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Physiological Investigation and Transcriptome Analysis of Polyethylene Glycol (PEG)-Induced Dehydration Stress in Cassava.

ABSTRACT: Cassava is an important tropical and sub-tropical root crop that is adapted to drought environment. However, severe drought stress significantly influences biomass accumulation and starchy root production. The mechanism underlying drought-tolerance remains obscure in cassava. In this study, changes of physiological characters and gene transcriptome profiles were investigated under dehydration stress simulated by polyethylene glycol (PEG) treatments. Five traits, including peroxidase (POD) activity, proline content, malondialdehyde (MDA), soluble sugar and soluble protein, were all dramatically induced in response to PEG treatment. RNA-seq analysis revealed a gradient decrease of differentially expressed (DE) gene number in tissues from bottom to top of a plant, suggesting that cassava root has a quicker response and more induced/depressed DE genes than leaves in response to drought. Overall, dynamic changes of gene expression profiles in cassava root and leaves were uncovered: genes related to glycolysis, abscisic acid and ethylene biosynthesis, lipid metabolism, protein degradation, and second metabolism of flavonoids were significantly induced, while genes associated with cell cycle/organization, cell wall synthesis and degradation, DNA synthesis and chromatin structure, protein synthesis, light reaction of photosynthesis, gibberelin pathways and abiotic stress were greatly depressed. Finally, novel pathways in ABA-dependent and ABA-independent regulatory networks underlying PEG-induced dehydration response in cassava were detected, and the RNA-Seq results of a subset of fifteen genes were confirmed by real-time PCR. The findings will improve our understanding of the mechanism related to dehydration stress-tolerance in cassava and will provide useful candidate genes for breeding of cassava varieties better adapted to drought environment.

SUBMITTER: Fu L

PROVIDER: S-EPMC4813147 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature

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Project description:Drought is a serious problem that causes losses in crop-yield every year, but the mechanisms underlying how roots respond to water deficit are difficult to study under controlled conditions. Methods for assaying root elongation and architecture, especially for seedlings, are commonly achieved on artificial media, such as agar, moistened filter paper, or in hydroponic systems. However, it has been demonstrated that measuring root characteristics under such conditions does not accurately mimic what is observed when plants are grown in soil. Morphological changes in root behavior occur because of differences in solute diffusion, mechanical impedance, exposure to light (in some designs), and gas exchange of roots grown under these conditions. To address such deficiencies, we developed a quantitative method for assaying seedling root lengths and germination in soil using a plate-based approach with wheat as a model crop. We also further developed the method to include defined water deficits stress levels using the osmotic properties of polyethylene glycol (PEG). Seeds were sown into soil-filled vertical plates and grown in the dark. Root length measurements were collected using digital photography through the transparent lid under green lighting to avoid effects of white light exposure on growth. Photographs were analyzed using the cross-platform ImageJ plugin, SmartRoot, which can detect root edges and partially automate root detection for extraction of lengths. This allowed for quick measurements and straightforward and accurate assessments of non-linear roots. Other measurements, such as root width or angle, can also be collected by this method. An R function was developed to collect exported root length data, process and reformat the data, and output plots depicting root/shoot growth dynamics. For water deficit experiments, seedlings were transplanted side-by-side into well-watered plates and plates containing PEG solutions to simulate precise water deficits.

Dataset Information

Physiological Investigation and Transcriptome Analysis of Polyethylene Glycol (PEG)-Induced Dehydration Stress in Cassava.

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