JPS6258934A - Mass propagation of potato by tissue culture - Google Patents
Mass propagation of potato by tissue cultureInfo
- Publication number
- JPS6258934A JPS6258934A JP19814085A JP19814085A JPS6258934A JP S6258934 A JPS6258934 A JP S6258934A JP 19814085 A JP19814085 A JP 19814085A JP 19814085 A JP19814085 A JP 19814085A JP S6258934 A JPS6258934 A JP S6258934A
- Authority
- JP
- Japan
- Prior art keywords
- buds
- medium
- culturing
- axillary
- seedlings
- 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.)
- Pending
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】
ビ)産業上の利用分野
本発明は、ヤマノイモ科植物ナガイモ
(Dioscorea opposita THUNB
あるいはDloscorea batatas DEC
NE ) (7)組織培養ニヨる大量増殖法に関する。DETAILED DESCRIPTION OF THE INVENTION B) Industrial application field
Or Dloscorea batatas DEC
(7) Relating to a method for mass propagation of tissue culture.
より詳しくは、植物の蔓より腋芽及び頂芽を含む茎の切
片を採取し、殺菌後無菌的に培養することにより、ひと
つの芽から多数の芽の形成を誘導した後、個々の芽を分
割し茎葉再生・発根を経て幼植物体を得ることにより、
ナガイモを大量増殖する方法に関する。本方法により、
ウィルスを除去した無病株等の優良系統を短期間の内に
遺伝的に均一な状態で大量に増殖することが可能となる
。More specifically, a section of the stem containing the axillary and apical buds is collected from the vine of the plant, sterilized and cultured aseptically to induce the formation of many buds from one bud, and then the individual buds are divided. By obtaining seedlings through stem and leaf regeneration and rooting,
Concerning a method for mass propagating Chinese potatoes. With this method,
It becomes possible to propagate large quantities of superior strains, such as disease-free strains from which viruses have been removed, in a genetically uniform state within a short period of time.
(ロ)従来技術及び問題点
ナガイモは他の多くのヤマノイモ科植物と同様、種子繁
殖を行わない栄養繁殖性植物である。通常の栽培では地
下部の担根体(云わゆる芋)を分割するか、蔓の葉液に
形成されるムカゴによって繁殖が図られるが、増殖率は
極めて低く、年間数十〜百倍程度である。(b) Prior Art and Problems Like many other plants of the Dioscoreaceae family, Japanese yam is a vegetatively propagated plant that does not reproduce by seeds. In normal cultivation, reproduction is attempted by dividing the underground root-bearing bodies (so-called potatoes) or by using the vines formed in the leaf sap of the vines, but the multiplication rate is extremely low, ranging from several tens to a hundred times more per year. .
;a織培養による増殖法として、茎頂(生長点)培養法
・カルス培養法等の方法による増殖の試みも多数あるが
、本植物種の培地上での生育は概して遅く、完全な個体
再生には長期間を要する上に、実用的増殖に充分な効率
を達成することは困難であった。さらに、これらのカル
スを経る手法では、培養中に染色体変異等を起こすこと
から再生植物体に遺伝的な変異が生じやすく、均一なり
ローン植物を大量に得る手段としては問題点が多い。;There have been many attempts at propagation using methods such as shoot apex (growing point) culture and callus culture, but the growth of this plant species on medium is generally slow and complete individual regeneration is not possible. In addition to requiring a long period of time, it has been difficult to achieve sufficient efficiency for practical propagation. Furthermore, these callus-based methods tend to cause genetic variations in regenerated plants due to chromosomal mutations during culturing, and there are many problems as a means of obtaining uniform lawn plants in large quantities.
日本の栽培種ナガイモの品種群であるながいも、いちょ
ういも、つくねいも、あるいは、やまといも等について
は、腋芽より培地上で多芽を形成させた上でこれを分割
し、それぞれを個体として再生させることにより多数の
幼植物を取得する手法については知られていない。For Japanese cultivars such as Nagaimo, Ichoimo, Tsukuneimo, and Yamatoimo, multiple buds are formed on the medium from the axillary buds, and then these are divided and each is regenerated as an individual. There is no known method for obtaining a large number of seedlings.
(/→ 発明の目的及び構成
発明者らは、ヤマノイモ科植物ナガイモの多芽形成条件
及び分割した芽からの幼植物再生条件について鋭意検討
した結果、サイトカイニンを10 M以上含有する培
地で培養すると高率の多芽形成が得られ、分割した芽を
植物生長ホルモンを含有しないか又は植物生長ホルモン
濃度の低い培地で培養することにより効率良く幼植物体
に再生できることを見いだし、本発明を完成した。(/→ Purpose and composition of the invention As a result of intensive studies on the conditions for multi-bud formation and the conditions for regeneration of seedlings from divided buds of the Dioscoreaceae family, the inventors found that when cultured in a medium containing 10 M or more of cytokinin, The present invention has been completed based on the discovery that a high rate of multi-bud formation can be obtained and that the split buds can be efficiently regenerated into seedlings by culturing in a medium that does not contain plant growth hormone or has a low concentration of plant growth hormone.
本発明は、ナガイモの腋芽又は頂芽を、培地上で無菌的
に培養することにより多芽を形成させた後、これらの多
芽を分割し個々の芽を培地に植継ぐことにより、多数の
幼植物体を得ることを特徴とするヤマノイモ科植物ナガ
イモの幼植物体の生産方法及びこれらの幼植物体を育成
することを特徴とするナガイモの増殖方法を提供する。The present invention involves forming multiple buds by aseptically culturing the axillary buds or apical buds of Japanese potato on a medium, and then dividing these multiple buds and transplanting the individual buds onto the medium. Provided are a method for producing young plants of Dioscoreaceae, a plant belonging to the Dioscoreaceae family, which is characterized by obtaining young plants, and a method for propagating the young plants, which is characterized by growing these young plants.
本発明の実施方法について、(1)材料とする植物の育
成、(2)腋芽及び頂芽の採取・殺菌、(3)多芽形成
の誘導、(4)多芽の分割・植継ぎ、(5)幼植物の馴
化・定植、の以上5段階に分け、以下、順を追って詳細
に記述する。Regarding the method of carrying out the present invention, (1) growing plants to be used as materials, (2) collecting and sterilizing axillary buds and apical buds, (3) inducing formation of multiple buds, (4) dividing/transplanting multiple buds, ( 5) Acclimation of young plants and planting, which are divided into the above five stages, will be described in detail below in order.
(1) 材料とする植物の育成
本発明が適用されるヤマノイモ科植物ナガイモとしては
、ながいも、いちょういも、つくねいも、あるいは、や
まといも等があげられる。これらの種芋を常法により切
断・消毒・催芽した後、ポットあるいは圃場に植付ける
。(1) Cultivation of plants used as materials Examples of the Dioscoreaceae plant to which the present invention is applied include long yam, ginkgo yam, Tsukune yam, and yam yam. These seed potatoes are cut, sterilized, and sprouted using conventional methods, and then planted in pots or fields.
当然ムカゴを利用することも可能である。Of course, it is also possible to use Mukago.
通常栽培の場合、切手切片の大きさは100〜250y
が良いとされるが、本方法の場合には蔓の採取が目的で
あるので、1片当り50〜100yで充分である。ナガ
イモの場合、種芋の大きさは品種によって異なるが、一
般にながいもで1000f以上、つくねいもで5001
程度の重量がある。従って、従来の方法である種芋の切
断により10〜20倍の増殖が実現される(第1段階増
殖)。In the case of normal cultivation, the size of the stamp section is 100 to 250y.
However, since the purpose of this method is to collect vines, 50 to 100 y per piece is sufficient. In the case of Nagaimo, the size of the seed potato varies depending on the variety, but in general, the size of the seed potato is 1000f or more for Nagaimo, and 5000F for Tsukuneimo.
It has some weight. Therefore, a 10 to 20-fold multiplication is achieved by cutting seed potatoes using the conventional method (first stage multiplication).
切手切片は切断直後切り口を消毒しておくのが望ましい
。消毒剤としては、ベンレイト、チウラム、キャブタン
、あるいはオーツサイド等を用い、水和剤浸漬処理ある
いは粉衣処理を行うが、簡便法として、消石灰の粉衣も
有効である。消毒後、切字切片を室温・暗所に保存し、
切り口の癒傷及び催芽を図るのが望ましい。通常約1カ
月でアズキ粒大の芽が切字切片の表面に形成され、これ
をポットあるいは圃場に植付ける。本方法の場合には、
切字切片の栄養分のみで充分i?−蔓が伸長するため、
ごく普通の土壌を用いれば良い。地上部に芽が出た後、
支柱を立て蔓を誘引することにより、蔓の伸長・茎葉の
繁茂を促進することができる。蔓が下垂すると葉液部の
生長点がムカゴ形成に向かう傾向があり、このような腋
芽は本方法には適当でないために、栽培管理上注意が必
要である。It is desirable to disinfect the cut end of the stamp section immediately after cutting. As a disinfectant, benlate, thiuram, cabtan, or oatside is used, and hydrating agent immersion treatment or powder coating treatment is performed, but as a simple method, slaked lime powder coating is also effective. After disinfection, store the cut sections at room temperature and in the dark.
It is desirable to heal the cut end and encourage germination. Usually, in about a month, sprouts the size of adzuki beans are formed on the surface of the cuttings, and these are planted in pots or fields. In the case of this method,
Is the nutrients in the cut section sufficient? - Because the vines elongate,
Just use ordinary soil. After sprouting above ground,
By erecting a support and attracting vines, it is possible to promote the elongation of the vines and the flourishing of the stems and leaves. When the vine droops, the growing points of the leaf sap tend to form bulrushes, and such axillary buds are not suitable for this method, so care must be taken in cultivation management.
(2)腋芽及び頂芽の採取、殺菌
高さ1〜2mに達した蔓を切断し、腋芽及び頂芽を採取
する。一般にヤマノイモ科植物では頂芽優勢が見られ、
頂芽が健全である間は腋芽あるいは側芽は伸長せず葉液
の部分に控えているのみである。鉦の切断の際、地上部
全部を切り取るのではなく一部腋芽を残しておくことに
より、切断後その部分から蔓が伸長し、再び腋芽の採取
を行うことができる。(2) Collection and sterilization of axillary buds and apical buds Cut the vines that have reached a height of 1 to 2 m and collect the axillary buds and apical buds. In general, apical dominance is observed in Dioscoreaceae plants,
While the apical bud is healthy, the axillary or lateral buds do not elongate and only remain in the leaf sap region. When cutting the gong, rather than cutting off the entire above-ground part, by leaving some of the axillary buds, the vines will grow from that part after cutting, and the axillary buds can be harvested again.
これにより、1切芋切片から通常200〜300の腋芽
を採取することが可能となる(第2段階増殖)。This makes it possible to usually collect 200 to 300 axillary buds from one potato section (second stage multiplication).
採取した蔓は実験室内で腋芽ごとに切り離す。最も望ま
しい形としては、腋芽の部分を茎に残したまま軸の上下
それぞれ51程度、さらに葉柄も5−程度を残して切断
する。このようなものは外植体として適度な大きさを持
ち扱い易い。採取した腋芽は、培養前に殺菌しておくの
が望ましい。殺菌法は通常、1%程度の次亜塩素酸ナト
リウム溶成に5〜10分浸漬した後、無菌条件下で滅菌
水による洗浄を8〜5回行う。この処理により、雑菌の
混入率を置床腋芽数当り5%以下に押えることができる
。The collected vines are separated into axillary buds in the laboratory. The most desirable form is to cut the axillary buds leaving about 51 cm above and below the stem, and also leave about 5 cm of the petiole. Such a plant has a suitable size and is easy to handle as an explant. It is desirable to sterilize the collected axillary buds before culturing. The sterilization method usually involves immersing the product in approximately 1% sodium hypochlorite solution for 5 to 10 minutes, followed by washing with sterile water 8 to 5 times under aseptic conditions. By this treatment, the contamination rate of various bacteria can be suppressed to 5% or less per number of axillary buds placed on the bed.
(3)多芽形成の誘導
殺菌した腋芽を、無菌条件下で適当な植物組織培養用培
地に置床する。培地はL8(Linsmaier &
8koog 1964 )あるいはMS(Murash
ige & Skoog 1962 )等の、市販の植
物組織培養用培地で充分であり、1〜5%、好ましくは
2〜4%濃度のショ糖及び0.5〜1.5%、好ましく
は0.8〜1.2%濃度の寒天を添加したものを用いる
。多芽を形成させる要因として最も重要なものは、植物
生長ホルモンのサイトカイニン類の添加である。サイト
カイニン類としては、カイネチン(Kinetin)6
−ベンジルアミノプリン(BA)、あるいは2−イソペ
ンテニルアデニン(2iP)等、市販のものを使用する
ことができる。多芽形成に有効なサイトカイニン類の添
加濃度は10 M以上であり、好ましくは10〜10
M@度が適している。培養器は試験管、三角フラス
コ等どのような形状のものを用いても良いが、直径60
■程度のプラスチックシャーレに10−程度の培地を入
れたものが作業効率上鏝も適している。培養条件として
は、温度は20〜ao”c、好ましくは23〜27°C
が適しており、日長は1000〜10000lux 、
好ましくは2000〜5000 luxで12〜24時
間程度の条件が適している。(3) Induction of multiple bud formation The sterilized axillary buds are placed on a suitable plant tissue culture medium under sterile conditions. The medium was L8 (Linsmaier &
8koog 1964) or MS (Murash
A commercially available plant tissue culture medium, such as G. ige & Skoog 1962), is sufficient, containing sucrose at a concentration of 1-5%, preferably 2-4% and 0.5-1.5%, preferably 0.8%. Agar added with a concentration of ~1.2% is used. The most important factor for forming multiple buds is the addition of cytokinins, which are plant growth hormones. As a cytokinin, kinetin 6
Commercially available products such as -benzylaminopurine (BA) or 2-isopentenyladenine (2iP) can be used. The added concentration of cytokinins effective for multi-bud formation is 10 M or more, preferably 10 to 10 M.
M @ degree is suitable. The incubator can be of any shape, such as a test tube or Erlenmeyer flask, but the incubator must have a diameter of 60 mm.
A trowel is also suitable for work efficiency, such as a plastic petri dish with a size of about 10 cm and a culture medium of about 10 mm. As for the culture conditions, the temperature is 20 to 27 degrees Celsius, preferably 23 to 27 degrees Celsius.
is suitable, and the photoperiod is 1000-10000lux,
Preferably, conditions of 2000 to 5000 lux and 12 to 24 hours are suitable.
以上のような条件で3〜4週間培養すると置床した腋芽
あるいは頂芽の生長点が肥大し、さらに各葉原基の基部
に新たな生長点が多数形成されるのが観察される。培養
7〜8週間で各々の生長点は数対の葉原基を伴った独立
の芽として生長し、ひとつの葉液部に形成された多芽と
しての様相を呈する。葉液当りの芽の形成数は通常10
〜40個である。(第3段階増殖)。When cultured under the above conditions for 3 to 4 weeks, the growing points of the axillary buds or apical buds placed on the bed become enlarged, and it is observed that many new growing points are formed at the base of each leaf primordium. After 7 to 8 weeks of culture, each growing point grows as an independent bud with several pairs of leaf primordia, and appears as multiple buds formed in a single leaf sap region. The number of buds formed per leaf sap is usually 10.
~40 pieces. (Third stage proliferation).
(4)多芽の分割・植継ぎ
多芽が形成された腋芽を培地より取り出し、無菌条件下
でメス等を用い、個々の芽を分割する。芽は各々、ひと
つの生長点と少なくとも2〜3対の葉原基が揃っていれ
ばその後の生育に支障はない。これらの芽を寒天培地上
に植継ぎ、茎葉の展開・発根を図る。培地は(3)と同
様に市販の植物組織培養用培地で充分であるが、植物生
長ホルモンはこの場合、無添加かあるいは10 M以
下の低濃度のサイトカイニン類及びオーキシン類を添加
する。(4) Division and transplantation of multiple buds The axillary buds on which multiple buds have been formed are removed from the medium, and individual buds are divided using a scalpel or the like under sterile conditions. As long as each bud has one growing point and at least 2 to 3 pairs of leaf primordia, there will be no problem with subsequent growth. These buds are transplanted onto an agar medium to develop stems and leaves and root. As in (3), a commercially available plant tissue culture medium is sufficient as the medium, but in this case no plant growth hormones are added, or cytokinins and auxins are added at a low concentration of 10 M or less.
オーキシン類としては、IAA(β−インドール酢酸)
、NAA(α−ナフタレンi[)あるいは2.4−D(
2,4−ジクロロフェノキシ酢酸)等があげられる。培
養条件は、前述と同様である。As auxin, IAA (β-indole acetic acid)
, NAA (α-naphthalene i[) or 2.4-D (
2,4-dichlorophenoxyacetic acid), etc. Culture conditions are the same as described above.
これらの分割された芽の約半数は、同培地上でl〜2カ
月培養することにより、それぞれ1〜2枚の展開した葉
と1〜2本の根を持つ幼植物体として再生することが可
能である。Approximately half of these divided buds can be cultured on the same medium for 1 to 2 months to regenerate as seedlings each with 1 to 2 expanded leaves and 1 to 2 roots. It is possible.
(5)幼植物体の馴化・定植
茎葉と根を再生した幼植物を有菌条件下に移し、外環境
に馴化すると共に健全な苗として育成する。最低成葉が
1枚展開し、1〜2閏の長さの根が1本再生すれば、幼
植物体として外環境に移すことが可能である。馴化は、
幼植物体の根についた寒天をよく洗い落し、バーミキュ
ライト、ピートモス、あるいはパーライト等の排水性の
良い適当な培土に移植し、湿潤な状態に保つことが望ま
しく、この操作により、90%以上の幼植物が活着する
。(5) Acclimatization and planting of seedlings The seedlings with regenerated stems, leaves and roots are transferred to germ-free conditions, acclimated to the outside environment, and grown as healthy seedlings. Once at least one adult leaf has developed and one root with a length of 1 to 2 leaps has been regenerated, the plant can be transferred to the outside environment as a young plant. Acclimation is
It is advisable to thoroughly wash the agar off the roots of the seedlings and transplant them into a suitable potting soil with good drainage, such as vermiculite, peat moss, or perlite, and keep it moist. Plants take root.
培土上で活着した幼植物は2週間程度で新たな茎葉を展
開し、土壌移植可能となる。The seedlings that have taken root on the soil will develop new stems and leaves in about two weeks and can be transplanted into the soil.
以下、実施例を挙げ、本発明を更に詳細に説明する。EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples.
実施例1
胃森県産ナガイモ栽培品種ガンクミジカの腋芽を採取し
、1%次亜塩素酸ナトリウム溶液中で10分間浸漬殺菌
後、無菌水で8回洗浄し以下の条件に従って培養した。Example 1 The axillary buds of Nagaimo cultivar Gankumijika produced in Stomori Prefecture were collected, sterilized by immersion in a 1% sodium hypochlorite solution for 10 minutes, washed eight times with sterile water, and cultured according to the following conditions.
培地は、シラ糖8%及び寒天1%を含みI) H5,8
に調整したMS培地を、口径40日、高さ30mのマヨ
ネーズビンにl〇−分注したものを使用した。植物生長
ホルモンは、サイトカイニンとしてBAを無添加及び1
0 M添加の2区、オーキシンとしてNAAを無添加
、10−6M添加、及び10 Mm加の8区、合せて
2x8=6区の濃度組合せを設けた。各ホルモン濃度区
につき培養ビン2本、腋芽6個を置床し、20”C,2
000lux 12時間照明で2力月間培養した。The medium contains 8% sila sugar and 1% agar I) H5,8
The MS medium adjusted to 100 ml was dispensed into mayonnaise bottles with a diameter of 40 days and a height of 30 m. Plant growth hormone contains BA as cytokinin without additives and 1
A total of 2 x 8 = 6 concentration combinations were provided: 2 groups with 0 M addition, and 8 groups with no NAA addition, 10 -6 M addition, and 10 Mm addition as auxin. Two culture bottles and six axillary buds were placed in each hormone concentration zone, 20"C, 2
The cells were cultured for 2 months under 000lux 12 hour illumination.
第1表 ナガイモ品種ガンクミジカ腋芽の培養2力月後
の形状0 茎葉伸長・発根 発根
カルス化IO多芽形成 多芽形成 カルス化
第1表に見られるように、ホルモン無添加培地では、腋
芽からの茎の伸長及び発根が容易に起こったが、in
vitro 培養条件下でも頂芽優勢が保たれ、伸長す
る茎は1本のみであった。サイトカイニンであるBAが
培地中に添加されると、この腋芽内の頂芽の伸長が抑制
され、葉原基基部に新たな生長点が形成された。オーキ
シンNAAの添加は効果なく、高調度ではカルス化が見
られ、培地の褐変も著しくなり生育の阻害が見られた。Table 1 Shape of axillary buds of Nagaimo cultivar Gankumijika after 2 months of culture 0 Stem and leaf elongation/rooting Rooting
Callus formation IO multi-bud formation Multi-bud formation Callus formation As seen in Table 1, stem elongation and rooting from axillary buds occurred easily in the hormone-free medium, but in
Apical dominance was maintained even under vitro culture conditions, with only one stem elongating. When BA, a cytokinin, was added to the medium, the elongation of the apical bud within this axillary bud was suppressed, and new growth points were formed at the base of the leaf primordium. Addition of auxin NAA had no effect, and at high concentrations, callus formation was observed, browning of the medium was also significant, and growth was inhibited.
実施例2
青森県産ナガイモ品種ガンクミジヵ及びトラクリの2品
種より腋芽を採取し、1%次亜塩素酸ナトリウム溶液中
で10分間浸漬殺菌後、無菌水で3回洗浄し以下の条件
に従って培養した。培地は、シ冒糖396及び寒天1%
を含み、pH5,8に調整したMS培地を直径60m5
.高さ10mのプラスチックシャーレに分注したものを
使用した。植物生長ホルモンは、サイトカイニンとして
BAを10 M添加した。各シャーレに腋芽3〜4個
を置床し、25”C,2000lux 24時間連続照
明で2力月間培養した。Example 2 Axillary buds were collected from two Japanese potato cultivars Gankumijika and Torakuri produced in Aomori Prefecture, sterilized by immersion in a 1% sodium hypochlorite solution for 10 minutes, washed three times with sterile water, and cultured according to the following conditions. The medium is 396 sulfur sugar and 1% agar.
60m5 in diameter containing MS medium adjusted to pH 5.
.. The sample was dispensed into a plastic Petri dish with a height of 10 m and used. As for the plant growth hormone, 10 M BA was added as cytokinin. Three to four axillary buds were placed in each petri dish and cultured for 2 months under 25"C, 2000 lux, 24 hour continuous lighting.
第2表 ナガイモ品種ガンクミジカ及びトラクリの品種
名 置床芽数 0−1010−2020−308()−
切平均芽形成数ガンクミジカ 112 34
42 30 6 16トツクリ
84 18 10 9 2
15第2表に見られるように、多芽形成は容易
かつ高率であり、しかも安定していた。また、品種開蓋
もそれほど大きなものではなく、2力月間の培養で腋芽
当り平均して15〜16個の独立した芽に分割すること
が可能であっtこ 。Table 2 Nagaimo varieties Gankumijika and Trakuri variety name Number of buds placed in bed 0-1010-2020-308()-
Cut-average bud formation number Ganku water deer 112 34
42 30 6 16 Totsukuri
84 18 10 9 2
15 As seen in Table 2, multiple bud formation was easy, high, and stable. In addition, the cultivar's opening size is not very large, and it is possible to divide each axillary bud into an average of 15 to 16 independent buds after two months of culture.
実施例8
ナガイモ品種ガンクミジカの腋芽8個を、実施例2に従
って殺菌・培養し、多芽を得た。Example 8 Eight axillary buds of the Japanese potato cultivar Gankumijika were sterilized and cultured according to Example 2 to obtain multiple buds.
形成された芽を分割し、それぞれを、直径18頷、長さ
130日の試験管に10−のM8ホルモンフリー寒天培
地を斜面状に分注した培地に植継ぎ、25°C116時
間照明で2力月間培゛養した。茎葉の再生及び発根の見
られた幼植物体を試験管より取り出し、培土ボンツルを
約100−人れた直径55■、高さ60ツのプラスチッ
クカップに移植し、25”C。The formed buds were divided and each was transplanted into a test tube with a diameter of 18 nods and a length of 130 days, in which 10-M8 hormone-free agar medium was dispensed in the form of a slant. I cultivated my strength for months. The seedlings that showed regeneration of stems and leaves and rooting were taken out from the test tubes and transplanted into plastic cups with a diameter of 55 cm and a height of 60 cm containing about 100 ml of culture soil, and then placed at 25"C.
24時間連続照明で1力月間馴化を行った。Acclimatization was performed for one month under continuous lighting for 24 hours.
第3表 ナガイモ品種ガンクミジカの多芽分割による増
殖$1 40 19
18$2 27 14
12第3表に見られるように、腋芽より形成した多芽
は30〜40個に分割することが可能で、それらのうち
の約半数を幼植物体として再生することができ、馴化後
の生育も順調であった。Table 3 Multiplication of Nagaimo cultivar Gankumijika by multi-bud division $1 40 19
18$2 27 14
12 As shown in Table 3, the multiple buds formed from the axillary buds can be divided into 30 to 40 pieces, and about half of them can be regenerated as seedlings, and the growth after acclimatization is was also going well.
に)発明の効果
本方法によるナガイモの増殖率は、第3段階増殖率10
倍、第2段階増殖率200倍、第3段階増殖率10倍と
、累計で最低限20.000倍の増殖率を達成すること
ができる。また、本方法による増殖過程はすべて植物本
来の持つ生長点形成能のみを利用したものであり、カル
ス由来の不定胚等を利用する方法と異なり、遺伝的突然
変異を最小限に押え得る方法である。従って本方法によ
り、ウィルスを除去した無病法あるいは選抜された優良
株等、特定の個体あるいは系統を、短期間のうちに遺伝
的に均一な状態で大量に増殖することが可能となる。2) Effect of the invention The multiplication rate of Japanese potato by this method is 3rd stage multiplication rate 10
It is possible to achieve a cumulative growth rate of at least 20,000 times, with a second stage growth rate of 200 times and a third stage growth rate of 10 times. In addition, the propagation process by this method uses only the plant's inherent ability to form growing points, and unlike methods that use callus-derived somatic embryos, it is a method that can minimize genetic mutations. be. Therefore, by this method, it is possible to propagate a large amount of specific individuals or strains in a genetically uniform state in a short period of time, such as virus-free disease-free or selected superior strains.
Claims (8)
上で無菌的に培養することにより多芽を形成させた後、
これらの多芽を分割し個々の芽を培地に植継ぎ培養する
ことにより多数の幼植物体を得ることを特徴とするヤマ
ノイモ科植物ナガイモ幼植物体の生産方法。(1) After forming multiple buds by aseptically culturing the axillary buds or apical buds of Dioscorea yam on a medium,
A method for producing young plants of Dioscoreaceae, a plant belonging to the family Dioscoreaceae, which comprises dividing these multiple buds and cultivating the individual buds in a culture medium to obtain a large number of young plants.
添加した培地を用い培養し多芽を形成することを特徴と
する特許請求の範囲第1項記載の方法。(2) Cytokinin from 10^-^4 to 10^-^6M
2. The method according to claim 1, which comprises culturing using the added medium to form multiple buds.
で12〜24時間の条件で培養し多芽を形成することを
特徴とする特許請求の範囲第2項記載の方法。(3) 20-30℃, photoperiod 1000-10000lux
3. The method according to claim 2, wherein the method is cultured for 12 to 24 hours to form multiple buds.
ーキシンを10^−^7M以下の濃度で添加した培地を
用い培養し幼植物体を得ることを特徴とする特許請求の
範囲第2項或いは第3項記載の方法。(4) Claims 2 or 3, characterized in that seedlings are obtained by culturing using a medium without the addition of plant hormones or with addition of cytokinin and auxin at a concentration of 10^-^7M or less. the method of.
上で無菌的に培養することにより多芽を形成させた後、
これらの多芽を分割し個々の芽を培地に植継ぎ培養する
ことにより多数の幼植物体を得、これらを育成し完全な
植物体として再生することを特徴とするヤマノイモ科植
物ナガイモの増殖方法。(5) After forming multiple buds by aseptically culturing the axillary buds or apical buds of Dioscorea yam on a medium,
A method for propagating yam, a plant belonging to the Dioscoreaceae family, characterized by dividing these multiple buds and culturing individual buds in a medium to obtain a large number of seedlings, which are then grown and regenerated as complete plants. .
添加した培地を用い培養し多芽を形成することを特徴と
する特許請求の範囲第5項記載の方法。(6) Cytokinin from 10^-^4 to 10^-^6M
6. The method according to claim 5, which comprises culturing using the added medium to form multiple buds.
で12〜24時間の条件で培養し多芽を形成することを
特徴とする特許請求の範囲第6項記載の方法。(7) 20-30℃, photoperiod 1000-10000lux
7. The method according to claim 6, wherein the method is cultured for 12 to 24 hours to form multiple buds.
ーキシンを10^−^7M以下の濃度で添加した培地を
用い培養し幼植物体を得ることを特徴とする特許請求の
範囲第6項或いは第7項記載の方法。(8) Claims 6 or 7, characterized in that seedlings are obtained by culturing using a medium without the addition of plant hormones or with addition of cytokinin and auxin at a concentration of 10^-^7M or less. the method of.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP19814085A JPS6258934A (en) | 1985-09-06 | 1985-09-06 | Mass propagation of potato by tissue culture |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP19814085A JPS6258934A (en) | 1985-09-06 | 1985-09-06 | Mass propagation of potato by tissue culture |
Publications (1)
Publication Number | Publication Date |
---|---|
JPS6258934A true JPS6258934A (en) | 1987-03-14 |
Family
ID=16386117
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP19814085A Pending JPS6258934A (en) | 1985-09-06 | 1985-09-06 | Mass propagation of potato by tissue culture |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS6258934A (en) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH01157313A (en) * | 1987-12-14 | 1989-06-20 | Ehime Pref Gov | Novel process for proliferating yam |
US5722705A (en) * | 1995-06-24 | 1998-03-03 | Nec Corporation | Lock structure for cover of electronic appliance |
JP2011083235A (en) * | 2009-10-16 | 2011-04-28 | Kinjirushi Kk | Method for forming multiple shoot from culture seedling and method for producing virus-free seed yam |
CN102860212A (en) * | 2012-10-10 | 2013-01-09 | 江苏省农业科学院 | Method for centrally quickly expanding breeding of yam seeds on seedbed by means of miniature Chinese yam blocks |
CN102870682A (en) * | 2012-10-26 | 2013-01-16 | 山东省农业科学院蔬菜研究所 | Culture medium for in-vitro induced regeneration plants of double-haploid stems of potatoes |
CN104350905A (en) * | 2014-10-17 | 2015-02-18 | 湖北理工学院 | Yam seedling cultivation method |
CN108990736A (en) * | 2018-07-07 | 2018-12-14 | 富川凯邦农资经营部 | A kind of Chinese yam implantation methods of high yield |
CN109937825A (en) * | 2019-04-08 | 2019-06-28 | 衡阳市蔬菜研究所 | A kind of sole potato extremely early mature High-quality Cultivation method |
-
1985
- 1985-09-06 JP JP19814085A patent/JPS6258934A/en active Pending
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH01157313A (en) * | 1987-12-14 | 1989-06-20 | Ehime Pref Gov | Novel process for proliferating yam |
JPH0342059B2 (en) * | 1987-12-14 | 1991-06-26 | ||
US5722705A (en) * | 1995-06-24 | 1998-03-03 | Nec Corporation | Lock structure for cover of electronic appliance |
GB2302611B (en) * | 1995-06-24 | 1998-05-13 | Nec Corp | Lock structure for cover of electronic appliance |
JP2011083235A (en) * | 2009-10-16 | 2011-04-28 | Kinjirushi Kk | Method for forming multiple shoot from culture seedling and method for producing virus-free seed yam |
CN102860212A (en) * | 2012-10-10 | 2013-01-09 | 江苏省农业科学院 | Method for centrally quickly expanding breeding of yam seeds on seedbed by means of miniature Chinese yam blocks |
CN102870682A (en) * | 2012-10-26 | 2013-01-16 | 山东省农业科学院蔬菜研究所 | Culture medium for in-vitro induced regeneration plants of double-haploid stems of potatoes |
CN104350905A (en) * | 2014-10-17 | 2015-02-18 | 湖北理工学院 | Yam seedling cultivation method |
CN104350905B (en) * | 2014-10-17 | 2016-04-20 | 湖北理工学院 | A kind of Chinese yam seedling-cultivating method |
CN108990736A (en) * | 2018-07-07 | 2018-12-14 | 富川凯邦农资经营部 | A kind of Chinese yam implantation methods of high yield |
CN109937825A (en) * | 2019-04-08 | 2019-06-28 | 衡阳市蔬菜研究所 | A kind of sole potato extremely early mature High-quality Cultivation method |
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