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Method for increasing saccharification efficiency of plants by down-regulating UXT (UDP-Xylose transporter) gene and application of method

A technology for saccharification efficiency and plant sugar, which is applied in the directions of botanical equipment and methods, biochemical equipment and methods, and applications, can solve problems such as affecting saccharification efficiency, and achieve the goal of reducing labor intensity, increasing relative content, and reducing xylose content. Effect

Active Publication Date: 2017-12-01
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Xylan improves the toughness and stress resistance of plants by cross-linking with cellulose and lignin in the secondary wall of plants, but the presence of xylan also has a great impact on the industrial application of lignocellulose , the cross-linked network structure formed by xylan around cellulose prevents the entry of cellulolytic enzymes and affects the saccharification efficiency

Method used

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  • Method for increasing saccharification efficiency of plants by down-regulating UXT (UDP-Xylose transporter) gene and application of method
  • Method for increasing saccharification efficiency of plants by down-regulating UXT (UDP-Xylose transporter) gene and application of method
  • Method for increasing saccharification efficiency of plants by down-regulating UXT (UDP-Xylose transporter) gene and application of method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Acquisition of T-DNA Line with mutation of AtUXT gene. Three AtUXT mutants were ordered from ABRC (Arabidopsis Biological Resource Center): uxt1 (SALK_086773), uxt2 (SALK_091753) and uxt3 (SALK_079036.37.50.x). The corresponding T-DNA Line identification primers were synthesized on the website of SALK (Salk Institute Genomic Analysis Laboratory, http: / / signal.salk.edu), as shown in Table 1. The three mutants were crossed, and double mutants uxt1uxt3 and uxt2uxt3 were firstly obtained, and these two double mutants were crossed to obtain a triple mutant uxt1uxt2uxt3. The uxt1uxt2uxt3 triple mutant grew short, had dark green leaves, and had a very low amount of seeds in pods ( figure 1 ).

[0034] Table 1 T-DNA Line Identification Primers

[0035] mutant

Embodiment 2

[0037] Preparation and observation of stem sections of Arabidopsis thaliana. Select the main stem of the plant that has grown for 6 to 7 weeks, take a 1 cm stem section 3 cm above the soil surface, embed the stem section with 3% (w / v) agarose, and slice it on a Leica VT1000S vibrating microtome with a thickness of 40 μm. Stain with blue for 1-2 minutes, place on a glass slide, observe and take pictures under an optical microscope. The wood fiber cell wall of wild-type Col-0 was significantly thicker than that of the uxt1uxt2uxt3 triple mutant, and the morphology of xylem vessels of wild-type Col-0 was normal ( figure 2 ), the uxt1uxt2uxt3 triple mutant has irregular vessel morphology and a high collapse ratio ( figure 2 ). Further, the slices were observed under a scanning electron microscope, and the slices were placed on the sample stage to be plated with gold, and then observed under a Hitachi Ion Sputter E-1010 (HITACHI) scanning electron microscope. It was found that...

Embodiment 3

[0039] Chemical immunological observation was performed on the slices used for optical microscope observation in Example 2. Spread 2 layers of filter paper in a Petri dish, soak it with deionized water, place a layer of Parafilm membrane on it, and place the slices on the Parafilm membrane. Then perform chemical immunostaining according to the following steps: use 0.1M phosphate buffer solution (PhosphateBuffer Solution, PBS; 1L: 0.218g KH 2 PO 4 , 1.463g K 2 HPO 4 , 29.22g NaCl, pH 7.2) to wash the slices for 5-10min; soak the slices in fresh 3% (w / v) skim milk for 1h, during which the skim milk was continuously blown; remove the skim milk, and wash the slices with PBS buffer for 5min; primary antibody Incubation: Incubate slices with 20-fold diluted rat anti-xylan antibody LM10 (Plantprobes) for 1 h, wash slices with PBS buffer 10 times to wash unbound primary antibody; secondary antibody incubation: use 50-fold diluted FITC- Goat anti-rat antibody (Zomanbio, Cat. Z1319)...

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Abstract

The invention discloses a method for increasing saccharification efficiency of plants by down-regulating a UXT (UDP-Xylose transporter) gene and an application of the method and belongs to the field of biomass energy. According to the method, the saccharification efficiency of the plants is increased by eliminating functions of the UXT gene or down-regulating the UXT gene. By means of the method, xylose content is reduced, relative cellulose content is increased correspondingly, by means of xylose content reduction, the saccharification efficiency of a mutant material is increased and the material disposal cost is reduced. Therefore, biomass characteristics of the material are changed fundamentally, the saccharification efficiency is increased radically, energy can be saved, and labor intensity can be reduced. The material obtained with the method can be applied to biomass transformation, pulp and papermaking industries.

Description

technical field [0001] The invention belongs to the field of biomass energy, and in particular relates to the use of biotechnology to down-regulate the content of uridine diphosphate xylose transporters (UXTs) in plants, so that plants can obtain higher saccharification efficiency. The invention relates to the application of UXTs down-regulated plants in various fields such as biomass conversion, pulp and paper industry; in particular, it relates to a method for improving plant saccharification efficiency by down-regulating UXT gene and its application. Background technique [0002] Biomass energy is the fourth largest energy source in the world, second only to oil, coal and natural gas. Because it comes from the photosynthesis of plants, it can be transformed in solid, liquid and gaseous states. It can be described as inexhaustible, renewable energy and environmentally friendly. The world today mainly uses traditional energy, which has limited storage, non-renewable energy...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
CPCC12N15/8213C12N15/8218C12N15/8245
Inventor 吴蔼民赵先海尚娜邓小梅刘念
Owner SOUTH CHINA AGRI UNIV
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