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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 