active cauliflower CHP eluted with a molecular weight of ~7000-140 000, although no specific sugar residue(s) co-eluted exactly with XAF activity. Cauliflower XAF activity survived cold alkali and warm dilute TFA (which break ester and glycofuranosyl linkages, respectively), but was inactivated by hot 2 m TFA (which breaks glycopyranosyl linkages). Cauliflower XAF activity was remarkably stable to diverse glycanases and glycosidases. CONCLUSIONS:XAFs are naturally occurring heat-stable polymers that specifically desorb (thereby activating) wall-bound XTHs. Their XAF activity considerably exceeds that of gum arabic and tamarind xyloglucan, and they were not identifiable as any major plant polysaccharide. We propose that XAF is a specific, minor, plant polymer that regulates xyloglucan transglycosylation in vivo, and thus wall assembly and restructuring.

Project description:Background and aimsAlthough xyloglucans are ubiquitous in land plants, they are less abundant in Poales species than in eudicotyledons. Poales cell walls contain higher levels of ?-1,3/1,4 mixed-linked glucans and arabinoxylans than xyloglucans. Despite the relatively low level of xyloglucans in Poales, the xyloglucan endotransglucosylase/hydrolase (XTH) gene family in rice (Oryza sativa) is comparable in size to that of the eudicotyledon Arabidopsis thaliana. This raises the question of whether xyloglucan is a substrate for rice XTH gene products, whose enzyme activity remains largely uncharacterized.MethodsThis study focused on OsXTH19 (which belongs to Group IIIA of the XTH family and is specifically expressed in growing tissues of rice shoots), and two other XTHs, OsXTH11 (Group I/II) and OsXTH20 (Group IIIA), for reference, and measurements were made of the enzymatic activities of three recombinant rice XTHs, i.e. OsXTH11, OsXTH20 and OsXTH19.Key resultsAll three OsXTH gene products have xyloglucan endohydrolase (XEH, EC 3·2·1·151) activity, and OsXTH11 has both XEH and xyloglucan endotransglycosylase (XET, EC 2·4·1207) activities. However, these proteins had neither hydrolase nor transglucosylase activity when glucuronoarabinoxylan or mixed-linkage glucan was used as the substrate. These results are consistent with histological observations demonstrating that pOsXTH19::GUS is expressed specifically in the vicinity of tissues where xyloglucan immunoreactivity is present. Transgenic rice lines over-expressing OsXTH19 (harbouring a Cauliflower Mosaic Virus 35S promoter::OsXTH19 cDNA construct) or with suppressed OsXTH19 expression (harbouring a pOsXTH19 RNAi construct) did not show dramatic phenotypic changes, suggesting functional redundancy and collaboration among XTH family members, as was observed in A. thaliana.ConclusionsOsXTH20 and OsXTH19 act as hydrolases exclusively on xyloglucan, while OsXTH11 exhibits both hydrolase and XET activities exclusively on xyloglucans. Phenotypic analysis of transgenic lines with altered expression of OsXTH19 suggests that OsXTH19 and related XTH(s) play redundant roles in rice growth.

Dataset Information

Petal abscission in rose is associated with the differential expression of two ethylene-responsive xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2.

Publications

Petal abscission in rose is associated with the differential expression of two ethylene-responsive xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2.

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