Method to obtain low triterpene/triterpenoid-containing natural rubber latex

Patent metadata

Field	Value
Publication number	US-2022010323-A1
Application number	US-201917291990-A
Country	US
Kind code	A1
Filing date	Nov 8, 2019
Priority date	Nov 8, 2018
Publication date	Jan 13, 2022
Grant date	—

Abstract

The invention provides a method of reducing or eliminating the content of triterpenes and/or triterpenoids of latex or rubber from a laticiferous plant or part thereof, comprising genetically modifying the laticiferous plant or part thereof, such that the activity of at least one oxidosqualene cyclase is reduced or eliminated, compared to the respective wild-type or untreated laticiferous plant or part thereof. The invention further provides a laticiferous plant or part thereof being genetically modified such that the activity of at least one oxidosqualene cyclase is reduced or eliminated, compared to the respective wild-type or untreated laticiferous plant or part thereof, and an RNAi being specifically directed against a mRNA transcript of at least one oxidosqualene cyclase, for use in a method of reducing or eliminating the content of triterpenes and/or triterpenoids of latex or rubber from a laticiferous plant or part thereof.

First claim

1 . Method of reducing or eliminating the content of triterpenes and/or triterpenoids of latex or rubber from a laticiferous plant or part thereof,
comprising genetically modifying the laticiferous plant or part thereof, such that the activity of at least one oxidosqualene cyclase is reduced or eliminated, compared to the respective wild-type or untreated laticiferous plant or part thereof.

2 . Method according to claim 1 , wherein the laticiferous plant or part thereof is genetically modified by at least one heterologous nucleotide sequence that is capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase,
preferably that is capable of reducing or eliminating the expression of the gene of the at least one oxidosqualene cyclase, and/or wherein the laticiferous plant or part thereof is genetically modified by subjecting it to and/or incubating it with agents, which are capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase.

3 . Method according to claim 2 , wherein the at least one heterologous nucleotide sequence encodes at least one RNAi that interferes with the mRNA encoding the at least one oxidosqualene cyclase, with the proviso that the at least one RNAi is capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase.

4 . Method according to claim 3 , wherein the at least one RNAi is specifically directed against the mRNA transcript of the at least one oxidosqualene cyclase,
preferably wherein the at least one RNAi comprises or consists of a nucleotide sequence according to SEQ ID NO: 1 and/or comprises or consists of a nucleotide sequence according to SEQ ID NO: 2 and/or comprises or consists of a nucleotide sequence being at least 60%, 70%, 80%, 90%, 95% or 99% identical to SEQ ID NO: 1 or SEQ ID NO: 2.

5 . Method according to any one of the preceding claims, wherein the laticiferous plant or part thereof is selected from the group of plant families consisting of Apocynaceae, Asteraceae, Euphorbiaceae and Moraceae.

6 . Method according to any one of the preceding claims, wherein the laticiferous plant or part thereof is selected from the group consisting of Agoseris glauca, Apocynum venetum, Chrysothamnus graveolens, Chrysothamnus nauseosus, Chrysothamnus viscidiflorus, Funtumia elastica, Parthenium argentaturn, Scorzonera acanthoclada, Scorzonera albicaulis, Scorzonera divaricate, Scorzonera hissaricata, Scorzonera racemosa, Scorzonera tau - saghyz, Scorzonera tragapogonoides, Scorzonera turkestania, Scorzonera virgate, Solidago canadensis scabra, Solidago fistulosa, Solidago leavenworthii, Solidago rigida, Sonchus oleraceus, Taraxacum brevicomiculaturn, Taraxacum hybemum, Taraxacum koksaghyz, Taraxacum megalorrhizon, Taraxacum officinale and crossings thereof,
preferably wherein the laticiferous plant or part thereof is selected from the group consisting of Parthenium argentatum, Chrysothamnus nauseosus, Taraxacum koksaghyz, Taraxacum officinale and crossings thereof, more preferably wherein the laticiferous plant or part thereof is Taraxacum koksaghyz.

7 . Method according to any one of the preceding claims, wherein the at least one oxidosqualene cyclase is selected from the group consisting of oxidosqualene cyclase 1 according to SEQ ID NO: 4, the oxidosqualene cyclase 2 according to SEQ ID NO: 6, the oxidosqualene cyclase 3 according to SEQ ID NO: 8, the oxidosqualene cyclase 4 according to SEQ ID NO: 10, the oxidosqualene cyclase 5 according to SEQ ID NO: 12, the oxidosqualene cyclase 6 according to SEQ ID NO: 14, the oxidosqualene cyclase 7 according to SEQ ID NO: 16, the oxidosqualene cyclase 8 according to SEQ ID NO: 18, the lupeol synthase according to SEQ ID NO: 20 and an amino acid sequence being at least 60%, 70%, 80%, 90%, 95% or 99% identical to SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18 or 20,
preferably wherein the at least one oxidosqualene cyclase is selected from the group consisting of the oxidosqualene cyclase 1 according to SEQ ID NO: 4, the oxidosqualene cyclase 2 according to SEQ ID NO: 6, the oxidosqualene cyclase 3 according to SEQ ID NO: 8 and the oxidosqualene cyclase 5 according to SEQ ID NO: 12 and an amino acid sequence being at least 60%, 70%, 80%, 90%, 95% or 99% identical to SEQ ID NOs: 4, 6, 8 or 12, more preferably wherein the at least one oxidosqualene cyclase is the oxidosqualene cyclase 1 according to SEQ ID NO: 4 or an amino acid sequence being at least 60%, 70%, 80%, 90%, 95% or 99% identical thereto.

8 . Method according to any one of the preceding claims, wherein the part of the laticiferous plant is selected from the group consisting of cells, tissues, organs, roots, stems, branches, leaves, exudates, latex and mixtures thereof.

9 . Laticiferous plant or part thereof being genetically modified such that the activity of at least one oxidosqualene cyclase is reduced or eliminated, compared to the respective wild-type or untreated laticiferous plant or part thereof.

10 . Laticiferous plant or part thereof according to claim 9 , wherein the laticiferous plant or part thereof is genetically modified by at least one heterologous nucleotide sequence that is capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase, preferably that is capable of reducing or eliminating the expression of the gene of the at least one oxidosqualene cyclase,
and/or wherein the laticiferous plant or part thereof is genetically modified by subjecting it to and/or incubating it with agents, which are capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase.

11 . Laticiferous plant or part thereof according to claim 9 or 10 , wherein the at least one heterologous nucleotide sequence encodes at least one RNAi that interferes with the mRNA encoding the at least one oxidosqualene cyclase with the proviso that the at least one RNAi is capable of reducing or eliminating the activity of the at least one oxidosqualene cyclase.

12 . Laticiferous plant or part thereof according to claim 11 , wherein the at least one RNAi is specifically directed against the mRNA transcript of the at least one oxidosqualene cyclase, preferably wherein the at least one RNAi comprises or consists of a nucleotide sequence according to SEQ ID NO: 1 and/or comprises or consists of a nucleotide sequence according to SEQ ID NO: 2 and/or comprises or consists of a nucleotide sequence being at least 60%, 70%, 80%, 90%, 95% or 99% identical to SEQ ID NO: 1 or SEQ ID NO: 2.

13 . Laticiferous plant or part thereof according to any one of claims 9 to 12 , wherein the laticiferous plant or part thereof is selected from the group of plant families consisting of Apocynaceae, Asteraceae, Euphorbiaceae and Moraceae,
preferably wherein the laticiferous plant or part thereof is selected from the group consisting of Agoseris glauca, Apocynum venetum, Chrysothamnus graveolens, Chrysothamnus nauseosus, Chrysothamnus viscidiflorus, Funtumia elastica, Parthenium argentatum, Scorzonera acanthoclada, Scorzonera albicaulis, Scorzonera divaricate, Scorzonera hissaricata, Scorzonera racemosa, Scorzonera tau - saghyz, Scorzonera tragapogonoides, Scorzonera turkestania, Scorzonera virgate, Solidago canadensis scabra, Solidago fistulosa, Solidago leavenworthii, Solidago rigida, Sonchus oleraceus, Taraxacum brevicomiculaturn, Taraxacum hybemum, Taraxacum koksaghyz, Taraxacum megalorrhizon, Taraxacum officinale and crossings thereof,
preferably wherein the laticiferous plant or part thereof is selected from the group consisting of Parthenium argentatum, Chrysothamnus nauseosus, Taraxacum koksaghyz, Taraxacum officinale and

Assignees

• Fraunhofer Ges Forschung

Inventors

• Schulze Gronover Christian
• Unland Kristina

Classifications

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12P5/007

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• A01H1/06 Primary

  Processes for producing mutations, e.g. treatment with chemicals or with radiation (specific mutations prepared by genetic engineering on plant cell or plant tissues C12N15/00 {; process for producing transgenic plants C12N15/82}) · CPC title

• C12Y504/99

  transferring other groups (5.4.99) · CPC title