JP6898637B2 - How to grow plants containing highly functional ingredients - Google Patents
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Description
本発明は、セファリオフォラ属の微生物の固体培地培養物を含む培養土を用い、カロテノイド、ビタミン類、ポリフェノール類であるβ−カロテン、レチノール当量、ビタミンE(α−トコフェロール)、ビタミンK、ビタミンC、クロロゲン酸、ポリフェノール等の高機能性成分を多く含む高機能性成分野菜を育成(製造)するための方法に関するものである。 The present invention uses culture soil containing a solid medium culture of a microorganism belonging to the genus Cefariophora, carotenoids, vitamins, polyphenols β-carotene, retinol equivalent, vitamin E (α-tocopherol), vitamin K, vitamin C, It relates to a method for growing (manufacturing) highly functional component vegetables containing a large amount of highly functional components such as chlorogenic acid and polyphenols.
食用とする目的で栽培する草本植物を野菜という。利用される部分によって葉菜 edible leaves、根菜 edible root、果菜 fruitsに分ける。葉菜は主として葉および柔らかな茎を食べるもので、キャベツ、ハクサイ、ホウレンソウ、根菜は主として根、ときとして地下茎を食べるもので、ダイコン、ニンジン、ハスなど、果菜は果実を食べるもので、トマト、キュウリ、エンドウマメなどである。野菜は食品として植物繊維、脂質、タンパク質、炭水化物、カルシウム、カリウムなどの無機塩類およびビタミンなどの各種栄養分の補給に必須の素材である。 Herbaceous plants cultivated for edible purposes are called vegetables. Divide into leafy vegetables edible leaves, root vegetables edible root, and fruit vegetables fruits according to the part used. Leafy vegetables mainly eat leaves and soft stems, cabbage, Chinese cabbage, spinach, root vegetables mainly eat roots, sometimes rhizomes, fruit vegetables such as radish, carrots, and hass, fruit vegetables, tomatoes, Cucumbers, radishes, etc. Vegetables are essential ingredients for supplementing various nutrients such as plant fibers, lipids, proteins, carbohydrates, inorganic salts such as calcium and potassium, and vitamins as foods.
非特許文献1によると、機能性を持った食品としては、食物繊維、ポリフェノール類、カロテノイド類が主要なものとされている。それらの研究成果としての機能性食品は、特定保健用食品として機能性が表示されて販売されるか、いわゆる健康食品として販売されている。新たな機能性表示制度の検討が規制改革の一環として消費者庁で実施され、2015年4月から、事業者が責任を持って自主的な機能性表示が可能とした制度(農林水産物も対象)がスタートした。「機能性食品」(非特許文献2参照)は、事業者の責任で、科学的根拠(臨床試験、研究レビュー)を基に商品パッケージに機能性を表示するものとして、消費者庁に届け出された食品であり、健常者や未病者の健康維持・増進に係る部位表現も範囲となった。
このように健康維持機能を野菜に求めようとする研究開発、施策が国内、海外でも積極的に進められ、野菜ジュースや野菜加工食品、野菜入り補助食品などを求める消費者の嗜好が強くなってきている。健康維持機能性成分(以下、機能性成分という。)を含有する野菜には、例えばニンジン、ホウレンソウ、パセリ、シュンギク、コマツナ、ニラ、カボチャ、ブロッコリー、サヤエンドウ、シソ、アスパラガスのように生体重当り100g中にβ−カロテンを600μg以上含んでいるもの、およびβ−カロテンの含有量が生体重当り100g中に600μg以下であっても、多く食べられているトマトやピーマンなどは緑黄色野菜と定義され、日常的に多く摂取することが推奨されている(非特許文献3及び4参照)。
According to Non-Patent
In this way, research and development and measures to require vegetables to maintain their health function are being actively promoted in Japan and overseas, and consumers' preference for vegetable juices, processed vegetable foods, and supplements containing vegetables is increasing. ing. Vegetables containing health maintenance functional ingredients (hereinafter referred to as functional ingredients) include, for example, carrots, spinach, parsley, shungiku, komatsuna, nira, pumpkin, broccoli, pods, perilla, and asparagus. Even if the content of β-carotene is 600 μg or more in 100 g and the content of β-carotene is 600 μg or less in 100 g per living weight, tomatoes and peppers that are eaten a lot are defined as green-yellow vegetables. , It is recommended to take a large amount on a daily basis (see Non-Patent
β−カロテンは人体内に摂取されるとビタミンAと同様の活性作用をもたらし、抗酸化作用により活性酸素を取り除き、血液が流れる作用や老化の防止やがんの増殖阻害、免疫機能の強化、体細胞の活性化などの機能性が知られている。葉菜類においては葉緑素(クロロフィル)とβ−カロテンは極めて高い相関を示すと知られているため、葉緑素含量が多い、すなわち葉色が濃い緑であるほどβ−カロテンの含量も高くなる傾向があることが知られている。
またβ−カロテンと同様に、カロテノイド色素に属するルテインも抗酸化物質で人体では目の網膜の中心にある黄斑に多く存在し、紫外線によって生じる活性酸素を不活性化して黄斑を保護し、加齢を原因とする黄斑変性にならないようにしているとされている。
When β-carotene is taken into the human body, it has the same active action as vitamin A, removes active oxygen by antioxidant action, prevents blood flow and aging, inhibits cancer growth, strengthens immune function, Functionality such as activation of somatic cells is known. In leafy vegetables, chlorophyll and β-carotene are known to show an extremely high correlation, so the higher the chlorophyll content, that is, the darker the leaf color, the higher the β-carotene content tends to be. Are known.
Like β-carotene, lutein, which belongs to the carotenoid pigment, is also an antioxidant and is abundant in the macula in the center of the retina of the eye in the human body. It inactivates active oxygen generated by ultraviolet rays to protect the macula and age. It is said that it prevents macular degeneration caused by.
近年、農業では自然界に存在している有効微生物を利用する試みが進められているが、植物の病害防除においてである。これらの病害防除に利用されている有効微生物は病原菌に対して拮抗性を持ち病原菌の増殖を抑制するものであり、化学合成農薬とは違い、自ら増殖するので防除効果が長続きする、農薬耐性菌の発生が抑えられる、元々自然界に存在する微生物であるので環境汚染の恐れが少ない等の利点がある。作物収穫を目的とした農園芸生産における上記有効微生物の利用技術は、広く知られている(特許文献1〜5)。
しかしながら、作物収穫を目的とした農園芸生産における上記有効微生物の利用技術は開発されていない。
In recent years, attempts have been made in agriculture to utilize effective microorganisms existing in the natural world, but in the control of plant diseases. Effective microorganisms used for controlling these diseases have antagonism against pathogens and suppress the growth of pathogens, and unlike chemically synthesized pesticides, they grow by themselves, so the control effect is long-lasting, and pesticide-resistant bacteria Since it is a microorganism that originally exists in the natural world, there are advantages such as less risk of environmental pollution. Techniques for utilizing the above-mentioned effective microorganisms in agricultural and horticultural production for the purpose of crop harvesting are widely known (
However, a technique for utilizing the above effective microorganisms in agricultural and horticultural production for the purpose of crop harvesting has not been developed.
上述のような背景のもと、日常、継続的に機能性成分を摂取するためには、野菜をはじめとする青果物から食物として無理なく食することが重要なことであり、野菜等の生産者にとっては栽培条件の改善により機能性成分の含有量を高めることは、一般消費者の健康維持、増進に寄与するだけではなく、野菜等の青果物の消費拡大や国際化が進む国内農業市場において競争力を付与することになると考えられる。
本発明は、β−カロテン、レチノール当量、ビタミンE(α−トコフェロール)、ビタミンK、ビタミンC、クロロゲン酸、ポリフェノール等の高機能性成分を多く含む高機能性成分野菜を育成(製造)するための方法を提供することを目的とする。
Against the background described above, in order to continuously ingest functional ingredients on a daily basis, it is important to eat vegetables and other fruits and vegetables reasonably as food, and producers of vegetables and the like. For me, increasing the content of functional ingredients by improving cultivation conditions not only contributes to the maintenance and promotion of the health of general consumers, but also competes in the domestic agricultural market where the consumption of fruits and vegetables such as vegetables is expanding and internationalization is advancing. It is thought that it will give power.
The present invention is for growing (manufacturing) highly functional component vegetables containing a large amount of highly functional components such as β-carotene, retinol equivalent, vitamin E (α-tocopherol), vitamin K, vitamin C, chlorogenic acid, and polyphenols. The purpose is to provide a method of.
本発明は、セファリオフォラ属の微生物、特に、受託番号NITE P−02438で寄託された微生物であるセファリオフォラ属(Cephaliophora sp.)xsd08001株の固体培地培養物を含む培養土を用いることにより、固体培地培養物を含まない培養土を用いる育成方法と比べて、植物固有の機能性成分を増加させることを特徴とする植物の育成方法を要旨としている。 The present invention uses a culture medium containing a culture medium containing a culture medium of the genus Cefariophora , particularly a strain of the genus Cefariophora (Cefariophora sp.) Xsd08001, which is a microorganism deposited under Accession No. NITE P-02438. The gist is a plant growing method characterized by increasing the functional components peculiar to the plant as compared with the growing method using a culture medium containing no substances.
また、本発明は、受託番号NITE P−02438で寄託された微生物であるセファリオフォラ属(Cephaliophora sp.)xsd08001株、およびその固体培地培養物を要旨としている。 The gist of the present invention is the Cefariophora sp. Xsd08001 strain, which is a microorganism deposited under accession number NITE P-02438, and a solid medium culture thereof.
本発明により、本発明は、β−カロテン、レチノール当量、ビタミンE(α−トコフェロール)、ビタミンK、ビタミンC、クロロゲン酸、ポリフェノール等の高機能性成分を多く含む高機能性成分野菜を育成(製造)するための方法を提供することができる。このようにして栽培条件の改善により野菜の機能性成分の含有量を高めることができる。機能性成分の含有量を高めた野菜から日常、継続的に機能性成分を食物として無理なく食することができる。
また、本発明により、高機能性成分を多く含む高機能性成分野菜を育成(製造)するための受託番号NITE P−02438で寄託された微生物であるセファリオフォラ属(Cephaliophora sp.)xsd08001株を提供することができる。
According to the present invention, the present invention grows highly functional component vegetables containing a large amount of highly functional components such as β-carotene, retinol equivalent, vitamin E (α-tocopherol), vitamin K, vitamin C, chlorogenic acid, and polyphenols ( A method for manufacturing) can be provided. In this way, the content of the functional component of the vegetable can be increased by improving the cultivation conditions. From vegetables with an increased content of functional ingredients, it is possible to eat the functional ingredients as food on a daily basis without difficulty.
In addition, the present invention provides a strain of the genus Cephaliophora sp. Xsd08001, which is a microorganism deposited under the accession number NITE P-02438 for growing (manufacturing) highly functional ingredient vegetables containing a large amount of highly functional ingredients. can do.
[xsd08001株の同定]
本発明者は土壌中から数多く単離した菌株の中に、その固体培地培養物を含む培養土を用いて育成すると、固体培地培養物を用いない育成方法と比べて、植物固有の機能性成分を増加させる微生物xsd08001株を見出した。
xsd08001株は、一般的に微生物の同定に用いられる近隣結合法(neighbor−joining method)により、ITS5SrRNA領域のシークエンスデータが即知であるCephaliophora sp.と96%の類似性を示した。
これをCephaliophora sp.xsd08001と命名した。
本出願に際し、Cephaliophora sp.xsd08001は、千葉県木更津市かずさ鎌足2−5−8所在の独立行政法人製品評価技術基盤機構 特許微生物寄託センター(NPMD)に2017年3月6日付で寄託し、AP−02438として受領書が発行された。微生物の生存確認試験が実施されて受託証が発行され、受託番号NITE P−02438として寄託された。
すなわち、本発明の実施例で使用した微生物は、Cephaliophora sp. xsd08001と命名され、受託番号NITE P−02438として寄託された微生物である。
[Identification of xsd08001 strain]
When the present inventor grows a large number of strains isolated from soil using a culture medium containing the solid medium culture, the present inventor has a functional component peculiar to the plant as compared with a growth method that does not use the solid medium culture. We have found a strain of microorganism xsd08001 that increases the number of strains.
The xsd08001 strain is found by the neighbor-joining method, which is commonly used to identify microorganisms, and the sequence data of the ITS5S rRNA region is immediately known from Cephaliophora sp. And showed 96% similarity.
This is referred to as Cephaliophora sp. It was named xsd08001.
In filing this application, Cephaliophora sp. xsd08001 was deposited on March 6, 2017 at the Patent Microorganisms Depositary Center (NPMD) of the Product Evaluation Technology Infrastructure Organization, located at 2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture, and received as AP-02438. It was issued. A microbial survival confirmation test was conducted, a certificate of acceptance was issued, and the deposit was made under the accession number NITE P-02438.
That is, the microorganism used in the examples of the present invention is Cephaliophora sp. It is a microorganism named xsd08001 and deposited under the accession number NITE P-02438.
[固体培地]
固体培地には、オカラ、竹粉、その他、オガクズ、フスマ、モミガラ、バガス、セルロースパウダー、セロビオース、コーヒー粕、デンプンなど、常用されている成分が用いられる。オカラまたは竹粉は元来、農業資材として使用されているものであり、本微生物を固体培地で培養した微生物を含む培養物を、そのまま培養土に用いることができる。
培養は乾燥した固体培地に適度な水分を加えて滅菌する。放冷後に固体培地上に微生物Cephaliophora sp.xsd08001の培養片をそっと置き、25度(室温で可)で約20日静置しておく。10日後には滅菌雰囲気下にて固体培地を混ぜ、全体に微生物が繁殖するようにした。
[Solid medium]
As the solid medium, commonly used ingredients such as okara, bamboo flour, and other commonly used ingredients such as okara, bran, fir, bagasse, cellulose powder, cellobiose, coffee grounds, and starch are used. Okara or bamboo powder is originally used as an agricultural material, and a culture containing a microorganism obtained by culturing this microorganism in a solid medium can be used as it is for potting soil.
The culture is sterilized by adding appropriate water to a dry solid medium. After allowing to cool, the microorganism Cephaliophora sp. Gently place the culture piece of xsd08001 and leave it at 25 ° C (room temperature is acceptable) for about 20 days. After 10 days, a solid medium was mixed in a sterilized atmosphere so that microorganisms could propagate throughout.
[育成の対象とする野菜]
本発明が育成の対象とする野菜は、特に限定されない。
野菜の青果栽培、つまり市場出荷や食用を目的とした栽培では、例えば、キャベツ、ハクサイ、ホウレンソウ等の葉部を食用とする葉菜類や、ダイコン、カブ、ニンジン、ネギ等の根茎部を食用とする根菜類では、花芽分化、抽だい、開花させることなく、植物学的には未熟な成長段階で収穫されるものである。また、ブロッコリー、カリフラワー等は、未熟な花蕾の段階で収穫されるものであり、開花させるには至らない。また、果実生産を目的とするキュウリ、ピーマン、ナス等の果菜類も未熟な果実を収穫するものである。
育成の対象とする野菜は、例えばホウレンソウ、フダンソウ、テンサイ等のアカザ科作物、レタス、サラダナ、シュンギク、ゴボウ等のキク科作物、キャベツ、ブロッコリー、ハクサイ、ダイコン、カブ等のアブラナ科作物、タマネギ、ネギ等のユリ科作物、ニンジン、セルリー、ミツバ等のセリ科作物、トマト、ナス、ピーマン、トウガラシ、トルバム、アカナス、タバコ等のナス科作物、キュウリ、メロン、スイカ、カボチャ、カンピョウ等のウリ科作物、スイートコーン等のイネ科作物、及び、エンドウ、ソラマメ、インゲン、ダイズ等のマメ科作物などを挙げることができる。
[Vegetables to be cultivated]
The vegetables to be cultivated by the present invention are not particularly limited.
In vegetable fruit and vegetable cultivation, that is, for market shipment and edible cultivation, for example, leafy vegetables such as cabbage, Chinese cabbage, and spinach that are edible, and root vegetables such as daikon, cub, carrot, and onion are edible. In root vegetables, they are harvested at a botanically immature growth stage without flower bud differentiation, drawing, or flowering. In addition, broccoli, cauliflower, etc. are harvested at the stage of immature flower buds and do not bloom. In addition, fruit vegetables such as cucumbers, peppers, and eggplants for the purpose of fruit production also harvest immature fruits.
Vegetables to be cultivated include, for example, cruciferous crops such as spinach, fudansou, and tensai, cruciferous crops such as lettuce, saladana, shungiku, and gobo, cruciferous crops such as cabbage, broccoli, hakusai, daikon, and cub, and onions. Cruciferous crops such as green onions, sericulture crops such as carrots, celery, and honey, cucurbitaceae crops such as tomatoes, eggplants, peppers, capsicum, tolbam, red mullet, and tobacco, cucurbitaceae such as cucumber, melon, watermelon, pumpkin, and citrus. Examples include crops, cruciferous crops such as sweet corn, and cruciferous crops such as pea, soramame, green beans, and soybeans.
[野菜の育成方法]
(1)慣行農法に基づいて、各地域において、農薬、肥料の投入量や散布回数等において相当数の生産者が実施している一般的な農法で育成した。
(2)有機農法モデルに基づいて、元肥のない土壌に有機堆肥を添加し、土壌中の有機物を栄養に作物を作る農法で育成した。
[How to grow vegetables]
(1) Based on the conventional farming method, it was cultivated by the general farming method practiced by a considerable number of producers in terms of the amount of pesticides and fertilizers input and the number of sprays in each area.
(2) Based on the organic farming model, organic compost was added to the soil without original fertilizer, and the crops were grown using the organic matter in the soil as nutrition.
[機能性成分含量の分析]
<ビタミンA(α-カロテン,β-カロテン)>
高速液体クロマトグラフ法(可視部吸光光度検出,ODS系順相カラム使用)にて、試料中のα-カロテン,β-カロテンの含量を求めた。試料溶液調整、測定条件は日本食品標準成分表分析マニュアル(七訂)に準じて行った。そのβ-カロテンの値よりレチノール当量を下式(1)にて求めた。
(1)β-カロテン値÷12=レチノール当量
(測定例)
試料を包丁で粗く切断し、4gを100ml容白色広口全量フラスコに量り取る。ピロガロール2g、水5ml、HAET混液(ヘキサン−アセトン−エタノール−トルエン 10:7:6:7)40ml及びエタノール20mlを加え、15分間振とうする。エタノールで定容し、超音波槽に10分間放置する。抽出液8mlを50ml容共栓褐色遠沈管に量り取り、エタノール8ml、60w/v%水酸化カリウム溶液1.6mlを加え、ガラス棒で時々かき混ぜながら70℃水浴中で30分間加熱けん化する。水冷後1w/v%塩化ナトリウム水溶液18.4mlとn-ヘキサン−2-プロパノール−酢酸エチル混液(9:1.5:1)10mlを加え。10分間振とう、遠心分離後上層(有機層)をナス型フラスコに移す。下層(水層)にn-ヘキサン−2-プロパノール−酢酸エチル混液10mlを加え、更に2回同様に抽出する。抽出液を40℃で減圧濃縮後エタノールに溶解して高速液体クロマトグラフに注入し、α-カロテン,β-カロテンのピーク面積を測定する。
(高速液体クロマトグラフ操作条件)
カラム:Inertsil ODS-P GL-Science Inc
移動相:メタノール85:エタノール15
流速:1.0ml/min
温度:40℃
測定波長:455nm
<ビタミンC(アスコルビン酸)>
高速液体クロマトグラフ法(可視部吸光光度検出,シリカゲル順相カラム使用)にて、試料中のアスコルビン酸含量を求めた。試料溶液調整、測定条件は日本食品標準成分表分析マニュアル(七訂)に準じて行った。
<ビタミンE(α-トコフェロール)>
高速液体クロマトグラフ法(蛍光検出,シリカゲル順相カラム使用)にて、試料中のα-トコフェロール含量を求めた。試料溶液調整、測定条件は日本食品標準成分表分析マニュアル(七訂)に準じて行った。
<ビタミンK(フィロキノン)>
高速液体クロマトグラフ法(蛍光検出,ODS系逆相カラム,白金カラム(還元カラム)使用)にて、試料中のフィロキノン含量を求めた。試料溶液調整、測定条件は日本食品標準成分表分析マニュアル(七訂)に準じて行った。
[Analysis of functional ingredient content]
<Vitamin A (α-carotene, β-carotene)>
The contents of α-carotene and β-carotene in the sample were determined by high performance liquid chromatography (visible part absorptiometry detection, using ODS-based normal phase column). Sample solution preparation and measurement conditions were performed according to the Standard Tables of Food Composition in Japan Analysis Manual (7th edition). The retinol equivalent was calculated from the β-carotene value by the following formula (1).
(1) β-carotene value ÷ 12 = retinol equivalent
(Measurement example)
Roughly cut the sample with a kitchen knife and weigh 4 g into a 100 ml white wide-mouthed volumetric flask. Add 2 g of pyrogallol, 5 ml of water, 40 ml of HAET mixture (hexane-acetone-ethanol-toluene 10: 7: 6: 7) and 20 ml of ethanol, and shake for 15 minutes. Volume with ethanol and leave in an ultrasonic bath for 10 minutes. Weigh 8 ml of the extract into a 50 ml capped brown centrifuge tube, add 8 ml of ethanol and 1.6 ml of 60 w / v% potassium hydroxide solution, and heat and saponify in a 70 ° C. water bath for 30 minutes with occasional stirring with a glass rod. After cooling with water, 18.4 ml of a 1 w / v% sodium chloride aqueous solution and 10 ml of an n-hexane-2-propanol-ethyl acetate mixture (9: 1.5: 1) were added. After shaking for 10 minutes and centrifuging, the upper layer (organic layer) is transferred to an eggplant-shaped flask. To the lower layer (aqueous layer), add 10 ml of a mixed solution of n-hexane-2-propanol-ethyl acetate, and extract twice in the same manner. The extract is concentrated under reduced pressure at 40 ° C., dissolved in ethanol, injected into a high performance liquid chromatograph, and the peak areas of α-carotene and β-carotene are measured.
(High performance liquid chromatograph operating conditions)
Column: Inertsil ODS-P GL-Science Inc
Mobile phase: Methanol 85:
Flow velocity: 1.0 ml / min
Temperature: 40 ° C
Measurement wavelength: 455 nm
<Vitamin C (ascorbic acid)>
The ascorbic acid content in the sample was determined by high performance liquid chromatography (visible part absorptiometry detection, using silica gel normal phase column). Sample solution preparation and measurement conditions were performed according to the Standard Tables of Food Composition in Japan Analysis Manual (7th edition).
<Vitamin E (α-tocopherol)>
The α-tocopherol content in the sample was determined by high performance liquid chromatography (fluorescence detection, using a silica gel normal phase column). Sample solution preparation and measurement conditions were performed according to the Standard Tables of Food Composition in Japan Analysis Manual (7th edition).
<Vitamin K (phylloquinone)>
The phylloquinone content in the sample was determined by high performance liquid chromatography (fluorescence detection, ODS-based reverse phase column, platinum column (reduction column) used). Sample solution preparation and measurement conditions were performed according to the Standard Tables of Food Composition in Japan Analysis Manual (7th edition).
[機能性成分の分析に使用する植物体の部位]
機能性成分の分析に使用する植物体の部位は発芽後から開花開始期までの栄養成長期間中の茎葉であれば特に限定されない。例えば、展開した子葉(一般に貝割れと称す)や展開途中や完全展開した成葉、開花開始前の花序部が使用できる。展開とは、生長点で分化した葉が生長して広がり葉身が受光する形態になることをいう。つまり、可食部である。
[Plant parts used for analysis of functional components]
The part of the plant body used for the analysis of the functional component is not particularly limited as long as it is a foliage during the vegetative growth period from the germination to the onset of flowering. For example, unfolded cotyledons (generally called shell cracks), inflorescences in the middle of unfolding or fully unfolded, and inflorescences before the start of flowering can be used. Expansion means that the leaves differentiated at the growth point grow and spread, and the leaf blades receive light. That is, it is an edible part.
本発明を実施例により詳細に説明する。本発明は、これらの実施例により限定されることはない。 The present invention will be described in detail by way of examples. The present invention is not limited to these examples.
固体培地(成分 オカラ又は竹粉、その他前述のものでも可)に適度の水分を加えて約20日間、約25度(室温も可)で培養した。その後、30度の乾燥機に入れて滅菌する。それを放冷後に固体培地上に微生物Cephaliophora sp.xsd08001の培養片をそっと置き、25℃(室温で可)で約20日間静置しておく。10日後には滅菌雰囲気下にて固体培地を混ぜ、全体に微生物が繁殖するようにした。培養後乾燥し、粉砕した。その粉砕したxsd08001株を含む固体培地の培養物を一定の割合で培養土に添加し野菜を育成した。 Appropriate water was added to a solid medium (ingredient okara or bamboo powder, and other above-mentioned ones are also acceptable), and the cells were cultured at about 25 ° C (room temperature is also acceptable) for about 20 days. Then, it is sterilized by putting it in a dryer at 30 degrees. After allowing it to cool, the microorganism Cephaliophora sp. The culture piece of xsd08001 is gently placed and allowed to stand at 25 ° C. (room temperature is acceptable) for about 20 days. After 10 days, a solid medium was mixed in a sterilized atmosphere so that microorganisms could propagate throughout. After culturing, it was dried and pulverized. A culture of a solid medium containing the crushed xsd08001 strain was added to the culture soil at a constant ratio to grow vegetables.
[実施例1]
サラダナを慣行農法モデルに基づいて育成した。
使用培養土:タキイ野菜の土(肥料内容 1リットルあたり N 280mg、P 320mg、K 300mg)670g/1鉢
試供種:サラダナ(岡山サラダナ)2株/1鉢、3反復
ハウス内温度:18〜25℃
固体培地の培養物添加量:1/0%
収穫物:図1参照
収穫したサラダナ地上部の生重量を図2に、これらの機能性成分の分析結果を図3に示す。
[Example 1]
Saladana was cultivated based on the conventional farming model.
Potting soil used: Takii vegetable soil (fertilizer content N 280 mg, P 320 mg,
Culture addition amount of solid medium: 1/0%
Harvested products: See FIG. 1. The raw weight of the above-ground part of the harvested saladna is shown in FIG. 2, and the analysis results of these functional components are shown in FIG.
結果は、図1に示したように、xsd08001株の固体培地培養物を添加した培養土で育成したサラダナの方が、見た目でも生長促進されていることがわかり、葉の数も多く、しかも1枚当たりの面積も大きい。収穫した地上部の生重量について比較したものが図2であって、固体培地培養物を添加した培養土で育成したサラダナの方が約1.5倍重い収量であり、そのサラダナの地上部100g当たりのβ-カロテン、ビタミンE、ビタミンC、ビタミンKの含有量も、無添加培養土のものに比べ、それぞれ約105%、127%、200%、130%と増加していることが図3に示されている。 As a result, as shown in FIG. 1, it was found that the saladna grown in the culture soil to which the solid medium culture of xsd08001 strain was added was visually promoted in growth, and the number of leaves was large, and 1 The area per sheet is also large. FIG. 2 shows a comparison of the raw weight of the harvested above-ground part, and the yield of saladana grown in the culture soil to which the solid medium culture was added was about 1.5 times heavier, and the above-ground part of the saladna was 100 g. The contents of β-carotene, vitamin E, vitamin C, and vitamin K per unit are also increased to about 105%, 127%, 200%, and 130%, respectively, as compared with those of the additive-free culture soil, respectively. It is shown in.
[実施例2]
サラダナを有機農法モデルに基づいて育成した。
使用培養土:そのきや培養土(NPK 元肥一切なし)670g/1鉢
堆肥:バイオダルマ pH7.0−7.2(N 3.4%、P2O5 6%、
K2O 3.6%、CaO 20%)主な成分は畜糞
試供種:サラダナ(岡山サラダナ) 2株/1鉢、3反復
ハウス内温度:18−25℃
固体培地培養物の添加量:1.0%
収穫物:図4参照
収穫したサラダナ地上部の生重量を図5に、これらの機能性成分の分析結果を図6に示す。
[Example 2]
Saladana was cultivated based on an organic farming model.
Use culture soil: Part-out Ya culture soil (NPK basal without any) 670g / 1 pot compost: Bio Dharma pH7.0-7.2 (N 3.4%, P 2 O 5 6%,
K 2 O 3.6%, CaO 20%) The main ingredients are livestock manure sample: Saladana (Okayama Saladana) 2 strains / 1 bowl, 3 repeats House temperature: 18-25 ° C
Addition amount of solid medium culture: 1.0%
Harvested products: See FIG. 4. The raw weight of the above-ground part of the harvested saladna is shown in FIG. 5, and the analysis results of these functional components are shown in FIG.
結果は、図4に示したように、xsd08001株の固体培地培養物を添加した培養土で育成したサラダナの方が、2倍以上に成長し、生長促進されていた。葉の数も多く、1枚当たりの面積も大きい。収穫した地上部の生重量について比較したものが図5であって、固体培地培養物を添加した培養土で育成したサラダナの方が約3.8倍重い収量であり、そのサラダナの地上部100g当たりのβ-カロテン、ビタミンE、ビタミンC、ビタミンKの含有量も、無添加培養土のものに比べ、それぞれ約121%、120%、111%、117%と増加していることが図6に示されている。 As a result, as shown in FIG. 4, the saladana grown in the culture soil to which the solid medium culture of xsd08001 strain was added grew more than twice and the growth was promoted. The number of leaves is large, and the area per leaf is also large. FIG. 5 shows a comparison of the raw weight of the harvested above-ground part, and the yield of saladana grown in the culture soil to which the solid medium culture was added was about 3.8 times heavier, and the above-ground part of the saladna was 100 g. The contents of β-carotene, vitamin E, vitamin C, and vitamin K per unit are also increased to about 121%, 120%, 111%, and 117%, respectively, as compared with those of the additive-free culture soil, respectively. It is shown in.
[実施例3]
固体培地培養物を含む培養土を用いて育成した野菜の安全性試験を行った。Cephaliophora sp.は糸状菌であるため、カビ毒を発生すると食中毒の原因となる。海外では残留基準値の設定はなく、日本国内でも同様だが、注意喚起として1800ppb−1200ppb以下とされている。
ロリトレムBの含量を調べた結果は図10であり、定量下限値以下であった。
[Example 3]
A safety test was conducted on vegetables grown using potting soil containing a solid medium culture. Cephaliophora sp. Since is a filamentous fungus, mycotoxins cause food poisoning. There is no residual standard value set overseas, and the same is true in Japan, but it is set to 1800 ppb-1200 ppb or less as a reminder.
The result of examining the content of lolitrem B was shown in FIG. 10, which was below the lower limit of quantification.
[実施例4]
ホウレンソウを慣行農法にてハウス内圃場にて育成した。
試供種:ホウレンソウ(トライ)
使用肥料:IB化成S1号
固体培地培養物の添加量:10g/m2
対照としては、培養物を添加せずに育成した。
収穫物:図7参照
収穫したホウレンソウ地上部の生重量を図8にこれらの機能性成分の分析結果を図9に示す。
[Example 4]
Spinach was cultivated in the field in the house by the conventional farming method.
Free sample: Spinach (try)
Fertilizer used: IB Kasei S1 solid medium Addition amount of culture: 10 g / m 2
As a control, the cells were grown without adding the culture.
Harvested product: See FIG. 7. The raw weight of the above-ground part of the harvested spinach is shown in FIG. 8, and the analysis result of these functional components is shown in FIG.
[実施例5]
トウモロコシを慣行農法にてハウス内圃場にて育成した。
試供種:ゴールドラッシュ88
使用肥料:IB化成S1号
固体培地培養物の添加量:10g/m2
対照としては、培養物を添加せずに育成した。
収穫物:図11参照
収穫した皮付きトウモロコシの生重量を図12に大きさを図13に示す。
[Example 5]
Corn was cultivated in the field in the house by the conventional farming method.
Free sample: Gold rush 88
Fertilizer used: IB Kasei S1 solid medium Addition amount of culture: 10 g / m 2
As a control, the cells were grown without adding the culture.
Harvest: See FIG. 11 The raw weight of the harvested corn with skin is shown in FIG. 12 and the size is shown in FIG.
[実施例6]
ナスを慣行農法にて露地栽培した。
試供種:千両ナス
使用肥料:サンライム、鶏フン
固体培地培養物の添加量:10g/m2
対照としては、培養物を添加せずに栽培した。
収穫物:4日間の収穫物を図14に示す。
4日間のナス収穫量とこれらの機能性成分の分析結果を図15に示す。
[Example 6]
Eggplants were cultivated outdoors by conventional farming methods.
Free sample: Senryo eggplant
Fertilizer used: sun lime, chicken droppings
Addition amount of solid medium culture: 10 g / m 2
As a control, the culture was cultivated without addition.
Harvest: The four-day harvest is shown in FIG.
FIG. 15 shows the eggplant yield for 4 days and the analysis results of these functional components.
[結果と考察]
有機農法で堆肥を使用した場合、化学肥料のように植物が根から吸収できる養分ではなく、地中において微生物等により分解された後のものしか吸収することができない。図1と図4を比較すると、図4の堆肥を使用した場合の方が、セファリオフォラ属を添加すると無添加の場合に比べて生長促進効果が2倍になり、よりその機能が働くことがわかった。
セファリオフォラ属は野菜の根部において、有機肥料を分解し、サラダナの生長が無添加に比べて著しく現れたものと思われる。
植物は地中に根を張った部分からのみ土壌成分を吸収することができるが、セファリオフォラ属は地中に広く菌糸を広げ、広範囲の土壌成分を自らの菌糸中に吸収、分解し、植物の根へその成分を送ると思われる。植物はセファリオフォラ属から得た土壌成分をもとに成長し、機能性成分を生成する。そして、その過程で生成した糖(炭素分)をセファリオフォラ属へ与え、セファリオフォラ属はより良い環境を求めて、広い範囲へと菌糸を伸ばしていく。このように本発明のセファリオフォラ属は植物と共生関係にあるエンドファイトではないかと考えられる。
[Results and discussion]
When compost is used in organic farming, it cannot absorb nutrients that plants can absorb from the roots like chemical fertilizers, but only those that have been decomposed by microorganisms in the ground. Comparing FIGS. 1 and 4, it was found that when the compost of FIG. 4 was used, the growth promoting effect was doubled when the genus Cefariophora was added, and the function worked more. It was.
It is considered that the genus Cefariophora decomposed organic fertilizers at the roots of vegetables, and the growth of saladana was remarkably exhibited as compared with the case where no addition was added.
Plants can absorb soil components only from the part that has roots in the ground, but the genus Cefariophora spreads mycelia widely in the ground and absorbs and decomposes a wide range of soil components into its own hyphae, and the plant's It seems to send the ingredient to the roots. The plant grows on the basis of soil components obtained from the genus Cefariophora and produces functional components. Then, the sugar (carbon content) produced in the process is given to the genus Cefariophora, and the genus Cephalophora grows hyphae over a wide range in search of a better environment. Thus, it is considered that the genus Cefariophora of the present invention is an endophyte having a symbiotic relationship with plants.
前述のような機能性成分を無理することなく摂取するためには、野菜をはじめとする青果物から食物として食することが重要であり、近年、品種改良によって高機能性野菜が市販されている。例えば、「坊ちゃんかぼちゃ」は、β−カロテンが従来の3−4倍、「こどもピーマンピー太郎」はβ−カロテンが従来の2倍である、また、「赤帯ゴーヤ」はビタミンCが従来の1.5倍、紫アスパラガスはグリーンアスパラの約10倍とされる。しかし、本発明のCephaliophora sp.の固体培養物を培養土に添加すると、品種改良をしていなくても、β−カロテン、レチノール当量、ビタミンE(α−トコフェロール)、ビタミンK、ビタミンC、クロロゲン酸、ポリフェノール等の高機能性成分を多く含む高機能性成分野菜を育成できる。
In order to ingest the above-mentioned functional ingredients without overdoing it, it is important to eat from fruits and vegetables such as vegetables as food, and in recent years, highly functional vegetables have been put on the market due to breeding. For example, "Bochan Pumpkin" has 3-4 times more β-carotene than before, "Children's Bell Pepper Pee Taro" has twice as much β-carotene as before, and "Red Belt Goya" has twice as much vitamin C as before. 1.5 times, purple asparagus is about 10 times that of green asparagus. However, the Cephaliophora sp. Of the present invention. When the solid culture of is added to the culture soil, high functionality such as β-carotene, retinol equivalent, vitamin E (α-tocopherol), vitamin K, vitamin C, chlorogenic acid, polyphenol, etc. It is possible to grow highly functional vegetables containing many ingredients.
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