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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 an important cereal crop, which is adversely affected by water deficit stress. The effect of induced stress can be reduced by the application of salicylic acid (SA). With the objective to combat drought stress in wheat, an experiment was conducted in greenhouse under hydroponic conditions. The treatments consisted of (a) no drought (DD0 = 0 MPa), mild drought (DD1 = -0.40 MPa) and severe drought (DD2 = -0.60 MPa) by applying PEG-8000, (b) two contrasting wheat varieties Barani-17 (drought tolerant) and Anaj-17 (drought-sensitive), and (c) foliar treatments of salicylic acid (0, 50 mM, 75 mM, and 100 mM). Evaluation of wheat plants regarding biochemical, physiological, and morphological attributes were rendered after harvesting of plants. Statistically, maximum shoot and root fresh and dry weights (18.77, 11.15 and 1.99, 1.81 g, respectively) were recorded in cultivar Barani-17 under no drought condition with the application of SA (100 mM). While, minimum shoot and root fresh and dry weights (6.65, 3.14 and 0.73, 0.61 g, respectively) were recorded in cultivar Anaj-2017 under mild drought stress without SA application. The maximum shoot length (68.0 cm) was observed in cultivar Barani-2017 under no drought condition with the application of SA (100 mM). While, maximum root length (59.67 cm) was recorded in cultivar Anaj-17 under moderate drought stress without application of SA. Further, minimum shoot length (28.67 cm) was recorded in Anaj-17 under moderate drought stress without SA application. Minimum root length (38.67 cm) was recorded in cultivar Barani-17 under no drought condition without SA application. Furthermore, maximum physio-biochemical traits, including membrane stability index (MSI), chlorophyl content, photosynthetic rates, stomatal conductance, antioxidant enzymatic activities and relative water content (RWC) were found highest in cultivar Barani-17 under no drought stress and SA application at 100 mM. However, minimum values of these traits were recorded in cultivar Anaj-17 under severe drought stress without SA application. Our results also demonstrated that under severe drought, application of SA at 100 mM significantly increased leaf nitrogen (N), phosphrus (P) and potassium (K) contents and cultivar Barani-17 demonstrated significantly higher values than Anaj-17. The obtained results also indicated that the cultivation of wheat under drought stress conditions noticeably declines the morphological, physiological, and biochemical attributes of the plants. However, the exogenous application of SA had a positive impact on wheat crop for enhancing its productivity.

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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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