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Transgenic plants with improved saccharification yields and methods of generating same

A plant and plant cell technology, applied in the directions of botanical equipment and methods, plant products, biochemical equipment and methods, etc., can solve the problem of reducing lignin, mechanical support, disease resistance and adverse effects of water transport, and increasing polysaccharide degradation. And other issues

Inactive Publication Date: 2013-08-07
YISSUM RES DEV CO OF THE HEBREWUNIVERSITY OF JERUSALEM LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, manipulation of lignin biosynthesis leads to reduced lignin and increased polysaccharide degradability, but also has adverse effects on mechanical support, disease resistance and water transport of (genetically) engineered plants [Halpin C et al., Tree Genetics & Genomes (2007) 3 (2): 101-110; Pedersen JF et al., Crop Science (2005) 45 (3): 812-819]

Method used

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  • Transgenic plants with improved saccharification yields and methods of generating same
  • Transgenic plants with improved saccharification yields and methods of generating same
  • Transgenic plants with improved saccharification yields and methods of generating same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0105] According to one embodiment of the present invention, the nucleic acid construct of the present invention comprises a polynucleotide encoding a heterologous acetylxypolynucleotide under the transcriptional control of a developmentally regulated promoter that is particularly active in plant cell walls after secondary cell wall deposition. Sugar esterase (AXE).

[0106] According to one embodiment of the invention, the nucleic acid construct of the invention comprises a polynucleotide encoding a heterologous glucuronide under the transcriptional control of a developmentally regulated promoter that is particularly active in plant cell walls after secondary cell wall deposition. Esterase (GE).

[0107] According to one embodiment of the present invention, the nucleic acid construct of the present invention comprises a polynucleotide encoding heterologous acetylxylan esterase (AXE) and a polynucleotide encoding heterologous glucuronyl esterase (GE), The above codes are all ...

Embodiment 1

[0264] Acetylxylan esterase (AXE) and glucuronylesterase (GE) cloning and transformation into tobacco plants

[0265] Promoter : Since overexpression of AX and GE in plants can reduce plant structural integrity and fitness, the inventors directed the expression of these enzymes to specific developmental stages such as secondary cell wall development or xylem cell development. Expression of a gene at a particular developmental stage can be achieved through a developmental-specific promoter. Examples of such promoters are, for example, promoters expressed only during secondary wall thickening, the CesA7 promoter and the 4CL-1 promoter. Examples of promoters expressed in xylem tissue development are the FRA8 promoter and the DOT1 promoter.

[0266] For expression during xylem development, AX and GE were fused to the FRA8 promoter (SEQ ID NO:21).

[0267] Constitutive overexpression of AX by the CaMV35S promoter was also tested.

[0268] Signal peptide: To direct AX and GE ...

Embodiment 2

[0275] Tobacco transformation and PCR to genomic DNA

[0276] Leaf disk transformation was performed with Nicotiana-SR1 plants as previously described [Block, M.D. et al., EMBO Journal (1984) 3:1681-1689]. For each binary vector, more than 15 independent tobacco transformants were generated, propagated in vitro and transferred to the greenhouse. Tobacco plants overexpressing AXEII under the regulation of the 35S promoter flowered earlier than plants expressing AXEII or AXEI under the regulation of the FRA8 promoter or wild-type plants (untransformed plants grown under the same culture conditions) , and exhibited different levels of engineered phenotypes, such as growth retardation and smaller stem diameters (data not shown). By Western blot analysis for nptII protein (data not shown) and by PCR on genomic DNA ( Figure 5A-Figure 5D ), where the presence of the transgene was confirmed using AX or GE specific primers (Table 1). Binary vectors were used as templates for positi...

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Abstract

A method of engineering a plant having reduced acetylation in a cell wall is disclosed. The method comprising expressing in the plant cell wall at least one isolated heterologous polynucleotide encoding an acetylxylan esterase (AXE) enzyme under the transcriptional control of a developmentally regulated promoter specifically active in the plant cell wall upon secondary cell wall deposit, thereby engineering the plant having reduced acetylation in the cell wall.

Description

technical field [0001] In some embodiments of the present invention, it relates to transgenic plants expressing acetylxylan esterase (AXE) and / or glucuronyl esterase (GE), more specifically but not limitedly, the present invention relates to the transgenic Plants are used in various applications such as for biomass conversion (eg, biofuel, hydrogen production), for feed and food applications, and for the pulp and paper industry. Background technique [0002] Consistent with the rapid growth of the world population, the quality of natural resources and the environment is declining. Current methods of energy consumption mainly based on fossil fuels are considered to be harmful to the environment and contribute to global warming. To address this growing problem, interest has increased in generating fuels from renewable resources, especially those from plant biomass. To date, most ethanol fuels have been produced from corn kernels or sugar cane, also known as "first generation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00C12N9/18
CPCC12N15/8255C12N15/8243C12N9/18C12N15/8246C12Y301/01072
Inventor 米龙·阿布拉姆松齐夫·沙尼奥戴德·绍塞尤夫
Owner YISSUM RES DEV CO OF THE HEBREWUNIVERSITY OF JERUSALEM LTD
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