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Cellulose-synthase-like enzymes and uses thereof

A compound, biosynthetic technology, applied in the direction of enzymes, enzymes, applications, etc., can solve problems such as unsatisfied

Pending Publication Date: 2021-06-15
YEDA RES & DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, there remains an unmet need for methods to generate these saponins in heterologous systems in order to provide commercial quantities of high value saponins such as but not limited to triterpene saponins

Method used

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  • Cellulose-synthase-like enzymes and uses thereof
  • Cellulose-synthase-like enzymes and uses thereof
  • Cellulose-synthase-like enzymes and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0870] Example 1: Materials and Methods of Examples 2-13

[0871] Plant material, treatment and generation of transgenic plants

[0872] Tomato (Solanum lycopersicum; cv. MicroTom) and potato (Solanum tuberosum; cv. Desiree) plants were collected as previously described (Itkin et al., 2001, supra). In potatoes, when the green parts begin to dry out, mature tubers are collected (stage 3), soiled, dried, and stored in complete darkness at 4°C.

[0873] The corresponding GAME9 DNA segments were introduced into the pK7GWIWG2(II) and pJCV52 binary vectors, respectively, to create GAME9 silencing (RNAi) and overexpression (OX) constructs. Transgenic lines were generated to silence and overexpress GAME9 in tomato and potato, and tissue extracts were prepared and analyzed according to the method of Itkin et al. (2011, supra).

[0874] Table 3 below describes the oligonucleotides used to generate the constructs herein. GAME4-silencing (RNAi; GAME4i), GAME4-overexpressing (GAME400e) ...

Embodiment 2

[0899] Example 2: Genes associated with SGA biosynthesis

[0900] To discover genes associated with SGA biosynthesis, co-expression analysis was performed using transcriptome data from tomato and potato plants. Co-expression of GAME1 / SGT1 (chromosome 7) and GAME4 (chromosome 12) together with GAME1 / SGT1 (chromosome 12) as bait in potato or tomato is presented in the form of a heat map in Tables 5-8 below. Genes highly co-expressed with GAME1 / SGT1 (chromosome 7) or GAME4 (chromosome 12) are shown in large font and bold.

[0901] Table 5: Accession numbers, putative proteins and co-expression r-values ​​- Tomato, Chromosome 7

[0902]

[0903]

[0904] Table 6: Accession numbers, putative proteins and co-expression r-values ​​- potato, chromosome 7

[0905]

[0906]

[0907] Table 7: Accession numbers, putative proteins and co-expression r-values ​​- tomato, chromosome 12

[0908]

[0909]

[0910] Table 8: Accession numbers, putative proteins and co-express...

Embodiment 3

[0921] Example 3: Functional analysis of GAME9 transcription factor

[0922] The corresponding GAME9 DNA segments were introduced into the pK7GWIWG2(II) and pJCV52 binary vectors, respectively, to create GAME9 silencing (RNAi) and overexpression (OX) constructs. Transgenic tomato and potato lines transformed with the corresponding GAME9 silencing and overexpression constructs were generated as previously described (Itkin et al., 2011, supra). Tissue extracts were prepared and analyzed as described in Itkin et al. (2011, supra). SEQ ID NO: 18 shows the sequence of the GAME9 RNAi silencing molecule. Metabolic profiling of steroidal alkaloids was performed using UPLC-TQ-MS on extracts from leaves and / or tubers from transgenic and wild-type tomato and / or potato plants. In potato tuber bark extracts obtained from a potato line in which the gene encoding GAME9 was silenced (GAME9-RNAi line), reductions in α-solanine and α-chaconine were observed (see Figure 5A and 5B ). Leaves...

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Abstract

Provided herein are genetically modified cells and genetically modified plants having increased or decreased expression of a cellulose synthase like G (CSLG) enzyme. These cells and plants may have an increased or decreased content a steroidal alkaloid, a steroidal saponin, or a triterpenoid saponin, compared to a corresponding unmodified cell or plant. Also provided herein are methods of producing a steroidal alkaloid, a steroidal saponin, or a triterpenoid saponin in a genetically modified cell, as well as methods of reducing the content of a steroidal alkaloid, a steroidal saponin, or a triterpenoid saponin in a cell of a plant or a plant part, and methods of increasing the content of a steroidal alkaloid, a steroidal saponin, or a triterpenoid saponin in a cell of a plant or a plant part.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims the benefit of US Patent Application No. 16 / 123,248, filed September 6, 2018, which is a continuation-in-part of US Patent Application No. 14 / 895,059, filed December 1, 2015, which U.S. Patent Application No. 14 / 895,059 is a national phase application of PCT International Application PCT / IL2014 / 050497, filed June 2, 2014, claiming No. 61 / IL2014 / 050497, filed June 5, 2013 the benefit of U.S. Provisional Application No. 831,164, all of which are incorporated herein by reference in their entirety; and this application also claims the benefit of Israeli Patent Application No. 268269, filed July 25, 2019, which is incorporated by reference in its entirety manner is incorporated herein. Background technique [0003] The plant kingdom produces thousands of different small compounds, often specific to the corresponding plant genus or family. Over 300,000 plant-specific metabolites (SMs) result in substan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N1/19C12N15/82C12N15/81C12N15/54A01H5/00A01H6/82A01H6/02A01H6/54
CPCC12N9/1051C12N9/1059C12N15/8218C12N15/8243C12N15/8247C12Y204/01017A01H6/82
Inventor A·阿哈罗尼P·索纳韦恩M·伊特金A·乔兹维亚克
Owner YEDA RES & DEV CO LTD
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