JP2009249301A - Plant growth-promoting agent - Google Patents
Plant growth-promoting agent Download PDFInfo
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本発明は植物生長促進剤に関し、特に野菜、穀物類等広範囲の植物の生長を促進し、収量向上、開花促進等の効果を有する植物生長促進剤に関する。 The present invention relates to a plant growth promoter, and more particularly to a plant growth promoter that promotes the growth of a wide range of plants such as vegetables and cereals and has effects such as yield improvement and flowering promotion.
植物の生長を促進させ、必要な野菜、穀物等の収量を向上させることは、食料増産の点から、光合成を向上させることにより地球温暖化の原因となるＣＯ2を減少させる点からも極めて重要であり、従来多くの手段が開発されてきた。このうち、植物ホルモンは植物の器官の一部に作用するものがほとんどであり、植物全体に作用し収量を増大させるものではなかった。また、遺伝子操作による植物の生長促進も研究されているが、その安全性については確立されていない。 Promoting plant growth and improving the yield of necessary vegetables and grains is extremely important from the viewpoint of increasing food production and reducing CO 2 that causes global warming by improving photosynthesis. In the past, many means have been developed. Of these, most of the plant hormones act on a part of plant organs, and do not act on the whole plant and increase the yield. In addition, research on the promotion of plant growth by genetic manipulation has been conducted, but its safety has not been established.
かかる観点から、自然界に存在する物質等の中から植物生長促進作用を有するものを見出すのが望ましく、本発明者は、シュードモナス マルギナリスに属する微生物が優れた植物生長促進作用を有することを見出し、特許出願した（特許文献１）。
しかしシュードモナス属に属する微生物は植物の病原となることもあることから、より安全性の高い植物生長促進剤が望まれていた。
従って、本発明は、安全性が高く、かつ優れた植物生長促進作用を有する天然物由来の植物生長促進剤を提供することを課題とする。
However, since microorganisms belonging to the genus Pseudomonas can be pathogenic to plants, a safer plant growth promoter has been desired.
Accordingly, an object of the present invention is to provide a plant growth promoter derived from a natural product having high safety and an excellent plant growth promoting action.
そこで、本発明者は、より安全性の高い優れた植物生長促進剤を開発すべく種々検討してきたところ、全く意外にもシュードモナス属に属する植物生長促進活性を有する微生物と、安全性が高いことは知られているが植物生長促進活性のないバチルス属微生物及び乳酸菌を混合培養したところ、バチルス属微生物や乳酸菌に植物生長促進活性という形質が導入されることを見出し、本発明を完成した。 Therefore, the present inventor has made various studies in order to develop a safer and superior plant growth promoter, and surprisingly, the microorganism has a plant growth promoting activity belonging to the genus Pseudomonas, and has high safety. As a result, when a mixed culture of Bacillus microorganisms and lactic acid bacteria that are known but have no plant growth promoting activity, a trait called plant growth promoting activity was introduced into the Bacillus microorganisms or lactic acid bacteria, and the present invention was completed.
すなわち、本発明は、バチルス属微生物又は乳酸菌を、シュードモナス属に属する植物生長促進活性を有する微生物と混合培養することを特徴とする。植物生長促進活性を有するバチルス属微生物又は乳酸菌の製造法を提供するものである。
また、本発明は、上記の方法により得られたバチルス属微生物又は乳酸菌を含有する植物生長促進剤を提供するものである。
That is, the present invention is characterized in that Bacillus microorganisms or lactic acid bacteria are mixed and cultured with microorganisms belonging to the genus Pseudomonas and having plant growth promoting activity. The present invention provides a method for producing Bacillus microorganisms or lactic acid bacteria having plant growth promoting activity.
Moreover, this invention provides the plant growth promoter containing the Bacillus microbe or lactic acid bacteria obtained by said method.
本発明によれば、食用に用いられていて安全性が高く、優れた植物生長促進活性を有するバチルス属微生物又は乳酸菌が容易に得られる。また、本発明の植物生長促進剤を用いれば、植物全体の生長が促進され、野菜、穀物等の収量増大を図ることができる。また、安全性も高い。 According to the present invention, a Bacillus microorganism or lactic acid bacterium that is used for food, has high safety, and has excellent plant growth promoting activity can be easily obtained. Moreover, if the plant growth promoter of this invention is used, the growth of the whole plant will be accelerated | stimulated and the yield of vegetables, grains, etc. can be aimed at. In addition, safety is high.
本発明においては、植物生長促進活性を有するシュードモナス属に属する微生物が有する植物生長促進活性を、混合培養することによりバチルス属微生物又は乳酸菌に導入させるものである。原料として用いる微生物は、シュードモナス属に属し、植物生長促進活性を有する微生物である。かかる微生物としては、シュードモナス マルギナリス（Pseudomonas marginalis）、シュードモナス ローデシア（Pseudomonas rhodesiae）等に属する微生物が挙げられる。より具体的には、シュードモナス エスピーＭ−１（ＦＥＲＭ Ｐ−１６４３７）（特開２０００−１９１４２１号記載）、シュードモナス ローデシアＭ−１（ＦＥＲＭ Ｐ−２１４４３）等が挙げられる。 In the present invention, the plant growth promoting activity of a microorganism belonging to the genus Pseudomonas having plant growth promoting activity is introduced into a Bacillus microorganism or lactic acid bacterium by mixed culture. The microorganism used as a raw material is a microorganism belonging to the genus Pseudomonas and having plant growth promoting activity. Examples of such microorganisms include microorganisms belonging to Pseudomonas marginalis, Pseudomonas rhodesiae, and the like. More specifically, Pseudomonas sp. M-1 (FERM P-16437) (described in JP-A No. 2000-191421), Pseudomonas rhodesia M-1 (FERM P-21443) and the like can be mentioned.
ここで、シュードモナス ローデシアＭ−１は、本発明者が土壌から分離した菌株であり、シュードモナス マルギナリス同様に植物生長促進活性を有することを確認している。 Here, Pseudomonas Rhodesia M-1 is a strain isolated by the present inventor from soil, and has been confirmed to have plant growth promoting activity like Pseudomonas marginalis.
一方、混合培養に供されるバチルス属に属する微生物としては、バチルス ズブチリス（Bacillus subtillis）が好ましく、特に納豆菌（バチルス ズブチリスvar. natto）が好ましい。また、乳酸菌としては、ラクトバチルス属に属する菌が好ましい。さらには、ラクトバチルス デルブリッキィ、ラクトバチルス ヘルベチカス、ラクトバチルス アシドフィルス、ラクトバチルス カゼイ、ラクトバチルス ブレビス、ラクトバチルス ブルガリカス、ラクトバチルス ブフネリ、ラクトバチルス プランタラム、ラクトバチルス ガッセリ、ラクトバチルス プレビス、ラクトコッカス ラクチス、ラクトコッカス クレモリス、ストレプトコッカス サーモフィルス、ビフィドバクテリウム属等が特に好ましい。 On the other hand, as a microorganism belonging to the genus Bacillus subjected to mixed culture, Bacillus subtillis is preferable, and Bacillus subtilis var. Natto is particularly preferable. Moreover, as lactic acid bacteria, bacteria belonging to the genus Lactobacillus are preferred. Furthermore, Lactobacillus delbriki, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus brevis, Lactobacillus bulgaricus, Lactobacillus buchneri, Lactobacillus plantarum, Lactobacillus gasseri, Lactobacillus previs, Lactococcus lactis Lactococcus cremolith, Streptococcus thermophilus, Bifidobacterium, etc. are particularly preferred.
これらの微生物を混合培養するには、両者の微生物が生育できる条件で行えばよい。例えば、両者が生育できる培地、土壌等で、１０〜３０℃で、５〜１０日間程度混合して培養すればよい。より具体的には、ＭＲＳ培地、ＴＳＡ培地、ＴＳＢ寒天培地等の培地中で好気的条件下に、１０〜３０℃で３〜７日間程度、特に３０℃で３日間混合培養すればよい。シュードモナス属と、バチルス属微生物又は乳酸菌との混合比は、等量混合がよい。 In order to culture these microorganisms in a mixed manner, the microorganisms may be grown under conditions that allow both microorganisms to grow. For example, what is necessary is just to mix and culture for about 5 to 10 days at 10-30 degreeC by the culture medium, soil, etc. in which both can grow. More specifically, mixed culture may be performed at 10 to 30 ° C. for about 3 to 7 days, particularly at 30 ° C. for 3 days under aerobic conditions in a medium such as MRS medium, TSA medium, and TSB agar medium. The mixing ratio of Pseudomonas and Bacillus microorganisms or lactic acid bacteria should be equal.
混合培養後に、バチルス属又は乳酸菌を培地から分離すれば、植物生長促進活性を有するバチルス属微生物又は乳酸菌が得られる。得られる菌は、植物生長促進活性を有している以外は、１６ＳｒＲＮＡはまったく同じ塩基配列であり、親株と同様の性質を保持していることから、安全性が高く、かつ植物生長促進活性剤として有用である。培地からの菌の分離は、バチルス属微生物又は乳酸菌の特性に基づいて行えばよい。 If Bacillus or lactic acid bacteria are separated from the medium after mixed culture, Bacillus microorganisms or lactic acid bacteria having plant growth promoting activity can be obtained. The obtained fungus has a highly safe and plant growth-promoting active agent because 16S rRNA has the same base sequence and retains the same properties as the parent strain, except that it has plant growth-promoting activity. Useful as. Separation of the bacteria from the medium may be performed based on the characteristics of the Bacillus microorganism or lactic acid bacteria.
かくして植物生長促進活性を付与されたバチルス属微生物の例としては、バチルス ズブチリスＭ−１（ＦＥＲＭ ＡＰ−２１４４１）、ラクトバチルス ブレビスＭ−１（ＦＥＲＭ ＡＰ−２１４４２）が挙げられる。 Examples of Bacillus microorganisms thus imparted with plant growth promoting activity include Bacillus subtilis M-1 (FERM AP-21441) and Lactobacillus brevis M-1 (FERM AP-21442).
得られた本発明バチルス属微生物又は乳酸菌は、通常の栄養培地で増殖させることができるが、例えば通常の有機物や塩類を含む栄養培地で前培養後、さらに例えば糖蜜のような栄養分を用いて１００倍に稀釈した水溶液を培地として本培養すれば、１０6〜１０7／mL程度の菌体数を得ることができるので、この培養液を滅菌して植物生長促進剤として用いることができる。 The obtained Bacillus microorganism or lactic acid bacterium of the present invention can be grown in a normal nutrient medium. For example, after pre-culturing in a nutrient medium containing normal organic matter and salts, a nutrient such as molasses is further used. When the aqueous solution diluted twice is used as the culture medium, the number of cells of about 10 6 to 10 7 / mL can be obtained. Therefore, this culture solution can be sterilized and used as a plant growth promoter.
本発明のバチルス属微生物又は乳酸菌の培養は、上記と同様に有機物や簡単な液体肥料を培地として行えばよい。 The culture of Bacillus microorganisms or lactic acid bacteria of the present invention may be carried out using organic matter or simple liquid fertilizer as a medium in the same manner as described above.
本発明で用いるバチルス属微生物又は乳酸菌は上記の方法で得られた培養物をそのまま用いてもよい。 For the Bacillus microorganism or lactic acid bacterium used in the present invention, the culture obtained by the above method may be used as it is.
本発明の植物生長促進剤は、バチルス属微生物又は乳酸菌のみでも十分効果を奏するものであるが、微生物資材なので肥料は通常通り用いる。農薬を用いてもさしつかえない。 Although the plant growth promoter of the present invention is sufficiently effective only with Bacillus microorganisms or lactic acid bacteria, since it is a microbial material, fertilizer is used as usual. You can use pesticides.
本発明の植物生長促進剤の剤型としては、液剤の他、粉末、粒剤等が挙げられ、これらは常法に従って製造することができる。 Examples of the dosage form of the plant growth promoter of the present invention include liquids, powders, granules and the like, and these can be produced according to a conventional method.
本発明植物生長促進剤を植物に適用するには、次の方法が例示される。
（a）土壌に灌注する方法
１０4〜１０8／mLの菌体数の液を６０℃、２４時間滅菌して用いる。灌注の時期は、植物によって適宜決定すればよいが、本葉が２〜５cm位の長さになる時期（播種後１２〜１６日）が好ましい。例えば、葉菜類等においては、成育を通しほぼ２０℃±５℃の場所で、１５cmのポットに土量７００〜８００ｇとし、１ポットに５００〜１０００倍液を５０mL注入すればよい。
（b）葉面散布法
（a）の液を用いて本葉２〜３cmの長さになる時期（播種後７〜１０日）に１回、５０〜１０００倍液を５０mL位散布する。
The following method is exemplified for applying the plant growth promoter of the present invention to plants.
(A) Method of irrigating the soil A liquid having 10 4 to 10 8 cells / mL is sterilized at 60 ° C. for 24 hours and used. The timing of irrigation may be appropriately determined depending on the plant, but the time (12 to 16 days after sowing) when the true leaves are about 2 to 5 cm long is preferable. For example, in leafy vegetables, the soil volume is 700 to 800 g in a 15 cm pot at a place of about 20 ° C. ± 5 ° C. through growth, and 50 mL of a 500 to 1000 times solution may be injected into one pot.
(B) Foliar spraying method 50 to 1000 times of the solution is sprayed once at a time (7 to 10 days after sowing) of the main leaf using the solution (a).
作物により異なるが、上記（a)、(b)いずれの方法でも、１回の処理でよい。生育促進効果は、１０〜１４日後位から著しく改善され、特に中後期の生育が著しい。そして処理後２０〜３０日後には、本発明品を用いない対照と比べて１．５〜２．５倍の増収となる。 Although it differs depending on the crop, any one of the above methods (a) and (b) may be performed once. The growth promoting effect is remarkably improved from about 10-14 days later, and the growth in the middle and late stages is particularly remarkable. And 20 to 30 days after the treatment, the sales increase by 1.5 to 2.5 times compared to the control without using the product of the present invention.
一般的にそれぞれの作物の生育適温での処理濃度は、５００〜１０００倍であり、生育適温外の高温では濃度を低くし（１０００〜１５００倍）、低温では少し高濃度（３００〜５００倍）が好ましい。 In general, the treatment concentration of each crop at a suitable temperature for growth is 500 to 1000 times, the concentration is lowered at a high temperature outside the optimum temperature for growth (1000 to 1500 times), and the concentration at a low temperature is slightly high (300 to 500 times). Is preferred.
本発明の植物生長促進剤の適用対象となる植物としては、例えば小松菜、ホウレンソウ、大型山東菜、野沢菜、広島菜、ハクサイ、ダイコン、キャベツ、カブ、カボチャ、ピーマン、トマト等の野菜類；イネ、大麦、小麦、ヒエ、トウモロコシ、アワ等の穀物類；コスモス、トレニア、キク、ガーベラ、パンジー、ラン、シャクヤク、チューリップ等の花卉類；アズキ、インゲン、大豆、落花生、ソラマメ、エンドウ等の豆類；コウライシバ、ベントグラス、ノシバ等の芝類；ネギ類；アルファルファ、クローバー、レンゲ等の牧草類等が挙げられる。 Examples of plants to which the plant growth promoting agent of the present invention is applied include vegetables such as komatsuna, spinach, large eastern vegetables, Nozawana, Hiroshima vegetables, Chinese cabbage, Japanese radish, cabbage, turnip, pumpkin, peppers, and tomatoes; Grains such as barley, wheat, barnyard millet, corn, millet; florets such as cosmos, torenia, chrysanthemum, gerbera, pansy, orchid, peonies, tulips; beans such as azuki bean, green beans, soybeans, peanuts, broad beans, peas; Turf such as bentgrass and wild buckwheat; leeks; pastures such as alfalfa, clover, and lotus.
これらのうち、花卉類に対しては、開花促進効果をも得ることができる。 Of these, flowering promotion effects can also be obtained for florets.
以下、実施例を挙げて本発明をさらに詳細に説明するが、本発明は、これらに限定されるものではない。 EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further in detail, this invention is not limited to these.
参考例１
土壌からシュードモナス ローデシアＭ−１（ＦＥＲＭ Ｐ−２１４４３）を得た。得られた菌株は、小松菜、ホウレンソウ等の植物生長を促進する作用を有していた。
Reference example 1
Pseudomonas Rhodesia M-1 (FERM P-21443) was obtained from the soil. The obtained strain had an action of promoting the growth of plants such as Japanese mustard spinach and spinach.
実施例１
市販納豆及び市販ヨーグルトを、それぞれＴＳＡ培地及びＭＲＳ培地で、２８℃、３日間培養し、納豆菌（Bucillus subtillis）と乳酸菌（Lactobacillus brevis）をそれぞれ得た。この乳酸菌をＴＳＢ培地で２５℃、５日間培養し、シュードモナス ローデシアＭ−１（ＦＥＲＭ ＡＰ−２１４４３）と混合培養した。
Example 1
Commercial natto and commercial yogurt were cultured in TSA medium and MRS medium at 28 ° C. for 3 days, respectively, to obtain natto bacteria (Bucillus subtillis) and lactic acid bacteria (Lactobacillus brevis). The lactic acid bacteria were cultured in a TSB medium at 25 ° C. for 5 days, and mixed and cultured with Pseudomonas Rhodesia M-1 (FERM AP-21443).
また、納豆菌は２５℃、２日間培養した。これと、シュードモナス ローデシアＭ−１をＴＳＢ培地で２０℃、３日間培養したものとを等量混合した液を、ＴＳＢ寒天培地に塗抹し３０℃、５日間培養した。 The natto bacteria were cultured at 25 ° C. for 2 days. A solution obtained by mixing equal amounts of Pseudomonas rhodesia M-1 cultured in TSB medium at 20 ° C. for 3 days was smeared on TSB agar medium and cultured at 30 ° C. for 5 days.
乳酸菌、納豆菌共にそれぞれ分離後さらに再分離を行った。納豆菌はＴＳＡ培地に、乳酸菌はＭＲＳ培地にそれぞれ塗抹培養し、形成されたコロニーについて形態及びグラム染色を行いそれぞれ試料分離菌と同一であることを確認した。確認菌をさらに１６ＳｒＲＮＡ塩基配列について同定を行った。その結果により両菌の塩基配列（１６ＳｒＲＮＡ）は納豆菌及び乳酸菌の塩基配列と一致した。 After separating both lactic acid bacteria and natto bacteria, they were further separated. The Bacillus natto was smeared on the TSA medium and the lactic acid bacteria were smeared on the MRS medium, and the formed colonies were morphologically and gram-stained to confirm that they were the same as the sample isolates. The confirmed bacteria were further identified for the 16S rRNA base sequence. As a result, the base sequences (16S rRNA) of both bacteria matched the base sequences of Bacillus natto and lactic acid bacteria.
実施例２
この混合培養後分離した乳酸菌及び納豆菌をそれぞれの培地に入れ、培養後滅菌したものをもとにして次の生育実験を行った。
Ａ．土壌
Ａ−１．鉢による実験
市販培養土又は、黒土、赤玉土（小粒）を等量混合させたものを（ＩＢ４８号又は普通化成肥料土壌７００ｇに１ｇ入れる。）を使用した。それぞれの土壌を５号鉢に入れる。
Ｂ．供試作物
小松菜、ホウレンソウ、大根（ショウゴイン）。
Ｃ．播種
それぞれの鉢に小松菜及びホウレンソウの種子を各１５粒ずつ播種し生育状態で本葉が２〜３cm位になったとき、同じ位の生育のもののみ３個体残し、それを処理に供する。
Example 2
The lactic acid bacteria and natto bacteria separated after this mixed culture were put in each medium, and the following growth experiment was performed based on the sterilized after culture.
A. Soil A-1. Experiments with pots Commercially-cultivated soil, or a mixture of equal amounts of black soil and red ball soil (small grains) (1 g in IB48 or 700 g of ordinary chemical fertilizer soil) was used. Put each soil in No. 5 pot.
B. Prototypes Komatsuna, spinach, radish (shogoin).
C. Seeding 15 seeds of komatsuna and spinach are seeded in each pot, and when the true leaves are about 2 to 3 cm in the grown state, leave only 3 individuals with the same growth and leave them for treatment.
Ｄ．処理
原液（培養液）を５００〜１０００倍に薄め灌注又は葉面散布にて５０mLずつ散布する。散布した日より起算して夏場（２０〜３０℃）は２０日後、冬場（２０〜１５℃）は３０日後に根を含めた全重量を測定し、対照と対比した。ここで対照としては、混合培養していない納豆菌又は乳酸菌を用いた。
D. Treatment The stock solution (culture solution) is diluted 500 to 1000 times and sprayed by 50 mL by irrigation or foliar spraying. The total weight including the roots was measured after 20 days in the summer (20 to 30 ° C.) and 30 days in the winter (20 to 15 ° C.) from the day of application, and compared with the control. As a control, natto or lactic acid bacteria that were not mixed and cultured were used.
その結果、混合培養して分離した納豆菌及び乳酸菌は、小松菜及びホウレンソウの全重量をそれぞれ約２．０倍及び１．５倍に増加させ、優れた植物生長促進活性を有することが判明した。 As a result, it was found that natto and lactic acid bacteria separated by mixed culture increased the total weight of komatsuna and spinach by about 2.0 times and 1.5 times, respectively, and had excellent plant growth promoting activity.
そこで、混合培養して分離した納豆菌をバチルス ズブチリスＭ−１株（ＦＥＲＭ Ｐ−２１４４１）と命名した。一方、混合培養して分離した乳酸菌をラクトバチルス ブレビスＭ−１（ＦＥＲＭ Ｐ−２１４４２）と命名した。 Therefore, the Bacillus natto isolated after mixed culture was named Bacillus subtilis M-1 strain (FERM P-21441). On the other hand, the lactic acid bacterium isolated by mixed culture was named Lactobacillus brevis M-1 (FERM P-21442).
実施例３（畑地での実験）
（１）小松菜、ダイコン（時無）（ほぼ１５〜３０℃）。
よく耕作された畑に１m2につき、苦土石灰１００ｇ、完熟堆肥２kg、配合肥料８０ｇを入れ平均に耕す。４穴の黒マルチを用い各穴に６粒の種子をまき、鉢と同じように本葉が２〜３cm位で生育同一のものを２ヶ体とり処理する。混合培養して得た分離株の５００倍液を５０mL灌注した。トンネルをしてべたかけ栽培用不織布をかける。
Example 3 (Experiment in a field)
(1) Komatsuna, Japanese radish (no time) (approximately 15 to 30 ° C.).
Well per 1m 2 in cultivation has been field, magnesia lime 100g, ripe compost 2kg, plow the average put a blended fertilizer 80g. Using a 4-hole black mulch, sow 6 seeds in each hole, and treat and treat 2 identically grown plants with about 2 to 3 cm of true leaves as in the pot. 50 mL of a 500-fold solution of the isolate obtained by mixed culture was irrigated. Tunnel and apply non-woven fabric for cultivation.
（２）サラダ菜ホウレンソウ
１m2苦土石灰１５０ｇ、完熟堆肥２kgにつき、有機配合肥料１２０ｇを均等に入れマルチ農法により不織布をかける。１穴に６ヶ播種し、６０穴とした。育成後２ヶ体生長の揃った物を選び、本葉が３〜４cm位になったとき、混合培養して得た分離株の５００倍液を５０mL灌注した。
(2) Salad vegetable spinach 1m 2 of 150g of clay lime and 2kg of fully-ripened compost, 120g of organic compound fertilizer is evenly added and a non-woven fabric is applied by a multi-agricultural method. Six seeds were seeded in one hole to make 60 holes. After the growth, the plants with the same growth were selected, and when the true leaves became about 3-4 cm, 50 mL of 500 times of the isolate obtained by mixed culture was irrigated.
（３）２５日後の生育の比較を行った。その結果、混合培養して得られた納豆菌及び乳酸菌ともに、小松菜に対しては１．９５倍、ダイコンに対しては２．５倍、ホウレンソウに対しては１．６５倍の植物生長促進活性を示した。 (3) The growth after 25 days was compared. As a result, both natto and lactic acid bacteria obtained by mixed culture are 1.95 times as much as Komatsuna, 2.5 times as long as Japanese radish, and 1.65 times as long as spinach. showed that.
実施例４
次に、実施例１で用いた納豆菌及び乳酸菌だけでなく、他の納豆菌及び乳酸菌に対しても、植物生長促進活性を有するシュードモナス属微生物の形質が導入されるか否かについての検討を行った。用いた菌は、次のとおりである。これらはいずれも独立行政法人製品評価技術基盤機構から入手した菌株である。
ａ．Bucillus subtillis NBRC13719株
ｂ．Bucillus subtillis NBRC13169株（納豆）
ｃ．Lactobacillus delbrueckii subsp. delbrueckii NBRC3202株
ｄ．Lactobacillus delbrueckii subsp. bulgaricus NBRC13953株（地中海ヨーグルト）
Example 4
Next, whether or not the traits of Pseudomonas sp went. The bacteria used are as follows. These are all strains obtained from the National Institute for Product Evaluation Technology.
a. Bucillus subtillis NBRC13719 strain b. Bucillus subtillis NBRC13169 strain (natto)
c. Lactobacillus delbrueckii subsp. Delbrueckii NBRC3202 strain d. Lactobacillus delbrueckii subsp. Bulgaricus NBRC13953 strain (Mediterranean yogurt)
（方法）
ａ．Bucillus subtillis NBRC13719株とシュードモナス ローデシア
ｂ．Bucillus subtillis NBRC13169株（納豆）とシュードモナス ローデシア
ｃ．Lactobacillus delbrueckii subsp. delbrueckii NBRC3202株とシュードモナス ローデシア
ｄ．Lactobacillus delbrueckii subsp. bulgaricus NBRC13953株（地中海ヨーグルト）とシュードモナス ローデシア
上記４組の検体をＴＳＢ培地でそれぞれ３０℃、７日間培養した。
培養後の培養液から次のようにして再分離を行った。
ａ．ＴＳＡ培地を用いてコロニー分離によりNBRC13719株を単離。
ｂ．ＴＳＡ培地を用いてコロニー分離によりNBRC13169株（納豆）を分離。
ｃ．ＭＲＳ寒天培地を用いてコロニー分離によりNBRC3202株を単離。
ｄ．ＭＲＳ寒天培地を用いてコロニー分離によりNBRC13953株（地中海ヨーグルト）を単離。
単離された検体はＴＳＢ培地を用いて３０℃、７２時間それぞれ培養し滅菌検液とした。
(Method)
a. Bucillus subtillis NBRC13719 strain and Pseudomonas rhodesia b. Bucillus subtillis NBRC13169 strain (natto) and Pseudomonas rhodesia c. Lactobacillus delbrueckii subsp. Delbrueckii NBRC3202 strain and Pseudomonas rhodesia d. Lactobacillus delbrueckii subsp. Bulgaricus NBRC13953 strain (Mediterranean yogurt) and Pseudomonas rhodesia The above four samples were each cultured in TSB medium at 30 ° C. for 7 days.
Re-separation was performed as follows from the culture broth after culturing.
a. NBRC13719 strain was isolated by colony separation using TSA medium.
b. NBRC13169 strain (natto) was isolated by colony separation using TSA medium.
c. NBRC3202 strain was isolated by colony separation using MRS agar medium.
d. NBRC13953 strain (Mediterranean yogurt) was isolated by colony separation using MRS agar medium.
The isolated specimens were cultured in TSB medium at 30 ° C. for 72 hours, respectively, and used as sterilized test solutions.
次に、実施例３と同様にして、サラダホウレンソウに対する生長促進活性を検討した。 Next, in the same manner as in Example 3, the growth promoting activity against salad spinach was examined.
培養土を１５cmポリビニール鉢に７００ｇ入れ、さらに粒状苦土石灰２０ｇ加える。１５粒同一大きさの種子を植えた。生育するにつれ同一大きさの苗を残し、１０日後同一大きさの苗（本葉２〜３cm位のもの）を３つ残した。このようなポリ鉢を各々１０ヶ、合計４０ヶ作る。各混合培養分離株の培養液を５００倍に薄めて灌注した。３０日後の生育重量（地上部のみ）測定した。 Put 700g of cultured soil in a 15cm polyvinyl pot and add 20g of granular clay lime. Fifteen seeds of the same size were planted. As seedlings grew, the same size seedlings were left, and after 10 days, three seedlings of the same size (2-3 cm true leaves) were left. Ten such plastic bowls are made, for a total of 40. The culture solution of each mixed culture isolate was diluted 500 times and irrigated. The growth weight after 30 days (only the above-ground part) was measured.
その結果、混合培養前の納豆菌を対照として、ＮＢＲＣ１３７１９株の混合培養分離株で１．６倍、ＮＢＲＣ１３１６９株の混合培養分離株で２．０倍、ＮＢＲＣ３２０２株の混合培養分離株で１．４倍、ＮＢＲＣ１３９５３株の混合培養分離株で１．６倍の生長促進活性が認められた。 As a result, natto bacteria before mixed culture were compared with 1.6 times for the mixed culture isolate of NBRC13719 strain, 2.0 times for the mixed culture isolate of NBRC13169 strain, and 1.4 times for the mixed culture isolate of NBRC3202 strain. 1.6 times the growth promoting activity was observed in the mixed culture isolate of NBRC13953.
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| JP2015504405A (en) * | 2011-11-04 | 2015-02-12 | テラジェン ホールディングス リミテッドＴｅｒｒａｇｅｎ Ｈｏｌｄｉｎｇｓ Ｌｉｍｉｔｅｄ | Microbial inoculum and fertilizer composition containing the same |
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