JP3759560B2

JP3759560B2 - Plant virus inoculation method

Info

Publication number: JP3759560B2
Application number: JP00484299A
Authority: JP
Inventors: 貞一佐藤; 正幸小湊; 春樹佐山; 英二石村
Original assignee: Nippon Del Monte Corp
Current assignee: Nippon Del Monte Corp
Priority date: 1999-01-12
Filing date: 1999-01-12
Publication date: 2006-03-29
Anticipated expiration: 2019-01-12
Also published as: JP2000201535A

Description

【０００１】
【発明の属する技術分野】
本発明は、碁盤格子状に小さな鉢（以下「セル」という）が数十〜数百連結した箱（以下「トレイ」という）の該セルに、小量（八〜九分目）の床土（培養土）を入れ、播種機により播種を行い、成形苗の生産を行うトレイ育苗法において、根生葉が重なり合って、地面に平たく放射状に拡がった苗（以下「ロゼット状態の苗」という）に、極めて簡単に植物ウイルスを１００％感染させることが可能な、植物ウイルスの接種方法に関する。
【０００２】
【従来の技術】
本発明者らは、植物体の表面に植物ウイルス液を噴霧した後、研磨剤を付着させたロ−ラーを圧接回転せしめ、該表面を被傷せしめると共に該植物体に植物ウイルスを接種する方法を開発し、特許出願した（特開平４−３３０００５）。
【０００３】
この方法によれば、綿棒、指、ガラス棒による接種法と比べ、接種に要する時間が著しく短縮され、またスプレーガンによる接種法に比べ、少ない植物ウイルス液（以下「接種液」ということがある）で高い感染率を得ることができる利点を有する。
【０００４】
しかしながら、この方法は、上記トレイ育苗法において、育苗途中の植物体の表面に植物ウイルス液を噴霧した後、該表面に対し、図４に示すように、把手１３、柄１２およびロ−ラ１１からなり、柄１２の一端部に把手１３を連結し、また柄１２の他端部をロ−ラ１１の軸受けとし、該軸受けにロ−ラ１１を回転自在に連結した植物ウイルス接種器の該ロ−ラ１１を、圧接回転しても、苗トレイ４の上端部とセル１４の床土表面との間隔が大きい状態で生育する植物体に対しては、ロ−ラ１１の外周面が当接できないため、被傷せしめることができず、植物ウイルスの感染率が低くなる欠点を有していた。
【０００５】
そして、根生葉が重なり合って、地面に平たく放射状に拡がる性質を有する苗（ロゼット状態の苗）は一般に育苗日数が比較的多くかかることから、灌水回数が多くなり、それにつれて苗トレイ４の各セル１４の床土１５表面がえぐりとられ、苗トレイ４の上端部と床土１５表面との間隔が大きくなり、育苗日数が多くなるにつれて円筒形ロ−ラ１１でセル穴１４内部の苗６ａの被傷作業が行いにくくなる。また、この方法は凹凸のある地面に植えた植物体に対して被傷せしめることが容易でない欠点を有する。
【０００６】
【発明が解決しようとする課題】
本発明は、凹凸のある地面に植えた植物体、特にトレイ育苗法により育苗されたロゼット状態の苗、に対して極めて簡単に被傷せしめ、植物ウイルスを１００％感染させることが可能な、植物ウイルスの接種方法を提供することを目的とする。
【０００７】
【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鋭意検討を重ねた結果、遂に本発明を完成した。
【０００８】
すなわち本発明は、（１）碁盤格子状に小さなセルが数十〜数百連結したトレイの該セルに培養土を入れ、播種を行い、植物体の成形苗の生産を行うトレイ育苗法において、該植物体の表面にブラシを当接し、該表面を被傷させ、該ブラシとしてブラシ幅が少なくとも２つのセルの上端開口部の合計幅を越えるものを使用することを特徴とする植物ウイルスの接種方法であり、（２）また本発明は、碁盤格子状に小さなセルが数十〜数百連結したトレイの該セルに培養土を入れ、播種を行い、植物体の成形苗の生産を行うトレイ育苗法において、該植物体の表面にブラシを当接し、該表面を被傷させ、その前または後において該表面に植物ウイルス液を接触させ、該ブラシとしてブラシ幅が少なくとも２つのセルの上端開口部の合計幅を越えるものを使用することを特徴とする植物ウイルスの接種方法であり、（３）また本発明は、碁盤格子状に小さなセルが数十〜数百連結したトレイの該セルに培養土を入れ、播種を行い、植物体の成形苗の生産を行うトレイ育苗法において、該植物体の表面に研磨剤の付着したブラシを当接し、該表面を被傷させ、その前または後において該表面に植物ウイルス液を接触させ、該ブラシとしてブラシ幅が少なくとも２つのセルの上端開口部の合計幅を越えるものを使用することを特徴とする植物ウイルスの接種方法である。
【０００９】
以下、本発明について詳細に説明する。
【００１０】
本発明の対象とする植物としては、任意の植物が挙げられるが、弱毒ウイルス接種によりウイルス防除効果が可能な作物、例えばトマト、ピ−マン、メロン、テンサイ、カボチャ、タバコ、ホウレン草、レタスなどの野菜類、リンドウ、ペチュニア、ベニバナ、キンギョソウ、百日草などの花卉類が好ましい。
【００１１】
また、本発明に用いるブラシとしては、把手あるいは柄のついた基面上に柔軟性があって弾力性に富む線条材を植え込んだものであれば任意のものが採用でき、例えば、塗装用ブラシ、歯磨き用ブラシ、洋服用ブラシ、たわし、あるいは柔軟性と弾力性に富む繊維状金属線を使用した剣山などが挙げられる。そして、本発明に用いるブラシは、そのブラシ幅が少なくとも２つのセルの上端開口部の合計幅を越えるものを使用する。これらはトレイ育苗法において苗床の大きさ、形、面積などに応じて、複数個連結使用することが好ましい。ブラシを複数連結して使用する場合は、同一の基面上に縦及び／又は横方向に必要個数固定し、各線状材の先端を揃えることが好ましい。
【００１２】
また、本発明のブラシに用いられる線状材は、外径０．０５〜０．２５ｍｍで、長さ１〜５ｃｍのものが好ましい。
【００１３】
植物体の表面に上記ブラシを当接すると、極めて簡単に該植物体の表面を被傷させることが可能となる。
当接は、植物体の表面に対して当てるだけでもよいが、当てた状態で擦る手段がより好ましい。当接は、１回でも可能であるが、２〜４回行うことが該植物体の表面を完全に被傷させる上で好ましい。
【００１４】
また本発明は、植物体の表面に研磨剤の付着したブラシを当接するときは、該植物体の表面に対し、より細かな傷を多量に形成できると共にウイルスの接種感染率を増大できるので好ましい。
【００１５】
研磨材としては、けいそう土（セライト）、ベントナイト、５００〜６００メッシュのカ−ボランダムなどが挙げられるが、特にセライトは軽く撹拌するだけで液に均一に分散し、また電動スプレ−を用いても噴口が詰まらないので好ましい。これらの粒径は５〜３５ミクロンが好ましく、また添加量は、液に１〜３重量％加える。
【００１６】
本発明を実施するには、植物体の表面にブラシを当接し、該表面を被傷させ、その前または後において該表面に植物ウイルス液を接触させる。
【００１７】
発明に使用される植物ウイルスとしては、植物特に作物に感染すると壊疽、モザイク、糸葉、矮化、黄変などの激しいウイルス症状を起こし収穫量が大幅に減る原因となるキュウリモザイクウイルス、その他トマト栽培におけるタバコモザイクウイルス、キユウリの栽培におけるキユウリ緑斑モザイクウイルス、柑橘類の栽培における柑橘トリステザウイルス（文献：亀谷満朗、１９８９「農業および園芸」６４巻、第１５９〜１６４頁、および栃原比呂志、１９８６「農業技術」４１巻（１１）、第１〜６頁参照）、ダイズモザイクウイルス、タバコネクロシスウイルス、タバコ茎えそウイルス、タバコ輪点ウイルス、タバコ矮化ウイルス、アルファルファモザイクウイルス、トマト黄化えそウイルス、ジャガイモＹウイルス、および上記各ウイルスの弱毒ウイルスなどが挙げられる。
【００１８】
上記植物ウイルスの弱毒ウイルスを、予め植物（苗）に接種感染しておくと、ウイルスの感染によって発生する上記植物ウイルス症状の被害を防ぐことができる利点を有する。
【００１９】
植物ウイルスを接種する時期は、目的とする植物の生育中の適宜な時期でよいが、弱毒ウイルスを利用してウイルス症状の被害を防ぐことが目的であれば、外界圃場に植える前の幼苗の時期が好ましく、とくに鉢上げ前のトレイでの育苗時期が集約的、効率的に接種できるので好ましい。
育苗トレイでの接種適期としては、（１）トマトなら子葉〜本葉２枚、（２）ピ−マンは子葉〜本葉２枚、（３）メロン、キュウリは子葉〜本葉１枚、（４）草丈、節間が短く苗がロゼット状態となる作物のペチュニア、トルコギキョウ、タバコでは４葉〜６葉、（５）リンドウは６葉〜８葉の頃が挙げられる。
【００２０】
植物ウイルス液は以下の方法により調製することが好ましい。
植物ウイルスの感染葉の搾汁液をガ−ゼで濾過し得られた液を滅菌水または適当な緩衝液（例えば、中性付近の０．１Ｍリン酸緩衝液）により、例えば感染葉の重量に対して５〜５０倍希釈して調製するか、或いは常法により得られた純化ウイルス液（例えばキュウリモザイクウイルス液）を滅菌水または適当な緩衝液により適宜の濃度、例えばウイルス濃度２５〜５００μｇ／ｍｌに希釈して調製する。
【００２１】
上記植物ウイルス液に、必要によりさらに着色料を添加して懸濁する。
着色料を添加すると、植物体の表面に植物ウイルス液を接触させた後、該ウイルス液が付着したか、否かの識別が容易となる利点を有する。
着色料としては、合成着色料、天然着色料など任意の着色料が挙げられるが、食用植物の場合は、食品用着色料を用いることが安全性の面で好ましく、またその色は青色もしくは赤色が植物体の表面の色（緑色）に対して付着の確認が容易となるので好ましい。
着色料の濃度（希釈濃度）は例えば合成着色料を使用するなら４００〜８００倍に薄める。
【００２２】
植物ウイルス液を接触させる手段としては、噴霧、浸漬、塗布、散水など任意の方法が挙げられるが、小量均一噴霧で接種効果が高い噴霧手段がいちばん好ましい。
噴霧手段としては、簡易ハンドスプレ−、動力噴霧機（例えば小型電動塗装噴霧機）、スプレ−ガンなどが挙げられる。
但し、簡易ハンドスプレ−を用いる場合、研磨材により噴霧口が詰まる場合があるので注意を要する。
小型電動塗装噴霧機は吐出量が５０〜５００ｍｌ／分程度であることが少量均一散布を行なううえで好ましい。
接種液の散布量は、トレイ育苗の場合（縦３０、横６０ｃｍのトレイの場合）は５〜１５ｍｌ、地床育苗の場合は、３．３ｍ²当り１００〜３００ｍｌが好ましい。
【００２３】
以下添付された図に従って、本発明をより具体的に説明する。
図１は、植物体（ロゼット状態の苗６ａ）の表面に植物ウイルス液５を小型電動塗装噴霧器２により付着させる状態を示し、また図２は該植物体の表面にブラシ８を当接し、該植物体の表面を被傷させる状態を示す。ブラシ８の右側の苗は被傷した状態を示している。
【００２４】
図１において、植物ウイルス液５を入れた小型電動塗装噴霧機２（例えば日本ワグナ−・スプレ−テック社製ミニペインタ−など）を用いて苗トレイ４のロゼット状態の苗６ａの表面に、予め着色料および研磨材を添加使用した植物ウイルス液５を噴霧し着色具合を見ながら満遍なく付着させる。
【００２５】
次に図２に示すように、苗トレイ４の苗６ａを、研磨材が付着したブラシ８を縦方向と横方向に数回擦る。
【００２６】
図３は、本発明の植物ウイルスの接種方法の概略説明図で、苗トレイ４のロゼット状態の苗６ａをブラシ８の毛１０で、各セル穴１４の内部の茎葉の隅々にいたるまで擦って接種していることを示す。
ブラシ８のそれぞれの毛１０がもつ弾力性、柔軟性、自由自在な動きにより、各セル穴１４から苗トレイ４の表面にはみでない苗６ａの地際部の茎葉に至るまで、苗全体の茎葉に噴霧付着した植物ウイルス液を満遍なく擦り付けて接種することができる。
また、苗トレイ４表面と床土１５表面との間隔（凹凸差）が大きい場合においても、ブラシ８の毛１０でセル穴１４内部の苗６ａを擦ることは容易である。図３において、９はブラシの把手（または柄）であり離れている苗に接種するときに手で掴んで用いる。
【００２７】
【実施例】
以下、実施例を示して本発明についてさらに詳細に説明する。
【００２８】
実施例１
（１）弱毒ウイルス接種対象となる作物の苗の育成例
先ず、トマト、ピ−マン、ペチュニア、タバコ用培養土としてホ−ネン社製の「セルトップミックス」とサカタのタネ社製の「ス−パ−ミックスＡ」を１：１に混合して作成し、リンドウ用培養土として細粒鹿沼土、細粒赤玉土、ピ−トモスを１：１：１に混合して作成し、それぞれの作物の苗に応じたヤンマ−農機社製の二種類の苗トレイ「ヤンマ−野菜トレイ２５−２００（セルの大きさ２５ｍｍ角×深さ４５ｍｍ）、ヤンマ−野菜トレイ３０−１２８（セルの大きさ３０ｍｍ角×深さ４５ｍｍ）」に充填し、それぞれの作物の種を播種した。
発芽させたあと、作物の苗に応じた適当な温度と採光条件のもと液肥を注入したり灌水を行う通常の育苗管理を行い、それぞれの苗を弱毒ウイルスを接種する生育適期まで育てた。接種適期は発芽が揃って生育して根鉢になって葉が数枚になるころであり、トレイ苗として出荷する５〜１０日前が適当である。この接種時期の苗について生育を調べて苗が本発明の接種に適合するかを調べた結果を示す。
【００２９】

【００３０】
表１の結果から、トマトとピ−マンの苗はセル穴内部から苗トレイ表面上に突き出て子葉と本葉は苗トレイ表面約３ｃｍ以上にあり、従来法の植物ウイルス接種方法（特開平４−３３０００５）の円筒形ロ−ラによる擦り付け接種を行う場合、苗トレイ表面上で充分に苗の茎葉を擦ることができる。
しかるに、苗がロゼット状態となるペチュニア、タバコ、リンドウはトレイ表面上の草丈が０．４〜１．２ｃｍでかなり低くく草丈の約半分はセル穴内部にあり、節間は極端に短く、葉数は折り重なって枚数が多く、従来の接種方法によるロ−ラではセル穴内部の茎葉に至るまで充分な擦り付けを行って接種することができない。
【００３１】
（２）ロゼット状態の苗における弱毒ウイルス接種結果の比較
キュウリモザイクウイルスの弱毒ウイルス（日本デルモンテ社製、「ＮＤＭ−１」）の純化液をリン酸緩衝液に濃度１００μｇ／ｍｌで希釈し、着色料（合成着色料、ブリリアントブル−ＦＣＦ）を５００倍で加えて接種液を作成した。
１２８穴トレイで４０日育苗した６〜７枚期のペチュニアに、本発明では接種液を簡易ハンドスプレ−で苗に噴霧し、茎葉が青色に着色する具合を見ながら少量均一噴霧を行い、その噴霧直後、けいそう土（セライト）を毛先に付着させたブラシ（富士ブラシ製造販売社製、ペンキ用刷毛、毛の長さ３．６ｃｍ、ブラシ幅７ｃｍ、ブラシの毛の外径約０．１ｍｍ、毛の素材は山羊）を四つ並列に並べて、図３のごとく作成したブラシ（ブラシ幅３０ｃｍ）で縦方向と横方法にセル穴内部の茎葉に至るまで苗全体の茎葉を満遍なく計３回擦った。
【００３２】
【比較例】
比較例１
上記実施例１と同様に接種液を噴霧した直後、けいそう土（セライト）が付着したロ−ラ（ニトムズ社製コロコロ、ロ−ラの長さ１６ｃｍ）で苗トレイの表面上に突き出ている苗の茎葉を計３回擦った。
【００３３】
【対照例】
対照例１
上記実施例１で得られる接種液を噴霧して、擦り操作は一切行わなかった。
【００３４】
対照例２
上記実施例１で得られる接種液にけいそう土（セライト）を２％混合し、その液に綿棒（ジョンソン・エンド・ジョンソン社製）を浸し、苗の葉の裏にあて板を差し当てて支えとし、それぞれ株の葉を６枚ずつ綿棒で擦り、綿棒は８株の接種を終える毎に交換した。
以上の接種操作を行った８日目に、それぞれの１２８穴トレイから調査株をランダムに取り、弱毒ウイルスを検出した。
【００３５】
上記実施例１、比較例、対照例１及び対照例２の結果をまとめて表２に示す。
【００３６】

【００３７】
表２の結果から、先ず比較例のロ−ラによる接種は、育苗トレイ表面からはみでた苗の葉がセル穴の角の先端とロ−ラに挟まり、ロ−ラの回転により葉が少し裂ける程度の擦り傷が３１％（４株／１３株中）発生した。
また、苗トレイのセル穴内部にある茎葉にはロ−ラは触れず、擦り操作が完全でなく感染率は８５％に止まった。
対照例１の接種液の噴霧のみによる接種では、苗を擦る操作が無いので全く感染させることができなかった。
対照例２では接種液を株当り六枚ずつ満遍なく綿棒で擦るので、接種液を多く必要とし、また接種時間を大幅に必要とした。
これに対して本発明は、苗に付着した少量の噴霧液を１２８穴トレイの縦幅（約３０ｃｍ）に応じたブラシで単純に数回擦るだけなので接種時間が少なく、育苗トレイの凹凸に対してブラシの柔軟性のある毛先の自由自在な動きによりセル穴内部にある茎葉に至るまで十分に擦ることができ、苗の擦り傷は極めて軽微で、接種損傷株発生数は無く、弱毒ウイルスを１００％感染させることができた。
従って、本発明は、少量の接種液量で、簡単な器具と簡便な方法により、迅速に、しかも集約的、効率的に擦り付け接種ができ、トレイ苗を満遍なく擦ることができ完全に植物ウイルスを感染させることができ、さらにブラシの柔軟性のある毛先の動きにより苗に損傷を与えずに接種できる。
【００３８】
（３）弱毒ウイルス接種株の葉の部位別ウイルス増殖比較
弱毒ウイルス接種後８日目の本発明と比較例のペチュニアを、上位葉、中位葉、下位葉にそれぞれ二枚ずつふり分け、その二枚ずつの葉をそれぞれ小袋に入れて揉み潰し、燐酸緩衝液を注入してセライトを混ぜ、その液に綿棒を浸したあと直ちにササゲ（品種：黒種３尺）の初生葉に一枚ずつに擦り付けて接種し、その四日後、それぞれの初生葉に斑点が発生した数を数え、本発明と比較例との弱毒ウイルス接種株の葉の部位別ウイルス増殖を、ペチュニアの株当りササゲ斑点発生数で比較した。
表３に本発明と比較例の結果を示す。
【００３９】

【００４０】
表３より、ロ−ラを用いて接種した比較例のササゲの斑点発生数は、下位葉が少なく、中位葉から上位葉になるにつれて斑点の発生が多くなった。これはロ−ラでは苗トレイの凹部分のセル穴内部の茎葉に至まで擦ることができず、苗の下部分の葉は弱毒ウイルスが増殖しずらかったことを示す。
これに対して、ブラシを用いて接種した本発明では、苗トレイの凹凸に対してブラシの柔軟性のある毛先の自由自在な動きによりセル穴内部にある茎葉を十分に擦り、下位葉、中位葉、上位葉のいずれからもほぼ同等の斑点が発生し、また、ペチュニア株当りササゲ斑点数の合計は比較例の倍となり、弱毒ウイルスがペチュニア体内に早く浸透して増殖したことを示す。
【００４１】
【本発明の効果】
本発明は、少量の植物ウイルス液と、簡易噴霧具やブラシなどの簡単な器具を用いて、苗に均一噴霧したあと数回苗を擦るだけの簡便な方法により、迅速に、効率的かつ集約的に大量の苗の接種作業を短時間に行える効果を有する。
また本発明は、トマト、ピ−マンのような育苗トレイ表面に葉が完全に突き出る作物のほか、ペチュニア、トルコキキョウ、タバコ、リンドウなどの節間が極度に短く葉が重なってロゼット状態になり育苗トレイの表面に葉が完全に出ていない苗においても、ブラシの柔軟性のある細い毛先の自由自在な動きにより、育苗トレイ表面の苗の先端の葉から、セル穴内部の地際部の茎葉に至るまで十分に擦って接種することができる効果を有する。
また苗を傷付けることなく接種でき、そのうえ植物ウイルスを早く植物体内に浸透させ、ほぼ完璧に感染させることできる効果も合わせて奏するものである。また本発明は平坦な地面に植えた植物体に対して植物ウイルスを接種する場合ばかりでなく、凹凸のある地面に植えた植物体に対しての接種ムラなく、植物ウイルスを接種し感染させることができる。
【図面の簡単な説明】
【図１】本発明の植物ウイルス液の噴霧状況を、略図的に表した概念図である。
【図２】本発明のブラシによる接種を、略図的に表した概念図である。
【図３】植物ウイルス液が付着した苗トレイのロゼット状苗を、本発明のブラシで擦って接種している状況の断面図である。
【図４】植物ウイルス液が付着した苗トレイのロゼット状苗を、従来法の接種方法（特開平４−３３０００５）のロ−ラで擦って接種している状況の断面図である。
【符号の説明】
２…電動噴霧器
４…苗トレイ
５…植物ウイルス液
６ａ…ロゼット状態の苗
８…ブラシ
９…ブラシの把手
１０…ブラシの毛
１１…円筒形ロ−ラ
１２…ロ−ラの柄
１３…ロ−ラの把手
１４…セル
１５…床土[0001]
BACKGROUND OF THE INVENTION
The present invention provides a small amount (8th to 9th minutes) of floor soil in a cell (hereinafter referred to as “tray”) in which several tens to hundreds of small bowls (hereinafter referred to as “cells”) are connected in a grid pattern. In the tray breeding method in which (cultured soil) is put in, seeded with a seeder, and molded seedlings are produced, seedlings (hereinafter referred to as “rosette state seedlings”) that have root leaves overlapped and spread radially on the ground The present invention relates to a method for inoculating a plant virus, which can very easily infect 100% of a plant virus.
[0002]
[Prior art]
The present inventors sprayed a plant virus solution on the surface of a plant body, and then pressed and rotated a roller to which an abrasive was attached to injure the surface and inoculate the plant body with a plant virus. Was developed and a patent application was filed (JP-A-4-330005).
[0003]
According to this method, the time required for inoculation is remarkably shortened compared to the method of inoculation with a cotton swab, finger, and glass rod, and there are also less plant virus solutions (hereinafter referred to as “inoculation solutions”) than inoculation methods with a spray gun. ) Has the advantage that a high infection rate can be obtained.
[0004]
However, in this method, after spraying the plant virus solution on the surface of the plant body in the middle of the seedling raising method, the handle 13, the handle 12 and the roller 11 are applied to the surface as shown in FIG. The handle 13 is connected to one end of the handle 12, the other end of the handle 12 is a bearing of the roller 11, and the roller 11 is rotatably connected to the bearing. Even if the roller 11 is pressed and rotated, the outer peripheral surface of the roller 11 is applied to a plant that grows with a large distance between the upper end of the seedling tray 4 and the floor soil surface of the cell 14. Since it could not be contacted, it could not be injured and had a drawback that the infection rate of plant viruses was low.
[0005]
The seedlings that have the property that the root leaves overlap and spread radially on the ground (seed in the rosette state) generally take a relatively large number of seedling days, so the number of watering times increases, and each cell in the seedling tray 4 accordingly. 14, the surface of the floor soil 15 is removed, the distance between the upper end of the seedling tray 4 and the surface of the floor soil 15 is increased, and the seedling 6a inside the cell hole 14 is formed by the cylindrical roller 11 as the number of seedling days increases. It will be difficult to perform the work. In addition, this method has a drawback that it is not easy to damage a plant planted on uneven ground.
[0006]
[Problems to be solved by the invention]
The present invention is a plant capable of very easily injuring a plant planted on uneven ground, in particular, a rosette seedling grown by a tray raising method, and infecting a plant virus 100%. The purpose is to provide a method for inoculating viruses.
[0007]
[Means for Solving the Problems]
As a result of intensive studies in order to solve the above problems, the present inventors have finally completed the present invention.
[0008]
That is, the present invention is (1) in a tray raising method for putting cultured soil into a cell of a tray in which several tens to several hundreds of small cells are connected in a grid pattern, sowing, and producing a molded seedling of a plant body. a brush on the surface of the plant contact, the surface is to be wound, brush width as the brush is a plant viruses characterized that you use those exceeding the total width of the top opening of the at least two cells It is an inoculation method, and (2) the present invention is to produce culture seedlings by putting culture soil into the cells of a tray in which several tens to several hundreds of small cells are connected in a grid pattern. in tray nursery, brush the contact with the surface of the plant, the surface is to be wound, by contacting the plant virus solution on the surface at that before or after, the upper end of the brush width as the brush are at least two cells Exceeds total width of opening A vaccination method of the plant viruses characterized that you use an object, (3) The present invention, the culture soil was placed in the cell tray a small cell has several tens to several hundreds coupled in a checkerboard lattice pattern, seeding was carried out, in the tray seedling method in which the production of molded seedlings plants, a brush attached abrasive on the surface of the plant contact, the surface is to be wound, plant surface in before or after contacting the virus solution, the brush width as the brush is vaccination method of the plant viruses characterized that you use those exceeding the total width of the top opening of the at least two cells.
[0009]
Hereinafter, the present invention will be described in detail.
[0010]
Examples of the plant targeted by the present invention include any plant, but crops capable of controlling the virus by inoculation with attenuated virus, such as tomato, pepper, melon, sugar beet, pumpkin, tobacco, spinach, lettuce, etc. Flowers, such as vegetables, gentian, petunia, safflower, snapdragon, zinnia are preferred.
[0011]
In addition, as the brush used in the present invention, any brush can be used as long as it has a flexible and elastic line material implanted on a handle or handle base. Examples include brushes, toothpaste brushes, clothes brushes, scrubbers, and swordsmen using fibrous metal wires that are highly flexible and elastic. The brush used in the present invention has a brush width exceeding the total width of the upper end openings of at least two cells. It is preferable to use a plurality of these in a tray raising method according to the size, shape, area, etc. of the nursery bed. When a plurality of brushes are connected and used, it is preferable to fix the required number in the vertical and / or horizontal direction on the same base surface and align the tips of the respective linear members.
[0012]
Moreover, the linear material used for the brush of the present invention preferably has an outer diameter of 0.05 to 0.25 mm and a length of 1 to 5 cm.
[0013]
When the brush is brought into contact with the surface of the plant body, the surface of the plant body can be very easily damaged.
The contact may be applied only to the surface of the plant body, but means for rubbing in the applied state is more preferable. The contact can be performed once, but it is preferable to perform the contact 2 to 4 times in order to completely damage the surface of the plant body.
[0014]
In addition, the present invention is preferable when a brush having an abrasive attached thereto is brought into contact with the surface of a plant because a large amount of finer scratches can be formed on the surface of the plant and the inoculation rate of virus can be increased. .
[0015]
Examples of abrasives include diatomaceous earth (celite), bentonite, 500-600 mesh carborundum, etc. Celite, in particular, can be evenly dispersed in the liquid just by lightly stirring, and using an electric spray. Is preferable because the nozzle hole is not clogged. These particle sizes are preferably 5 to 35 microns, and the addition amount is 1 to 3% by weight to the liquid.
[0016]
In order to carry out the present invention, a brush is brought into contact with the surface of a plant body, the surface is damaged, and a plant virus solution is brought into contact with the surface before or after that.
[0017]
Plant viruses used in the invention include cucumber mosaic virus and other tomatoes that cause severe viral symptoms such as gangrene, mosaic, thread, hatching and yellowing when infected with plants, especially crops. Tobacco Mosaic Virus in Cultivation, Cucumber Green Spot Mosaic Virus in Cultivation of Citrus, Citrus Tristeza Virus in Cultivation of Citrus (Reference: Mitsuru Kameya, 1989 “Agriculture and Horticulture”, Vol. 64, pp. 159-164, and Hiroshi Tochihara, 1986 “Agricultural Technology”, Vol. 41 (11), pp. 1-6), soybean mosaic virus, tobacco necrosis virus, tobacco stem virus, tobacco ringspot virus, tobacco dwarf virus, alfalfa mosaic virus, tomato yellowing Esovirus, potato Y virus, and above Such as attenuated virus of the virus, and the like.
[0018]
When a plant (seedling) is inoculated and infected with the attenuated virus of the plant virus in advance, it has an advantage that damage of the plant virus symptoms caused by the virus infection can be prevented.
[0019]
The plant virus can be inoculated at an appropriate time during the growth of the target plant, but if the aim is to prevent the damage of viral symptoms using attenuated viruses, the seedlings before planting in the external field can be used. The season is preferred, and the seedling raising time in the tray before potting is particularly preferred because it can be inoculated intensively and efficiently.
Appropriate time for inoculation in the seedling tray includes (1) cotyledons to two true leaves for tomatoes, (2) Peman to cotyledons to two true leaves, (3) melon and cucumber to cotyledons to one true leaf, ( 4) Plant height, short internodes, and crops where seedlings are in a rosette state. Petunia, Eustoma and tobacco have 4 to 6 leaves, and (5) Gentian has 6 to 8 leaves.
[0020]
The plant virus solution is preferably prepared by the following method.
The juice obtained by filtering the squeezed solution of plant virus infected leaves with gauze is sterilized with sterilized water or an appropriate buffer solution (for example, 0.1M phosphate buffer solution near neutrality), for example, to the weight of the infected leaves. The purified virus solution (for example, cucumber mosaic virus solution) obtained by diluting 5 to 50 times with an appropriate concentration, for example, a virus concentration of 25 to 500 μg / Prepare by diluting to ml.
[0021]
If necessary, a coloring agent is added to the plant virus solution and suspended.
Addition of a coloring agent has an advantage that it becomes easy to identify whether or not the virus solution has adhered after contacting the plant virus solution to the surface of the plant body.
Examples of the colorant include arbitrary colorants such as a synthetic colorant and a natural colorant. In the case of an edible plant, it is preferable to use a food colorant in terms of safety, and the color is blue or red. Is preferable because it is easy to confirm adhesion to the surface color (green) of the plant body.
If the colorant concentration (dilution concentration) is, for example, a synthetic colorant, it is diluted 400 to 800 times.
[0022]
The means for contacting the plant virus solution may be any method such as spraying, dipping, coating, watering, etc., but spraying means with a small amount of uniform spray and high inoculation effect is the most preferable.
Examples of the spraying means include a simple hand spray, a power sprayer (for example, a small electric paint sprayer), and a spray gun.
However, when a simple hand spray is used, care must be taken because the spraying port may be clogged with the abrasive.
The small electric paint sprayer preferably has a discharge amount of about 50 to 500 ml / min in order to uniformly spray a small amount.
The amount of the inoculum sprayed is preferably 5 to 15 ml in the case of tray seedlings (in the case of 30 cm in length and 60 cm in width), and 100 to 300 ml per 3.3 m ^{2 in} the case of ground raising seedlings.
[0023]
Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings.
FIG. 1 shows a state in which a plant virus solution 5 is attached to the surface of a plant body (roselet seedling 6a) by a small electric paint sprayer 2, and FIG. The state which damages the surface of a plant body is shown. The seedling on the right side of the brush 8 shows a damaged state.
[0024]
In FIG. 1, the surface of the seedling 6a in the rosette state of the seedling tray 4 is preliminarily applied to the seedling tray 4 by using a small electric paint sprayer 2 (for example, a mini painter manufactured by Nippon Wagner Spray Tech Co., Ltd.) containing the plant virus solution 5. The plant virus solution 5 added with coloring agents and abrasives is sprayed and adhered uniformly while observing the coloration.
[0025]
Next, as shown in FIG. 2, the seedlings 6a of the seedling tray 4 are rubbed several times in the vertical and horizontal directions with the brush 8 to which the abrasive is attached.
[0026]
FIG. 3 is a schematic explanatory view of the plant virus inoculation method of the present invention. The seedling 6a in the rosette state of the seedling tray 4 is rubbed with the hair 10 of the brush 8 until it reaches every corner of the foliage inside each cell hole 14. Show that you are inoculated.
Due to the elasticity, flexibility, and free movement of each hair 10 of the brush 8, the entire seedlings from the cell holes 14 to the foliage of the ground part of the seedling 6 a that is not on the surface of the seedling tray 4 can be obtained. It can be inoculated by rubbing the plant virus solution sprayed on the foliage evenly.
Further, even when the distance between the seedling tray 4 surface and the floor soil 15 surface (unevenness difference) is large, it is easy to rub the seedlings 6 a inside the cell holes 14 with the bristles 10 of the brush 8. In FIG. 3, 9 is a handle (or handle) of a brush, and is used by grasping with a hand when inoculating a remote seedling.
[0027]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples.
[0028]
Example 1
(1) Example of growing seedlings of crops to be inoculated with attenuated virus First, “Cell Top Mix” manufactured by Hohnen as a culture soil for tomatoes, peppers, petunia and tobacco, -Purmix A "was prepared by mixing 1: 1, and fine kanuma soil, fine red bean soil, and peat moss were mixed 1: 1: 1 as gentian culture soil. Two kinds of seedling trays “Yanma-vegetable tray 25-200 (cell size 25 mm square × depth 45 mm) made by Yanma Agricultural Machinery Co., Ltd. according to crop seedlings”, Yanma-vegetable tray 30-128 (cell size 30 mm square × depth 45 mm) ”and seeded each crop seed.
After germination, normal seedling management was performed by injecting liquid fertilizer and irrigating under appropriate temperature and lighting conditions according to the seedlings of the crops, and each seedling was grown to a suitable growth period for inoculating the attenuated virus. The appropriate time for inoculation is when germination is complete and grows to form root pots and several leaves, and 5-10 days before shipping as tray seedlings is appropriate. The result of having investigated growth about the seedling of this inoculation time, and having investigated whether a seedling matched the inoculation of this invention is shown.
[0029]

[0030]
From the results in Table 1, the tomato and pepper seedlings protrude from the inside of the cell hole onto the seedling tray surface, and the cotyledons and true leaves are about 3 cm or more in the seedling tray surface. -330005), the seedling foliage can be sufficiently rubbed on the seedling tray surface.
However, petunia, tobacco, and gentian, where the seedlings are in a rosette state, have a plant height of 0.4 to 1.2 cm on the tray surface and are very low, about half of the plant height is inside the cell hole, the internodes are extremely short, and the leaves The number of folds overlaps and the number of sheets is large, and the conventional inoculation method cannot be inoculated with sufficient rubbing until the foliage inside the cell hole.
[0031]
(2) Comparison of attenuated virus inoculation results in seedlings in rosette state Purified solution of cucumber mosaic virus attenuated virus (“DDM-1”, manufactured by Nippon Del Monte) was diluted in phosphate buffer at a concentration of 100 μg / ml and colored An inoculum was prepared by adding a material (synthetic colorant, Brilliant Blue-FCF) at 500 times.
In the present invention, spray seedlings with 6 to 7 stage petunias grown in a 128-hole tray for 40 days with a simple hand spray and spray a small amount uniformly while watching how the leaves and leaves turn blue. Immediately after spraying, a brush with diatomaceous earth (celite) attached to the hair tip (manufactured by Fuji Brush Manufacturing and Sales Co., Ltd., paint brush, hair length 3.6 cm, brush width 7 cm, brush hair outer diameter of about 0.1 mm). 4 mm of hair material (goat) is arranged in parallel, and the total number of stems and leaves of the entire seedling is reached with the brush (brush width 30 cm) as shown in FIG. I rubbed.
[0032]
[Comparative example]
Comparative Example 1
Immediately after spraying the inoculum in the same manner as in Example 1 above, a roller with diatomaceous earth (Celite) adhered (Nittoms CoroCoro, roller length 16 cm) protrudes onto the surface of the seedling tray. The seedling foliage was rubbed a total of 3 times.
[0033]
[Control example]
Control Example 1
The inoculum obtained in Example 1 was sprayed and no rubbing operation was performed.
[0034]
Control Example 2
2% of diatomaceous earth (Celite) is mixed in the inoculum obtained in Example 1 above, a cotton swab (manufactured by Johnson & Johnson) is immersed in the liquid, and a pad is placed against the back of the seedling leaves. As a support, six leaves of each strain were rubbed with a cotton swab, and the cotton swabs were replaced every time eight strains were inoculated.
On the 8th day after the above inoculation operation, the investigation strain was randomly picked from each 128-well tray to detect the attenuated virus.
[0035]
The results of Example 1, Comparative Example, Control Example 1 and Control Example 2 are summarized in Table 2.
[0036]

[0037]
From the results in Table 2, first, inoculation with the roller of the comparative example, the leaves of the seedling protruding from the seedling tray surface are sandwiched between the tip of the corner of the cell hole and the roller, and the leaf is slightly broken by the rotation of the roller. A moderate degree of scratching occurred in 31% (out of 4 strains / 13 strains).
Further, the rollers were not touched on the foliage inside the cell hole of the seedling tray, the rubbing operation was not complete, and the infection rate was only 85%.
In the inoculation only by spraying the inoculum of Control Example 1, since there was no operation for rubbing the seedlings, no infection was possible.
In Control Example 2, six inoculums were uniformly rubbed with cotton swabs per strain, so that a large amount of inoculum was required and inoculation time was significantly required.
In contrast, in the present invention, a small amount of sprayed liquid adhering to the seedling is simply rubbed several times with a brush corresponding to the vertical width (about 30 cm) of the 128-hole tray, so the inoculation time is short, and the unevenness of the seedling tray is reduced. With the flexible movement of the brush's flexible bristles, it can be rubbed to the foliage inside the cell hole, the seedlings are extremely scratched, the number of inoculated strains is not generated, and the attenuated virus is removed. 100% infection was possible.
Therefore, the present invention can be rubbed quickly, intensively and efficiently with a simple instrument and a simple method with a small amount of inoculum, and the tray seedlings can be rubbed evenly, and the plant virus can be completely removed. It can be infected and can be inoculated without damaging the seedlings due to the flexible movement of the brush tip.
[0038]
(3) Comparison of the virus growth by leaves of the attenuated virus inoculated strain The present invention and the petunia of the comparative example on the 8th day after the attenuated virus inoculation were sprinkled into the upper leaf, the middle leaf and the lower leaf, respectively, Put each leaf in a sachet, crush it, inject phosphate buffer, mix with celite, dip a cotton swab into the solution, and immediately put it on the primary leaves of cowpea (variety: black 3). Four days later, count the number of spots on each primary leaf, and count the number of leaves of the attenuated virus-inoculated strains according to the present invention and comparative examples to determine the growth of viruses per petunia. Compared by number.
Table 3 shows the results of the present invention and comparative examples.
[0039]

[0040]
From Table 3, as for the number of spots of the cowpea of the comparative example inoculated with a roller, the number of spots was increased as the number of the lower leaves decreased and the middle leaves changed to the upper leaves. This indicates that the roller could not rub the stems and leaves inside the cell hole corresponding to the concave portion of the seedling tray, and that the leaves of the lower part of the seedling were difficult to propagate the attenuated virus.
On the other hand, in the present invention inoculated with a brush, the leaves in the cell hole are sufficiently rubbed by the free movement of the flexible hair tip of the brush against the irregularities of the seedling tray, the lower leaves, Approximately the same spots occur from both the middle and upper leaves, and the total number of cowpea spots per petunia strain is double that of the comparative example, indicating that the attenuated virus penetrated and grew rapidly in the petunia. .
[0041]
[Effect of the present invention]
The present invention uses a small amount of plant virus solution and a simple device such as a simple spraying tool or a brush to spray seedlings uniformly, and then swiftly, efficiently and aggregate by a simple method of rubbing the seedlings several times. In effect, a large amount of seedling can be inoculated in a short time.
In addition to crops with leaves protruding completely on the surface of the seedling tray such as tomatoes and peppers, the present invention is extremely short between pet nodes, such as petunia, turkey, tobacco, and gentian. Even in the seedlings where the leaves are not completely exposed on the surface of the seedling tray, the free movement of the brush's flexible thin hair tips allows the edge part of the seedlings on the surface of the seedling tray to move from the leaves of the seedling to the ground part inside the cell hole. It has the effect that it can be inoculated by thoroughly rubbing until it reaches the foliage.
In addition, it can be inoculated without damaging the seedlings, and also has the effect of allowing the plant virus to penetrate the plant body quickly and infect it almost completely. In addition, the present invention not only inoculates a plant virus planted on a flat ground but also inoculates and infects a plant virus without inoculation unevenness on a plant planted on uneven ground. Can do.
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a conceptual diagram schematically showing a spraying state of a plant virus solution of the present invention.
FIG. 2 is a conceptual diagram schematically showing inoculation with the brush of the present invention.
FIG. 3 is a cross-sectional view of a situation where a rosette-like seedling in a seedling tray to which a plant virus solution is attached is rubbed with the brush of the present invention and inoculated.
FIG. 4 is a cross-sectional view of a situation where a rosette-like seedling in a seedling tray to which a plant virus solution is attached is inoculated by rubbing with a roller of a conventional inoculation method (Japanese Patent Laid-Open No. 4-330005).
[Explanation of symbols]
2 ... Electric sprayer 4 ... Seedling tray 5 ... Plant virus solution 6a ... Rosette seedling 8 ... Brush 9 ... Brush handle 10 ... Brush hair 11 ... Cylindrical roller 12 ... Roller handle 13 ... Roh La handle 14 ... cell 15 ... floor soil

Claims

Checkerboard lattice-like small cell put culture soil in several tens to several hundreds linked the cell tray, perform seeding, the tray seedling method in which the production of molded seedlings plants, a brush on the surface of the plant abut, said surface is the wound, inoculation method of the plant virus brush width as the brush, characterized in that you use those exceeding the total width of the top opening of the at least two cells.

Checkerboard lattice-like small cell put culture soil in several tens to several hundreds linked the cell tray, perform seeding, the tray seedling method in which the production of molded seedlings plants, a brush on the surface of the plant abut, said surface is the wound, that you use the ones that before or after contacting the plant virus solution on the surface, the brush width as the brush exceeding the total width of the top opening of the at least two cells A plant virus inoculation method characterized by the above.

Checkerboard lattice-like small cell put culture soil in several tens to several hundreds linked the cell tray, perform seeding, the tray seedling method in which the production of molded seedlings plants, abrasive surface of the plant The surface of the brush is contacted, the surface is damaged, and the plant virus liquid is contacted with the surface before or after the brush, and the brush width exceeds the total width of the upper end openings of at least two cells. inoculation method of plant viruses characterized by that you use.

The inoculation method of the plant virus in any one of Claims 1-3 using the brush which planted many linear materials whose outer diameter is 0.05-0.25 mm on a base surface.

The plant virus inoculation method according to any one of claims 1 to 3, wherein the plant body is a vegetable seedling, a flower seedling seedling or a fruit tree seedling in a rosette state.

The inoculation method of the plant virus in any one of Claims 1-5 using the plant virus liquid which added the coloring agent as a plant virus liquid.