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A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco.

ABSTRACT: BACKGROUND AND AIMS: Multiple copies of genes encoding the catalytic subunit (c) of protein phosphatase 2A (PP2A) are commonly found in plants. For some of these genes, expression is up-regulated under water stress. The aim of this study was to investigate expression and characterization of TaPP2Ac-1 from Triticum aestivum, and to evaluate the effects of TaPP2Ac-1 on Nicotiana benthamiana in response to water stress. METHODS: TaPP2Ac-1 cDNA was isolated from wheat by in silico identification and RT-PCR amplification. Transcript levels of TaPP2Ac-1 were examined in wheat responding to water deficit. Copy numbers of TaPP2Ac-1 in wheat genomes and subcellular localization in onion epidermal cells were studied. Enzyme properties of the recombinant TaPP2Ac-1 protein were determined. In addition, studies were carried out in tobacco plants with pCAPE2-TaPP2Ac-1 under water-deficit conditions. KEY RESULTS: TaPP2Ac-1 cDNA was cloned from wheat. Transcript levels of TaPP2Ac-1 in wheat seedlings were up-regulated under drought condition. One copy for this TaPP2Ac-1 was present in each of the three wheat genomes. TaPP2Ac-1 fused with GFP was located in the nucleus and cytoplasm of onion epidermis cells. The recombinant TaPP2Ac-1 gene was over-expressed in Escherichia coli and encoded a functional serine/threonine phosphatase. Transgenic tobacco plants over-expressing TaPP2Ac-1 exhibited stronger drought tolerance than non-transgenic tobacco plants. CONCLUSIONS: Tobacco plants with pCAPE2-TaPP2Ac-1 appeared to be resistant to water deficit, as shown by their higher capacity to maintain leaf relative water content, leaf cell-membrane stability index, water-retention ability and water use efficiency under water stress. The results suggest that the physiological role of TaPP2Ac-1 is related to drought stress response, possibly through its involvement in drought-responding signal transduction pathways.

SUBMITTER: Xu C

PROVIDER: S-EPMC2802960 | biostudies-literature | 2007 Mar

REPOSITORIES: biostudies-literature

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A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco.

Xu Chongyi C Jing Ruilian R Mao Xinguo X Jia Xiaoyun X Chang Xiaoping X

Annals of botany 20070201 3

<h4>Background and aims</h4>Multiple copies of genes encoding the catalytic subunit (c) of protein phosphatase 2A (PP2A) are commonly found in plants. For some of these genes, expression is up-regulated under water stress. The aim of this study was to investigate expression and characterization of TaPP2Ac-1 from Triticum aestivum, and to evaluate the effects of TaPP2Ac-1 on Nicotiana benthamiana in response to water stress.<h4>Methods</h4>TaPP2Ac-1 cDNA was isolated from wheat by in silico ident ...[more]

PMID: 17272305

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Project description:Eukaryotic type 2A protein phosphatases (protein phosphatase 2A, PP2A) consist of a scaffold subunit A, a regulatory subunit B, and a catalytic subunit C. Little is known about the roles of PP2Ac proteins that are involved in plant responses to necrotrophic fungal pathogens. Sharp eyespot, caused by the necrotrophic fungus Rhizoctonia cerealis, is a destructive disease of wheat (Triticum aestivum), an important staple food crop. Here, we isolated TaPP2Ac-4D from wheat, which encodes a catalytic subunit of the heterotrimeric PP2A, and characterized its properties and role in plant defense response to R. cerealis. Based on the sequence alignment of TaPP2Ac-4D with the draft sequences of wheat chromosomes from the International Wheat Genome Sequencing Consortium (IWGSC), it was found that TaPP2Ac-4D gene is located on the long arm of the wheat chromosome 4D and has two homologs assigned on wheat chromosomes 4A and 4B. Sequence and phylogenetic tree analyses revealed that the TaPP2Ac protein is a typical member of the PP2Ac family and belongs to the subfamily II. TaPP2Ac-4B and TaPP2Ac-4D displayed higher transcriptional levels in the R. cerealis-susceptible wheat cultivar Wenmai 6 than those seen in the resistant wheat line CI12633. The transcriptional levels of TaPP2Ac-4B and TaPP2Ac-4D were significantly elevated in wheat R. cerealis after infection and upon H2O2 treatment. Virus-induced gene silencing results revealed that the transcriptional knockdown of TaPP2Ac-4D and TaPP2Ac-4B significantly increased wheat resistance to R. cerealis infection. Meanwhile, the transcriptional levels of certain pathogenesis-related (PR) and reactive oxygen species (ROS)-scavenging enzyme encoding genes were increased in TaPP2Ac-silenced wheat plants. These results suggest that TaPP2Ac-4B and TaPP2Ac-4D negatively regulate defense response to R. cerealis infection possibly through modulation of the expression of certain PR and ROS-scavenging enzyme genes in wheat. This study reveals a novel function of the plant PP2Ac genes in plant immune responses.

Project description:Wheat is the most widely grown and consumed crop because of its economic and social benefits. This crop is more important globally for food and feed, and its productivity is particularly vulnerable to abiotic factors. In this study, 40 wheat genotypes were studied to access the drought tolerance level using completely randomized design (CRD) in 250 ml disposable cups through morpho-physiological attributes at seedling stage. The wheat germplasm was tested under normal and two drought stress level D1 (50% field capacity) and D2 (75% field capacity) for different seedling attributes such as germination percentage (GP), chlorophyll content (CC), shoot length (SL), root length (RL), shoot fresh weight (SFW), root fresh weight (RFW), seedling fresh weight (SDFW), shoot dry weight (SDW), root dry weight (RDW), relative water content (RWC), root/shoot ratio (RS), and seedling dry weight (SeDW). The results of analysis of variance (ANOVA) and spider analysis indicate that significant amount of genetic variation was present and behavior of studied germplasm showed different behavior in different environment. The correlation analysis showed that root length has significantly positive association with root/shoot ratio, dry weight, and fresh weight while negatively correlated with shoot length and relative water content. Based on the positively associated traits, the studied genetic material would improve genetic gain for drought tolerance. The multivariate analysis showed that out 13 principal components only five PCs were significant and has eigenvalue > 1, cumulatively showed 82.33, 83.07, and 97.34% of total variation under normal, D1 and D2 conditions, respectively. Significantly, the result of spider graph and multivariate analysis showed that genotypes G47, G48, G65, G68, and G80 performed well in all drought stress conditions and considered as drought-tolerant genotypes. The best performing genotypes can be used in future breeding programs. The selection on the bases of studied attributes is effective for development of drought-tolerant and high-yielding varieties for sustainable food security.

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A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco.

Publications

A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco.

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