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Transgenic trees having reduced xylan content

A transgenic and genetic technology, applied in the direction of genetic engineering, glycosyltransferase, plant gene improvement, etc., can solve problems such as reduction and short stature

Inactive Publication Date: 2016-06-08
SWETREE TECHOLOGIES AB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the above discussion, the problem with trees with reduced xylan content is that they are stunted and / or produce xylem with shrunken and / or thin cell walls

Method used

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  • Transgenic trees having reduced xylan content
  • Transgenic trees having reduced xylan content
  • Transgenic trees having reduced xylan content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1: Cloning of wood-specific promoters from the GT43 gene family.

[0144] To identify good promoters from the GT43 gene family that are highly expressed in developing wood, transcript levels of all seven genes in plant aboveground tissues were quantified by qRT-PCR. All 7 promoters were cloned and the WP (SEQ ID NO: 23 to 30) and 35S promoters were cloned into the GATEWAY cassette vector, followed by the GUS gene for easy detection, resulting in 8 different constructs. The sequences of the cloned promoters are shown as SEQ ID NO:23 to 30.

[0145] Nine pGT43::GUS fusion constructs were stably expressed in hybrid aspen, and stems of two-month-old greenhouse-grown trees were analyzed histochemically for GUS activity.

[0146] Staining for GT43A promoter, GT43B promoter and WP promoter was found in developing xylem and phloem fibers. In contrast, the activity of the GT43C, GT43D and GT43E promoters was present in the phloem parenchyma. For comparison, the acti...

Embodiment 2

[0149] Example 2: Transcript levels of all 7 GT43 genes in non-transgenic hybrid poplar.

[0150] Transcript levels of all seven GT43 genes in plant tissues of hybrid aspen (Populustremula x tremuloides) were quantified in plant shoot tissues by qRT-PCR. The results are summarized in Table I above.

[0151] The mRNA transcripts of all seven genes were quantified in plant above-ground tissues by qRT-PCR. Based on non-normalized expression, which can compare the expression levels of different genes, the genes GT43C and GT43E were found to be most highly expressed in developing wood, followed by GT43B and GT43A. The expression of GT43F and GT43G was not detected in this tissue.

[0152] Among the highly expressed genes, three genes GT43A, GT43B and GT43C were found to be specifically expressed in developing wood, xylem and syringa. The expression of GT43A and GT43B was more wood-specific than that of GT43C.

Embodiment 3

[0153] Example 3: Transgenic Lines in Hybrid Poplar

[0154]Transgenic lines were generated in hybrid aspens designed to silence each GT43 clade using clade-specific sequences of the genes most important for each clade: GT43B for clade I, GT43E from clade II, and GT43C for clade III. These fragments were then combined to silence either two clades or all three clades at a time. All 7 types of inverted repeats were driven by either the CMV35S promoter or the WP promoter, resulting in 14 constructs that were transferred to clone T89.

[0155] About 20 independent lines were pre-screened in vitro for each construct to select the three most impactful lines, which were several times higher than WT, and were grown in the greenhouse.

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Abstract

The invention relates to a method for producing a transgenic tree having reduced xylan content compared to a wild-type tree of the same species, said method comprising reducing expression of one or more genes from the glycosyltransferase 43 (GT43), family in the said transgenic tree, whereby growth properties, mechanical properties, and / or saccharification properties are improved in the said transgenic tree.

Description

technical field [0001] The present invention relates to the field of improving tree properties. In particular, the present invention relates to methods of down-regulating the glycosyltransferase 43 gene in the xylan biosynthetic machinery while enhancing growth characteristics, improving saccharification characteristics and mechanical characteristics. The invention also relates to the expression vectors, transgenic trees and wood used. Such trees and wood can be used for afforestation and bioenergy purposes. Background technique [0002] As an abundant raw material for renewable biofuels, wood is an excellent energy source. The cell walls of higher plants comprise polysaccharides such as cellulose, hemicellulose, pectin and polyphenolic compounds such as lignin. All of these cell wall components constitute lignocellulosic biomass, which is a rich source of energy. [0003] Xylan is the major hemicellulose in hardwood, constituting about 20% of woody biomass. The length ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/00A01H5/00C12N9/10
CPCC12N15/8218C12N15/8246C12Y204/01C12N15/1137C12N2310/14
Inventor 埃瓦·J·梅莱罗维奇莱夫·琼森克里斯蒂娜·拉特克芭芭拉·捷列比涅茨尚德拉·维内斯特兰德
Owner SWETREE TECHOLOGIES AB