JPH09294495A - Proliferation of germ-free sweet potato seedling - Google Patents
Proliferation of germ-free sweet potato seedlingInfo
- Publication number
- JPH09294495A JPH09294495A JP10975996A JP10975996A JPH09294495A JP H09294495 A JPH09294495 A JP H09294495A JP 10975996 A JP10975996 A JP 10975996A JP 10975996 A JP10975996 A JP 10975996A JP H09294495 A JPH09294495 A JP H09294495A
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- Japan
- Prior art keywords
- sweet potato
- seedlings
- culture
- plantlets
- sterile
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、無菌サツマイモ苗
の増殖方法に関する。さらに詳しくは、組織培養法によ
って実質的に無菌的にサツマイモの小植物体を培養して
増殖する方法に関する。この方法によって得られた無菌
サツマイモ苗は、培養苗として利用することができる。TECHNICAL FIELD The present invention relates to a method for growing sterile sweet potato seedlings. More specifically, it relates to a method of culturing and propagating sweet potato plantlets substantially aseptically by a tissue culture method. The sterile sweet potato seedlings obtained by this method can be used as cultured seedlings.
【０００２】[0002]
【従来の技術】サツマイモは、食糧（カロリー源）、食
料（野菜）、加工原料（デンプン、糖、アルコール飲料
など）、アルコール燃料用原料などとして利用される多
用途植物である。しかも、寒冷地から熱帯地までの地域
的な制約がなく、傾斜地、砂地、イネの栽培に適さない
土壌での栽培が可能で、古来より飢餓作物、救荒作物と
呼ばれているように、干害、高温、強風（台風）などの
気象災害、および酸性土壌耐性が比較的高いという特色
をもつ。このため、将来生起すると予想される食糧、エ
ネルギー・資源および環境の三大問題を同時並行的に解
決するのに重要な役割を演じる植物であると評価されて
いる。BACKGROUND OF THE INVENTION Sweet potato is a versatile plant used as food (calorie source), food (vegetable), processing raw material (starch, sugar, alcoholic beverage, etc.), alcohol fuel raw material, and the like. Moreover, there is no regional restriction from cold regions to tropical regions, and it can be cultivated on slopes, sandy soils, and soils that are not suitable for rice cultivation. It is characterized by high temperature, weather disasters such as strong winds (typhoons), and relatively high resistance to acidic soil. Therefore, it is evaluated as a plant that plays an important role in simultaneously solving the three major food, energy / resource and environmental problems that are expected to occur in the future.
【０００３】このように、将来重要な役割を演じると目
されるサツマイモを増産するには、現在のところ、さし
穂苗、組織培養（マイクロプロパゲーション）による苗
（以下、培養苗ともいう）、タネイモからの萌芽苗など
による方法が実用化されている。しかし、将来は栽培土
壌の整備、栽培規模の拡大、自動移植機による苗の移植
などの栽培技術の進歩により、苗の需要が大幅に拡大す
ることが予想される。大幅に拡大した苗の需要に応える
には、従来のさし穂苗、タネイモからの萌芽苗によって
は対応しきれない。そこで、培養苗が注目されることに
なる。As described above, in order to increase the production of sweet potatoes, which are expected to play an important role in the future, at present, cutting seedlings and seedlings by tissue culture (micropropagation) (hereinafter also referred to as cultured seedlings) , A method using sprouted seedlings from Tanemo has been put to practical use. However, it is expected that the demand for seedlings will greatly increase in the future due to the improvement of cultivation techniques such as the maintenance of cultivation soil, the expansion of cultivation scale, and the transplanting of seedlings by an automatic transplanter. To meet the demand for significantly expanded seedlings, conventional cuttings and sprouted seedlings from Taneimo cannot be used. Therefore, the cultured seedlings will be paid attention.
【０００４】組織培養による苗（培養苗）は、組織培養
器内で植物細胞、組織ないし小植物体を培養して得た苗
であり、遺伝的により優れた品質の植物苗を提供できる
ので、世界的に普及しつつある。しかし、培養苗は従来
の実生苗、さし穂苗、タネイモからの萌芽苗に比較し
て、コストが高いという欠点があり、現在までのとこ
ろ、その普及は、主に、ラン、イチゴ、カーネーショ
ン、観葉植物など、一部の高級施設園芸植物に限られて
いる。Seedlings (cultured seedlings) obtained by tissue culture are seedlings obtained by culturing plant cells, tissues or plantlets in a tissue culture vessel, and can provide genetically superior quality plant seedlings. It is spreading worldwide. However, compared to conventional seedlings, cuttings, and sprouting seedlings from Tanemo, cultured seedlings have the drawback of high cost, and so far, their spread has been mainly in orchids, strawberries, and carnations. , Limited to some high-end horticultural plants such as houseplants.
【０００５】培養苗のコストが高くなる第一の理由は、
その生産方法が多分に手工業的であり、生産コストの６
５〜７０％を人件費が占めることによる。その第二の理
由は、組織培養期間中における植物細胞、組織ないし小
植物体の成長が遅いことにあり、第三の理由は、それに
よって得られる培養苗のうち、実際に商品価値がありそ
の後の馴化過程において、良好に活着ないし生育するも
のの歩留りが低いことにある。The first reason why the cost of cultured seedlings is high is
The production method is probably handicraft, and the production cost is 6
Personnel expenses account for 5 to 70%. The second reason is that the growth of plant cells, tissues or plantlets during the tissue culture period is slow, and the third reason is that among the cultured seedlings obtained by that, there is actually commercial value and In the acclimation process of, the good yield or growth of the product is low.
【０００６】一方、従来、組織培養中の小植物体は光合
成能力をかなり失い、組織培養中の小植物体を一貫して
独立栄養生長させ、それによって培養苗を生産するよう
なことは実用的に不可能と考えられていた。このような
固定観念が定着した原因は、植物の組織培養の技術が、
本質的に従属栄養生長である微生物や動物の細胞ないし
組織の培養技術に準拠して発達したものであることを挙
げることができる。[0006] On the other hand, conventionally, it is practical that plantlets in tissue culture lose their photosynthetic ability considerably, and plantlets in tissue culture are consistently autotrophically grown, thereby producing cultured seedlings. Was considered impossible. The reason why such stereotypes have taken root is that plant tissue culture technology
It can be mentioned that it has been developed in accordance with the culturing technology of microorganisms or animal cells or tissues that are essentially heterotrophic growth.
【０００７】従来、植物の組織培養技術においては、小
植物体を生長させるには培地中に糖（ショ糖、グルコー
ス、フラクトースなど）を炭素源として加え、従属栄養
的生長の側面の強い混合栄養生長させるのが一般的な常
識となっていた。このような従来技術において、その生
産性を高め、生産コストを低減するための研究の方向
は、いずれも従属栄養的な生長・増殖を極限まで効率化
し、かつ、省力化する方向において推進されてきた。Conventionally, in plant tissue culture technology, in order to grow a plantlet, sugar (sucrose, glucose, fructose, etc.) is added to the medium as a carbon source, and a strong mixed nutrition in the aspect of heterotrophic growth. It was common sense to grow. In such conventional technologies, research has been conducted in order to increase productivity and reduce production cost in the direction of maximizing the efficiency of heterotrophic growth and proliferation and saving labor. It was
【０００８】例えば、糖を含む液体培地中に小植物体を
浸漬して、さらにその培養器内に酸素を供給し、培養器
を震とうまたは回転させるなどして、糖および酸素など
の小植物体への吸収を高める方法などは、上記のような
方向の研究開発の典型的な手法として挙げることができ
る。For example, by immersing the plantlets in a liquid medium containing sugar and further supplying oxygen into the incubator to shake or rotate the incubator, plantlets such as sugar and oxygen A method of enhancing absorption into the body can be mentioned as a typical method of research and development in the above-mentioned direction.
【０００９】そして、上記のような従来法においては、
その培地中に多量の糖類を添加するため、培地は培養期
間中に外部からの雑菌による汚染を受け易く、このため
培養期間中に外部からの雑菌汚染により、培養中の小植
物体が腐死するというトラブルが発生するので、これを
防止するために培養器や、多数の培養器を収納する培養
室などの培養環境を、実質的に無菌状態に維持するよう
厳重な取扱を必要としていた。Then, in the conventional method as described above,
Since a large amount of saccharides is added to the medium, the medium is easily contaminated by external bacteria during the culture period, so that the external plant contamination during the culture period causes the plantlets in the culture to rot. Therefore, in order to prevent this, strict handling is required to maintain the culture environment such as the incubator and the culture room accommodating a large number of incubators in a substantially sterile state.
【００１０】しかも、このような培養器で、従属栄養的
な生長・増殖を主体として生産された培養苗を、試験
管、フラスコまたはビーカに移植し、独立栄養生長条件
での馴化過程に移行させるには、培養苗に環境の激変を
与えることになり、馴化過程における活着率の低下や、
生育の遅れの一因になるものと考えられる。Moreover, the cultured seedlings produced mainly by heterotrophic growth / proliferation in such an incubator are transplanted to a test tube, a flask or a beaker, and transferred to an acclimatization process under autotrophic growth conditions. Will cause a drastic change in the environment of the cultured seedlings, a decrease in the survival rate during the acclimation process,
It is considered to contribute to the delay in growth.
【００１１】[0011]
【発明が解決しようとする課題】本発明者はかかる状況
にあり、上記のような固定観念にとらわれることなく、
サツマイモの培養苗を能率的に提供すべく鋭意検討した
結果、本発明を完成するに至った。本発明は、次の諸課
題を解決することを目的とする。 １．遺伝的に優れた品質の無菌サツマイモ苗を能率的に
提供すること。 ２．培養期間中に雑菌による汚染の少ない無菌サツマイ
モ苗の増殖方法を提供すること。 ３．環境の激変を与えることなく馴化過程に移行でき、
馴化過程における活着率が良好で、その後の生育も良好
な無菌サツマイモ苗を提供すること。The present inventor is in such a situation, and is not restricted by the above-mentioned fixed idea.
As a result of intensive studies to provide cultured seedlings of sweet potato efficiently, the present invention has been completed. An object of the present invention is to solve the following problems. 1. To efficiently provide sterile sweet potato seedlings of genetically excellent quality. 2. To provide a method for growing a sterile sweet potato seedling that is less contaminated by various bacteria during the culture period. 3. You can shift to the acclimatization process without giving a drastic change in the environment,
To provide aseptic sweet potato seedlings that have a high survival rate in the acclimation process and that also grow well thereafter.
【００１２】[0012]
【課題を解決するための手段】上記課題を解消するため
に、本発明では、組織培養法によってサツマイモの小植
物体を培養して無菌サツマイモ苗を増殖する方法におい
て、支持体に液体成分を支持させた無糖培地を使用し、
無菌状態で増殖することを特徴とする無菌サツマイモ苗
の増殖方法を提供する。[Means for Solving the Problems] In order to solve the above problems, in the present invention, a liquid component is supported on a support in a method of culturing sweet potato plantlets by a tissue culture method to grow sterile sweet potato seedlings. Using the sugar-free medium
Provided is a method for growing a sterile sweet potato seedling, which is characterized by growing in a sterile state.
【００１３】[0013]
【発明の実施の形態】以下、本発明を詳細に説明する。
従来の培養苗の増殖方法においては、前記した通り、培
地内に多量の糖を添加しているために、雑菌による汚染
が生起し易く、これを防止するために培養器の栓の部分
を半密閉状態にするのが通常である。そして、このよう
な半密閉状態にすることによって、当然のことながら、
外部から炭酸ガスを含有した空気の供給が抑制されるこ
とを意味し、培養器内の光合成が不可能な状態が一層強
化される。その結果、従来の培養苗の増殖方法において
は、実質的に独立栄養生長による培養は不可能である。BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.
In the conventional method for growing cultured seedlings, as described above, since a large amount of sugar is added to the medium, contamination by miscellaneous bacteria is likely to occur, and in order to prevent this, the stopper portion of the incubator is half It is usually sealed. And by making such a semi-sealed state, of course,
This means that the supply of air containing carbon dioxide from the outside is suppressed, and the state in which photosynthesis in the incubator is impossible is further strengthened. As a result, in the conventional method for growing cultured seedlings, culturing by autotrophic growth is substantially impossible.
【００１４】これに対して、本発明方法では、組織培養
法によってサツマイモの小植物体を培養して無菌サツマ
イモ苗を増殖する方法において、液体成分を支持させた
無糖培地を使用し、無菌状態で培養することを必須と
し、一貫して完全な独立栄養生長条件下で組織培養を行
うものである。なお、本発明においてサツマイモの小植
物体とは、サツマイモの細胞組織を培養して得られた、
葉を有する節(node)、および、この植物体の茎部につい
て、複数の節を節単位で切断し、液体成分を支持させた
無糖培地を使用して、これに節挿し増殖培養させること
により、茎部が伸び、発根し、複数の節および葉が生じ
たものを言う。On the other hand, according to the method of the present invention, a sugar-free medium supporting a liquid component is used in a sterile state in a method of cultivating a sweet potato plantlet by culturing a sweet potato plantlet by a tissue culture method. It is essential to cultivate the tissue in the following manner, and the tissue culture is consistently performed under complete autotrophic growth conditions. Incidentally, the sweet potato plantlets in the present invention, obtained by culturing the sweet potato cell tissue,
For nodes having leaves and stems of this plant, cutting multiple nodes in node units, using a sugar-free medium supporting liquid components, and performing node-cultivation and growth culture The stems are elongated and rooted, resulting in multiple nodes and leaves.
【００１５】液体成分を支持させる培地としては、無菌
状態としたロックウールまたはバーミキュライトが好ま
しい。これら培地を無菌状態にするには、培地として使
用するロックウールまたはバーミキュライトを加圧滅菌
する。これらの無菌状態の培地を使用することにより、
成長速度を従来の寒天を培地として使用した光混合栄養
培養の場合に比較し、１．５〜２倍程度に向上させるこ
とができる。As the medium for supporting the liquid component, rock wool or vermiculite in a sterile state is preferable. To make these media aseptic, rock wool or vermiculite used as the media is autoclaved. By using these sterile media,
The growth rate can be improved by about 1.5 to 2 times as compared with the case of photomixture nutrient culture using conventional agar as a medium.
【００１６】上記培地に含ませる液体成分は、糖分を含
まないもの（無糖）であることを必須とする。無糖培地
を使用することにより、小植物体は、従属栄養的生長で
はなく、一貫して完全な独立栄養生長条件下で組織培養
を行うことができる。液体成分は無糖であれば特に制限
がなく、従来から知られている基本培地、例えばホーグ
ランド・アノン、ムラシゲ・スグーク、ホワイト、リン
スマイヤー・スグーク、ニッチ・ニッチなどが挙げられ
る。It is essential that the liquid component contained in the medium is one that does not contain sugar (sugar-free). By using a sugar-free medium, plantlets can undergo tissue culture under consistently complete autotrophic growth conditions, rather than heterotrophic growth. The liquid component is not particularly limited as long as it is sugar-free, and conventionally known basic media such as Hoagland Anon, Murashige Sugouk, White, Rinsmeier Sugouk, niche niche and the like can be mentioned.
【００１７】培養器は、従来の従属栄養的生長の生長・
増殖に使用されていた試験管、三角フラスコなどに代え
て、滅菌されたセル成形苗箱を使用するのが好適であ
る。培養する際の環境は、セル成形苗箱を個別に調節し
てもよいが、多数のセル成形苗箱を大型の培養室に収容
して一挙に調節するのが能率的で好ましい。The incubator is used for conventional heterotrophic growth.
It is preferable to use a sterilized cell-shaped seedling box in place of the test tube, Erlenmeyer flask, etc. used for the propagation. The environment for culturing may be adjusted individually for the cell-molded seedling boxes, but it is efficient and preferable to accommodate a large number of cell-molded seedling boxes in a large-sized culturing chamber at once.
【００１８】本発明方法では、サツマイモの小植物体を
培養して増殖する期間中、培養器および／または培養室
には炭酸ガスを施用するものとし、炭酸ガス濃度を４０
０〜３０００μｍｏｌ／ｍｏｌに保持するのが好まし
い。培養器および／または培養室内の炭酸ガス濃度が４
００μｍｏｌ／ｍｏｌ未満では、小植物体の光合成が必
ずしも十分ではなく、３０００μｍｏｌ／ｍｏｌ以上で
は小植物体に障害が生ずることがあり、いずれも好まし
くない。このように培養器および／または培養室内に炭
酸ガスを施用することにより、独立栄養生長を促進する
ことができ、培養苗を培養器外に移植したり実栽培土壌
に移植して、独立栄養生長条件にさらしても、環境の激
変がないので、馴化過程における活着率が良好で、その
後の生育も良好な培養苗を得ることができる。In the method of the present invention, carbon dioxide is applied to the incubator and / or the culture chamber during the period in which the sweet potato plantlets are cultured and propagated, and the carbon dioxide concentration is 40%.
It is preferable to keep it at 0 to 3000 μmol / mol. Carbon dioxide concentration in the incubator and / or culture chamber is 4
If it is less than 00 μmol / mol, the photosynthesis of the plantlets is not always sufficient, and if it is 3000 μmol / mol or more, the plantlets may be damaged, which is not preferable. By applying carbon dioxide gas to the incubator and / or the culture chamber in this way, autotrophic growth can be promoted, and the cultured seedlings can be transplanted to the outside of the incubator or to the actual cultivation soil for autotrophic growth. Since the environment does not change drastically even when exposed to the conditions, it is possible to obtain a cultured seedling having a good survival rate in the acclimation process and good growth thereafter.
【００１９】本発明方法でサツマイモの小植物体を増殖
培養する際には、セル成形苗箱を増殖培養室に多数配置
し、例えば温度、光の種類、光の強度、光照射方向、明
期・暗期の周期、相対湿度などの増殖培養環境を同一条
件として、培養する。増殖培養に適する最適条件は、適
宜、実験によって確認することにより選ぶことができ
る。When the sweet potato plantlets are grown and cultivated by the method of the present invention, a large number of cell-shaped seedling boxes are arranged in a growth culture chamber, for example, temperature, kind of light, light intensity, light irradiation direction, light period.・ Incubate under the same growth and culture environment such as dark cycle and relative humidity. The optimum conditions suitable for growth culture can be appropriately selected by confirming through experiments.
【００２０】本発明方法では、サツマイモの小植物体を
セル成形苗箱（トレイ）を使用し、当初から一貫して独
立栄養生長させるので、培地の調製が容易で、増殖培養
工程の省力化が達成される。また、増殖培養して得た小
植物体は、実栽培土壌に移植する際に環境の激変を与え
ないで馴化過程に移行でき、馴化過程における活着率が
向上し、その後の生育も良好となる。このように、本発
明方法は多量の無菌サツマイモ苗を得るのに極めて好適
である。In the method of the present invention, the sweet potato plantlets are used in cell-shaped seedling boxes (tray) to consistently allow autotrophic growth from the beginning, so that the medium can be easily prepared and labor saving in the growth culture step can be achieved. To be achieved. In addition, the plantlets obtained by proliferating and culturing can be transferred to the acclimation process without causing a drastic change in the environment when transplanted to actual cultivated soil, the survival rate in the acclimation process is improved, and the subsequent growth is also good. . Thus, the method of the present invention is extremely suitable for obtaining a large amount of sterile sweet potato seedlings.
【００２１】本発明方法では、サツマイモの小植物体を
セル成形苗箱を使用して同一条件下で増殖培養させるこ
とができるので、各セル成形苗箱で生育させた無菌サツ
マイモ苗は、茎部の径・長さ、発根状態、葉数をほぼ均
一とすることができる。従って、本発明方法で得られた
無菌サツマイモ苗は、実栽培土壌に移植する際に自動移
植機による移植が容易になり、大規模栽培にサツマイモ
を収穫する目的の苗として利用され、実栽培土壌に移植
・育成される。In the method of the present invention, since sweet potato plantlets can be grown and cultured under the same conditions using cell-shaped seedling boxes, the sterilized sweet potato seedlings grown in each cell-shaped seedling box are The diameter, length, rooting state, and number of leaves can be made almost uniform. Therefore, the aseptic sweet potato seedlings obtained by the method of the present invention are easily transplanted by an automatic transplanter when transplanted to a real cultivated soil, and are used as seedlings for the purpose of harvesting sweet potatoes in a large-scale cultivation. Will be transplanted and trained.
【００２２】[0022]
【実施例】以下、本発明を実施例に基いて詳細に説明す
るが、本発明はその趣旨を超えない限り、以下に記載の
例に限定されるものではない。EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to the examples described below unless departing from the gist.
【００２３】［実施例１］口径１６mm、深さ２５mmのセ
ルに、無菌化したロックウールを詰めたポリカーボネー
ト製のセル成形苗箱（トレイ）（みのる産業社製）３５
個を、縦１９０mm、横１４５mm、高さ８１mmの透明なポ
リスチレン製の培養器（Baumgartner Papiers SA社製）
内に設置した。各々のセルトレイに、糖類を含まないホ
ーグランド・アノンの液体培地を、培養器当たり１８０
ミリリットル宛支持させて培地とした。[Example 1] Cell-made seedling box (tray) made of polycarbonate (sterilized by Minoru Sangyo Co., Ltd.) 35 made of polycarbonate in which cells having a diameter of 16 mm and a depth of 25 mm are filled with sterilized rock wool.
A piece of transparent polystyrene incubator (Baumgartner Papiers SA) measuring 190 mm in length, 145 mm in width and 81 mm in height.
Installed inside. Each cell tray was filled with saccharide-free Hogland Anone broth at 180 per well.
The medium was supported by supporting milliliters.
【００２４】従来法にしたがって、光混合栄養培養した
サツマイモ（Ipomoea batatas(L.)Lam.品種：ベニアズ
マ）を、葉１枚を含む単節に切り分け、各セルトレイの
上記培地上にその単節を置床し、上記培養器に通気性フ
ィルタを設け、培養試験開始から７日目までは通気性フ
ィルタを透明フィルムで覆い、８日目以降は透明フィル
ムを取り培養器内を気体が通るようにして、合計２８日
間光独立栄養培養で、増殖培養させた。増殖培養させる
際の環境条件は、明期の温度を２４〜２８±１℃、暗期
の温度を２０±１℃、明期の相対湿度を６０〜８０％、
暗期の相対湿度を８０〜９０％、白色蛍光ランプの照射
強度を１００μｍｏｌ／ｍ2／ｓｅｃ、照射時間を１６
時間／日、炭酸ガス濃度を１０００〜１１００μｍｏｌ
／ｍｏｌ（以下、「高濃度」と言う）とした。培養２８
日後に、小植物体の生体重、乾物重、乾物率、葉数およ
び葉面積および葉部乾物重当たりのクロロフィル濃度
（以下、「クロロフィル濃度」と言う）を測定した。そ
の結果を表−１に示す。According to the conventional method, sweet potato (Ipomoea batatas (L.) Lam. Variety: Venezuma) cultivated by photo-mix nutrient feeding was cut into single nodes containing one leaf, and the single nodes were placed on the above medium of each cell tray. Place it on the floor, install a breathable filter on the incubator, cover the breathable filter with a transparent film from the 7th day after the start of the culture test, and remove the transparent film from the 8th day onwards to allow gas to pass through the incubator. For a total of 28 days, the cells were proliferated by photoautotrophic culture. The environmental conditions for the growth culture are as follows: the light period temperature is 24 to 28 ± 1 ° C., the dark period temperature is 20 ± 1 ° C., and the light period relative humidity is 60 to 80%.
The relative humidity in the dark period is 80 to 90%, the irradiation intensity of the white fluorescent lamp is 100 μmol / m 2 / sec, and the irradiation time is 16
Time / day, carbon dioxide concentration of 1000 to 1100 μmol
/ Mol (hereinafter referred to as "high concentration"). Culture 28
After a day, the fresh weight, dry matter weight, dry matter rate, leaf number and leaf area, and chlorophyll concentration per leaf dry matter weight (hereinafter referred to as "chlorophyll concentration") of the plantlets were measured after a day. The results are shown in Table-1.
【００２５】［実施例２］実施例１に記載の例におい
て、無菌化したロックウールに代えて無菌化したバーミ
キュライトを使用した他は、同例におけると同様の手順
で合計２８日間光独立栄養培養で、増殖培養させた。培
養２８日後に、小植物体の生体重、乾物重、乾物率、葉
数および葉面積および葉部乾物重当たりのクロロフィル
濃度などを、常法に従って測定した。その結果を表−１
に示す。[Example 2] In the example described in Example 1, except that sterilized vermiculite was used instead of sterilized rock wool, photoautotrophic culture was performed for a total of 28 days by the same procedure as in the same example. Then, it was proliferated and cultured. After 28 days of culturing, the fresh weight, dry matter weight, dry matter rate, number of leaves and leaf area, and chlorophyll concentration per leaf dry matter weight of the plantlets were measured according to conventional methods. Table 1 shows the results.
Shown in
【００２６】［比較例１］実施例１に記載の例におい
て、無菌化したロックウールに代えて糖類を含まない寒
天を使用した他は、同例におけると同様の手順で合計２
８日間光独立栄養培養で、増殖培養させた。培養２８日
後に、小植物体の生体重、乾物重、乾物率、葉数および
葉面積および葉部乾物重当たりのクロロフィル濃度を測
定した。その結果を表−１に示す。[Comparative Example 1] In the example described in Example 1, except that sugar-free agar was used in place of the sterilized rock wool, a total of 2 was obtained by the same procedure as in the same example.
Proliferation culture was carried out by photoautotrophic culture for 8 days. After 28 days of culturing, the fresh weight, dry matter weight, dry matter rate, number of leaves and leaf area, and chlorophyll concentration per leaf dry matter weight of the plantlets were measured. The results are shown in Table-1.
【００２７】［実施例３］実施例１に記載の例におい
て、白色蛍光ランプの照射強度を１７０μｍｏｌ／ｍ2
／ｓｅｃと変更した他は、同例におけると同様の手順で
合計２８日間光独立栄養培養で、増殖培養させた。培養
２８日後に、小植物体の生体重、乾物重、乾物率、葉数
および葉面積および葉部乾物重当たりのクロロフィル濃
度を測定した。その結果を表−１に示す。[Example 3] In the example described in Example 1, the irradiation intensity of the white fluorescent lamp was 170 μmol / m 2.
/ Sec, except that the procedure was the same as in the same example, and the cells were proliferated by photoautotrophic culture for a total of 28 days. After 28 days of culturing, the fresh weight, dry matter weight, dry matter rate, number of leaves and leaf area, and chlorophyll concentration per leaf dry matter weight of the plantlets were measured. The results are shown in Table-1.
【００２８】［実施例４］実施例２に記載の例におい
て、白色蛍光ランプの照射強度を１７０μｍｏｌ／ｍ2
／ｓｅｃと変更した他は、同例におけると同様の手順で
合計２８日間光独立栄養培養で、増殖培養させた。培養
２８日後に、小植物体の生体重、乾物重、乾物率、葉数
および葉面積および葉部乾物重当たりのクロロフィル濃
度を測定した。その結果を表−１に示す。Example 4 In the example described in Example 2, the irradiation intensity of the white fluorescent lamp was 170 μmol / m 2.
/ Sec, except that the procedure was the same as in the same example, and the cells were proliferated by photoautotrophic culture for a total of 28 days. After 28 days of culturing, the fresh weight, dry matter weight, dry matter rate, number of leaves and leaf area, and chlorophyll concentration per leaf dry matter weight of the plantlets were measured. The results are shown in Table-1.
【００２９】［比較例２］実施例３に記載の例におい
て、無菌化したロックウールに代えて糖類を含まない寒
天を使用した他は、同例におけると同様の手順で合計２
８日間光独立栄養培養で、増殖培養させた。培養２８日
後に、小植物体の生体重、乾物重、乾物率、葉数および
葉面積および葉部乾物重当たりのクロロフィル濃度を測
定した。その結果を表−１に示す。[Comparative Example 2] In the example described in Example 3, a total of 2 was obtained by the same procedure as in the same example except that sugar-free agar was used in place of the sterilized rock wool.
Proliferation culture was carried out by photoautotrophic culture for 8 days. After 28 days of culturing, the fresh weight, dry matter weight, dry matter rate, number of leaves and leaf area, and chlorophyll concentration per leaf dry matter weight of the plantlets were measured. The results are shown in Table-1.
【００３０】［比較例３］比較例１に記載の例におい
て、通気性フィルタのない培養器を用い、液体成分とし
てショ糖を２０ｇ／ｌ含有するムラシゲ・スグークを使
用し、白色蛍光ランプの照射強度を５０μｍｏｌ／ｍ2
／ｓｅｃとし、培養試験開始から合計２８日間光混合栄
養培養で、増殖培養させた。培養器の炭酸ガス濃度は、
３５０〜５００μｍｏｌ／ｍｏｌ（以下、「低濃度」と
言う）とした。培養２８日後に、小植物体の乾物重、生
体重、乾物率、葉面積および葉部乾物重当たりのクロロ
フィル濃度を測定した。その結果を表−１に示す。[Comparative Example 3] In the example described in Comparative Example 1, an incubator without a breathable filter was used, Murashige Suguuk containing 20 g / l of sucrose as a liquid component was used, and irradiation with a white fluorescent lamp was performed. Strength of 50 μmol / m 2
/ Sec, and a total of 28 days from the start of the culture test were propagated by photo-mixed nutrient culture. The carbon dioxide concentration of the incubator is
It was set to 350 to 500 μmol / mol (hereinafter referred to as “low concentration”). After 28 days of culturing, the dry matter weight, fresh weight, dry matter rate, leaf area and chlorophyll concentration per leaf dry matter of the plantlets were measured. The results are shown in Table-1.
【００３１】[0031]
【表１】 [Table 1]
【００３２】表−１より、次のことが明らかである。 (1) 全乾物重について見ると、光独立栄養培養区（実施
例１〜実施例４）の値が光混合栄養培養区（比較例３）
の値の１．４（実施例１）〜２．１倍（実施例３）であ
る。 (2) 全生体重について見ると、光独立栄養培養区（実施
例１〜実施例４）の値が光混合栄養培養区（比較例３）
の値の１．３（実施例１）〜１．９倍（実施例３）であ
る。 (3) 乾物率について見ると、光独立栄養培養区（実施例
１〜実施例４）と光混合栄養培養区（比較例３）とでは
有意差がある。The following is clear from Table-1. (1) Regarding the total dry matter weight, the values in the photoautotrophic culture (Examples 1 to 4) are the values in the photomixture nutrient culture (Comparative Example 3).
Is 1.4 times (Example 1) to 2.1 times (Example 3). (2) Regarding the total fresh weight, the values of the photoautotrophic culture (Examples 1 to 4) are the values of the photomixture nutrient culture (Comparative Example 3).
The value is 1.3 (Example 1) to 1.9 times (Example 3). (3) Regarding the dry matter rate, there is a significant difference between the photoautotrophic culture section (Examples 1 to 4) and the photomixture nutrient culture section (Comparative Example 3).
【００３３】(4) 葉面積について見ると、光独立栄養培
養区（実施例１〜実施例４）の値が光混合栄養培養区
（比較例３）の値の１．６（実施例１）〜２．４倍（実
施例３）である。培地として寒天を用いた区（比較例１
と比較例２）では、他の試験区に比べて葉色が薄く、葉
部に養分欠乏と思われる症状が認められた。 (5) 液体成分を支持する媒体として、ロックウールまた
はバーミキュライトを使用した試験区は、乾物重、生体
重、乾物率および葉面積は、寒天を支持体として使用し
た試験区のものと同等であることから、ロックウールま
たはバーミキュライトを支持体として使用することがで
きる。(4) Regarding the leaf area, the value of the photoautotrophic culture (Examples 1 to 4) is 1.6 (Example 1) which is the value of the photomixture nutrient culture (Comparative Example 3). Up to 2.4 times (Example 3). Section using agar as the medium (Comparative Example 1
In Comparative Example 2), the leaf color was lighter than that of the other test plots, and the symptoms of nutrient deficiency were observed in the leaves. (5) The test area using rockwool or vermiculite as the medium for supporting the liquid component has the same dry matter weight, fresh weight, dry matter rate and leaf area as those of the test area using agar as the support. Therefore, rockwool or vermiculite can be used as a support.
【００３４】[0034]
【発明の効果】本発明は、次のような特別に有利な効果
を奏し、その産業上の利用価値は極めて大である。 １．本発明方法では、無菌状態の培地を使用することに
より、遺伝的に優れた品質の無菌サツマイモ苗を、従来
の培養法に比し１．５〜２倍程度の成長速度で、能率的
に得ることができる。 ２．本発明方法では、支持体に液体成分を支持させた無
糖培地を使用するので、培養期間中に雑菌による汚染の
極めて少ない無菌サツマイモ苗を得ることができる。 ３．本発明方法では、当初から一貫して独立栄養生長さ
せるので、培地の調製が容易で、増殖培養工程の省力化
が可能である。また、増殖培養して得た無菌サツマイモ
苗は、実栽培土壌に移植する際に環境の激変を与えない
で馴化過程に移行でき、馴化過程における活着率が向上
し、その後の生育も良好となる。 ４．本発明方法で得られた無菌サツマイモ苗は、実栽培
土壌に移植する際に自動移植機による移植が容易にな
り、大規模栽培にサツマイモを収穫する目的の苗として
好適である。INDUSTRIAL APPLICABILITY The present invention has the following special advantageous effects and its industrial utility value is extremely large. 1. In the method of the present invention, by using a sterile medium, sterile sweet potato seedlings of genetically excellent quality can be efficiently obtained at a growth rate of about 1.5 to 2 times that of conventional culture methods. be able to. 2. In the method of the present invention, since a sugar-free medium in which a liquid component is supported on a support is used, it is possible to obtain a sterile sweet potato seedling that is extremely free from contamination by miscellaneous bacteria during the culture period. 3. According to the method of the present invention, autotrophic growth is consistently performed from the beginning, so that the medium can be easily prepared and labor for the growth culture step can be saved. In addition, aseptic sweet potato seedlings obtained by growth culture can be transferred to the acclimation process without causing drastic changes in the environment when transplanted to actual cultivation soil, the survival rate in the acclimation process is improved, and the subsequent growth is also good. . 4. The aseptic sweet potato seedlings obtained by the method of the present invention are suitable for seedlings for the purpose of harvesting sweet potatoes for large-scale cultivation, because transplantation by an automatic transplanter becomes easy when transplanting to actual cultivation soil.
Claims (4)
体を培養して無菌サツマイモ苗を増殖する方法におい
て、支持体に液体成分を支持させた無糖培地を使用し、
無菌状態で増殖することを特徴とする無菌サツマイモ苗
の増殖方法。1. A method for cultivating sweet potato plantlets by tissue culture to grow sterile sweet potato seedlings, using a sugar-free medium in which a liquid component is supported on a support,
A method for growing aseptic sweet potato seedlings, which is characterized by growing in a sterile state.
ュライトである請求項１記載の無菌サツマイモ苗の増殖
方法。2. The method for growing a sterile sweet potato seedling according to claim 1, wherein the support is rock wool or vermiculite.
で行う請求項１または請求項２記載の無菌サツマイモ苗
の増殖方法。3. The method for growing a sterile sweet potato seedling according to claim 1, wherein the plantlets are cultured and grown in a cell-molded seedling box.
ス濃度を４００〜３０００μｍｏｌ／ｍｏｌに保持す
る、請求項１ないし請求項３いずれか１項に記載の無菌
サツマイモ苗の増殖方法。4. The method for growing aseptic sweet potato seedlings according to claim 1, wherein carbon dioxide gas is applied during the culture period to maintain the carbon dioxide gas concentration at 400 to 3000 μmol / mol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10975996A JP3877800B2 (en) | 1996-04-30 | 1996-04-30 | Propagation method of aseptic sweet potato seedlings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10975996A JP3877800B2 (en) | 1996-04-30 | 1996-04-30 | Propagation method of aseptic sweet potato seedlings |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH09294495A true JPH09294495A (en) | 1997-11-18 |
| JP3877800B2 JP3877800B2 (en) | 2007-02-07 |
Family
ID=14518525
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10975996A Expired - Lifetime JP3877800B2 (en) | 1996-04-30 | 1996-04-30 | Propagation method of aseptic sweet potato seedlings |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP3877800B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1048203A2 (en) | 1999-04-27 | 2000-11-02 | Nisshinbo Industries, Inc. | Method for producing plantlets of sweet potato |
| JP2015037385A (en) * | 2013-08-18 | 2015-02-26 | 国立大学法人島根大学 | Cultivation method of sweet potato |
| CN116098060A (en) * | 2020-06-10 | 2023-05-12 | 甘肃省农业科学院经济作物与啤酒原料研究所（甘肃省农业科学院中药材研究所） | Rooting culture method for angelica sinensis tissue culture seedlings |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102257958B (en) * | 2011-05-17 | 2013-06-05 | 北方民族大学 | Medium maturity virus-free potato basic seedling culture medium and preparation method thereof |
| CN102257959B (en) * | 2011-05-17 | 2013-06-05 | 北方民族大学 | Culture medium for late maturing potato detoxified basic seedling and preparation method thereof |
| CN102972355B (en) * | 2012-12-05 | 2014-12-10 | 华南农业大学 | Method for breeding, culturing and storing plant nematodes by using sweet potato calluses |
| CN103548684A (en) * | 2013-10-31 | 2014-02-05 | 定西市凯凯生态园植物快繁有限公司 | Inoculation method of tissue culture seedlings of potatoes |
-
1996
- 1996-04-30 JP JP10975996A patent/JP3877800B2/en not_active Expired - Lifetime
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1048203A2 (en) | 1999-04-27 | 2000-11-02 | Nisshinbo Industries, Inc. | Method for producing plantlets of sweet potato |
| EP1048203A3 (en) * | 1999-04-27 | 2001-11-28 | Nisshinbo Industries, Inc. | Method for producing plantlets of sweet potato |
| JP2015037385A (en) * | 2013-08-18 | 2015-02-26 | 国立大学法人島根大学 | Cultivation method of sweet potato |
| CN116098060A (en) * | 2020-06-10 | 2023-05-12 | 甘肃省农业科学院经济作物与啤酒原料研究所（甘肃省农业科学院中药材研究所） | Rooting culture method for angelica sinensis tissue culture seedlings |
Also Published As
| Publication number | Publication date |
|---|---|
| JP3877800B2 (en) | 2007-02-07 |
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