Virus-resistant tobacco and breeding method therefor

Patent metadata

Field	Value
Publication number	US-11659807-B2
Application number	US-201916284600-A
Country	US
Kind code	B2
Filing date	Feb 25, 2019
Priority date	Aug 26, 2016
Publication date	May 30, 2023
Grant date	May 30, 2023

Abstract

In order to provide tobacco resistant to a virus, tobacco in accordance with the present invention is arranged such that: (i) a translation initiation factor eIF(iso)4E protein which is non-functional with respect to a virus is produced or (ii) expression of a translation initiation factor eIF(iso)4E gene is suppressed; and (a) a translation initiation factor eIF4E2 protein, which is non-functional with respect to a virus, is produced or (b) expression of a translation initiation factor eIF4E2 gene is suppressed.

First claim

The invention claimed is:

1. A virus-resistant tobacco plant including a plurality of mutations in the genome of the virus-resistant tobacco plant, the plurality of mutations including:
a nonsense mutation in an exon of a translation initiation factor eIF(iso)4E gene, the nonsense mutation causing a production of a translation initiation factor eIF(iso)4E protein with eIF(iso)4E gene function suppressed as compared with a wild type for the virus-resistant tobacco plant and
a nonsense mutation in an exon of a translation initiation factor eIF4E2 gene, the nonsense mutation causing a production of a translation initiation factor eIF4E2 protein with eIF4E2 gene function suppressed as compared with a wild type for the virus-resistant tobacco plant,
an eIF(iso)4E gene including eIF(iso)4E-S and eIF(iso)4E-T genes, and an eIF4E2 gene including eIF4E2-S and eIF4E2-T genes,
the translation initiation factor eIF(iso)4E protein being at least eIF(iso)4E-T protein, and
the translation initiation factor eIF4E2 protein being at least eIF4E2-S protein, wherein the eIF4E2-S gene encodes the eIF4E2-S protein which has a sequence identity of 95% or higher with respect to an amino acid sequence represented by SEQ ID NO: 2,
the eIF4E2-T gene encodes the eIF4E2-T protein which has a sequence identity of 95% or higher with respect to an amino acid sequence represented by SEQ ID NO: 5,
the eIF(iso)4E-S gene encodes the eIF(iso)4E-S protein which has a sequence identity of 97% or higher with respect to an amino acid sequence represented by SEQ ID NO: 8, and
the eIF(iso)4E-T gene encodes the eIF(iso)4E-T protein which has a sequence identity of 97% or higher with respect to an amino acid sequence represented by SEQ ID NO: 11,
wherein the virus is at least both of (i) a strain of Potato virus Y, wherein said strain breaks virus resistance of Virgin A mutant of a tobacco and (ii) the Potato virus Y, and
wherein the virus-resistant tobacco plant is a Nicotiana plant and the eIF4E2-S, eIF4E2-T, eIF(iso)4E-S, and eIF(iso)4E-T genes are each a gene existing in a genome of the Nicotiana plant,
wherein the wild type for the virus resistant tobacco plant is the same as said virus resistant tobacco plant, except that said wild type for the virus resistant tobacco plant does not have said nonsense mutations, and
wherein the wild type has
(i) a wild-type translation initiation factor eIF(iso)4E-S gene which encodes a translation initiation factor eIF(iso)4E-S protein consisting of an amino acid sequence represented by SEQ ID NO: 8; and
(ii) a wild-type translation initiation factor eIF(iso)4E-T gene which encodes a translation initiation factor eIF(iso)4E-T protein consisting of an amino acid sequence represented by SEQ ID NO: 11.

2. The virus-resistant tobacco plant as set forth in claim 1 , wherein:
the translation initiation factor eIF(iso)4E protein includes both of eIF(iso)4E-T protein and eIF(iso)4E-S protein.

3. The virus-resistant tobacco plant as set forth in claim 1 , wherein:
the translation initiation factor eIF4E2 protein includes both of eIF4E2-S protein and eIF4E2-T protein.

4. The virus-resistant tobacco plant as set forth in claim 1 , wherein the nonsense mutation is any one of the following mutations (1) through (4):
(1) a mutation in which C of codon CAA is substituted by T; (2) a mutation in which C of codon CGA is substituted by T; (3) a mutation in which C of codon CAG is substituted by T; and (4) a mutation in which G (either one or both of two Gs) of codon TGG is substituted by A.

5. The virus-resistant tobacco plant as set forth in claim 1 , wherein the virus-resistant tobacco plant further has resistance to a virus belonging to the genus Umbravirus.

6. The virus-resistant tobacco plant as set forth in claim 5 , wherein the virus belonging to the genus Umbravirus is Tobacco bushy top virus.

7. A method for producing a virus-resistant tobacco plant, comprising the step of:
producing tobacco plant resistant to a virus by (i) introducing a nonsense mutation into an exon of a translation initiation factor eIF(iso)4E gene, the nonsense mutation causing production of a translation initiation factor eIF(iso)4E protein with eIF(iso)4E gene function suppressed as compared with a wild type for the virus-resistant tobacco plant, and (ii) introducing a nonsense mutation into an exon of a translation initiation factor eIF4E2 gene, the nonsense mutation causing production of a translation initiation factor eIF4E2 with eIF4E2 gene function suppressed as compared with a wild type for the virus-resistant tobacco plant,
the eIF(iso)4E gene including eIF(iso)4E-S and eIF(iso)4E-T genes, and the eIF4E2 gene including eIF4E2-S and eIF4E2-T genes,
the translation initiation factor eIF(iso)4E protein being at least eIF(iso)4E-T protein, and
the translation initiation factor eIF4E2 protein being at least eIF4E2-S protein, wherein
the eIF4E2-S gene encodes the eIF4E2-S protein which has a sequence identity of 95% or higher with respect to an amino acid sequence represented by SEQ ID NO: 2,
the eIF4E2-T gene encodes the eIF4E2-T protein which has a sequence identity of 95% or higher with respect to an amino acid sequence represented by SEQ ID NO: 5,
the eIF(iso)4E-S gene encodes the eIF(iso)4E-S protein which has a sequence identity of 97% or higher with respect to an amino acid sequence represented by SEQ ID NO: 8, and
the eIF(iso)4E-T gene encodes the eIF(iso)4E-T protein which has a sequence identity of 97% or higher with respect to an amino acid sequence represented by SEQ ID NO: 11,
wherein the virus is at least both of (i) a strain of Potato virus Y, wherein said strain breaks virus resistance of Virgin A mutant of a tobacco and (ii) the Potato virus Y,
wherein the virus-resistant tobacco plant is a Nicotiana plant and the eIF4E2-S, eIF4E2-T, eIF(iso)4E-S, and eIF(iso)4E-T genes are each a gene existing in a genome of the Nicotiana plant,
wherein the wild type for the virus resistant tobacco plant is the same as said virus resistant tobacco plant, except that said wild type for the virus resistant tobacco plant does not have said nonsense mutations, and
wherein the wild type has
(i) a wild-type translation initiation factor eIF(iso)4E-S gene which encodes a translation initiation factor eIF(iso)4E-S protein consisting of an amino acid sequence represented by SEQ ID NO: 8; and
(ii) a wild-type translation initiation factor eIF(iso)4E-T gene which encodes a translation initiation factor eIF(iso)4E-T protein consisting of an amino acid sequence represented by SEQ ID NO: 11.

8. The method as set forth in claim 7 , wherein:
the translation initiation factor eIF(iso)4E protein includes both of eIF(iso)4E-T protein and eIF(iso)4E-S protein.

9. The method as set forth in claim 7 , wherein:
the translation initiation factor eIF4E2 protein includes both of eIF4E2-S protein and eIF4E2-T protein.

10. The method as set forth in claim 7 , wherein the nonsense mutation is one or more any one of the following mutations (1) through (4): (1) a mutation in which C of codon CAA is substituted by T; (2) a mutation in which C of codon CGA is substituted by T; (3) a mutation in which C of codon CAG is substituted by T; and (4) a mutation in which G (either one or both of two Gs) of codon TGG is substituted by A.

11. The method as set forth in claim 7 , wherein the virus is a virus belonging to the genus Potyvirus.

12. The method as set forth in claim 11 , wherein the virus belonging to the genus Potyvirus is at least one of (i) a strain of Potato virus Y which strain breaks virus resistance of Virgin A mutant of tobacco and (ii) the Potato virus Y.

13. A method f

Assignees

• Japan Tobacco Inc

Inventors

• Takakura Yoshimitsu
• Shinjo Akira
• Udagawa Hisashi
• Koga Kazuharu

Classifications

• A01H6/823 Primary

  Nicotiana, e.g. tobacco · CPC title

• A01H5/12 Primary

  Leaves · CPC title

• A01H1/06

  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

• C12N15/8283

  for virus resistance · CPC title

• A24B1/00

  Preparation of tobacco on the plantation · CPC title