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      AtCesA8-driven OsSUS3 expression leads to largely enhanced biomass saccharification and lodging resistance by distinctively altering lignocellulose features in rice

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          Abstract

          Background

          Biomass recalcitrance and plant lodging are two complex traits that tightly associate with plant cell wall structure and features. Although genetic modification of plant cell walls can potentially reduce recalcitrance for enhancing biomass saccharification, it remains a challenge to maintain a normal growth with enhanced biomass yield and lodging resistance in transgenic plants. Sucrose synthase (SUS) is a key enzyme to regulate carbon partitioning by providing UDP-glucose as substrate for cellulose and other polysaccharide biosynthesis. Although SUS transgenic plants have reportedly exhibited improvement on the cellulose and starch based traits, little is yet reported about SUS impacts on both biomass saccharification and lodging resistance. In this study, we selected the transgenic rice plants that expressed OsSUS3 genes when driven by the AtCesA8 promoter specific for promoting secondary cell wall cellulose synthesis in Arabidopsis. We examined biomass saccharification and lodging resistance in the transgenic plants and detected their cell wall structures and wall polymer features.

          Results

          During two-year field experiments, the selected AtCesA8:: SUS3 transgenic plants maintained a normal growth with slightly increased biomass yields. The four independent transgenic lines exhibited much higher biomass enzymatic saccharification and bioethanol production under chemical pretreatments at P < 0.01 levels, compared with the controls of rice cultivar and empty vector transgenic line. Notably, all transgenic lines showed a consistently enhanced lodging resistance with the increasing extension and pushing forces. Correlation analysis suggested that the reduced cellulose crystallinity was a major factor for largely enhanced biomass saccharification and lodging resistance in transgenic rice plants. In addition, the cell wall thickenings with the increased cellulose and hemicelluloses levels should also contribute to plant lodging resistance. Hence, this study has proposed a mechanistic model that shows how OsSUS3 regulates cellulose and hemicelluloses biosyntheses resulting in reduced cellulose crystallinity and increased wall thickness, thereby leading to large improvements of both biomass saccharification and lodging resistance in transgenic rice plants.

          Conclusions

          This study has demonstrated that the AtCesA8:: SUS3 transgenic rice plants exhibited largely improved biomass saccharification and lodging resistance by reducing cellulose crystallinity and increasing cell wall thickness. It also suggests a powerful genetic approach for cell wall modification in bioenergy crops.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s13068-017-0911-0) contains supplementary material, which is available to authorized users.

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          Most cited references53

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          Hemicelluloses.

          Hemicelluloses are polysaccharides in plant cell walls that have beta-(1-->4)-linked backbones with an equatorial configuration. Hemicelluloses include xyloglucans, xylans, mannans and glucomannans, and beta-(1-->3,1-->4)-glucans. These types of hemicelluloses are present in the cell walls of all terrestrial plants, except for beta-(1-->3,1-->4)-glucans, which are restricted to Poales and a few other groups. The detailed structure of the hemicelluloses and their abundance vary widely between different species and cell types. The most important biological role of hemicelluloses is their contribution to strengthening the cell wall by interaction with cellulose and, in some walls, with lignin. These features are discussed in relation to widely accepted models of the primary wall. Hemicelluloses are synthesized by glycosyltransferases located in the Golgi membranes. Many glycosyltransferases needed for biosynthesis of xyloglucans and mannans are known. In contrast, the biosynthesis of xylans and beta-(1-->3,1-->4)-glucans remains very elusive, and recent studies have led to more questions than answers.
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            Sucrose synthase affects carbon partitioning to increase cellulose production and altered cell wall ultrastructure.

            Overexpression of the Gossypium hirsutum sucrose synthase (SuSy) gene under the control of 2 promoters was examined in hybrid poplar (Populus alba x grandidentata). Analysis of RNA transcript abundance, enzyme activity, cell wall composition, and soluble carbohydrates revealed significant changes in the transgenic lines. All lines showed significantly increased SuSy enzyme activity in developing xylem. This activity manifested in altered secondary cell wall cellulose content per dry weight in all lines, with increases of 2% to 6% over control levels, without influencing plant growth. The elevated concentration of cellulose was associated with an increase in cell wall crystallinity but did not alter secondary wall microfibril angle. This finding suggests that the observed increase in crystallinity is a function of altered carbon partitioning to cellulose biosynthesis rather than the result of tension wood formation. Furthermore, the augmented deposition of cellulose in the transgenic lines resulted in thicker xylem secondary cell wall and consequently improved wood density. These findings clearly implicate SuSy as a key regulator of sink strength in poplar trees and demonstrate the tight association of SuSy with cellulose synthesis and secondary wall formation.
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              Three distinct rice cellulose synthase catalytic subunit genes required for cellulose synthesis in the secondary wall.

              Several brittle culm mutations of rice (Oryza sativa) causing fragility of plant tissues have been identified genetically but not characterized at a molecular level. We show here that the genes responsible for three distinct brittle mutations of rice, induced by the insertion of the endogenous retrotransposon Tos17, correspond to CesA (cellulose synthase catalytic subunit) genes, OsCesA4, OsCesA7 and OsCesA9. Three CesA genes were expressed in seedlings, culms, premature panicles, and roots but not in mature leaves, and the expression profiles were almost identical among the three genes. Cellulose contents were dramatically decreased (8.9%-25.5% of the wild-type level) in the culms of null mutants of the three genes, indicating that these genes are not functionally redundant. Consistent with these results, cell walls in the cortical fiber cells were shown to be thinner in all the mutants than in wild-type plants. Based on these observations, the structure of a cellulose-synthesizing complex involved in the synthesis of the secondary cell wall is discussed.
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                Author and article information

                Contributors
                luckyfcf@163.com
                fengsq@mail.hzau.edu.cn
                hjfjiayoua@yeah.net
                wyt@mail.hzau.edu.cn
                905885754@qq.com
                340163951@qq.com
                lqwang@mail.hzau.edu.cn
                yuant@mail.hzau.edu.cn
                xiatao@mail.hzau.edu.cn
                jingyanglee@163.com
                xiwen.cai@ndsu.edu
                +86-27-87281765 , lpeng@mail.hzau.edu.cn , http://bbrc.hzau.edu.cn
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                16 September 2017
                16 September 2017
                2017
                : 10
                : 221
                Affiliations
                [1 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, Biomass and Bioenergy Research Centre, , Huazhong Agricultural University, ; Wuhan, China
                [2 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, National Key Laboratory of Crop Genetic Improvement, , Huazhong Agricultural University, ; Wuhan, China
                [3 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, College of Plant Science and Technology, , Huazhong Agricultural University, ; Wuhan, China
                [4 ]ISNI 0000 0004 1790 4137, GRID grid.35155.37, College of Life Science and Technology, , Huazhong Agricultural University, ; Wuhan, China
                [5 ]ISNI 0000 0000 9835 1415, GRID grid.453499.6, HaiKou Experimental Station, Chinese Academy of Tropical Agricultural Sciences, ; Haikou, 570102 China
                [6 ]ISNI 0000 0001 2293 4611, GRID grid.261055.5, Department of Plant Science, , North Dakota State University, ; Fargo, ND USA
                Author information
                http://orcid.org/0000-0002-8739-3470
                Article
                911
                10.1186/s13068-017-0911-0
                5603028
                28932262
                3ec13e6f-add7-4741-a308-83491862a03d
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 28 June 2017
                : 8 September 2017
                Funding
                Funded by: the Fundamental Research Funds for the Central Universities of China
                Award ID: 2662015PY173
                Award ID: 2662015PY018
                Award Recipient :
                Funded by: the National Transgenic Project
                Award ID: 2009ZX08009-119B
                Award Recipient :
                Funded by: the National Science Foundation of China
                Award ID: 31670296
                Award Recipient :
                Funded by: the Huazhong Agricultural University Scientific & Technological Self-innovation Foundation
                Award ID: 2011SC01
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Biotechnology
                sucrose synthase,transgenic rice,biomass saccharification,yeast fermentation,lodging resistance,cell wall,cellulose crystallinity

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