Cotton GalT1 Encoding a Putative Glycosyltransferase Is Involved in Regulation of Cell Wall Pectin Biosynthesis during Plant Development

Qin, Lixia; Rao, Yue; Li, Long; Huang, Junfeng; Xu, Wen‐Liang; Li, Xuebao

doi:10.1371/journal.pone.0059115

Cited by 32 publications

(22 citation statements)

References 46 publications

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“…Then, the constructed GhRAV1:eGFP fusion gene was cloned into pBI121 vector, replacing the GUS gene. Tobacco leaf epidermal cells were injected with the GhRAV1:eGFP construct by Agrobacterium -mediated DNA transfer as described previously [ 20 ]. The cells transient-expressing GhRAV1 : eGFP gene under the control of CaMV 35S promoter were selected for detecting GFP fluorescence on a SP5 Meta confocal laser microscope (Leica, Germany) with a filter set of 488 nm for excitation and 506–538 nm for emission, after the injected tobacco leaves were cultured on MS medium for 72 h. SP5 software (Leica, Germany) was employed to record and process the digital images taken [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

“…Chlorophyll content was determined on a fresh-weight basis. Total chlorophyll from 0.1 g leaves of ten-day-old seedlings with or without NaCl treatment was extracted with 95% ethanol, and chlorophyll concentrations were calculated by the method described earlier [ 20 ] using spectrophotometer (TU-1901) and microplate reader (Bioteck, USA). Differences in chlorophyll content between wild type and the transgenic seedlings were evaluated using a P value generated by a one-sided t-test.…”

Section: Methodsmentioning

confidence: 99%

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Overexpression of Cotton RAV1 Gene in Arabidopsis Confers Transgenic Plants High Salinity and Drought Sensitivity

Zhou

et al. 2015

PLoS ONE

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RAV (related to ABI3/VP1) protein containing an AP2 domain in the N-terminal region and a B3 domain in the C-terminal region, which belongs to AP2 transcription factor family, is unique in higher plants. In this study, a gene (GhRAV1) encoding a RAV protein of 357 amino acids was identified in cotton (Gossypium hirsutum). Transient expression analysis of the eGFP:GhRAV1 fusion genes in tobacco (Nicotiana tabacum) epidermal cells revealed that GhRAV1 protein was localized in the cell nucleus. Quantitative RT-PCR analysis indicated that expression of GhRAV1 in cotton is induced by abscisic acid (ABA), NaCl and polyethylene glycol (PEG). Overexpression of GhRAV1 in Arabidopsis resulted in plant sensitive to ABA, NaCl and PEG. With abscisic acid (ABA) treatment, seed germination and green seedling rates of the GhRAV1 transgenic plants were remarkably lower than those of wild type. In the presence of NaCl, the seed germination and seedling growth of the GhRAV1 transgenic lines were inhibited greater than those of wild type. And chlorophyll content and maximum photochemical efficiency of the transgenic plants were significantly lower than those of wild type. Under drought stress, the GhRAV1 transgenic plants displayed more severe wilting than wild type. Furthermore, expressions of the stress-related genes were altered in the GhRAV1 transgenic Arabidopsis plants under high salinity and drought stresses. Collectively, our data suggested that GhRAV1 may be involved in response to high salinity and drought stresses through regulating expressions of the stress-related genes during cotton development.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Overexpression of Cotton RAV1 Gene in Arabidopsis Confers Transgenic Plants High Salinity and Drought Sensitivity

Zhou

et al. 2015

PLoS ONE

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“…We designed gene‐specific primers for each gene and qRT‐PCR analyses do show that the expression levels of CotAD_70785, CotAD_68102, CotAD_43585 and CotAD_16822 were significantly reduced in GhGalT1 ‐silenced lines (Figure S10b). These data illustrate that the above five genes might function redundantly (or partially redundantly) to encode similar enzyme activity, and that cross‐silencing them led to the most prominent fiber phenotype; however, when heterologously expressed in Arabidopsis, GhGalT1 can promote plant growth (Qin et al ., ). These seemingly contrasting phenotypes may result from off‐target effects of the ectopic expression of GhGalT1 , or because the putative functional enzymatic complex formed in Arabidopsis may not be optimal because key interacting partners to a specific cell type (such as fiber cell) may be absent in Arabidopsis.…”

Section: Discussionmentioning

confidence: 97%

“…To date, no cotton glycosyltransferase linked to the biosynthesis of the AGP glycan moiety has been characterized in vivo . We previously identified a gene ( GhGalT1 ) encoding a putative galactosyltransferase (GalT) in cotton and found that the heterologous expression of GhGalT1 in Arabidopsis affected plant growth and development (Qin et al ., ). Here, we provide genetic, cellular and biochemical evidence that GhGalT1 is a galactosyltransferase involved in AGP β‐1,3‐galactan backbone biosynthesis, and proffer a mechanism by which AGPs participate in controlling the fiber development of cotton.…”

Section: Introductionmentioning

confidence: 97%

The cotton β‐galactosyltransferase 1 (GalT1) that galactosylates arabinogalactan proteins participates in controlling fiber development

et al. 2017

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Arabinogalactan proteins (AGPs) are highly glycosylated proteins that play pivotal roles in diverse developmental processes in plants. Type-II AG glycans, mostly O-linked to the hydroxyproline residues of the protein backbone, account for up to 95% w/w of the AGP, but their functions are still largely unclear. Cotton fibers are extremely elongated single-cell trichomes on the seed epidermis; however, little is known of the molecular basis governing the regulation of fiber cell development. Here, we characterized the role of a CAZy glycosyltransferase 31 (GT31) family member, GhGalT1, in cotton fiber development. The fiber length of the transgenic cotton overexpressing GhGalT1 was shorter than that of the wild type, whereas in the GhGalT1-silenced lines there was a notable increase in fiber length compared with wild type. The carbohydrate moieties of AGPs were altered in fibers of GhGalT1 transgenic cotton. The galactose: arabinose ratio of AG glycans was higher in GhGalT1 overexpression fibers, but was lower in GhGalT1-silenced lines, compared with that in the wild type. Overexpression of GhGalT1 upregulates transcript levels of a broad range of cell wall-related genes, especially the fasciclin-like AGP (FLA) backbone genes. An enzyme activity assay demonstrated that GhGalT1 is a β-1,3-galactosyltransferase (β-1,3-GalT) involved in biosynthesis of the β-1,3-galactan backbone of the type-II AG glycans of AGPs. We also show that GhGalT1 can form homo- and heterodimers with other cotton GT31 family members to facilitate AG glycan assembly of AGPs. Thus, our data demonstrate that GhGalT1 influences cotton fiber development via controlling the glycosylation of AGPs, especially FLAs.

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“…Extraction of cell wall residues (i.e., alcohol-insoluble residues [AIRs]) was conducted as described previously Qin et al, 2013). Cell wall crystalline cellulose amount was measured according to Kim et al (Kim et al, 2013).…”

Section: Cell Wall Extraction and Crystalline Cellulose Analysismentioning

confidence: 99%

Cotton GhMYB7 is predominantly expressed in developing fibers and regulates secondary cell wall biosynthesis in transgenic Arabidopsis

Huang

Chen

et al. 2016

Sci. China Life Sci.

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The secondary cell wall in mature cotton fibers contains over 90% cellulose with low quantities of xylan and lignin. However, little is known regarding the regulation of secondary cell wall biosynthesis in cotton fibers. In this study, we characterized an R2R3-MYB transcription factor, GhMYB7, in cotton. GhMYB7 is expressed at a high level in developing fibers and encodes a MYB protein that is targeted to the cell nucleus and has transcriptional activation activity. Ectopic expression of GhMYB7 in Arabidopsis resulted in small, curled, dark green leaves and also led to shorter inflorescence stems. A cross-sectional assay of basal stems revealed that cell wall thickness of vessels and interfascicular fibers was higher in transgenic lines overexpressing GhMYB7 than in the wild type. Constitutive expression of GhMYB7 in Arabidopsis activated the expression of a suite of secondary cell wall biosynthesis-related genes (including some secondary cell wall-associated transcription factors), leading to the ectopic deposition of cellulose and lignin. The ectopic deposition of secondary cell walls may have been initiated before the cessation of cell expansion. Moreover, GhMYB7 was capable of binding to the promoter regions of AtSND1 and AtCesA4, suggesting that GhMYB7 may function upstream of NAC transcription factors. Collectively, these findings suggest that GhMYB7 is a potential transcriptional activator, which may participate in regulating secondary cell wall biosynthesis of cotton fibers.cotton (Gossypium hirsutum), fiber development, MYB transcription factor, secondary cell wall (SCW) biosynthesis, ectopic gene expression

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Cotton GalT1 Encoding a Putative Glycosyltransferase Is Involved in Regulation of Cell Wall Pectin Biosynthesis during Plant Development

Cited by 32 publications

References 46 publications

Overexpression of Cotton RAV1 Gene in Arabidopsis Confers Transgenic Plants High Salinity and Drought Sensitivity

Overexpression of Cotton RAV1 Gene in Arabidopsis Confers Transgenic Plants High Salinity and Drought Sensitivity

The cotton β‐galactosyltransferase 1 (GalT1) that galactosylates arabinogalactan proteins participates in controlling fiber development

Cotton GhMYB7 is predominantly expressed in developing fibers and regulates secondary cell wall biosynthesis in transgenic Arabidopsis

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