JPH02222627A - Production of banana seedling by culture of plant tissue - Google Patents
Production of banana seedling by culture of plant tissueInfo
- Publication number
- JPH02222627A JPH02222627A JP1043201A JP4320189A JPH02222627A JP H02222627 A JPH02222627 A JP H02222627A JP 1043201 A JP1043201 A JP 1043201A JP 4320189 A JP4320189 A JP 4320189A JP H02222627 A JPH02222627 A JP H02222627A
- Authority
- JP
- Japan
- Prior art keywords
- ppm
- banana
- medium
- buds
- cell
- 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.)
- Granted
Links
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- 239000007788 liquid Substances 0.000 claims abstract description 21
- 238000012258 culturing Methods 0.000 claims abstract description 11
- 238000013019 agitation Methods 0.000 claims abstract description 9
- 238000005273 aeration Methods 0.000 claims abstract description 8
- 235000020415 coconut juice Nutrition 0.000 abstract description 19
- 235000015097 nutrients Nutrition 0.000 abstract description 5
- 238000005286 illumination Methods 0.000 abstract description 3
- 230000003247 decreasing effect Effects 0.000 abstract 1
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- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 14
- 229960001669 kinetin Drugs 0.000 description 14
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- 229960000367 inositol Drugs 0.000 description 6
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000012882 rooting medium Substances 0.000 description 6
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 6
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 5
- 239000004471 Glycine Substances 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 238000004382 potting Methods 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 4
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 4
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 4
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 4
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 4
- 229960000344 thiamine hydrochloride Drugs 0.000 description 4
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 4
- 239000011747 thiamine hydrochloride Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 3
- 235000021015 bananas Nutrition 0.000 description 3
- 235000001968 nicotinic acid Nutrition 0.000 description 3
- 229960003512 nicotinic acid Drugs 0.000 description 3
- 239000011664 nicotinic acid Substances 0.000 description 3
- 230000000644 propagated effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 101100136092 Drosophila melanogaster peng gene Proteins 0.000 description 2
- -1 Nicotinic acid hydrochloride Pyridoxine Hydrochloride Chemical compound 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 2
- 229960000686 benzalkonium chloride Drugs 0.000 description 2
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000003864 humus Substances 0.000 description 2
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 description 2
- 239000010451 perlite Substances 0.000 description 2
- 235000019362 perlite Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000019157 thiamine Nutrition 0.000 description 2
- 229960003495 thiamine Drugs 0.000 description 2
- 239000011721 thiamine Substances 0.000 description 2
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 2
- 239000010455 vermiculite Substances 0.000 description 2
- 235000019354 vermiculite Nutrition 0.000 description 2
- 229910052902 vermiculite Inorganic materials 0.000 description 2
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 description 1
- 101100352919 Caenorhabditis elegans ppm-2 gene Proteins 0.000 description 1
- 235000002568 Capsicum frutescens Nutrition 0.000 description 1
- 235000013162 Cocos nucifera Nutrition 0.000 description 1
- 244000060011 Cocos nucifera Species 0.000 description 1
- 244000304337 Cuminum cyminum Species 0.000 description 1
- 235000007129 Cuminum cyminum Nutrition 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000272168 Laridae Species 0.000 description 1
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- 240000000905 Nymphoides indica Species 0.000 description 1
- 235000017590 Nymphoides indica Nutrition 0.000 description 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 241000270666 Testudines Species 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
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- 238000012136 culture method Methods 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
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- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
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- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000000554 iris Anatomy 0.000 description 1
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- 238000009630 liquid culture Methods 0.000 description 1
- 210000003126 m-cell Anatomy 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
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- 229960002715 nicotine Drugs 0.000 description 1
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 230000008659 phytopathology Effects 0.000 description 1
- 229940072033 potash Drugs 0.000 description 1
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 1
- 235000015320 potassium carbonate Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、液体培地における細胞組織の培養によるバナ
ナの苗の生産方法に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method for producing banana seedlings by culturing cell tissue in a liquid medium.
本発明により生産されたバナナの苗は、爲株に由来する
植物病がない健全なものであって、バナナの栽培に利用
することができる。The banana seedlings produced according to the present invention are healthy and free from plant diseases caused by the seedlings, and can be used for banana cultivation.
バナナは熱帯から亜熱帯にかけて広く分布するパシ!つ
科の多年生草本植物であって、その実をそのまま食用に
供する生食用バナナ、その実を料物体を観賞に供する1
11葉植物としてのバナナかある。Bananas are widely distributed from the tropics to the subtropics! Banana is a perennial herbaceous plant belonging to the family, and its fruit is edible as is, and its fruit is used for ornamental purposes.1
There is a banana as an 11-leaf plant.
これまでのバナナの繁殖は、爲株の塊茎から発生する吸
芽(5ucker )をそのまま生長して、現株とする
か、または吸芽な摘出してこれを移植して生長させ、親
株とする無性繁殖によっていたが、爲株が病害虫に侵さ
れていると、その病害虫が無性繁殖による子株にそのま
ま移行して、子株も病害虫に侵されたものとなること、
および吸芽の発生が不定期的であり、その数も限られて
いることにより、大量のバナナの苗を生産することがで
きないことを避けることができない。Up until now, bananas have been propagated by growing suckers (5uckers) that develop from the tubers of the seedlings and using them as the current plant, or by removing the suckers and transplanting them to grow and use them as the parent plant. It was propagated asexually, but if the seedlings are infected by pests, the pests will be transferred to the asexually propagated offspring, and the offspring will also be infected by pests.
Also, the occurrence of suckers is irregular and the number thereof is limited, making it impossible to produce a large number of banana seedlings.
これまでに、バナナの塊茎より発生する吸芽の茎頂部の
植物組織を人工の培地に植え付け、培養して、バナナの
苗を得ることが報告され、〔ペルグおよびブスタマンテ
;フィトバソロジー(Berg and Bust
amaat* : Phytopathology
) N6411 g320〜322 (1974年
) ) 、 !!りl’)(7)茎頂部の植物組織か、
ら、複数のバナナの苗を生産することが報告され〔ジエ
イ・シー・ベツセイおよびジエイ・ニー・リベラ:ター
リアルバ(J、C,Vessey& J、A、Rlv
era : Turrlalbm) jJE31巻第
162〜163頁(1981年)〕〔ニス・ニス・クロ
ナラエルおよびニー・デイ−・クリコニアン:ホードサ
イエンス(S、S、Cronauer &A、D、Kr
1konlan : Hort、5elenee )第
19巻第2号第234〜235頁(1984年)〕、さ
らにバナナの植物細胞組織を固体培地で培養して、シュ
ートを形成し、これを肢体培地において振とう培養をし
て、マルチプルシュートを形成することが報告されてい
る〔エフ・ジエイ・ノバーク、アール・アフザ、ブイ・
ファドビブリア、ティー・ヘルメリン、エイチ櫓ブルン
ナーおよびビーードニニ:ヌック・チック・ビトロ・カ
ルト・プラント会インプロプ(F、J、NovmkJ、
Afts++V、Phadvlbulya+T+Her
melin、H,Brunoer & B+Donln
1 : Nuel。Up to now, it has been reported that banana seedlings are obtained by planting and culturing the plant tissue of the shoot apex of suckers that develop from banana tubers in an artificial medium. Bust
amaat*: Phytopathology
) N6411 g320-322 (1974) ),! ! (7) Plant tissue at the top of the stem,
[J. C. Vessey & J. A. Rlv.
era: Turrralbm) JE 31, pp. 162-163 (1981)] [Nis Nis Cronauer and N.D. Krikonian: Horde Science (S, S, Cronauer & A, D, Kr
1konlan: Hort, 5elenee) Vol. 19, No. 2, pp. 234-235 (1984)], banana plant cell tissues were further cultured on a solid medium to form shoots, which were cultured with shaking in a limb medium. It has been reported that multiple shoots are formed by
Fadobibria, Tee Hermelin, H Yagura Brunner and Biednini: Nook Chic Vitro Cult Plant Kai Improp (F, J, NovmkJ,
Afts++V, Phadvlbulya+T+Her
Melin, H. Brunoer & B+Donln
1: Nuel.
Tech、Vitro、Cu1t、Plant、Imp
rov、 ) 第167〜174頁(1986年)〕。Tech, Vitro, Cult, Plant, Imp
rov, ) pp. 167-174 (1986)].
本発明者らは、aI姿の細胞組織の培養について永年研
究を続けているが、その研究において、バナナの細胞組
織を液体培地における通気撹拌により培養すると、その
バナナの細胞組織における芽の分化およびシュートの形
成を促進し、これを発根すると、定植をすることができ
るバナナの苗を多数生産しうろこと、およびバナナの吸
芽の細胞組織をココナツツ・ウォーターを含む培地で培
養すると、充分に肥大した細胞組織が得られるが、この
肥大した細胞組織をココナツツ・ウォーターを含まない
培地で培養すると、多数の芽の分化した細胞組織が得ら
れることを見出し、これらの知見に基づいて本発明に到
達した。The present inventors have been conducting research on the cultivation of cell tissues in the aI form for many years, and in their research, they found that when banana cell tissues were cultured in a liquid medium with aeration and agitation, bud differentiation in the banana cell tissues and By promoting the formation of shoots and rooting them, a large number of banana seedlings that can be planted can be produced. When the scales and cell tissues of banana suckers are cultured in a medium containing coconut water, An enlarged cell tissue is obtained, and it has been discovered that when this enlarged cell tissue is cultured in a medium containing no coconut water, a cell tissue with a large number of differentiated buds can be obtained.Based on these findings, the present invention has been made. Reached.
本発明の目的は、バナナの健全な苗を多数生産すること
ができるバナナの苗の生産方法を提供することにあり、
詳しくは、多数の健全なバナナの苗を高い生産性におい
て生産することができるバナナの苗の生産方法を提供す
ることにある。An object of the present invention is to provide a method for producing banana seedlings that can produce a large number of healthy banana seedlings.
Specifically, the purpose is to provide a method for producing banana seedlings that can produce a large number of healthy banana seedlings with high productivity.
本発明は、バナナの細胞組織を培養し、そのシュートを
発根して、バナナの苗を生産する方法において、バナナ
の細胞組織を液体培地において通気撹拌により培養して
、芽の分化およびシュートの形成を増加し、それによっ
て多数の健全なバナナの苗を得ることを特徴とするバナ
ナの苗の生産方法である。The present invention provides a method for producing banana seedlings by culturing banana cell tissues and rooting the shoots. A method for producing banana seedlings characterized by increasing formation and thereby obtaining a large number of healthy banana seedlings.
本発明のバナナの細胞組織の培養によるバナナの苗の生
産方法において、バナナの細胞組織なココナツツ・ウォ
ーターを含む培地で培養して、細胞組織を肥大し、その
肥大した細胞組織をココナツツ・ウォーターを含まない
培地で培養して、芽の分化した細胞組織とし、その芽の
分化した細胞組織を液体培地で通気撹拌により培養して
、多数の健全なバナナの苗を生産することができる。In the method of producing banana seedlings by culturing banana cell tissues of the present invention, banana cell tissues are cultured in a medium containing coconut water to enlarge the cell tissues, and the enlarged cell tissues are injected with coconut water. A large number of healthy banana seedlings can be produced by culturing in a medium containing no banana to produce differentiated cell tissues of buds, and culturing the differentiated cell tissues of the buds in a liquid medium with aeration and agitation.
本発明のバナナのll[[I胞組織の培養によるバナナ
の苗の生産方法において、細胞組織の分化した芽から生
長したシュートを、栄養諒の濃度を半減した培地におい
て培養して、シュートを発根し、それによって定植する
ことができる小植物体(苗)を生産することができる。In the method for producing banana seedlings by culturing banana tissue of the present invention, shoots grown from buds with differentiated cell tissue are cultured in a medium containing half the concentration of nutrients, and the shoots are generated. It is possible to produce plantlets (seedlings) that can take root and thereby be planted.
(発明の詳細な説明〕
バナナの欄の生えぎわに、吸芽(5ucker )が生
えてくる。吸芽はバナナの塊茎から生えてくる植物体で
ある。(Detailed Description of the Invention) A sucker (5ucker) grows at the growth edge of the banana column. A sucker is a plant that grows from a banana tuber.
本発明では、@場より吸芽を採取し、これを充分に水洗
した後、外皮の一部を剥ぎ、茎頂部位を中心に含む植物
体片を摘出する。その植物体片を、殺菌剤C例えば、塩
化ベンザルコニウム水浴液、次亜塩素醍ナトリウム水溶
液およびエタノール)により殺菌し、クリーンベンチ内
において、実体輩微鏡を使用し、その植物体片から茎頂
部および茎頂周囲の葉原茎を含むバナナの細胞組織を摘
出する。この細胞組織は本発明の植物組織の培養による
バナナの苗の生産方法におけるバナナの細胞組織である
。In the present invention, suckers are collected from the field, thoroughly washed with water, a part of the outer skin is peeled off, and plant pieces containing mainly the shoot apex are extracted. The plant pieces are sterilized with a disinfectant C (e.g., benzalkonium chloride bath solution, sodium hypochlorite aqueous solution, and ethanol), and in a clean bench, using a physical microscope, the plant pieces are removed from the stems. The banana cell tissue, including the top and leaf progenitors around the shoot apex, is removed. This cell tissue is a banana cell tissue in the method for producing banana seedlings by culturing plant tissue of the present invention.
バナナのm胞組識は、ココナツツ・ウォーターを含む初
代@池に植え付け、 1000〜3000ルツクスの
24時間の照明下に、 20〜30℃において2〜3週
間培養する。ココナツツ・ウォーターを含む初代培地は
、ムラシゲ・スクーグ培地の無機塩類に、ミオイノシト
ール100〜TOOpp■ (好ましくは300〜50
0 p9鳳)、ニコチン#0.2〜0.711PII
(好ましくは0.4〜O・a ppm)、塩酸ピリド
キシン0.2〜0.7 ppm (好ましくは0.4〜
0.6pp■)、塩酸チアミン0.5〜2.Oppm
(好ましくは!60〜1.59p謹)、グリシン1〜3
ppm(好ましくは1.5〜2.59p諷)、ココナツ
ツ・ウォーター50〜25Qd/j! (好ましくは
100〜20〇−/l)およびシ!IM30000〜5
0000 pp醜 (好ましくは35000〜4500
0 p9菖)を加え、さらにベンジルアデニン1〜16
ppm (好ましくは465〜11−3 pp+a
)またはカイネチンr〜22ppm(好ましくは4.3
〜15.0 ppm)、および寒天粉末0・7〜1.0
%(重量)を加えて得た固形培地を使用する。ココナツ
ツ・ウォーターは、ヤシの実の空洞に入っている水であ
る。バナナの細胞組織は、培養開始から約!週間で白色
から黄緑色に変り、約3週間で5H程度に肥大した細胞
組織の集合体になる。Banana m cell tissues are planted in primary ponds containing coconut water and cultured for 2 to 3 weeks at 20 to 30°C under 24 hour illumination of 1000 to 3000 lux. The primary medium containing coconut water contains 100~TOOpp of myo-inositol (preferably 300~50~
0 p9 Otori), Nicotine #0.2-0.711 PII
(preferably 0.4-0.a ppm), pyridoxine hydrochloride 0.2-0.7 ppm (preferably 0.4-0.a ppm)
0.6pp■), thiamine hydrochloride 0.5-2. Oppm
(preferably! 60-1.59p), glycine 1-3
ppm (preferably 1.5-2.59p), coconut water 50-25Qd/j! (preferably 100 to 200-/l) and shi! IM30000~5
0000 pp ugly (preferably 35000-4500
0 p9 irises) and further benzyladenine 1-16
ppm (preferably 465-11-3 pp+a
) or kinetin r ~ 22 ppm (preferably 4.3
~15.0 ppm), and agar powder 0.7~1.0
Use a solid medium obtained by adding % (by weight). Coconut water is water contained in the hollow space of a coconut. Banana cell tissue grows from the start of culture to approx. The color changes from white to yellow-green within a week, and in about 3 weeks it becomes an aggregate of cellular tissue that has enlarged to around 5H.
この肥大した細胞組織の集合体は、ココナツツ・ウォー
ターを含まない増殖培地に植え付け、24時間の照明下
に20〜30℃において2〜3週間培養するりココナツ
ツ・ウォーターを含まない増殖培地は、前記の初代培地
からココナツツ・ウォーターを除いた培地である。肥大
した細胞組織の集合体は、多数の芽を分化し、多数の芽
を付けた細胞組織の集合体となるが、その芽からシュー
トを形成しているものもある。This enlarged cell tissue aggregate is planted in a growth medium that does not contain coconut water and cultured for 2 to 3 weeks at 20 to 30°C under 24-hour light. This is a medium obtained by removing coconut water from the primary medium of . The enlarged cell tissue aggregate differentiates into many buds and becomes a cell tissue aggregate with many buds, and some of these buds form shoots.
多数の芽を付けた細胞組織の集合体は、少なくとも1(
II(好ましくは2〜3個)の芽を含む細胞組織に分割
し、通気式ジャーファーメンタ−中の液体培地に入れ、
肢体培地ll当り0.3〜1.OtZ分(好ましくは0
.4〜0.7//分)の空気を送って通気撹拌しながら
、24時間の照明下に、20〜30℃において培養する
。この液体培地は、前記の増殖培地においてベンジルア
デニンまたはカイネチンの濃度を、ベンジルアデニン0
.2〜2.2 lap論 (好ましくは0・6〜1.5
ppm)またはカイネチン0.1〜2.1 ppm
(好ましくは0.4〜!、5pp■)に変更し、寒天を
除いたものであって、ココナツツ・ウォーターは含まな
いものである。A collection of cell tissues with numerous buds is composed of at least one (
Divide into cell tissues containing II (preferably 2 to 3) buds and place them in a liquid medium in an aerated jar fermentor.
0.3 to 1.0% per liter of limb culture medium. OtZ min (preferably 0
.. The cells are cultured at 20 to 30° C. under illumination for 24 hours while aerating and agitating the cells by blowing air at a rate of 4 to 0.7/min). This liquid medium changes the concentration of benzyladenine or kinetin in the growth medium to 0 to 0.
.. 2-2.2 lap theory (preferably 0.6-1.5
ppm) or kinetin 0.1-2.1 ppm
(preferably 0.4~!, 5pp■), agar is removed, and coconut water is not included.
芽を付けた細胞組織の集合体は、液面付近で上下動を繰
り返しながら芽を増殖するとともに、芽からシュートを
形成し、そのシュートが伸長する。The aggregate of cell tissues with buds moves up and down near the liquid surface, multiplying the buds, forming shoots from the buds, and elongating the shoots.
このジャーファーメンタ−における通気撹拌培養は、2
0〜40日間継読することができ、それによって伸長し
たシュートを形成したm胞組職の集合体が得られる。The aeration agitation culture in this jar fermenter is 2
It can be continued for 0 to 40 days, thereby obtaining a collection of m-cell tissue that has formed an elongated shoot.
伸長したシュートを形成した細胞組織の集合体からシュ
ートをその基部から摘出し、そのシュートを発根培地に
植え付け、24時間の照明の下に20〜30℃において
3〜4週間培養すると、シュートの基部に根が出てきて
伸長し、小M物体(plantlet )が得られる。The shoot is removed from the base of the aggregate of cell tissue that formed the elongated shoot, planted in a rooting medium, and cultured for 3 to 4 weeks at 20 to 30°C under 24-hour light. Roots emerge from the base and elongate, yielding a small M object (plantlet).
発根培地は、ココナツツ・ウォーターならびに練物ホル
モンのベンジルアデニンお上びカイネチンを含まず、他
の栄養源の濃度を薄くした@地であって、前記の初代培
地の組成において、ココナツツ・ウォーター、ベンジル
アデニンお上びカイネチンを除くが、寒天の量は初代培
地と同じ0.7〜1.0%であり、その他の無機塩類お
よびとクミン類は1〜4倍(好ましくは約2倍)に希釈
したものである1発根培地で得られた小植物体は植物体
に根が付いた苗であるが、これを1IInの外界の環境
に耐え得るものとするために、馴化する。The rooting medium does not contain coconut water or the refined hormones benzyladenine and kinetin, and has a dilute concentration of other nutrients, and in the composition of the primary medium, coconut water, benzyl Adenine and kinetin are excluded, but the amount of agar is the same as the primary medium, 0.7 to 1.0%, and other inorganic salts and cumin are diluted 1 to 4 times (preferably about 2 times). The plantlets obtained using the 1 rooting medium are seedlings with roots attached to the plants, which are acclimatized in order to be able to withstand the external environment of 1IIn.
プランタ−に水はけのよい土壌を入れ、このプランタ−
に小植物体を植え付け、このプランタ−をlθ〜30″
Cの温度、60〜90%相対湿度および5000〜30
000ルツクスの照明の条件下に置いて2〜3a間栽培
すると、小植物体から2〜3校の葉が出てきて、根、茎
および葉を具えた植物体になる。この小植物体の栽培に
おいて、少量のチッ禦、リン酸、カリおよび微量要素を
含む植物養液を施用する。プランタ−に入れる水はけの
良い土壌は、バーミキュライト、パーライト、赤玉土、
應沼土、腐葉土および砂などの単品または混合土壌を使
用する。Fill the planter with well-drained soil and use this planter.
Plant the plantlets in the
Temperature of C, 60-90% relative humidity and 5000-30
When cultivated for 2 to 3 hours under lighting conditions of 000 lux, 2 to 3 leaves will emerge from the plantlet, forming a plant with roots, stems, and leaves. In cultivating this plantlet, a plant nutrient solution containing a small amount of chili, phosphoric acid, potash and trace elements is applied. Well-drained soil to put in the planter is vermiculite, perlite, akadama soil,
Use single or mixed soils such as swamp soil, humus, and sand.
この植物体は、同様の水はけのよい土壌を入れたポット
に植栽し、そのポットに同様の植物養液を施用し、通常
のネットハウスにそのポットを置いて2〜3ケ月m培す
ると、その地上部が20〜30mに伸長してバナナの苗
が得られる。このバナナの苗は圃場に定植して、バナナ
を収穫することができるものである。This plant is planted in a pot filled with the same well-drained soil, applied with the same plant nutrient solution, placed in a regular net house, and cultivated for 2 to 3 months. The above-ground part grows to 20 to 30 meters and banana seedlings are obtained. These banana seedlings can be planted in a field and harvested as bananas.
以下において、試験例および実施例により本発明をさら
に詳しく説明する。In the following, the present invention will be explained in more detail by means of test examples and examples.
試験例
バナナの吸芽の細胞の組織培養によるバナナの苗の育成
において、バナナの培養細胞のマルチプルシュートの形
成における培養方法の影響について試験を行なった。Test Example In growing banana seedlings by tissue culture of banana sucker cells, a test was conducted to examine the influence of the culture method on the formation of multiple shoots of cultured banana cells.
(1)試料のmtl (芽の分化したllllN組鱈の
集合体の形成)
沖縄の在来種のシマバナナ(小笠原諸島原産)の吸芽を
圃場で採取し、水洗した後、外皮を剥ぎ取り、その茎頂
部を中心とする墓頂部岨織〔20jIjICタテ)X2
0關(ヨコ)X40+w(高す)〕を切り取った。この
茎頂部組繊は、70%エタノール水溶液、0・2%塩化
ベンザルコニウム水溶液および1%次亜塩se#ナトリ
ウム水溶液に、順次浸漬、撹拌して、殺菌した後、滅菌
水により洗浄した。その茎頂部組織は、クリーンベンチ
内で、実体顕微鏡を用いて、茎頂および葉原基を含む円
形組織片〔0,8關(径)Xlsua(長す)〕を摘出
した。(1) Sample mtl (Formation of aggregate of llllN cod with differentiated buds) Suckers of Shimabanana, a native species of Okinawa (originating from the Ogasawara Islands), were collected in the field, washed with water, and the outer skin was peeled off. Grave apex centering around the stem apex [20jIjIC vertical) X2
0 (width) x 40 + w (height)] was cut out. The shoot apical fibers were sterilized by being immersed and stirred in a 70% ethanol aqueous solution, a 0.2% benzalkonium chloride aqueous solution, and a 1% hypochlorite se# sodium aqueous solution, and then washed with sterilized water. A circular tissue piece [0.8 mm (diameter) Xlsua (long)] containing the shoot apex and leaf primordia was excised from the shoot apex tissue using a stereomicroscope in a clean bench.
その円形組織片を初代培地15−を入れた試験管(25
m径)に植え付け、2500ルツクスの24時間龍量子
に、25層2℃において21日間培養して、黄緑化し、
5鴎φに肥大した細胞組織片を得た・
(初代@地の組成)
MgSOか7HO
CaC1e2HO
370pH諷
440 ppm
1900 ppm
1650 11$11
!?Oppm
27.8 pp諺
37.3 pP鵬
22.3 ppm
8・6 ppm
0.025 Ppm
にNo
NHNO
NHPO
FeSO−7H0
Na EDTA
Mn50 ・ 4HO
ZnSO壷 7H0
CuSO・ 58 0
CoC1−6)1 0 0
.025 ppmにI
O,83++9鵬HBO
6,299■
Na MoO繍 2H00,25ppmミオイノシト
ール 400 ppmニコチン酸
0.5 ppm塩酸ピリドキシン
0.5 pH箇塩酸チアミン l・3
ppmグリシン 2・o pp論
シvs 1140000 Ppm
ココナツツ・ウォーター 150d/lベンジルア
デニン 6.7 pHml寒天
700099■
その肥大したlllIgI組織片を、増殖培地10−を
入れた試験管(25sw径)に植え付け、前記と同じ照
明下に、25層2℃において、18日間培養して、多数
の芽の分化した細胞組織の集合体を得た。The circular tissue piece was placed in a test tube (25
m diameter), and cultured in 25 layers at 2°C for 21 days at 2500 lux for 24 hours to turn yellowish green.
A piece of cell tissue enlarged to 5mm diameter was obtained. (Composition of the primary site) MgSO or 7HO CaC1e2HO 370pH 440 ppm 1900 ppm 1650 11$11! ? Oppm 27.8 pp Proverbs 37.3 pP Peng 22.3 ppm 8.6 ppm 0.025 Ppm No NHNO NHPO FeSO-7H0 Na EDTA Mn50 4HO ZnSO Bottle 7H0 CuSO 58 0 Co C1-6) 1 0 0
.. 025 ppm I
O,83++9PengHBO
6,299■ Na MoO embroidery 2H00, 25ppm myo-inositol 400 ppm nicotinic acid
0.5 ppm pyridoxine hydrochloride
0.5 pH Thiamine hydrochloride l・3
ppm glycine 2・o pp theory vs 1140000 Ppm coconut water 150d/l benzyladenine 6.7 pHml agar
700099■ The enlarged lllIgI tissue piece was planted in a test tube (25sw diameter) containing 10-mL of growth medium, and cultured in 25 layers at 2°C for 18 days under the same lighting as above to differentiate a large number of buds. A collection of cell tissues was obtained.
(増殖培地の組成)
MgSO・ 7■ 0 3
70 pp鳳CaC1・ 2H0
N03
NHN。(Composition of growth medium) MgSO・7■03
70 pp Otori CaC1・2H0 N03 NHN.
にHPO
FeSO・ 7H0
Na EDTA
Mn50 ・411 0
ZnSO−7HO
4゛2
CuSO−5H0
CoC1−680
にl
BQ
Ha MoO−2H0
ミオイノシトール
ニコチン酸
塩酸ピリドキシン
塩酸チアミン
グリジン
シ目糖
ベンジルアデニン・
440 ppm
1900 ppm
1650 pp+m
170 ppm
27.8 pp膳
37.3 p9鵬
22・3 Ppm
8+6 99層
0.025 ppw+
0.025 pp諺
0.83 pHl1
6+2 ppm
0・25 ppm
400 ppm
0.5pp議
0・599■
1.3 pp鳳
2、Opp議
40QO099鳳
6φ7 99層
寒天 7000 pllll
(注)増殖培地は、初代培地において、ココナツツ・ウ
ォーターを使用しないものである。HPO FeSO・7H0 Na EDTA Mn50・411 0 ZnSO-7HO 4゛2 CuSO-5H0 CoC1-680 BQ Ha MoO-2H0 Myo-inositol Nicotinic acid hydrochloride Pyridoxine Hydrochloride Thiamine Glysine Sugar benzyladenine・440 ppm 1900ppm 1650 pp+m 170 ppm 27.8 pp Zen 37.3 p9 Peng 22.3 Ppm 8+6 99 layer 0.025 ppw+ 0.025 pp proverb 0.83 pHl1 6+2 ppm 0.25 ppm 400 ppm 0.5pp proverb 0.59 9■ 1.3 pp 2, Opp 40QO099 6φ7 99-layer agar 7000 pllll (Note) The growth medium is a primary medium without coconut water.
(2)試験方法
(2−A)II体@地による試験
(2−^−1)マルチプルシュートの形成前記の(1)
で得た芽の分化した細胞組織の集合体を、芽2個を含む
細胞組織に分割し、その分割細胞組織を、前記の(1)
の増殖培地10−を入れた試験管(25m径)50本に
植え付け、前記の(1)と同じ照明下に、25±2℃に
おいて18日間培養して、マルチプルシュートを形成し
た芽を有する細胞組織片を形成し、各試験管における細
胞組織の芽の数および2個以上に伸長したシュートの数
をカウントした。(2) Test method (2-A) Test using II body @ ground (2-^-1) Formation of multiple shoots (1) above
The aggregate of differentiated cell tissues of the buds obtained in step 1 is divided into cell tissues containing two buds, and the divided cell tissues are divided into cell tissues as described in (1) above.
Cells with buds that formed multiple shoots were planted in 50 test tubes (25 m diameter) containing 10 - of growth medium and cultured for 18 days at 25 ± 2°C under the same lighting as in (1) above. Tissue pieces were formed, and the number of cell tissue buds and the number of shoots that had elongated to two or more in each test tube were counted.
(2−A−2)シュートの発根
前記の(2−A−1)で形成したシュートを、その基部
において切断し、その細胞組織から分層し、そのシュー
トを発根培地IO−を入れた試験管(25錦径)に植え
付け、前記の(1)と同じ照明下に、25±2℃におい
て4遍間培養して、根の長さが5個以上、また地上部の
長さが4cs以上の小植物体を形成した。(2-A-2) Rooting of shoots The shoots formed in (2-A-1) above are cut at their bases, separated from the cell tissue, and the shoots are filled with rooting medium IO-. Plant it in a test tube (25 brocade diameter) and culture it for 4 cycles at 25±2℃ under the same lighting as in (1) above, until it has 5 or more roots and the length of the above-ground part. Plantlets of 4 cs or more were formed.
(発根培地の組成)
MgSO・7H0
CaCIII2H0
185ppm
220 ppm
950 ppm
825 ppm
s ppm
13.9 p9霞
!8φ65 9p8
11.15 ppm
4・3 ppm
0−0125 ppm
0.0125 ppm
0.415 ppm
3・1 ppm
0.125 119論
20G +1911
にNo3
HNO
KII PO
FeSO番 7H0
Na EDTA
Mn5Oψ 4H0
ZnSO−7H0
CuSO−5H0
CoCj 令 6HO
に!
BO
Na MoO画 28 0
ミオイノシトール
ニコチン酸 0.25 ppm塩酸ピ
リドキシン 0.25 T)9膿塩酸チア
ミン 0.65 ppmグリシン
1.0 ppm2g li
2G000 ppm寒天 8000
pp腸
活性炭 tooo ppm(注)発
根培地は、初代培地において、ココナツツ・ウォーター
およびベンジルアデニンを使用せず、寒天の濃度を高く
シ、また活性炭を使用するが、それ以外の改修は、その
1度を%にしたものである。(Composition of rooting medium) MgSO・7H0 CaCIII2H0 185 ppm 220 ppm 950 ppm 825 ppm s ppm 13.9 p9 Kasumi! 8φ65 9p8 11.15 ppm 4・3 ppm 0-0125 ppm 0.0125 ppm 0.415 ppm 3・1 ppm 0.125 119 theory 20G +1911 No. 3 HNO KII PO FeSO No. 7H0 Na EDTA Mn5Oψ 4H0 ZnSO-7H0 CuSO- 5H0 CoCj order 6HO! BO Na MoO drawing 28 0 Myo-inositol nicotinic acid 0.25 ppm Pyridoxine hydrochloride 0.25 T) 9 Pus Thiamine hydrochloride 0.65 ppm Glycine
1.0 ppm2g li
2G000 ppm agar 8000
pp intestinal activated carbon too ppm (Note) The rooting medium does not use coconut water or benzyladenine in the primary medium, has a high concentration of agar, and uses activated carbon, but other modifications are as follows: The degree is expressed as a percentage.
(2−A−3)小植物体の馴化、鉢上げおよび育苗
前記の(2−A−2)で形成した小植物体を試験管から
取り出し、水道水により充分洗浄した。(2-A-3) Acclimation, Potting, and Seedling Growth of Plantlets The plantlets formed in (2-A-2) above were taken out from the test tubes and thoroughly washed with tap water.
バーミキュライトおよびパーライトの混合土壌(t :
1)をプランタ−(18m(タテ)X40cR(ヨコ
)X14m(深す)〕に入れ、これに、前記の水洗した
小植物体20本を植え付けた。このプランタ−をガラス
aXに入れ、18〜25℃の温度、60〜90%の相′
M湿度および30000ルツクスの24時間の照明下に
おいて、小植物体の馴化を行なった。3週間後には、新
芽が2〜3枚に展開し、小植物体は生き生きとしていた
。Mixed soil of vermiculite and perlite (t:
1) was placed in a planter (18 m (vertical) x 40 cR (horizontal) x 14 m (depth)], and 20 of the above-mentioned washed plantlets were planted in it. This planter was placed in a glass ax, and 25℃ temperature, 60-90% phase'
The plantlets were acclimated under M humidity and 30,000 lux for 24 hours. Three weeks later, two or three new shoots had developed, and the plantlets were lively.
赤玉土および腐葉土の混合土II (2: 1)の入っ
た鉢に、この小植物体を植え付け、通常のネットハウス
において育苗を行なった。80日後に、地上部が257
!11内外の苗が得られた。The plantlets were planted in pots containing mixed soil II (2:1) of Akadama soil and humus, and seedlings were raised in a regular net house. After 80 days, the above ground part will be 257
! 11 seedlings were obtained.
この苗は、圃場において、充分定植可能なものであった
。These seedlings were fully capable of being planted in the field.
(2−8)液体増殖培III(カイネチン)による試験
(2−8−1)マルチプルシュートの形成前記の(1)
で得た芽の分化した細胞組織の集合体を、芽2個を含む
細胞組織に分割し、その分割細胞組織を、液体増殖培地
Cカイネチン)!0〇−を入れた300−容エルレンマ
イヤーフラスコ5個に、1フラスコ当り4個づつ植え付
け、前記の(1)と同じ照明下に、100 rpmの振
とうをしながら、25重2℃において18日間培養して
、マルチプルシュートを形成した芽を有する細胞組織片
を形成し、各フラスコにおける細胞組織片の芽の数およ
び2clL以上に伸長したシ為−トの数をカウントした
。(2-8) Test using liquid growth medium III (kinetin) (2-8-1) Formation of multiple shoots (1) above
The aggregate of differentiated cell tissues of the buds obtained was divided into cell tissues containing two buds, and the divided cell tissues were added to the liquid growth medium C kinetin)! Plant 4 seeds per flask in five 300-capacity Erlenmeyer flasks containing 0. The cells were cultured for 18 days to form cell tissue fragments having buds forming multiple shoots, and the number of buds and the number of sheets elongated to 2 clL or more in the cell tissue fragments in each flask were counted.
〔液体増N培ft1l(カイネチン)の組成〕Mg5O
・7HO370ppm
CaCI C2HO440ppm
NO
NHNO
NHPG
FeSO・ 7H0
Na EDTA
M!150 ・4H0
ZnSO・ 7H0
CuSO−5HQ
CoC1φ 6H0
にI
BO
$900111111
1650 ppm
27・8 ppm
37.3 ppm
22.3 ppm
am6 ppm
0.025 9911
0自0259p亀
0・83 ppm
6・2 ppm
Na MoOj 2)1 0
0.25 pp諺ミオイノシトール
400 H1■ニコチン酸 0.
511p諺塩酸ピリドキシン 0.5 p
p飄塩酸チアミン 1.3 ppmグ
リシン 2.Opp鳳シ!1 @
40000 pp■カイネチン
1.!ppH(注)液体増殖培地c
カイネチン)は、前記の(1)の初代培地において、コ
コナツツ・ウォーターおよび寒天を使用しないこと、お
よびベンジルアデニンの代りにカイネチント1 ppm
を使用したこと以外は、初代培地ご同じ組成のMIl+
である。[Composition of liquid enriched N culture ft1l (kinetin)] Mg5O
・7HO370ppm CaCI C2HO440ppm NO NHNO NHPG FeSO・7H0 Na EDTA M! 150 ・4H0 ZnSO・ 7H0 CuSO-5HQ CoC1φ 6H0 to IBO $900111111 1650 ppm 27・8 ppm 37.3 ppm 22.3 ppm am6 ppm 0.025 9911 0 self 0259p Turtle 0.83 ppm 6.2 ppm Na MoOj 2) 1 0
0.25 pp proverb myo-inositol
400 H1 ■ Nicotinic acid 0.
511p Proverb Pyridoxine Hydrochloride 0.5p
Thiamine hydrochloride 1.3 ppm Glycine 2. Opp Otoshi! 1 @
40000 pp■Kinetin
1. ! ppH (note) liquid growth medium c
kinetin) in the primary medium of (1) above, do not use coconut water or agar, and add 1 ppm kinetin instead of benzyladenine.
MIl+ with the same composition as the primary medium except that MIL+ was used.
It is.
(2−8−2)シュートの発根
前記の細胞組織片を使用し、前記の(2−A−2)と同
様にして、シュートの発根を行なったが、前記の(2−
A−2)と同様の小植物体を得た。(2-8-2) Rooting of shoots Using the cell tissue pieces described above, shoots were rooted in the same manner as in (2-A-2).
A plantlet similar to A-2) was obtained.
(2−8−3)小植物体の馴化、鉢上げおよび育苗
前記の小植物体を使用し、前記の(2−A−3)と同様
にして、その小M物体の1b化、鉢上げおよび育苗を行
なフたが、amに移植することができるバナナの苗を得
た。(2-8-3) Acclimatization, potting, and seedling raising of plantlets Using the above plantlet, convert the small M object into 1b and potting it in the same manner as in (2-A-3) above. Then, seedlings were raised to obtain banana seedlings that can be transplanted to am.
(2−c)液体増殖培地(ベンジルアデニン)による試
験
(2−C−1)マルチプルシュートの形成前記の(1)
で得た芽の分化した[l泡Ifl鵬の集合体を、芽2個
を含む細胞組織に分割し、その分!IIB胞組織を、液
体増殖培地(ベンジルアデニン)100−を入れた30
〇−容エルレンマイヤーフラスコ5個に、1フラスコ当
り3個づつ植え付け、前記のc凰) と同じ照明下に、
100 rpmの振どうをしながら、25重2℃におい
て18日間培養して、マルチプルシュートを形成した芽
を有する細胞組織片を形成し、各フラスコにおける細胞
組織片の芽の数および2α以上に伸長したシュートの数
をカウントした。(2-c) Test using liquid growth medium (benzyladenine) (2-C-1) Formation of multiple shoots (1) above
The differentiated bud aggregates obtained were divided into cell tissues containing two buds, and the amount! The IIB cell tissue was grown for 30 minutes in liquid growth medium (benzyladenine).
Plant the seeds in 5 Erlenmeyer flasks, 3 per flask, under the same lighting as above.
While shaking at 100 rpm, cells were cultured in 25 layers at 2°C for 18 days to form cell tissue pieces with buds that formed multiple shoots, and the number of cell tissue pieces in each flask and their elongation to 2α or more were determined. The number of shots made was counted.
〔液体増殖培地 Mg5ON7H0 CaC1−2H0 NO NHN。[Liquid growth medium Mg5ON7H0 CaC1-2H0 NO NHN.
にHPo
F@SOll7H0
Na EDTA
Mn5O−480
Z+xSOも 7H0
CuSOj 5H0
CoCI 116HO
にI
(ベンジルアデニン)の組呟〕
370 ppm
440 ppm
1900 ppm
1650 [1謙
170 ppm
27.8 ppm
37.3 99膳
22・3 ppm
8・6 ppm
0.025 ppm
0.02529m
0.83 99重
6.2 pp嘗
0.25 PP謹
400 ppm
0・5 ppm
0・5 ppm
l・3 ppm
2拳o ppm
BO
Na Mo0 ・ 2H0
ミオイノシトール
ニコチン酸
塩酸ピリドキシン
塩酸チアミン
グリジン
シ!I m40000 ppm
ベンジフレアデニン 0.9 ppwr(
注)液体増殖培地(ベンジルアデニン)は、前記(2−
B−1)の液体増殖層tmcカイネチン)におけるカイ
ネチンの代りにベンジルアデニン0.9 ppmを使用
したこと以外は、液体増殖@地(カイネチン)と同じ培
地である。HPo F@SOll7H0 Na EDTA Mn5O-480 Z+xSO also 7H0 CuSOj 5H0 CoCI 116HO I (benzyladenine) combination] 370 ppm 440 ppm 1900 ppm 1650 [1 Ken170 ppm 27.8 ppm 37.3 99 meals 22・3 ppm 8・6 ppm 0.025 ppm 0.02529m 0.83 99 weight 6.2 ppm 0.25 PP 400 ppm 0・5 ppm 0・5 ppm l・3 ppm 2 fist o ppm BO Na Mo0 ・2H0 Myo-inositol nicotinic acid hydrochloride pyridoxine hydrochloride thiamine glycine! I m40000 ppm Bendiflaedenine 0.9 ppwr (
Note) The liquid growth medium (benzyladenine) is as described above (2-
The medium is the same as the liquid growth layer (Kinetin) except that 0.9 ppm of benzyladenine was used instead of kinetin in the liquid growth layer (TMC kinetin) in B-1).
(2−C−23シユートの発根
前記の細胞組織片を使用し、前記の(2−A−2)と同
様にして、シュートの発根を行なったが、前記(2−A
−2)と同様の小植物体を得た。(2-C-23 Rooting of shoots Using the cell tissue pieces described above, rooting of shoots was carried out in the same manner as in (2-A-2) above.
A plantlet similar to -2) was obtained.
(2−C−3)小植物体の劇化、鉢上げおよび育苗
前記の小植物体を使用し、前記の(2−A−3)と同様
にして、その小植物体の−σ化、鉢上げおよび育苗を行
なったが、圃場に移植することができるバナナの苗を得
た。(2-C-3) Dramaticization of plantlets, potting up, and raising seedlings Using the above plantlets, -σization of the plantlets in the same manner as in (2-A-3) above, After raising pots and raising seedlings, we obtained banana seedlings that can be transplanted to the field.
(2−f+)通気撹拌培養による試験
(2−D−1)マルチプルシュートの形成前記の(1)
で得た芽の分化した側胞組繊の集合体を、芽3個を含む
M胞組繊片に分割し、その分割細胞組織片20個を、液
体増殖層all(ベンジルアデニン) 3tを入れた4
を容ジャーファーメンタ−に植え付け、前記の(1)と
同じ照明下に、培1I111当り0.5t/分の通気撹
拌をしながら、25±2℃において40日間培養して、
マルチプルシュートを形成した芽を有する細胞組織片を
形成し、それぞれのam岨纏片の芽の数および2礪以上
に伸長したシュートの数をカウントした。(2-f+) Test by aerated agitation culture (2-D-1) Formation of multiple shoots (1) above
The aggregate of differentiated lateral tissue fibers of the buds obtained was divided into M cell tissue fragments containing 3 buds, and 20 of the divided cell tissue fragments were added with 3T of liquid proliferative layer ALL (benzyladenine). 4
were planted in a jar fermenter, and cultured at 25±2°C for 40 days under the same lighting as in (1) above, with aeration and agitation of 0.5 t/min per culture medium,
Cell tissue pieces having buds that formed multiple shoots were formed, and the number of buds and the number of shoots that had elongated to 2 cm or more in each piece were counted.
(2−o−2)シュートの発根
前記の1III胞岨識片を使用し、前記の(2−A−2
)と同様にして、シュートの発根を行なったが、前記の
(2−A−2)と同様の小植物体を得た。(2-o-2) Rooting of shoots Using the above-mentioned 1III rooting fragment, the above-mentioned (2-A-2)
) The shoots were rooted in the same manner as in (2-A-2), and the same plantlets as in (2-A-2) were obtained.
(2−D−3)小植物体の馴化、鉢上げおよび育苗
前記の小植物体を使用!ノ、前記の(2−八−3)と同
様にして、その小植物体の馴化、鉢上げおよび育苗を行
なったが、圃場に移植することができるバナナの苗を得
た。(2-D-3) Acclimatization of plantlets, potting up and raising seedlings Use the above plantlets! (2) The plantlet was acclimatized, potted, and raised in the same manner as in (2-8-3) above, and banana seedlings that could be transplanted to the field were obtained.
(3)試験の結果 第1表に示すとおりであった。(3) Test results It was as shown in Table 1.
(以下余白〕
(4)考察
第1表によると、バナナの吸芽から摘出した細胞組織の
増殖を液体培地で行なうと、細胞組織に付いている芽が
増殖することがわかる。1導胞組織に付いている芽が伸
長してシュートを形成し、そのシュートを発根すると、
バナナの苗になるから、芽の増殖は、細胞組織から易導
されるバナナの苗の数を増加する。(Margin below) (4) Discussion According to Table 1, when the cell tissue extracted from banana suckers is grown in a liquid medium, the buds attached to the cell tissue proliferate.1.Gull tissue When the buds attached to the plant elongate and form a shoot, and the shoot takes root,
Since it becomes a banana seedling, the proliferation of buds increases the number of banana seedlings that can be easily derived from the cell tissue.
バナナの吸芽から摘出した細胞組織の増殖を液体培地に
おける通気撹拌培養で行なうと、増殖した芽から伸長す
るシュートの数を増加することがわかる。そのシュート
を発根すると、バナナの苗になるから、シュートの比率
の増加は、バナナの苗の5導を促進することになり、そ
のことはバナナ0苗の形成に必要な期間を短縮すること
になる。It has been found that when cell tissue extracted from banana suckers is grown in a liquid medium with aeration and agitation, the number of shoots extending from the grown buds increases. When the shoots are rooted, they become banana seedlings, so an increase in the shoot ratio will promote the growth of banana seedlings, which will shorten the period required for the formation of banana seedlings. become.
したがって、液体培養における通気撹拌培養は、バナナ
の苗の生産を向上することがわかる。Therefore, it can be seen that aeration agitation culture in liquid culture improves the production of banana seedlings.
手続補正書 平成 元年11月15日Procedural amendment November 15, 1989
Claims (1)
して、バナナの苗を生産する方法において、バナナの細
胞組織を液体培地において通気撹拌により培養すること
を特徴とする植物組織の培養によるバナナの苗の生産方
法。(1) A method for producing banana seedlings by culturing banana cell tissues and rooting the shoots, which is characterized in that the banana cell tissues are cultured in a liquid medium with aeration and agitation. How to produce banana seedlings.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1043201A JPH02222627A (en) | 1989-02-27 | 1989-02-27 | Production of banana seedling by culture of plant tissue |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1043201A JPH02222627A (en) | 1989-02-27 | 1989-02-27 | Production of banana seedling by culture of plant tissue |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH02222627A true JPH02222627A (en) | 1990-09-05 |
JPH0452734B2 JPH0452734B2 (en) | 1992-08-24 |
Family
ID=12657317
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP1043201A Granted JPH02222627A (en) | 1989-02-27 | 1989-02-27 | Production of banana seedling by culture of plant tissue |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH02222627A (en) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101869076A (en) * | 2010-07-06 | 2010-10-27 | 广东省农业科学院作物研究所 | Method for inoculating stem tip of banana sucker |
CN103444535A (en) * | 2013-09-05 | 2013-12-18 | 广东省农业科学院果树研究所 | Novel method for increasing tissue culture and rapid propagation coefficients of banana |
CN104885935A (en) * | 2015-05-12 | 2015-09-09 | 广西壮族自治区农业科学院甘蔗研究所 | Photoautotrophic rooting method for banana test-tube plantlets |
CN104982285A (en) * | 2015-06-17 | 2015-10-21 | 广东省农业科学院果树研究所 | Method for rapid propagation of banana seedlings based on suckers |
CN106212281A (en) * | 2016-07-28 | 2016-12-14 | 广西陆川县乌坭坡珍珠番石榴专业合作社 | A kind of method for tissue culture improving Fructus Musae survival rate |
CN106416777A (en) * | 2016-09-29 | 2017-02-22 | 河口云山农业科技有限公司 | Banana flower removing method |
CN111543324A (en) * | 2020-06-04 | 2020-08-18 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55118319A (en) * | 1979-03-07 | 1980-09-11 | Kyowa Hakko Kogyo Kk | Mass breeding of plant seedlings |
-
1989
- 1989-02-27 JP JP1043201A patent/JPH02222627A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55118319A (en) * | 1979-03-07 | 1980-09-11 | Kyowa Hakko Kogyo Kk | Mass breeding of plant seedlings |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101869076A (en) * | 2010-07-06 | 2010-10-27 | 广东省农业科学院作物研究所 | Method for inoculating stem tip of banana sucker |
CN103444535A (en) * | 2013-09-05 | 2013-12-18 | 广东省农业科学院果树研究所 | Novel method for increasing tissue culture and rapid propagation coefficients of banana |
CN104885935A (en) * | 2015-05-12 | 2015-09-09 | 广西壮族自治区农业科学院甘蔗研究所 | Photoautotrophic rooting method for banana test-tube plantlets |
CN104885935B (en) * | 2015-05-12 | 2017-03-01 | 广西壮族自治区农业科学院甘蔗研究所 | Banana plantlet in vitro photoautotrophy rooting method |
CN104982285A (en) * | 2015-06-17 | 2015-10-21 | 广东省农业科学院果树研究所 | Method for rapid propagation of banana seedlings based on suckers |
CN106212281A (en) * | 2016-07-28 | 2016-12-14 | 广西陆川县乌坭坡珍珠番石榴专业合作社 | A kind of method for tissue culture improving Fructus Musae survival rate |
CN106416777A (en) * | 2016-09-29 | 2017-02-22 | 河口云山农业科技有限公司 | Banana flower removing method |
CN111543324A (en) * | 2020-06-04 | 2020-08-18 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
CN111543324B (en) * | 2020-06-04 | 2021-09-03 | 云南省农业科学院农业环境资源研究所 | Banana tissue culture method suitable for DNA material requirements of BioNano technology |
Also Published As
Publication number | Publication date |
---|---|
JPH0452734B2 (en) | 1992-08-24 |
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