Dataset Information

Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Vacuole-Targeted Thermotoga maritima BglB Related to Elevated Levels of Liberated Hormones.

ABSTRACT: The hyperthermostable ?-glucosidase BglB of Thermotoga maritima was modified by adding a short C-terminal tetrapeptide (AFVY, which transports phaseolin to the vacuole, to its C-terminal sequence). The modified ?-glucosidase BglB was transformed into tobacco (Nicotiana tabacum L.) plants. We observed a range of significant phenotypic changes in the transgenic plants compared to the wild-type (WT) plants. The transgenic plants had faster stem growth, earlier flowering, enhanced root systems development, an increased biomass biosynthesis rate, and higher salt stress tolerance in young plants compared to WT. In addition, programed cell death was enhanced in mature plants. Furthermore, the C-terminal AFVY tetrapeptide efficiently sorted T. maritima BglB into the vacuole, which was maintained in an active form and could perform its glycoside hydrolysis function on hormone conjugates, leading to elevated hormone [abscisic acid (ABA), indole 3-acetic acid (IAA), and cytokinin] levels that likely contributed to the phenotypic changes in the transgenic plants. The elevation of cytokinin led to upregulation of the transcription factor WUSCHELL, a homeodomain factor that regulates the development, division, and reproduction of stem cells in the shoot apical meristems. Elevation of IAA led to enhanced root development, and the elevation of ABA contributed to enhanced tolerance to salt stress and programed cell death. These results suggest that overexpressing vacuole-targeted T. maritima BglB may have several advantages for molecular farming technology to improve multiple targets, including enhanced production of the ?-glucosidase BglB, increased biomass, and shortened developmental stages, that could play pivotal roles in bioenergy and biofuel production.

SUBMITTER: Nguyen QA

PROVIDER: S-EPMC4642495 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Vacuole-Targeted Thermotoga maritima BglB Related to Elevated Levels of Liberated Hormones.

Nguyen Quynh Anh QA Lee Dae-Seok DS Jung Jakyun J Bae Hyeun-Jong HJ

Frontiers in bioengineering and biotechnology 20151109

The hyperthermostable β-glucosidase BglB of Thermotoga maritima was modified by adding a short C-terminal tetrapeptide (AFVY, which transports phaseolin to the vacuole, to its C-terminal sequence). The modified β-glucosidase BglB was transformed into tobacco (Nicotiana tabacum L.) plants. We observed a range of significant phenotypic changes in the transgenic plants compared to the wild-type (WT) plants. The transgenic plants had faster stem growth, earlier flowering, enhanced root systems devel ...[more]

PMID: 26618153

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Project description:Transgenic tobacco (N. tabacum cv. Xanthi nc) expressing Capsicum chinense CchGLP gene that encodes an Mn-SOD, constitutively produces hydrogen peroxide that increase endogenous ROS levels. Previous studies using these plants against geminivirus infections as well as drought stress confirmed that CchGLP expression conferred resistance against biotic and abiotic stresses. Cadmium (Cd) and Aluminium (Al) contamination in soils are a major ecological concern since they are two of the most widespread toxic elements in terrestrial environments. Trying to explore additional possible tolerance to another stresses in these plants, the aim of this work was to analyse the response to cadmium and aluminium salts during germination and early stages of plantlet development and a differential transcriptome of microRNAs (miRNAs) expression in expressing CchGLP transgenic lines and an azygote non-CchGLP expressing line. Plants were grown in vitro with addition of CdCl2 and AlCl3 at three different concentrations: 100, 300 and 500 μM and 50, 150 and 300 μM, respectively. The results showed higher tolerance to Cd and Al salts evaluated in two CchGLP-expressing transgenic lines L8 and L26 in comparison with the azygous non-CchGLP expressing line L1. Interestingly, L8 under Al stress presented vigorous roots and development of radicular hairs in comparison with azygous control (L1). Differentially expressed miRNAs in the comparison between L8 and L1 were associated with up and down-regulation of target genes related with structural molecule activity and ribosome constituents, as well as down-regulation in proton-transporting V-type ATPase (Vacuolar ATPase or V-ATPase). Moreover, KEGG analysis of the target genes for the differentially expressed miRNAs, led to identification of genes related with metabolic pathways and biosynthesis of secondary metabolites. One possible explanation of the tolerance to Cd and Al displayed in the transgenic tobaccos evaluated, might involve the fact that several down-regulated miRNAs, were found associated with target genes expressing V-ATPase. Specifically, miR7904-5p was down regulated and related with the up-regulation of one V-ATPase. The expression levels of these genes was confirmed by qRT-PCR assays, thus suggesting that a cation transport activity driven by the V-ATPases-dependent proton motive force, might significantly contribute as one mechanism for Cd and Al detoxification by vacuolar compartmentation in these transgenic tobacco plants.

Dataset Information

Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Vacuole-Targeted Thermotoga maritima BglB Related to Elevated Levels of Liberated Hormones.

Publications

Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Vacuole-Targeted Thermotoga maritima BglB Related to Elevated Levels of Liberated Hormones.

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