Spatially modified gene expression in plants

A plant and gene technology, applied in the direction of cells modified by introducing foreign genetic material, genetic engineering, sugar derivatives, etc., can solve the problems of decreased biomass yield and difficult to obtain crops, etc.

Active Publication Date: 2013-11-20
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, most attempts to reduce the lignin content of plants have resulted in severe reductions in biomass yield (Voelker et al., 2010; Shadle et al., 2007; Franke et al., 2002), and as such, it is not easy to obtain Crops with declining nutrients

Method used

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  • Spatially modified gene expression in plants
  • Spatially modified gene expression in plants
  • Spatially modified gene expression in plants

Examples

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preparation example Construction

[0242] F. Preparation of recombinant expression vector

[0243] Once the promoter sequence and the coding sequence of the gene of interest (e.g., lignin biosynthesis enzymes, xylan biosynthesis enzymes, or transcription factors regulating secondary cell wall production) are obtained, the sequences can be used to prepare An expression cassette expressing the gene of interest. Typically, plant transformation vectors include one or more cloned plant coding sequences (genomic or cDNA) encoding proteins of interest (such as transcription factors) under the transcriptional control of 5' and 3' regulatory sequences. Vectors also typically contain a dominant selectable marker. In typical embodiments, such plant transformation vectors also contain a promoter of interest (e.g., a vessel-specific promoter as described herein or a promoter whose expression is regulated by a transcription factor that regulates secondary cell wall production), a transcription initiation Initiation sites, ...

Embodiment 1

[0265] Example 1: Secondary cell wall deposition in reengineered plants

[0266] This study combined 2 strategies for overcoming cell wall recalcitrance and filling fiber cells with cell wall polymers without altering plant development. The first protocol allows reducing lignin in places other than vessels, while the second protocol specifically increases cell wall deposition in woody tissues. This combinatorial strategy strategy uses synthetic biology to fine-tune lignin biosynthesis and establish new feedback loops to reengineer the control of secondary cell wall deposition.

[0267] Materials and methods

[0268] Plasmid construction

[0269] The protein coding regions of C4H (ref3) gene (AT2G30490), F5H (At4g36220) and CADc gene (AT3G19450) were amplified from Arabidopsis cDNA, and the translational start of VND6 gene (At5g62380) was amplified with appropriate primers (see Table 1). The 2756 bp region 5' upstream of the start site was pVND6 as genomic DNA.

[0270] Tab...

Embodiment 2

[0308] Example 2. Engineering in Arabidopsis (dicots) and Brachypodium (monocots) Modified Positive Feedback Loop

[0309] Figure 27 A cell wall deposition positive feedback loop is explained. Cell wall densification is based on the establishment of an artificial positive feedback loop to enhance the expression of fiber-specific transcription factors. It is established by expressing new copies of fiber-specific transcription factors (eg, NST1 ) under the control of inducible promoters downstream of xylan or cellulose biosynthesis. This protocol is compatible with xylan and lignin engineering strategies.

[0310] Figure 31 A shows UV images of wild-type Arabidopsis (dicots) and stem cross-sections of wild-type Arabidopsis genetically modified to contain the pCesA4:NST1 expression construct. Establishment of a positive feedback loop involving a secondary cell wall cellulose promoter (pCesA4) and a secondary cell wall transcription factor (NST1) enhances secondary cell ...

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Abstract

The invention provides methods of engineering plants having lignin deposition or xylan deposition that is substantially localized to the vessels of xylem tissue in the plant. The invention also provides methods of engineering plants to increase production of a desired biosynthetic product, e.g., to have increased secondary cell wall deposition or increased wax / cutin accumulation. The engineered plants of the present invention can be used for bioenergy production, e.g., by improving the density and the digestibility of biomass derived from the plant and to improve water usage requirements.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 61 / 437,569, filed January 28, 2011, which is incorporated herein by reference for all purposes. [0003] Statement Concerning Rights to Inventions Made Under Federally Sponsored Research and Development [0004] This invention was made with Government support under Contract No. DE-AC02-05CH11231 awarded by the US Department of Energy. The US Government has certain rights in this invention. Background technique [0005] Plant cell walls are the only source of cellulose for the paper industry and are a promising source of sugars for lignocellulosic biofuels. The use of plants to convert solar energy into transportable and storable energy will have a positive impact on the environment, as the use of plants can help to drastically reduce the use of fossil-derived fuels, reduce carbon emissions into the atmosphere, and even promote carbon sequestration. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C12N5/14
CPCC12N15/8246C12N15/8226C12N15/8255C12N15/8247C12N15/8217C12N15/8257C12N15/8261Y02A40/146
Inventor 多米尼克·洛克亨利克·韦彼·斯盖勒
Owner RGT UNIV OF CALIFORNIA
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