US-12618080-B2 - Solanaceous plant and solanaceous plant cell having resistance to tomato spotted wilt virus, and method for producing solanaceous plant

US12618080B2US 12618080 B2US12618080 B2US 12618080B2US-12618080-B2

Abstract

The present invention addresses the problem of providing: a solanaceous plant and a solanaceous plant cell that have resistance to tomato spotted wilt virus (TSWV) and that have the property of inhibiting TSWV infection, the property of suppressing the propagation of TSWV after infection, and/or the property of suppressing the onset of the symptoms of TSWV infection; and a method for producing the solanaceous plant. The present invention provides a solanaceous plant that has resistance to TSWV, in which at least one gene selected from the group consisting of receptor-like kinase RLK genes and homologous genes thereof has a mutation, and in which as a result of the mutation, either the expression of the gene having the mutation is suppressed or a protein encoded by the gene having the mutation is non-functional with respect to TSWV.

Inventors

Hiroki Atarashi
Kenji Nakahara

Assignees

KIKKOMAN CORPORATION
NATIONAL UNIVERSITY CORPORATION HOKKAIDO UNIVERSITY

Dates

Publication Date: 20260505
Application Date: 20201207
Priority Date: 20191224

Claims (8)

1 . A solanaceous plant having a mutation in a receptor-like kinase RLK gene having a cDNA sequence comprising the nucleotide sequence as set forth in SEQ ID NO:1, wherein the solanaceous plant is obtained by introducing genomic gene mutation to the genome of the solanaceous plant by genome editing techniques, wherein the RLK gene is mutated to have a cDNA sequence comprising the nucleotide sequence as set forth in SEQ ID NO: 10 or SEQ ID NO: 11, wherein the mutation either inhibits expression of the mutated gene or makes a protein encoded by the mutated gene to be non-functional for tomato spotted wilt virus, and wherein the protein which is non-functional for the virus is either a protein which cannot be used by the virus during infection and replication, or a protein which reduces the infection and replication of the virus, as compared to a control solanaceous plant without the introduction of the mutation, and wherein the solanaceous plant has virus resistance against the tomato spotted wilt virus.
2 . The solanaceous plant according to claim 1 which is a tomato.
3 . A part of the solanaceous plant according to claim 1 .
4 . A processed material of the solanaceous plant according to claim 1 or a part thereof, wherein the processed material comprises at least one cell of the solanaceous plant having the mutation in the receptor-like kinase RLK gene.
5 . A method for producing a tomato spotted wilt virus resistant solanaceous plant, the method comprising: selecting the receptor-like kinase RLK gene having a cDNA sequence comprising the nucleotide sequence as set forth in SEQ ID NO:1; introducing a mutation into the selected gene in a genome of a solanaceous plant by genome editing techniques, wherein the mutation is introduced so that the RLK gene has a cDNA sequence comprising the nucleotide sequence as set forth in SEQ ID NO: 10 or SEQ ID NO: 11, wherein the introduced mutation is either a mutation inhibiting an expression of the mutated gene or a mutation making a protein encoded by the mutated gene to be non-functional for tomato spotted wilt virus, and wherein the protein which is non-functional for the virus is either a protein which cannot be used by the virus during infection and replication, or a protein which reduces the infection and replication of the virus, as compared to a control solanaceous plant without the introduction of the mutation; and selecting the solanaceous plant showing lower infection rate for the tomato spotted wilt virus, as compared to control solanaceous plant without the introduction of the mutation, wherein the infection rate is based on the presence or absence of tomato spotted wilt virus symptoms judged by observation, and/or the presence or absence of a gene of the tomato spotted wilt virus detected by PCR, on day 20 or more post tomato spotted wilt virus inoculation.
6 . A tomato spotted wilt virus resistant solanaceous plant obtained by the production method according to claim 5 .
7 . A method for producing a bred progeny of a tomato spotted wilt virus resistant solanaceous plant, the method comprising: self-pollination or cross-pollination of either the tomato spotted wilt virus resistant solanaceous plant obtained by the production method according to claim 5 or a progeny of the tomato spotted wilt virus resistant solanaceous plant.
8 . A tomato spotted wilt virus resistant solanaceous plant obtained by the production method of claim 7 .

Description

CROSS REFERENCE TO RELATED APPLICATIONS This is the U.S. national stage of application No. PCT/JP2020/045381, filed on Dec. 7, 2022. Priority under 35 U.S.C. § 119(a) and 35 U.S.C. § 365(b) is claimed from Japanese Application No. 2019-232766 filed Dec. 24, 2019, the disclosure of which is also incorporated herein by reference. TECHNICAL FIELD The present invention relates to a solanaceous plant resistant to tomato spotted wilt virus, a solanaceous plant cell, and a method for producing the solanaceous plant. BACKGROUND ART In accordance with the increase in commercial distribution of agricultural products, viral diseases, which have been localized in the past, begun to spread throughout the world. Representatives of such viruses are viruses of genus Tospovirus belonging to the family Bunyaviridae and of genus Begomovirus belonging to the family Geminiviridae. Tomato spotted wilt virus (hereinafter, frequently abbreviated to “TSWV”) is a very important virus causing scientific and economic impact and is ranked in top five among various plant viruses. Tomato spotted wilt virus is first discovered in 1915 and is one of the viruses having relatively long history. However, study of this virus have been delayed due to difficulty in purifying a complete virion. Number of studies began to finally increase in 1990's, and now, the virus is classified as a type virus of the genus Orthotospovirus of the family Tospoviridae (see, for example, NPL 1). In the family Bunyaviridae, there are four other genera in addition to the genus Tospovirus, and the viruses of these other genera infect animals. Therefore, this family is taxonomically very special virus group which include animal viruses and plant viruses in the same family (see, for example, NPL 2 and NPL 3). TSWV is a spherical virus having an envelope with a diameter of about 100 nm, and having a trisegmented, closed circular filament-shaped nucleocapsid therein. The viral genome is single-stranded RNA which is also triple-segmented, and goes through genetic translation basically as a minus strand. A large factor causing worldwide outbreaks of viruses of the genus Tospovirus, such as TSWV, and of the genus Begomovirus of the family Geminiviridae, such as tomato yellow leaf curl virus (TYLCV), is the extension of the distribution of insect vectors in accordance with the globalization of the commercial distribution. The important insect vectors for both viruses have expanded their distribution by attaching to agricultural products including flowers. TSWV is mainly transmitted by Thysanoptera, very small insects of about 1 mm in body length. Various species of Thysanoptera, such as soybean thrips, onion thrips, western flower thrips, and flower thrips, are known to mediate the virus. Thrips is capable of acquiring TSWV only in larval stage by sap-sucking, and although larva before eclosion also has infectivity, the TSWV infection generally occurs through sap-sucking by adult insects. At the beginning, thrips mediating TSWV were native species, such as flower thrips and thrips setosus, but western flower thrips, known as an important vector for TSWV, entered Japan in 1990 from overseas. In accordance with the expansion of western flower thrips by its ability to move long distances, occurrence of TSWV changed drastically. As Thysanoptera prefers pollen, viruses of genus Tospovirus, including TSWV, cause drastic damages to various flowers and also to vegetables cultivated next to flower fields. Host spectrum of TSWV is very broad, including 900 or more plants, and nowadays, worldwide occurrence of TSWV is seen mainly in vegetables and flowering plants, including tomato, bell pepper, tobacco, melon, chrysanthemum, dahlia, and eustoma (see, for example, NPL 4). Further, TSWV is able to infect weeds of Asteraceae, Polygonaceae, and the like, and some plants overwinter and become infection source next year. Once the virus of the genus Tospovirus infect a plant, they are likely to be localized and masked in plants, thus, these viruses are difficult to remove. So far, the measures for preventing the TSWV disease are either by breeding resistant varieties, complete extermination of thrips, or early removal of plants infected with TSWV. Thrips is a very small insect with a length of 1 mm or less, and the greenhouse must be covered with fine-mesh net to control the invasion of thrips. However fine-mesh net may cause increasing temperature in the greenhouses, this control is actually hesitated. In addition, as thrips prefers pollen and dives into perianth, pesticides is not effective. As a plant breeding conferring TSWV resistance, for example, in 1998, resistant gene Sw-5 was isolated from wild-relatives tomato Solanum peruvianum L. having resistance to TSWV, and was introgressed into general cultivars (see, for example, NPL 5). Sw-5 was a promising resistant gene in tomato, however resistance-overcoming strains of TSWV emerged in different places worldwide before becoming popular