JP2851506B2 - Method for producing entomopathogenic fungi and medium for growing entomopathogenic fungi - Google Patents
Method for producing entomopathogenic fungi and medium for growing entomopathogenic fungiInfo
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- JP2851506B2 JP2851506B2 JP5054450A JP5445093A JP2851506B2 JP 2851506 B2 JP2851506 B2 JP 2851506B2 JP 5054450 A JP5054450 A JP 5054450A JP 5445093 A JP5445093 A JP 5445093A JP 2851506 B2 JP2851506 B2 JP 2851506B2
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- medium
- entomopathogenic
- conidia
- mannitol
- yeast extract
- Prior art date
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Description
【０００１】[0001]
【産業上の利用分野】本発明は、昆虫病原糸状菌の製造
方法及び昆虫病原糸状菌用培地に関する。The present invention relates to a method for producing entomopathogenic fungi and a medium for entomopathogenic fungi.
【０００２】[0002]
【従来の技術】ボーベリア（Beauveria)、メタリジュー
ム(Metarhizium) 、バーティシリューム( Verticilliu
m) 、ペーシロミセス（Pae-cilomyces)、エントモフト
ラ(Entomophthora) 等の昆虫病原糸状菌類は、昆虫に寄
生することにより昆虫を死に到らしめる糸状菌で、鱗翅
目昆虫・鞘翅目昆虫・半翅目昆虫・アザミウマ目昆虫等
に特に病原性が高いことで知られている。2. Description of the Related Art Beauveria, Metarhizium, Verticilliu
m), insect pathogenic fungi such as Pae-cilomyces and Entomophthora are fungal fungi that parasitize insects and cause the death of insects. -It is known to be particularly pathogenic to thrips and the like.
【０００３】又、害虫の殺虫剤に対する感受性低下の問
題等から、生物農薬として、総合害虫管理技術の１資源
として重要視され続けている。[0003] In addition, due to the problem of reduced sensitivity of pests to pesticides, etc., they have been regarded as important as resources of integrated pest management technology as biological pesticides.
【０００４】更に、昆虫病原糸状菌類は、昆虫以外には
感染せず、人畜、魚介、鳥類に無害であることから、無
公害の殺虫剤として利用が探求されてきた。Furthermore, entomopathogenic filamentous fungi do not infect insects other than insects and are harmless to humans, livestock, seafood, and birds. Therefore, their use as non-polluting insecticides has been sought.
【０００５】[0005]
【発明が解決しようとする問題点】ところで、上記の病
原菌は特に分生胞子が感染することによって、より高い
病原性が得られることがわかっており、また保存性に関
しても分生胞子が優れることが明らかとなっている。However, it has been found that the above-mentioned pathogens can obtain higher pathogenicity especially when conidia are infected, and that conidia are superior in preservation. Is clear.
【０００６】この分生胞子の増殖には、元来液体振盪培
養により短菌糸を増殖させ、この短菌糸を分生胞子形成
培地に接種し、寒天による培養・液体培地による静置培
養・ポリウレタンや不織布による培養・フスマや穀類を
用いた培養等の方法が用いられている。For the growth of the conidiospores, short hyphae are originally grown by liquid shaking culture, and the short hyphae are inoculated into a conidiospore-forming medium, and cultured on agar, static culture on a liquid medium, polyurethane, or the like. Methods such as culturing with nonwoven fabric and culturing with bran or cereals are used.
【０００７】しかし、大量の分生胞子を得るためには培
地の組成がコスト高となると共に生産量が良好でないと
いう問題点があった。However, in order to obtain a large amount of conidia, there have been problems that the composition of the medium is expensive and the production is not good.
【０００８】本発明は、上記問題点の改善を図った昆虫
病原糸状菌の製造方法及び昆虫病原糸状菌用培地を提供
することを目的としている。An object of the present invention is to provide a method for producing an entomopathogenic filamentous fungus and a medium for the entomopathogenic filamentous fungus which have improved the above problems.
【０００９】[0009]
【課題を解決するための手段】上記目的を達成するため
に、本発明の昆虫病原糸状菌の製造方法は、昆虫病原糸
状菌を酵母エキス及びマンニトールを主成分とした培地
で培養し、培養物より昆虫病原糸状菌を採取するもので
ある。Means for Solving the Problems To achieve the above object, a method for producing an entomopathogenic filamentous fungus according to the present invention comprises culturing an entomopathogenic filamentous fungus in a medium containing yeast extract and mannitol as main components. It collects entomopathogenic fungi.
【００１０】又、本発明の昆虫病原糸状菌用培地は、水
1000ml中に下記成分 酵母エキス 2.5 ｇ〜 10 ｇ マンニトール 10 ｇ〜 20 ｇ 寒天 0 ｇ〜 15 ｇ が、少なくとも溶解されたものである。[0010] The medium for entomopathogenic fungi of the present invention comprises water.
The following components yeast extract 2.5 g to 10 g Mannitol 10 g to 20 g Agar 0 g to 15 g are dissolved at least in 1000 ml.
【００１１】又、本発明の昆虫病原糸状菌の製造方法に
おける昆虫病原糸状菌は、ボーベリア、メタリジュー
ム、バーティシリューム、ペーシロミセス、エントモフ
トラの内のいずれか１つである。In addition, the entomopathogenic fungus in the method for producing an entomopathogenic fungus of the present invention is any one of Beauveria, Metallurgium, Verticillium, Paecilomyces, and Entomoftra.
【００１２】[0012]
【実施例】次に、具体的に、昆虫病原糸状菌の製造方法
及び昆虫病原糸状菌用培地の一実施例を示して、説明す
る。Next, an embodiment of a method for producing an entomopathogenic filamentous fungus and a culture medium for an entomopathogenic filamentous fungus will be specifically described.
【００１３】本発明者らは前記問題点を解決するための
検討過程においてマンニトール及び酵母エキスを用いた
ところ実用に供し得る殺虫剤を製造するに足る分生胞子
が安定的に得られることを見い出し、本発明を完成し
た。The inventors of the present invention have found that when mannitol and yeast extract are used in the course of study for solving the above problems, conidia sufficient to produce a practically usable insecticide can be stably obtained. Thus, the present invention has been completed.
【００１４】即ち、昆虫病原糸状菌の製造方法は、昆虫
病原糸状菌類の分生胞子をマンニトール10ｇ〜20ｇ（水
1000ml中）、酵母エキス2.5 ｇ〜10ｇ（水1000ml中）、
無機塩類無添加・pH7.0(無調整) 、寒天 0ｇ〜 15 ｇ
（水1000ml中）により培養するものである。なお、昆虫
病原糸状菌類の栽培方法及び害虫に対する病原性の確認
は、下記方法で行なった。That is, a method for producing an entomopathogenic filamentous fungus comprises the steps of preparing a conidia of an entomopathogenic filamentous fungus by adding 10 g to 20 g of mannitol (water).
In 1000 ml), 2.5 g to 10 g of yeast extract (in 1000 ml of water),
No inorganic salts added, pH 7.0 (no adjustment), agar 0g ~ 15g
(In 1000 ml of water). In addition, the cultivation method of insect pathogenic fungi and the confirmation of pathogenicity against insect pests were performed by the following methods.
【００１５】a)昆虫病原糸状菌類の栽培方法 寒天培地により培養した昆虫病原糸状菌（例えば、ボー
ベリア）類より、分生胞子または菌糸をかきとり、液体
培地を入れた１００〜５００mlフラスコで、２５℃、３
〜５日間振盪培養する。この培養液を1:1 〜1:20の割合
で寒天培地または液体培地と混合し、減菌した適当な培
養容器に流し込み、２５℃前後で５〜１４日培養する。
分生胞子が形成されたら分生胞子を採集、または分生胞
子を培地ごと乾燥し粉砕して、低温で保存する。A) Cultivation method of entomopathogenic filamentous fungi Conidiospores or hyphae are scraped from entomopathogenic filamentous fungi (for example, Beauveria) cultured on an agar medium and placed in a 100-500 ml flask containing a liquid medium at 25 ° C. , 3
Shake and culture for ~ 5 days. This culture solution is mixed with an agar medium or liquid medium at a ratio of 1: 1 to 1:20, poured into an appropriate sterilized culture vessel, and cultured at about 25 ° C. for 5 to 14 days.
When conidia are formed, the conidia are collected, or the conidia are dried together with the medium, crushed, and stored at a low temperature.
【００１６】b)害虫に対する病原性の確認 コナガ及びミナミキイロアザミウマに対して本菌を浸漬
処理及び散布処理することで病原性の確認を行う。次
に、実施例により詳細に説明する。B) Confirmation of pathogenicity against pests Pathogenicity is confirmed by immersion and spraying of the present bacterium on Japanese moth and Thrips palmi. Next, an example will be described in detail.
【００１７】（実施例１） 基本となる培地の選定 まず、培地７種を用い、各培地における分生子形成数の
比較検討を行った。以下、培養は、無菌的に行なった。 分生胞子の培養方法 例えばマンニトール・酵母エキス寒天培地により培養し
た昆虫病原糸状菌類（例えば、ボーベリア）より、分生
胞子または菌糸をかきとり、特に限定はしないが、例え
ばマンニトール・酵母エキス液体培地を入れた１００ml
フラスコで、２５℃、３日間振盪培養する。この培養液
を下記の組成の培地に1:２０の割合で混合し、減菌した
シャーレに流し込み、２５℃で１０日培養する。Example 1 Selection of Basic Medium First, seven types of mediums were used to compare the number of conidia formed in each medium. Hereinafter, the culture was performed aseptically. Conidia spore culture method For example, scraping conidia or hyphae from entomopathogenic fungi (for example, Beauveria) cultured on a mannitol / yeast extract agar medium, for example, but not limited thereto, for example, adding a mannitol / yeast extract liquid medium 100ml
Incubate in a flask at 25 ° C. for 3 days with shaking. This culture solution is mixed with a medium having the following composition at a ratio of 1:20, poured into a sterilized petri dish, and cultured at 25 ° C. for 10 days.
【００１８】分生胞子数の測定 培養物を凍結乾燥機にかけ、分生胞子の舞たちをなるべ
く減らして、適当な界面活性剤0.1%ほどを溶かした水に
いれ、適当にホモジナイズして分生胞子を均一に懸濁さ
せる。この懸濁液から１滴を採り、血球計算盤にて分生
胞子数を測定する。分生子数が多すぎた場合には適宜希
釈してから測定を行う。Determination of the number of conidiospores The culture was freeze-dried, the conidiospores were reduced as much as possible, and the resulting mixture was placed in water containing about 0.1% of an appropriate surfactant and homogenized appropriately. Suspend the spores evenly. One drop is taken from this suspension, and the number of conidia is measured using a hemocytometer. If the number of conidia is too large, measure appropriately after dilution.
【００１９】[0019]
【表１】 [Table 1]
【００２０】[0020]
【表２】 [Table 2]
【００２１】気中菌糸の達観調査の結果を表わす。（-
〜 +++= 無〜多) 表１、表２より、昆虫病原糸状菌の培地として、マンニ
トール・酵母エキスが最適であることが判明した。2 shows the results of an objective survey of aerial hyphae. (-
From Table 1 and Table 2, it was found that mannitol / yeast extract is most suitable as a medium for entomopathogenic filamentous fungi.
【００２２】（実施例２） 炭素源の違いによる分生胞子形成量 分生胞子の培養方法 例えばマンニトール・酵母エキス寒天培地により培養し
た昆虫病原糸状菌類（例えば、ボーベリア）より、分生
胞子または菌糸をかきとり、特に限定はしないが、例え
ばマンニトール・酵母エキス液体培地を入れた１００ml
フラスコで、２５℃、３日間振盪培養する。 この培養
液を1:２０の割合で炭素源を変え窒素源を酵母エキスと
した寒天培地と混合し、減菌したシャーレに流し込み、
２５℃で１０日培養する。(Example 2) Conidiospore formation amount depending on the difference of carbon source Culture method of conidia spores For example, conidia or hyphae from entomopathogenic fungi (for example, Beauveria) cultured on mannitol / yeast extract agar medium 100 ml containing, for example, but not limited to, a mannitol / yeast extract liquid medium.
Incubate in a flask at 25 ° C. for 3 days with shaking. This culture solution was mixed with an agar medium using a yeast extract as a nitrogen source while changing the carbon source at a ratio of 1:20, and poured into a sterilized petri dish,
Incubate at 25 ° C for 10 days.
【００２３】分生胞子数の測定 培養物を凍結乾燥機にかけ、分生胞子の舞たちをなるべ
く減らして、適当な界面活性剤0.1%ほどを溶かした水に
いれ、適当にホモジナイズして分生胞子を均一に懸濁さ
せる。この懸濁液から１滴を採り、血球計算盤にて分生
胞子数を測定する。分生子数が多すぎた場合には適宜希
釈してから測定を行う。Determination of the number of conidiospores The culture was freeze-dried to reduce the number of conidiospores as much as possible, put in water containing about 0.1% of an appropriate surfactant, and homogenized appropriately to condense. Suspend the spores evenly. One drop is taken from this suspension, and the number of conidia is measured using a hemocytometer. If the number of conidia is too large, measure appropriately after dilution.
【００２４】[0024]
【表３】 [Table 3]
【００２５】（実施例３） 窒素源の違いによる分生胞子形成量 分生胞子の培養方法 実施例１と同様に、特に限定はしないが、例えばマンニ
トール・酵母エキス寒天培地により培養した昆虫病原糸
状菌類より、分生胞子または菌糸をかきとり、特に限定
はしないが、例えばマンニトール・酵母エキス液体培地
をいれた１００mlフラスコで、２５℃、３日間振盪培養
する。この培養液を１：２０の割合で炭素源をマンニト
ールとし窒素源を変えた寒天培地と混合し、減菌したシ
ャーレに流し込み、２５℃で１０日培養する。Example 3 Conidiospore Formation Amount Due to Difference in Nitrogen Source Culture Method of Conidia Spores As in Example 1, for example, insect pathogenic filaments cultured on a mannitol / yeast extract agar medium From the fungi, conidia or hypha is scraped off, and the cells are shake-cultured in a 100 ml flask containing, for example, but not limited to, a mannitol / yeast extract liquid medium at 25 ° C. for 3 days. This culture solution is mixed at a ratio of 1:20 with an agar medium in which the carbon source is mannitol and the nitrogen source is changed, poured into a sterilized petri dish, and cultured at 25 ° C. for 10 days.
【００２６】分生胞子数の測定 実施例１と同様に、測定を行う。 分生胞子の培養方法 例えばマンニトール・酵母エキス寒天培地により培養し
た昆虫病原糸状菌類（例えば、ボーベリア）より、分生
胞子または菌糸をかきとり、特に限定はしないが、例え
ばマンニトール・酵母エキス液体培地を入れた１００ml
フラスコで、２５℃、３日間振盪培養する。この培養液
を下記の組成の培地に1:２０の割合で混合し、減菌した
シャーレに流し込み、２５℃で１０日培養する。Measurement of the number of conidiospores The measurement is performed in the same manner as in Example 1. Conidia spore culture method For example, scraping conidia or hyphae from entomopathogenic fungi (for example, Beauveria) cultured on a mannitol / yeast extract agar medium, for example, but not limited thereto, for example, adding a mannitol / yeast extract liquid medium 100ml
Incubate in a flask at 25 ° C. for 3 days with shaking. This culture solution is mixed with a medium having the following composition at a ratio of 1:20, poured into a sterilized petri dish, and cultured at 25 ° C. for 10 days.
【００２７】分生胞子数の測定 培養物を凍結乾燥機にかけ、分生胞子の舞たちをなるべ
く減らして、適当な界面活性剤0.1%ほどを溶かした水に
いれ、適当にホモジナイズして分生胞子を均一に懸濁さ
せる。この懸濁液から１滴を採り、血球計算盤にて分生
胞子数を測定する。分生子数が多すぎた場合には適宜希
釈してから測定を行う。Measurement of the number of conidiospores The culture was freeze-dried, the number of conidiospores was reduced as much as possible, and the resultant was placed in water containing about 0.1% of an appropriate surfactant and homogenized appropriately to condense. Suspend the spores evenly. One drop is taken from this suspension, and the number of conidia is measured using a hemocytometer. If the number of conidia is too large, measure appropriately after dilution.
【００２８】[0028]
【表４】 [Table 4]
【００２９】（実施例４） 寒天の添加の有無の検討 分生胞子の培養方法 実施例１と同様に、例えばマンニトール・酵母エキス寒
天培地により培養した昆虫病原糸状菌類（例えば、ボー
ベリア）より、分生胞子または菌糸をかきとり、特に限
定はしないが、例えばマンニトール・酵母エキス液体培
地を入れた１００mlフラスコで、２５℃、３日間振盪培
養する。この培養液を下記の組成の培地に1:２０の割合
で混合し、減菌したシャーレに流し込み、２５℃で１０
日培養する。(Example 4) Investigation of the presence or absence of addition of agar A method for culturing conidiospores In the same manner as in Example 1, for example, a culture was performed using an entomopathogenic filamentous fungus (eg, Beauveria) cultured on a mannitol / yeast extract agar medium. The viable spores or hypha are scraped off, and the cells are shake-cultured in a 100 ml flask containing, for example, but not limited to, a mannitol / yeast extract liquid medium at 25 ° C. for 3 days. This culture solution was mixed with a medium having the following composition at a ratio of 1:20, poured into a sterilized petri dish, and incubated at 25 ° C. for 10 hours.
Incubate for a day.
【００３０】分生胞子数の測定 培養物を凍結乾燥機にかけ、分生胞子の舞たちをなるべ
く減らして、適当な界面活性剤0.1%ほどを溶かした水に
いれ、適当にホモジナイズして分生胞子を均一に懸濁さ
せる。この懸濁液から１滴を採り、血球計算盤にて分生
胞子数を測定する。分生子数が多すぎた場合には適宜希
釈してから測定を行う。Determination of the number of conidiospores The culture was freeze-dried to reduce the number of conidiospores as much as possible, put in water containing about 0.1% of an appropriate surfactant, and homogenized appropriately to condense. Suspend the spores evenly. One drop is taken from this suspension, and the number of conidia is measured using a hemocytometer. If the number of conidia is too large, measure appropriately after dilution.
【００３１】[0031]
【表５】 [Table 5]
【００３２】YM寒天は蒸留水１リットルに対して寒天15
g を添加、その他は、0g （実施例５） 無機塩類添加の違いによる分生胞子形成量 分生胞子の
培養方法 実施例１と同様に、特に限定はしないが、例えばマンニ
トール・酵母エキス寒天培地により培養した昆虫病原糸
状菌類より、分生胞子または菌糸をかきとり、特に限定
はしないが、例えばマンニトール・酵母エキス液体培地
をいれた１００mlフラスコで、２５℃、３日間振盪培養
する。この培養液を１：２０の割合で炭素源をマンニト
ールとし窒素源を酵母エキスとして無機塩類(KH2PO4、Na
Cl、MgSO47H2O)の添加の有無を変えた寒天培地と混合
し、減菌したシャーレに流し込み、２５℃で１０日培養
する。[0032] YM agar is used for 15 liters of agar per liter of distilled water.
g was added, and the others were 0 g. (Example 5) The amount of conidiospore formation due to the difference in the addition of inorganic salts Method for culturing conidia as in Example 1, but not particularly limited, for example, mannitol / yeast extract agar medium The conidia or hyphae are scraped off from the entomopathogenic fungi cultured by the above method, and the culture is carried out with shaking in a 100 ml flask containing, for example, but not limited to, a mannitol / yeast extract liquid medium at 25 ° C. for 3 days. This culture solution was treated at a ratio of 1:20 using mannitol as a carbon source, yeast extract as a nitrogen source and inorganic salts (KH 2 PO 4 , Na
The mixture is mixed with an agar medium with or without the addition of Cl, MgSO 4 7H 2 O), poured into a sterilized petri dish, and cultured at 25 ° C. for 10 days.
【００３３】分生胞子数の測定 実施例１と同様に、測定を行う。Measurement of the number of conidiospores The measurement is performed in the same manner as in Example 1.
【００３４】[0034]
【表６】 [Table 6]
【００３５】（実施例６） ｐＨの違いによる分生胞子形成量 分生胞子の培養方法 実施例１と同様に、特に限定はしないが、例えばマンニ
トール・酵母エキス寒天培地により培養した昆虫病原糸
状菌類より、分生胞子または菌糸をかきとり、特に限定
はしないが、例えばマンニトール・酵母エキス液体培地
を入れた１００mlフラスコで、２５℃、３日間振盪培養
する。この培養液を１：２０の割合で炭素源をマンニト
ールとし窒素源を酵母エキスとした寒天培地を１Ｎ塩酸
および水酸化ナトリウム により希望するpHに調整したものと
混合し、減菌したシャーレに流し込み、２５℃で１０日
間培養する。Example 6 Conidial Spore Formation Amount Due to Difference in pH Culture Method of Conidia Spores As in Example 1, for example, but not limited to, entomopathogenic fungi cultured on a mannitol / yeast extract agar medium. The conidia or mycelium is scraped off, and the cells are shake-cultured in a 100 ml flask containing, for example, but not limited to, a mannitol / yeast extract liquid medium at 25 ° C. for 3 days. This culture was mixed with an agar medium containing mannitol as a carbon source and yeast extract as a nitrogen source at a ratio of 1:20 adjusted to a desired pH with 1N hydrochloric acid and sodium hydroxide, and poured into a sterilized petri dish. Incubate at 25 ° C for 10 days.
【００３６】分生胞子数の測定 実施例１と同様に、測定を行う。Measurement of the number of conidiospores The measurement is performed in the same manner as in Example 1.
【００３７】[0037]
【表７】 [Table 7]
【００３８】（実施例７） マンニトール及び酵母エキスの添加量と分生胞子形成量 分生胞子の培養方法 実施例１と同様に、特に限定はしないが、例えばマンニ
トール・酵母エキス寒天培地により培養した昆虫病原糸
状菌類より、分生胞子または菌糸をかきとり、特に限定
はしないが、例えばマンニトール・酵母エキス液体培地
を入れた１００mlフラスコで、２５℃、３日間振盪培養
する。この培養液を１：２０の割合で炭素源のマンニト
ールと窒素源の酵母エキスとの添加量をそれぞれ変えた
寒天培地と混合し、減菌したシャーレに流し込み、２５
℃で１０日培養する。(Example 7) The amount of mannitol and yeast extract added and the amount of conidia formed The method of culturing conidia as in Example 1 was carried out, for example, but not limited to, on a mannitol-yeast extract agar medium. The conidia or hypha is scraped off from the entomopathogenic fungi, and the cells are shake-cultured in a 100 ml flask containing, for example, but not limited to, a mannitol / yeast extract liquid medium at 25 ° C. for 3 days. This culture solution was mixed at a ratio of 1:20 with an agar medium in which the addition amounts of mannitol as a carbon source and yeast extract as a nitrogen source were respectively changed, and the mixture was poured into a sterilized petri dish.
Incubate at 10 ° C for 10 days.
【００３９】分生胞子数の測定 実施例１と同様に、測定を行う。Measurement of the number of conidiospores The measurement is performed in the same manner as in Example 1.
【００４０】[0040]
【表８】 [Table 8]
【００４１】（実施例８）上記の最適培地を用いて培養
した昆虫病原糸状菌類の分生胞子を、特に限定はしない
が例えばTWEEN80 のような分生胞子の生存に影響のほと
んどない界面活性剤約0.1%水溶液で希釈して分生胞子の
濃度がおよそ1.0 ×108 /ml になるようにする。Example 8 Conidia of entomopathogenic filamentous fungi cultured using the above-mentioned optimal medium are not particularly limited, and are, for example, surfactants such as TWEEN80 that have little effect on the survival of conidia. Dilute with about 0.1% aqueous solution to give a conidia concentration of about 1.0 × 10 8 / ml.
【００４２】この懸濁液にだいこん葉を浸漬し、風乾後
にだいこん葉２枚をシャーレ内にいれ、コナガ３齢幼虫
１０頭を接種する。反復は３回で行い、処理４日後に殺
虫率を求め、１０日後に羽化率を求めて効果を判断す
る。A Japanese radish leaf is immersed in this suspension, and after air-drying, two Japanese radish leaves are placed in a petri dish and inoculated with ten third instar larvae. The repetition is performed three times, the insecticidal rate is determined 4 days after the treatment, and the emergence rate is determined 10 days after the treatment to determine the effect.
【００４３】[0043]
【表９】 [Table 9]
【００４４】[0044]
【表１０】 [Table 10]
【００４５】（実施例９）実施例８と同様に最適培地を
用いて培養した昆虫病原糸状菌類の分生胞子を、特に限
定はしないが例えばTWEEN80 のような分生胞子の生存に
影響のほとんどない界面活性剤約0.1%水溶液で希釈して
分生胞子の濃度がおよそ1.0 ×108 /ml になるようにす
る。Example 9 Conidia of entomopathogenic filamentous fungi cultured in an optimal medium in the same manner as in Example 8 are not particularly limited, but have almost no effect on the survival of conidia such as TWEEN80. Dilute with about 0.1% aqueous solution of detergent to make the concentration of conidia approximately 1.0 × 10 8 / ml.
【００４６】0.5%寒天にきゅうり葉を葉裏を上にして浮
かべ、雌成虫５匹を接種し、ガラススプレーにて懸濁液
を散布後、蓋をする。反復は６回で行い、処理３日後に
死亡虫率を求め効果を判断する。Cucumber leaves are floated on 0.5% agar with the leaf back facing up, 5 adult females are inoculated, the suspension is sprayed with a glass spray, and the lid is closed. The repetition is performed six times, and the mortality rate is determined three days after the treatment to determine the effect.
【００４７】[0047]
【表１１】 [Table 11]
【００４８】[0048]
【発明の効果】本発明の昆虫病原糸状菌の製造方法は、
昆虫病原糸状菌を酵母エキス及びマンニトールを主成分
とした培地で培養し、培養物より昆虫病原糸状菌を採取
するものであるから、昆虫病原糸状菌類の分生胞子をき
わめて効率的に生産することが可能となった。The method for producing an entomopathogenic fungus of the present invention comprises:
The entomopathogenic filamentous fungi are cultured in a medium containing yeast extract and mannitol as the main components, and the entomopathogenic filamentous fungi are collected from the culture. Became possible.
【００４９】又、本発明の昆虫病原糸状菌用培地は、水
1000ml中に下記成分 酵母エキス 2.5 ｇ〜 10 ｇ マンニトール 10 ｇ〜 20 ｇ 寒天 0 ｇ〜 15 ｇが、少なくとも溶解さ
れたものであるから、昆虫病原糸状菌類の分生胞子をき
わめて効率的に生産することが可能となった。Further, the medium for entomopathogenic filamentous fungi of the present invention comprises water
The following components in 1000 ml Yeast extract 2.5 g to 10 g Mannitol 10 g to 20 g Agar 0 g to 15 g is at least dissolved, so that conidia of entomopathogenic fungi can be produced very efficiently. It became possible.
【００５０】又、本発明の昆虫病原糸状菌の製造方法に
おける昆虫病原糸状菌は、ボーベリア、メタリジュー
ム、バーティシリューム、ペーシロミセス、エントモフ
トラの内のいずれか１つである。In addition, the entomopathogenic fungus in the method for producing an entomopathogenic fungus of the present invention is any one of Beauveria, Metallium, Verticillium, Pesilomies, and Entomoftra.
【００５１】なお、かつ難防除害虫であるコナガ及びミ
ナミキイロアザミウマに対する病原性を認めた。In addition, pathogenicity was observed against the hard-to-control pests of the Japanese moth and Thrips palmi.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 山 下 慶 晃 静岡県静岡市春日２丁目12番25号 トモ ノ農薬株式会社内 (58)調査した分野(Int.Cl.6，ＤＢ名) C12N 1/14 ＢＩＯＳＩＳ（ＤＩＡＬＯＧ) ＷＰＩ（ＤＩＡＬＯＧ)──────────────────────────────────────────────────続 き Continuation of the front page (72) Inventor Yoshiaki Yamashita 2-12-25 Kasuga, Shizuoka City, Shizuoka Prefecture Inside Tomono Agrochemical Co., Ltd. (58) Field surveyed (Int.Cl. 6 , DB name) C12N 1/14 BIOSIS (DIALOG) WPI (DIALOG)
Claims (3)
トールを主成分とした培地で培養し、培養物より昆虫病
原糸状菌を採取することを特徴とする昆虫病原糸状菌の
製造方法。1. A method for producing an entomopathogenic filamentous fungus, comprising culturing an entomopathogenic filamentous fungus in a medium containing yeast extract and mannitol as main components, and collecting the entomopathogenic filamentous fungus from the culture.
状菌増殖用培地。2. A medium for growing entomopathogenic fungi, wherein at least 2.5 g to 10 g of yeast extract, 10 g to 20 g of mannitol, and 0 g to 15 g of agar are dissolved in 1000 ml of water.
ューム、バーティシリューム、ペーシロミセス、エント
モフトラの内のいずれか１つである請求項１記載の昆虫
病原糸状菌の製造方法。3. The method for producing an entomopathogenic filamentous fungus according to claim 1, wherein the entomopathogenic filamentous fungus is any one of Beauveria, Metallismum, Verticillium, Paecilomyces, and Entomoftra.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5054450A JP2851506B2 (en) | 1993-03-16 | 1993-03-16 | Method for producing entomopathogenic fungi and medium for growing entomopathogenic fungi |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5054450A JP2851506B2 (en) | 1993-03-16 | 1993-03-16 | Method for producing entomopathogenic fungi and medium for growing entomopathogenic fungi |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH06261742A JPH06261742A (en) | 1994-09-20 |
| JP2851506B2 true JP2851506B2 (en) | 1999-01-27 |
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|---|---|---|---|
| JP5054450A Expired - Fee Related JP2851506B2 (en) | 1993-03-16 | 1993-03-16 | Method for producing entomopathogenic fungi and medium for growing entomopathogenic fungi |
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Families Citing this family (3)
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|---|---|---|---|---|
| ES2134174B1 (en) * | 1998-01-30 | 2000-05-01 | Univ Alicante | PROCEDURE FOR THE PRODUCTION OF ENTOMOPATHOGEN FUNGI FROM THE USE OF THE ALMOND MESOCARD. |
| JP2006265226A (en) * | 2005-02-28 | 2006-10-05 | Sumitomo Chemical Co Ltd | Insecticidal oil-based formulation containing insecticidal filamentous fungus |
| JP2007145718A (en) * | 2005-11-24 | 2007-06-14 | Sumitomo Chemical Co Ltd | Method for applying microorganism having pest controlling ability |
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