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      Comparison of Leaf Proteomes of Cassava ( Manihot esculenta Crantz) Cultivar NZ199 Diploid and Autotetraploid Genotypes

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          Abstract

          Cassava polyploid breeding has drastically improved our knowledge on increasing root yield and its significant tolerance to stresses. In polyploid cassava plants, increases in DNA content highly affect cell volumes and anatomical structures. However, the mechanism of this effect is poorly understood. The purpose of the present study was to compare and validate the changes between cassava cultivar NZ199 diploid and autotetraploid at proteomic levels. The results showed that leaf proteome of cassava cultivar NZ199 diploid was clearly differentiated from its autotetraploid genotype using 2-DE combined MS technique. Sixty-five differential protein spots were seen in 2-DE image of autotetraploid genotype in comparison with that of diploid. Fifty-two proteins were identified by MALDI-TOF-MS/MS, of which 47 were up-regulated and 5 were down-regulated in autotetraploid genotype compared with diploid genotype. The classified functions of 32 up-regulated proteins were associated with photosynthesis, defense system, hydrocyanic acid (HCN) metabolism, protein biosynthesis, chaperones, amino acid metabolism and signal transduction. The remarkable variation in photosynthetic activity, HCN content and resistance to salt stress between diploid and autotetraploid genotypes is closely linked with expression levels of proteomic profiles. The analysis of protein interaction networks indicated there are direct interactions between the 15 up-regulation proteins involved in the pathways described above. This work provides an insight into understanding the protein regulation mechanism of cassava polyploid genotype, and gives a clue to improve cassava polyploidy breeding in increasing photosynthesis and resistance efficiencies.

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          Most cited references47

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          Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.

          An improved procedure for staining of proteins following separation in polyacrylamide gels is described which utilizes the colloidal properties of Coomassie Brilliant Blue G-250 and R-250. The new method is based on addition of 20% v/v methanol and higher concentrations of ammonium sulfate to the staining solution previously described. The method combines the advantage of much shorter staining time with high sensitivity, a clear background not requiring destaining, stepwise staining, and stable fixation after staining. The method has been applied to staining of polyacrylamide gels after sodium dodecyl sulfate-electrophoresis and isoelectric focusing in carrier ampholyte-generated pH gradients.
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            The Cassava Genome: Current Progress, Future Directions

            The starchy swollen roots of cassava provide an essential food source for nearly a billion people, as well as possibilities for bioenergy, yet improvements to nutritional content and resistance to threatening diseases are currently impeded. A 454-based whole genome shotgun sequence has been assembled, which covers 69% of the predicted genome size and 96% of protein-coding gene space, with genome finishing underway. The predicted 30,666 genes and 3,485 alternate splice forms are supported by 1.4 M expressed sequence tags (ESTs). Maps based on simple sequence repeat (SSR)-, and EST-derived single nucleotide polymorphisms (SNPs) already exist. Thanks to the genome sequence, a high-density linkage map is currently being developed from a cross between two diverse cassava cultivars: one susceptible to cassava brown streak disease; the other resistant. An efficient genotyping-by-sequencing (GBS) approach is being developed to catalog SNPs both within the mapping population and among diverse African farmer-preferred varieties of cassava. These resources will accelerate marker-assisted breeding programs, allowing improvements in disease-resistance and nutrition, and will help us understand the genetic basis for disease resistance.
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              Arabidopsis type I metacaspases control cell death.

              Metacaspases are distant relatives of animal caspases found in protozoa, fungi, and plants. Limited experimental data exist defining their function(s), despite their discovery by homology modeling a decade ago. We demonstrated that two type I metacaspases, AtMC1 and AtMC2, antagonistically control programmed cell death in Arabidopsis. AtMC1 is a positive regulator of cell death and requires conserved caspase-like putative catalytic residues for its function. AtMC2 negatively regulates cell death. This function is independent of the putative catalytic residues. Manipulation of the Arabidopsis type I metacaspase regulatory module can nearly eliminate the hypersensitive cell death response (HR) activated by plant intracellular immune receptors. This does not lead to enhanced pathogen proliferation, decoupling HR from restriction of pathogen growth.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                11 April 2014
                : 9
                : 4
                : e85991
                Affiliations
                [1 ]Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Ministry of Agriculture for Germplasm Resources Conservation and Utilization of Cassava, Hainan, China
                [2 ]Analysis and Testing Center, Jiangsu University, Jiangsu, China
                [3 ]Proteomics Core Facility, Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Manoa, Hawaii, United States of America
                [4 ]Guangxi Sub-tropical Crop Research Institute, Nanning, China
                [5 ]Genetic Resources and Biotechnology, Embrapa, Brazil
                University of South Florida College of Medicine, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: FA JF JL QXL KL SC. Performed the experiments: FA JF JL KL WZ FW SC. Analyzed the data: FA JF JL QXL KL WZ FW LJCBC SC. Contributed reagents/materials/analysis tools: FA JF JL KL WZ FW LJCBC SC. Wrote the paper: FA QXL KL SC.

                Article
                PONE-D-13-35008
                10.1371/journal.pone.0085991
                3984080
                24727655
                c2a23456-2213-47d3-abbf-893040a828d2
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 25 August 2013
                : 3 December 2013
                Page count
                Pages: 13
                Funding
                This work was supported by a Major Project of Chinese National Programs for Fundamental Research and Development Grants (2010CB126606), National Scientific and Technological Programs in Rural Fields (2012AA101204-2), the Earmarked Fund for Modern Agro-industry Technology Research System (nycytx-17) and the Initial Fund of High-level Creative Talents in Hainan Province. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Agriculture
                Agronomy
                Plant Breeding
                Crops
                Biochemistry
                Proteomics
                Protein Abundance
                Plant Biochemistry
                Cell Biology
                Molecular Cell Biology
                Plant Cell Biology
                Developmental Biology
                Plant Growth and Development
                Ecology
                Plant Ecology
                Plant-Environment Interactions
                Organisms
                Plants
                Plant Science
                Plant Anatomy
                Leaves
                Ecology and Environmental Sciences

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                Uncategorized

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