Transformed plants accumulating mono- and/or sesquiterpenes

A technology of plants and plant materials, applied in the fields of plant products, plant genetic improvement, plant equipment and methods, etc.

Inactive Publication Date: 2008-06-25
UNIV OF KENTUCKY RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prior art has so far not addressed this issue, which has mainly implicated recombinant organisms with altered properties in the MVP or MEP pathway and the observed accumulation of specific terpene end-products

Method used

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  • Transformed plants accumulating mono- and/or sesquiterpenes
  • Transformed plants accumulating mono- and/or sesquiterpenes
  • Transformed plants accumulating mono- and/or sesquiterpenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1: Selection of plant material

[0105] Homozygous lines of the "Shanxi" tobacco line containing a truncated hamster hydroxymethyl-glutaryl-CoA reductase gene were bred according to the method described in Chappell et al. (hereinafter referred to as 14-8) and an F2 segregating sibling line lacking the transgene (hereinafter referred to as 14-2). In summary, the hamster gene first characterized by Chin et al. (1984) was modified by removing nucleotides 28-1023 to obtain a truncated form of the gene, referred to as delta-227HMGR. The BamH1 / Sst1 fragment of the truncated HMGR cDNA was isolated and inserted into the intermediate plastid vector pKYLX 61, and the EcoR1 / Cla1 fragment of this construct was subsequently inserted into the corresponding pKYLX71 site to generate the pKYLX71-truncated HMGR gene construct. Schardl et al. (1987) developed the Ti-plasmid series pKYLX61 and pKYLX71.

[0106] The pKYLX71-HMGR plasmid was transferred into Agrobacterium tumefac...

Embodiment 2-5

[0110] Embodiment 2-5: Construction of recombinant and plant transformation vector

[0111] Selection The hygromycin selection marker (Hajdukiewicz et al., 1994) was used to generate the selection marker for transformed plants. New vectors were designed with appropriate recombinant cloning sites as described by Hartley et al. (2000).

Embodiment 2

[0112] Example 2: Formation of pBDON vector

[0113] Figure 8 , the pBI101 vector (Invitrogen, Carlsbad, CA) was digested with restriction enzymes Sph1 and Sst1 and purified by agarose gel (Sambrook et al., 1989) to isolate the DNA corresponding to the plasmid vector (excluding the RB border and NPTII gene cassette) Fragment (1). Simultaneously, the attp recombinant cassette including the ccdb gene and the chloramphenicol resistance gene was amplified from the pDON221 vector (Invitrogen, Carlsbad, CA) with primers Attp1-SstI-FW and Attp2-SphI-RV using standard PCR conditions ( 2). The PCR-amplified DNA fragment was digested with Sph1 / Sst1, gel purified and ligated into the corresponding sites of the similarly digested pBI101 vector described above to produce the intermediate pBattp vector (3).

[0114] The hygromycin gene cassette was prepared using a two-step method. First use PCR primers HPT-NotI-FD and HPT-Xba1-RV that hygromycin gene (HPT) and CaUTR (termination seq...

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Abstract

The present invention relates to plant expressing transgenes and to plants transformed to comprise additional copies of genes, said genes encoding at least a HMGR- CoA reductase and a terpene synthase. The invention further claims methods for preparing the plants, and a method for producing terpenes. The present thus provides a reliable and cost effective platform for generating any terpene, in particular any mono-and / or sesquiterpene of interest. For example, the skilled person may use any gene encoding a sesquiterpene synthase for accumulating the respective sesquiterpene in the plant of the present invention.

Description

technical field [0001] The present invention relates to transformed plants. In particular it relates to transformed plants accumulating monoterpenes and / or sesquiterpenes. The plants were transformed to overexpress HMG-CoA reductase (HMGR) and terpene synthase (TS) and optional prenyl transferase (PRT), or to contain additional genes encoding these enzymes. The present invention further relates to a method of preparing the plant, a method of altering the content of a specific terpene in a plant, a method of producing a terpene and the use of the above gene in the production of transgenic plants. Background technique [0002] Terpenes and terpenoids are found in most organisms. Their increasing commercial importance is linked to the variable range surrounding the biological activity and functionality of the different terpenes. As a result, many vitamins, hormones, insect repellants, drugs, flavorings and fragrances are found contained in such a huge class of compounds, all...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/60C12N15/53A01H5/00C07C62/00C07C35/22
CPCC12N15/8243C12N9/1085C12N9/0006
Inventor 乔·夏贝尔巫水钦米歇尔·沙尔克安东尼·克拉克
Owner UNIV OF KENTUCKY RES FOUND
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