Deprecated: The each() function is deprecated. This message will be suppressed on further calls in /home/zhenxiangba/zhenxiangba.com/public_html/phproxy-improved-master/index.php on line 456
JP4336844B2 - Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho - Google Patents
[go: Go Back, main page]

JP4336844B2 - Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho - Google Patents

Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho Download PDF

Info

Publication number
JP4336844B2
JP4336844B2 JP2003015270A JP2003015270A JP4336844B2 JP 4336844 B2 JP4336844 B2 JP 4336844B2 JP 2003015270 A JP2003015270 A JP 2003015270A JP 2003015270 A JP2003015270 A JP 2003015270A JP 4336844 B2 JP4336844 B2 JP 4336844B2
Authority
JP
Japan
Prior art keywords
sweet potato
virus
attenuated
mosaic virus
attenuated virus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
JP2003015270A
Other languages
Japanese (ja)
Other versions
JP2004222613A (en
Inventor
山崎修一
英世 後藤
薫 花田
淳一 酒井
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
National Agriculture and Food Research Organization
Oita Prefectural Government
Original Assignee
National Agriculture and Food Research Organization
Oita Prefectural Government
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by National Agriculture and Food Research Organization, Oita Prefectural Government filed Critical National Agriculture and Food Research Organization
Priority to JP2003015270A priority Critical patent/JP4336844B2/en
Publication of JP2004222613A publication Critical patent/JP2004222613A/en
Application granted granted Critical
Publication of JP4336844B2 publication Critical patent/JP4336844B2/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Images

Landscapes

  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Description

【0001】
【発明の属する技術分野】
本発明は、サツマイモ斑紋モザイクウイルスの弱毒ウイルス(10-O)、該弱毒ウイルス(10-O)を用いるサツマイモ帯状粗皮病の防除方法及びサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショに関する。
【0002】
【従来の技術】
カンショ栽培の阻害要因として、サツマイモ斑紋モザイクウイルス強毒系統の感染による帯状粗皮病の発生があり、これによる品質の低下や販売単価の下落が生産者に深刻な打撃を与えている。
【0003】
サツマイモ帯状粗皮病の防除対策は長い間困難を極めてきたが、その理由として、SPFMV-Sの塊根伝染性がある。これにより、病原ウイルスが感染したカンショを種イモとすると、それから得られた苗がほとんど全てウイルスに感染し、生産したいもの大部分が発病してしまう。
【0004】
サツマイモ帯状粗皮病に効果の高い農薬はなく、対策法として、ウイルスフリー化したカンショ苗(以下ウイルスフリー苗)の栽培により被害軽減を図っている。しかし、ウイルスフリー苗は、ウイルス抵抗性を持たないため、アブラムシの媒介により病原ウイルスに容易に再感染してしまう。そのため、苗の有効期間も1〜2作と短期間で利用不能となり、定期的な苗の更新が必要となることから、生産者は、少なくとも隔年で苗を更新(購入)しなくてはならず、負担が大きい。
【0005】
【発明が解決しようとする課題】
本発明は、ウイルスフリー苗に予防接種してサツマイモ帯状粗皮病に対する抵抗力を付け発病を抑える弱毒ウイルス(10-O)、該弱毒ウイルスを用いて長期的な防除効果を可能とするサツマイモ帯状粗皮病の防除法及びサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショを提供することを目的とする。
【0006】
【課題を解決するための手段】
本発明者らは、上記課題を解決するため、自然界より得たウイルス症状の軽微なカンショ塊根の中から、サツマイモ斑紋モザイクウイルスの弱毒ウイルスの選抜を重ねた結果、カンショ塊根に対するウイルス症状が極めて軽微であり、ウイルスフリー苗に予防接種しておくことによりサツマイモ帯状粗皮病の発生を防ぐことができる弱毒ウイルス(10-O)の単離に成功し、本発明を完成した。
【0007】
即ち、請求項1記載の発明は、カンショに帯状粗皮病を発症せず、サツマイモ斑紋モザイクウイルス強毒系統に干渉効果を有することを特徴とするサツマイモ斑紋モザイクウイルスに属する弱毒ウイルス(10-O)であり、請求項2記載の発明は、上記の弱毒ウイルス(10-O)を、カンショに接種することを特徴とするサツマイモ帯状粗皮病の防除法であり、また、請求項3記載の発明は、上記の弱毒ウイルス(10-O)を接種して得られるサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショであり、請求項4記載の発明は、上記の弱毒ウイルス(10-O)を接種して得られるサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショの一部を栄養繁殖して得られるサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショである。
【0008】
【発明の実施の形態】
以下、本発明を詳細に説明する。
『弱毒ウイルスの作出方法(弱毒株の探索)』
弱毒ウイルスは自然界からの選択分離により作出した。また、分離源(弱毒株)は次のように得た。平成7〜11年度に大分県大野郡4町2村の現地カンショ畑において、カンショ約5万株について病徴調査を実施し、塊根の症状の軽微なカンショ249株を採集した。これらの株についてサツマイモ斑紋モザイクウイルス強毒系統及び普通系統の外被蛋白質領域及び3'末端側非翻訳領域の共通配列を元にしたプライマーによるRT-PCRに供試した。その結果、全ての株でサツマイモ斑紋モザイクウイルスの感染が確認された。そこで、これらにサツマイモ斑紋モザイクウイルス強毒系統を接木接種によりチャレンジ接種し、6代にわたり病徴調査を実施したところ、なお発病しない干渉効果を有している株を得たため、これを弱毒株として弱毒ウイルスの分離源とした。
【0009】
『弱毒ウイルスの作出方法(弱毒ウイルスの選択分離)』
弱毒株は、それ自身干渉効果を有しているものの、複数のサツマイモ斑紋モザイクウイルスの系統に重複感染していたため、干渉効果を有する弱毒ウイルスを特定するために、これらの系統を全身感染宿主であるアサガオを用いた希釈分離法により分離した。すなわち、弱毒株にアサガオを接木し、分離した各ウイルスは、分離確認のため、サツマイモ斑紋モザイクウイルス各系統及びサツマイモウイルスGのNIb蛋白質領域及び外被蛋白質領域の共通配列を元にしたプライマーと2種類の制限酵素(BamHI、EcoRI)を用いたRFLP-RT-PCRによる系統識別法に供試した。その結果、弱毒株からは、普通系統、JC系統と同様のバンドパターンを示す2系統が分離された。これらをカンショウイルスフリー株に接種して症状を検討すると共に、接種株に強毒系統をチャレンジ接種して干渉効果の有無を確認したところ、普通系統と同様のバンドパターンを示した1系統のみが干渉効果を有しており、実用的な弱毒ウイルス(10-O)として選抜した(図1、9)。
【0010】
『弱毒ウイルスの特性』
弱毒ウイルス(10-O)はひも状を呈する植物ウイルスであり、サツマイモ斑紋モザイクウイルスに属する。また、全塩基配列を解読したところ、塩基からなり、5'側1.2Kbpが強毒系統と、3'側9.2Kbpが普通系統と相同性が高く、これまで報告のあるサツマイモ斑紋モザイクウイルスの各系統とは異なる系統であった(図2〜8)。カンショ及びアサガオで全身感染する。アサガオでは、容易に汁液接種で増殖できるが、カンショでは汁液接種で感染しないため、カンショウイルスフリー株に感染させる場合は、アサガオもしくは、カンショ弱毒ウイルス感染株(弱毒株)からの接木接種により感染させる必要がある。一方で、このウイルスはカンショで塊根伝染性が認められるため、一度感染させると蔓や塊根を媒介として次代以降もカンショ体内に維持される。
病原ウイルスであるサツマイモ斑紋モザイクウイルス強毒系統との識別は、段落番号0009に示したRFLP-RT-PCRによる系統識別法により容易に区別できる(図1)。6代にわたる病徴調査や系統識別法による検討等の結果、弱毒ウイルス(10-O)の病原力は安定して弱く、変異しにくい弱毒ウイルスであることが確認された。
【0011】
『弱毒ウイルスのクローニングとシーケンス』
弱毒ウイルス(10-O)に感染したサツマイモ葉から全RNAを抽出した。この弱毒ウイルスは、段落番号0009に示したRFLP-RT-PCRによる系統識別法によりサツマイモ斑紋モザイクウイルス普通系統と極めてよく似た性質を持つことが明らかとなっていたため(図1)、普通系統の塩基配列を参考にして合成したDNAプライマーを用いてRT-PCRを行い、弱毒ウイルス(10-O)のゲノムの全長をカバーするcDNA断片を複数得た。これらをpBluescript SKII+(STRATAGENE社)のEcoRV部位に導入し、クローニングした。得られたクローンを普通系統の塩基配列を参考にして合成したDNAプライマーを用いてDNAシークエンサー(ABI373A)で塩基配列を決定した(図3〜8)。
【0012】
『弱毒ウイルスの接種方法と感染苗の利用方法』
カンショへの弱毒ウイルス(10-O)の接種方法は、弱毒ウイルス(10-O)に感染したアサガオを穂木とする接木接種を用いておこなう。また、穂木用のアサガオは、弱毒ウイルスに感染したアサガオ葉0.1gに緩衝液(中性付近の0.1Mリン酸緩衝液)を10ml加え作出した磨砕液を葉1枚あたり100μlずつカーボランダム法により汁液接種し、21℃、16時間日長下で14日間育苗したものを用いた。一方、接木は、腹接ぎにより行い、21℃、16時間日長下で14日間の育苗すると、弱毒ウイルス(10-O)はカンショに全身感染する。
弱毒ウイルス(10-O)に感染したカンショ苗(弱毒ウイルス感染苗)を実際に利用する場合は、最初に接木接種すれば、ウイルスの塊根伝染性により接種株からの採苗により感染苗を得ることができる。特に次年度以降は、苗を保存するか、収穫した塊根を貯蔵し、それから採苗することにより、接種をすることなく容易に感染苗を得ることができる。
【0013】
【実施例1】
弱毒ウイルス(10-O)の防除効果試験について説明する。
段落番号0012に従い弱毒ウイルス(10-O)を接種したカンショに、接種14日後にサツマイモ斑紋モザイクウイルス強毒系統をチャレンジ接種し、さらに21℃、16時間日長下で14日間育苗したポット苗から採苗し、表の耕種概要により農業技術センター畑地利用部圃場(大分県三重町)に挿苗し、塊根を収穫して病徴調査を行ったところ、弱毒ウイルス(10-O)をあらかじめ接種しておいたカンショは発病せず、塊根の品質低下も認められなかった。しかし、弱毒ウイルス(10-O)をあらかじめ接種していなかったカンショでは、発病して塊根表皮に帯状の細かなひび割れが生じるほか、退色、くびれ等が生じ、外観を損ない、著しく品質がが低下した。
【0014】
また、弱毒ウイルス(10-O)をあらかじめ接種しておいたカンショは、次代(図9)、三代においても発病せず、塊根の品質低下も認められなかった。
さらに、このカンショは、ウイルスフリー苗と同程度の収量が得られた。
【0015】
【実施例2】
弱毒ウイルス(10-O)の現地実証試験について説明する。
段落番号0012の苗を大分県内の農家(三重町1件、大野町2件)のカンショ圃場に挿苗したところ、いずれの場合も弱毒ウイルス(10-O)をあらかじめ接種しておいたカンショは発病せず、塊根の品質低下も認められなかった(図10)。
また、収量についてもウイルスフリー苗と同程度の量が得られた。
【0016】
【本発明の効果】
本発明の弱毒ウイルス(10-O)は、自然界に揉まれて存在したサツマイモ斑紋モザイクウイルスの系統から選抜しているので変異しにくく安全な弱毒ウイルスである。また、弱毒ウイルス(10-O)をカンショに接種し、一般栽培圃場において栽培した場合、防除効果を有し、直ちに実用化できる弱毒ウイルスである。その効果は、当代だけでなく生産された塊根を母本として利用することにより次代以降も継続されるため、苗の更新期間が延長され、苗代にかかる生産者の負担を軽減させることができる。さらに、ウイルスを媒介するアブラムシ防除の農薬や、アブラムシ忌避資材なども要しないことから、環境汚染の問題がなく、栽培経費の大幅な削減効果が期待できるものである(図11)。
【図面の簡単な説明】
【図1】RFLP-RT-PCRによる弱毒ウイルス(10-O)の解析
【図2】弱毒ウイルス(10-O)とサツマイモ斑紋モザイクウイルス2系統との塩基配列の相同性の比較
【図3】弱毒ウイルス(10-O)の塩基配列(1〜1980)
【図4】弱毒ウイルス(10-O)の塩基配列(1981〜3960)
【図5】弱毒ウイルス(10-O)の塩基配列(3961〜5940)
【図6】弱毒ウイルス(10-O)の塩基配列(5941〜7980)
【図7】弱毒ウイルス(10-O)の塩基配列(7981〜9960)
【図8】弱毒ウイルス(10-O)の塩基配列(9961〜10820)
【図9】弱毒ウイルス(10-O)の干渉効果
【図10】弱毒ウイルス(10-O)の現地実証試験
【図11】弱毒ウイルス(10-O)の防除効果写真
【符号の説明】
使用した制限酵素
1〜4:BamHI 5〜8:EcoRI
解析したウイルス
4,8:10-O 1,5:SPFMV-S 2,6:SPFMV-O 3,7:SPFMV-JC
サイズマーカー
M:λ/HindIII・HincII
発病度={Σ(発病指数※×同株数)/(3×全株数)}×100
※発病指数:0:無症,1:わずかな退色,2:退色、わずかな粗皮症状
3:激しい粗皮症状や退色、くびれ
A ウイルスフリー株
B 10-O感染株にSPFMV-Sを接種
C ウイルスフリー株にSPFMV-Sを接種
[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an attenuated virus (10-O) of sweet potato mottled mosaic virus, a method for controlling sweet potato stripy skin disease using the attenuated virus (10-O), and a sweet potato mottled mosaic virus highly virulent resistant potato.
[0002]
[Prior art]
As an inhibitory factor in sweet potato cultivation, the occurrence of banded bark disease caused by infection with a sweet potato mottled mosaic virus virulent strain has caused serious damage to producers due to the decline in quality and the decline in sales unit price.
[0003]
Measures to control sweet potato banding have been extremely difficult for a long time because of the tuberculosis infectivity of SPFMV-S. As a result, when a sweet potato infected with a pathogenic virus is used as a seed potato, almost all the seedlings obtained from it are infected with the virus, and most of the plants that are desired to be produced are ill.
[0004]
There are no pesticides that are highly effective against sweet potato striped skin disease, and as a countermeasure, damage is reduced by growing virus-free sweet potato seedlings (hereinafter virus-free seedlings). However, since virus-free seedlings do not have virus resistance, they are easily reinfected with a pathogenic virus through aphids. Therefore, the effective period of seedlings will be unavailable in a short period of 1 to 2 crops, and regular seedling renewal will be required, so producers must renew (purchase) seedlings at least every other year. The burden is large.
[0005]
[Problems to be solved by the invention]
The present invention relates to an attenuated virus (10-O) that vaccinates virus-free seedlings and thereby suppresses the onset of disease by causing resistance to sweet potato striped skin disease, and a sweet potato striped skin that enables a long-term control effect using the attenuated virus. An object of the present invention is to provide a method for controlling diseases and a sweet potato strain resistant to sweet potato mottle mosaic virus.
[0006]
[Means for Solving the Problems]
In order to solve the above problems, the present inventors have repeatedly selected attenuated viruses of sweet potato mottled mosaic virus from sweet potato roots with mild viral symptoms obtained from nature. Thus, the present inventors have succeeded in isolating an attenuated virus (10-O) that can prevent the occurrence of sweet potato striped skin disease by immunizing virus-free seedlings, thereby completing the present invention.
[0007]
That is, the invention according to claim 1 is an attenuated virus (10-O) belonging to the sweet potato mottled mosaic virus, characterized in that it does not develop a bandy skin disease in sweet potato and has an interference effect on the virulent strain of sweet potato mottled mosaic virus. The invention according to claim 2 is a method for controlling sweet potato striped skin disease characterized by inoculating sweet potato with the attenuated virus (10-O), and the invention according to claim 3 A sweet potato mottle mosaic virus highly resistant strain resistant sweet potato obtained by inoculating the attenuated virus (10-O), and the invention according to claim 4 inoculates the attenuated virus (10-O). This is a sweet potato mottled mosaic virus virulent resistant potato obtained by vegetative propagation of a portion of the sweet potato mottled mosaic virus virulent resistant potato.
[0008]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail.
"How to create attenuated viruses (search for attenuated strains)"
Attenuated viruses were created by selective isolation from nature. Moreover, the isolation source (attenuated strain) was obtained as follows. From 1995 to 1999, symptom survey was conducted on about 50,000 strains of local sweet potatoes in 4 towns and 2 villages in Ono-gun, Oita Prefecture, and 249 strains of mild sweet potato with mild tuber symptoms were collected. These strains were subjected to RT-PCR using primers based on the consensus sequence of the coat protein region and the 3 'terminal untranslated region of the sweet potato mottle mosaic virus virulent strain and the normal strain. As a result, infection of sweet potato mottled mosaic virus was confirmed in all strains. Therefore, we challenged these plants with a sweet potato mottled mosaic virus virulent strain by grafting, and conducted a disease symptom survey for 6 generations. As a result, we obtained a strain that had an interference effect that did not cause disease. A source of attenuated virus was used.
[0009]
“Method of creating attenuated virus (selection and separation of attenuated virus)”
Although the attenuated strains themselves had an interference effect, they were co-infected with multiple sweet potato mottle mosaic virus strains. It isolate | separated by the dilution separation method using a certain morning glory. In other words, each virus isolated after grafting morning glory on an attenuated strain was separated from the primer based on the common sequence of the NIb protein region and coat protein region of each sweet potato mottle mosaic virus strain and sweet potato virus G for confirmation of separation. This was subjected to a line identification method by RFLP-RT-PCR using various types of restriction enzymes (BamHI, EcoRI). As a result, two strains showing the same band pattern as the normal strain and JC strain were isolated from the attenuated strain. While inoculating these into a kansho virus-free strain and examining the symptoms, when the inoculated strain was challenged with a virulent strain and checked for the presence or absence of interference effects, only one strain that showed the same band pattern as the normal strain was found. It was selected as a practical attenuated virus (10-O) having an interference effect (FIGS. 1 and 9).
[0010]
"Characteristics of attenuated viruses"
The attenuated virus (10-O) is a plant virus having a string shape and belongs to the sweet potato mottle mosaic virus. In addition, when the entire nucleotide sequence was deciphered, each of the sweet potato mottled mosaic viruses reported so far consisted of bases, the 5 ′ side 1.2 Kbp is highly toxic and the 3 ′ side 9.2 Kbp is highly homologous to the normal strain. It was a system different from the system (FIGS. 2 to 8). Systemic infection with sweet potato and morning glory. In morning glory, it can easily grow by inoculation with sap, but in sweet potato, it does not infect by sap inoculation. Therefore, when infecting kancho virus-free strains, infect by grafting from morning glory or kansho attenuated virus-infected strain (attenuated strain). There is a need. On the other hand, since this virus has tuberculosis infectivity in sweet potato, it is maintained in the sweet potato after the next generation through vines and tuberous roots once infected.
The pathogenic virus can be easily distinguished from the highly virulent strain of sweet potato mottled mosaic virus by the line identification method by RFLP-RT-PCR shown in paragraph 0009 (FIG. 1). As a result of symptom surveys over six generations and studies using strain identification methods, it was confirmed that the attenuated virus (10-O) has a stable and weak pathogenicity and is not easily mutated.
[0011]
"Cloning and sequencing of attenuated viruses"
Total RNA was extracted from sweet potato leaves infected with attenuated virus (10-O). This attenuated virus has been shown to have very similar properties to the normal line of sweet potato mottle mosaic virus by the line identification method by RFLP-RT-PCR shown in paragraph 0009 (Fig. 1). RT-PCR was performed using DNA primers synthesized with reference to the nucleotide sequence, and a plurality of cDNA fragments covering the entire genome of the attenuated virus (10-O) were obtained. These were introduced into the EcoRV site of pBluescript SKII + (STRATAGENE) and cloned. The base sequence was determined with a DNA sequencer (ABI373A) using DNA primers obtained by synthesizing the obtained clone with reference to the base sequence of a normal strain (FIGS. 3 to 8).
[0012]
"Methods for inoculating attenuated viruses and using infected seedlings"
The inoculation method of sweet potato virus (10-O) to sweet potato is carried out using graft inoculation using morning glory infected with attenuated virus (10-O). As for the morning glory for hogi, the carborundum method of 100 μl per leaf of milled solution made by adding 10 ml of buffer solution (0.1M phosphate buffer solution near neutrality) to 0.1 g of morning glory leaves infected with attenuated virus After inoculating the sap, the seedlings were grown for 14 days at 21 ° C. for 16 hours. On the other hand, grafting is performed by abdominal attachment, and when the seedlings are grown for 14 days at 21 ° C. for 16 hours, the attenuated virus (10-O) is systemically infected with sweet potato.
When actually using sweet potato seedlings (attenuated virus-infected seedlings) infected with the attenuated virus (10-O), the infected seedlings can be obtained by seeding from the inoculated strain due to the infectivity of the tuberous roots by inoculating the graft first. be able to. In particular, after the next fiscal year, infected seedlings can be easily obtained without inoculation by storing seedlings or storing harvested tuberous roots and then collecting seedlings.
[0013]
[Example 1]
The control effect test of attenuated virus (10-O) is explained.
From potato seedlings inoculated with sweet potato mottle mosaic virus virulent strain 14 days after inoculation into sweet potato inoculated with attenuated virus (10-O) according to paragraph 0012, and further grown for 14 days at 21 ° C for 16 hours We collected seedlings, planted seedlings in the Agricultural Technology Center field use section field (Miemachi, Oita Prefecture) according to the summary of cultivation in the table, harvested tuberous roots and conducted symptom surveys, then inoculated with attenuated virus (10-O) in advance The dried sweet potato did not become sick and the quality of tuberous root was not deteriorated. However, in sweet potatoes that had not been pre-inoculated with attenuated virus (10-O), it became sick and caused fine strip-like cracks in the tuberous epidermis, as well as fading and constriction, which deteriorated the appearance and markedly reduced the quality. did.
[0014]
In addition, sweet potato pre-inoculated with the attenuated virus (10-O) did not develop disease in the next generation (FIG. 9) and the third generation, and no deterioration in tuberous root quality was observed.
Furthermore, this sweet potato yielded the same yield as virus-free seedlings.
[0015]
[Example 2]
The field demonstration test of attenuated virus (10-O) will be explained.
When the seedlings of paragraph number 0012 were planted in potato fields of farmers in Oita prefecture (one in Mie Town and two in Ono Town), the potatoes that had been pre-inoculated with the attenuated virus (10-O) in both cases became ill No degradation of tuberous root was observed (FIG. 10).
In addition, the yield was comparable to that of virus-free seedlings.
[0016]
[Effect of the present invention]
The attenuated virus (10-O) of the present invention is a safe attenuated virus that is difficult to mutate because it is selected from the strain of sweet potato mottle mosaic virus that has been found in nature. In addition, when an attenuated virus (10-O) is inoculated into sweet potatoes and cultivated in a general cultivation field, the attenuated virus has a controlling effect and can be put into practical use immediately. The effect is continued not only in the present generation but also in subsequent generations by using the produced tuberous root as a mother plant, so that the renewal period of the seedling can be extended and the burden on the producer related to the seedling can be reduced. Furthermore, since no aphid pesticides or aphid repellent materials that mediate viruses are required, there is no problem of environmental pollution, and a significant reduction in cultivation costs can be expected (FIG. 11).
[Brief description of the drawings]
[Fig. 1] Analysis of attenuated virus (10-O) by RFLP-RT-PCR [Fig. 2] Comparison of nucleotide sequence homology between attenuated virus (10-O) and two sweetpotato mottle mosaic viruses [Fig. 3] Attenuated virus (10-O) base sequence (1-1980)
FIG. 4 Base sequence of attenuated virus (10-O) (1981 to 3960)
FIG. 5: Base sequence of attenuated virus (10-O) (3961-5940)
FIG. 6: Base sequence of attenuated virus (10-O) (5941-7980)
[Fig. 7] Base sequence of attenuated virus (10-O) (7981-9960)
[Fig. 8] Base sequence of attenuated virus (10-O) (9961-10820)
[Fig. 9] Interference effect of attenuated virus (10-O) [Fig. 10] Field demonstration test of attenuated virus (10-O) [Fig. 11] Photo of control effect of attenuated virus (10-O) [Explanation of symbols]
Restriction enzymes used 1-4: BamHI 5-8: EcoRI
Analyzed viruses 4, 8: 10-O 1, 5: SPFMV-S 2, 6: SPFMV-O 3, 7: SPFMV-JC
Size marker M: λ / HindIII / HincII
Disease severity = {Σ (morbidity index * × number of same strains) / (3 × total number of strains)} × 100
* Disease index: 0: asymptomatic, 1: slight fading, 2: fading, slight rough skin symptom 3: severe rough skin or fading, constriction A virus free strain B 10-O inoculated with SPFMV-S C virus Inoculate free strains with SPFMV-S

Claims (4)

カンショに帯状粗皮病を発症せず、サツマイモ帯状粗皮病の病原ウイルスであるサツマイモ斑紋モザイクウイルス強毒系統に干渉効果を有し、図3〜8に記載した塩基配列を含むことを特徴としたサツマイモ斑紋モザイクウイルスに属する弱毒ウイルス(10-O)Sweet potato characterized in that it does not develop stripy skin disease in sweet potato, has an interference effect on sweet potato mottled mosaic virus virulent strain, which is a pathogenic virus of sweet potato striping skin disease, and contains the base sequences described in FIGS. Attenuated virus belonging to mottle mosaic virus (10-O) 請求項1記載のサツマイモ斑紋モザイクウイルスに属する弱毒ウイルス(10-O)を、カンショに接種することを特徴とするサツマイモ帯状粗皮病の防除法A method for controlling sweet potato stripy skin disease characterized by inoculating sweet potato with the attenuated virus (10-O) belonging to the sweet potato mottle mosaic virus according to claim 1 請求項1記載のサツマイモ斑紋モザイクウイルスに属する弱毒ウイルスを接種して得られるサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショSweet potato mottle mosaic virus resistant potato strain obtained by inoculating the attenuated virus belonging to the sweet potato mottle mosaic virus according to claim 1 請求項3記載のサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショの一部を栄養繁殖して得られるサツマイモ斑紋モザイクウイルス強毒系統抵抗性カンショSweet potato mottled mosaic virus virulent strain resistant sweet potato obtained by vegetative propagation of a portion of the sweet potato mottled mosaic virus virulent strain resistant potato of claim 3
JP2003015270A 2003-01-23 2003-01-23 Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho Expired - Fee Related JP4336844B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2003015270A JP4336844B2 (en) 2003-01-23 2003-01-23 Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2003015270A JP4336844B2 (en) 2003-01-23 2003-01-23 Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho

Publications (2)

Publication Number Publication Date
JP2004222613A JP2004222613A (en) 2004-08-12
JP4336844B2 true JP4336844B2 (en) 2009-09-30

Family

ID=32903076

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2003015270A Expired - Fee Related JP4336844B2 (en) 2003-01-23 2003-01-23 Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho

Country Status (1)

Country Link
JP (1) JP4336844B2 (en)

Also Published As

Publication number Publication date
JP2004222613A (en) 2004-08-12

Similar Documents

Publication Publication Date Title
JP7370333B2 (en) TBRFV resistant tomato plants
Gonsalves et al. Papaya ringspot virus
Grimova et al. Apple mosaic virus
IL281310A (en) Resistance to tolcndv in melons
Alishiri et al. Prevalence of tobacco mosaic virus in Iran and evolutionary analyses of the coat protein gene
Reddy Distinct Begomoviruses Closely Related to Cassava Mosaic Viruses Cause Indian Jatropha Mosaic Disease" DS Aswatha Narayana, KT Rangaswamy," KS Shankarappa,“MN Maruthi,'CN. Lakshminarayana Reddy,'AR Rekha and" KV Keshava Murthy" Department of Plant Pathology, University of Agricultural Sciences
MX2014010474A (en) Tswv resistant capsicum plants.
CN112626093A (en) Tomato bacterial wilt resistance gene Sl alpha-KGDH E2 and application thereof
Chain et al. Evaluation of the durability of the Barley yellow dwarf virus-resistant Zhong ZH and TC14 wheat lines
US12414514B2 (en) Tolerance to ToLCNDV in cucumber
CASTAGNONE‐SERENO et al. Differential expression of root‐knot nematode resistance genes in tomato and pepper: evidence with Meloidogyne incognita virulent and avirulent near‐isogenic lineages
Islam et al. Characterization of progenies from intergeneric hybridization between Oryza sativa L. and Porteresia coarctata (Roxb.) Tateoka
Venkataravanappa et al. Molecular evidence for association of Tobacco curly shoot virus and a betasatellite with curly shoot disease of common bean (Phaseolus vulgaris L.) from India
JP4336844B2 (en) Sweet Potato Mottle Mosaic Virus Attenuated Virus, Control Method of Sweet Potato Stripe Peeling Disease Using the Attenuated Virus, and Sweet Potato Mottle Mosaic Virus Virulent Line Resistance Kansho
Zhang et al. A leucine-rich repeat receptor-like protein kinase enhances tomato resistance to Phelipanche aegyptiaca
Hajimorad et al. Evaluation of seed transmissibility of Soybean vein necrosis-associated virus in two soybean cultivars grown under field conditions
Rajasekharam Biological and molecular characterization and management of watermelon bud necrosis virus
CN114786474B (en) Burn virus resistance gene
CN116732220A (en) SNP locus related to resistance of tomato brown wrinkle fruit virus, TBR11 gene and application
Jamous et al. Biological and Molecular Characterization of Tomato brown rugose fruit virus (ToBRFV) on Tomato Plants in the State of Palestine
Mallor et al. Resistance to Melon necrotic spot virus in Cucumis melo L.‘Doublon’artificially inoculated by the fungus vector Olpidium bornovanus
JP5128169B2 (en) Plant plague control agent, control method and plant disease resistant plant
US20240065191A1 (en) Genetic basis for pythium resistance
Hore et al. Crop wild relatives of major cereals and millets: an overview
JPH11276178A (en) Control method of satellite RNA, acupuncture mosaic virus attenuated virus, and cucumber mosaic virus, and plants resistant to cucumber mosaic virus

Legal Events

Date Code Title Description
A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20030124

A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A821

Effective date: 20030124

A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20030407

A621 Written request for application examination

Free format text: JAPANESE INTERMEDIATE CODE: A621

Effective date: 20060119

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20081111

A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20090309

TRDD Decision of grant or rejection written
A01 Written decision to grant a patent or to grant a registration (utility model)

Free format text: JAPANESE INTERMEDIATE CODE: A01

Effective date: 20090519

A01 Written decision to grant a patent or to grant a registration (utility model)

Free format text: JAPANESE INTERMEDIATE CODE: A01

A61 First payment of annual fees (during grant procedure)

Free format text: JAPANESE INTERMEDIATE CODE: A61

Effective date: 20090611

R150 Certificate of patent or registration of utility model

Free format text: JAPANESE INTERMEDIATE CODE: R150

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20120710

Year of fee payment: 3

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20120710

Year of fee payment: 3

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20130710

Year of fee payment: 4

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20130710

Year of fee payment: 4

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20130710

Year of fee payment: 4

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20130710

Year of fee payment: 4

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

S533 Written request for registration of change of name

Free format text: JAPANESE INTERMEDIATE CODE: R313533

S533 Written request for registration of change of name

Free format text: JAPANESE INTERMEDIATE CODE: R313533

R350 Written notification of registration of transfer

Free format text: JAPANESE INTERMEDIATE CODE: R350

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

LAPS Cancellation because of no payment of annual fees