JPH0956285A - Preservation and regeneration of plant shoot apex - Google Patents
Preservation and regeneration of plant shoot apexInfo
- Publication number
- JPH0956285A JPH0956285A JP23931895A JP23931895A JPH0956285A JP H0956285 A JPH0956285 A JP H0956285A JP 23931895 A JP23931895 A JP 23931895A JP 23931895 A JP23931895 A JP 23931895A JP H0956285 A JPH0956285 A JP H0956285A
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- JP
- Japan
- Prior art keywords
- plant
- shoot apex
- shoot
- low temperature
- apex
- 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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- Cultivation Of Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は植物の成長点培養に
よる発芽育成等に使用する植物茎頂の保存方法及び再生
方法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preserving and regenerating plant shoot apices used for germination and growth of plants by growing point culture.
【０００２】[0002]
【従来の技術】一般に種子では純粋な固定種を維持する
ことが困難な植物は、圃場環境あるいは施設、制御環境
のもとでその種を維持、保存していく必要がある。この
他に、組織培養による継代保存法あるいは凍結保存法も
可能となっているが、大部分は圃場での栽培により保存
されている。しかし、この方法は病虫害の発生、災害等
による消失の危険も大きく、且つその維持に多大な労力
と経費を必要とする。2. Description of the Related Art In general, it is necessary to maintain and preserve the seeds of plants which are difficult to maintain pure fixed seeds in seeds under the field environment, facility or controlled environment. In addition to this, a subculture preservation method by tissue culture or a cryopreservation method is also possible, but most of them are preserved by cultivation in a field. However, this method has a large risk of occurrence of pests and damages and disappearance due to disasters, and its maintenance requires a great deal of labor and cost.
【０００３】一方、培養法により維持保存する方法は、
講談社「サイエンティフィック」の「植物遺伝資源集
成」第１巻Ｐ．８５に記載されているように、茎頂等を
培養し、物理的あるいは化学的方法により継代培養によ
り維持する方法である。これにより植物体は圃場での場
合と比べ安定的に維持可能であるが、定期的に植え替え
をする必要がある。しかも、これら培養による方法は、
長期的にわたって継代培養を続けていると変異が起こる
危険性も指摘されている。On the other hand, the method of maintaining and preserving by the culture method is
Kodansha "Scientific""Plant Genetic Resource Assembly" Vol. 85, the shoot apex and the like are cultured and maintained by subculture by a physical or chemical method. As a result, the plant can be maintained more stably than in the case of the field, but it is necessary to replant regularly. Moreover, these culture methods are
It has been pointed out that there is a risk of mutations if subculture is continued for a long period of time.
【０００４】また、本出願人の出願に係る特願平６−１
５９２６８号「ワサビ及びササユリ茎頂等の保存再生方
法」の明細書及び図面に記載されているように、ガラス
化法あるいは緩速予備凍結法による植物の茎頂の超低温
保存法が提案されている。この方法は、半永久的に省ス
ペースでの保存が可能な方法であるが、用いる茎頂が１
ｍｍ程度と小さいため、一度に大量の茎頂を処理できな
いという欠点を持っている。したがって、より省力的、
効率的で遺伝的に安定した長期保存法の開発が要望され
ている。Further, Japanese Patent Application No. 6-1 filed by the present applicant.
As described in the specification and drawings of No. 59268 "Preservation and Regeneration Method of Wasabi and Sasayuri Shoot Apex, etc.", an ultracryopreservation method of plant shoot apices by a vitrification method or a slow prefreezing method has been proposed. . This method is a method that can be stored semi-permanently in a space-saving manner.
Since it is as small as mm, it has a drawback that it cannot process a large amount of shoot tips at one time. Therefore, more labor-saving,
Development of an efficient and genetically stable long-term storage method is desired.
【０００５】[0005]
【発明が解決しようとする課題】植物体の長期的な保存
は、現時点では主として圃場で栽培する方法と培養によ
り試験管内で維持する方法が採られている。前者は、品
種、系統等を圃場で栽培しながら株分けにより維持保存
していく方法であるが、ウイルス等の病害に感染しやす
い欠点を持っている。また、天災あるいは人災により圃
場が壊滅する危険性を常に持っている。For the long-term storage of plants, at present, a method of mainly cultivating in a field and a method of maintaining in a test tube by culture are adopted. The former is a method of maintaining and preserving cultivars, strains, etc. by dividing them into strains while cultivating them in the field, but they have the drawback of being easily infected by diseases such as viruses. In addition, there is always a risk that the fields will be destroyed by natural disasters or man-made disasters.
【０００６】一方、後者は、天候に左右されず、病虫害
の問題もなく遺伝的に同じ形質のクローンを維持できる
ため有効な方法と言える。しかし、定期的な分割、培地
更新が不可欠であり、これに要する労力、経費も長期間
になるほど膨大なものとなる。また、植物体として維持
保存するため、それらを入れた培養容器の占有面積は決
して小さいものでなく、それだけ電気代等のコストが高
くなる結果となる。さらに、保存中の変異も無視できな
い。On the other hand, the latter can be said to be an effective method because it is possible to maintain clones of the same trait genetically without being affected by weather and without the problem of pests and diseases. However, periodic division and medium renewal are indispensable, and the labor and cost required for this become enormous as they become longer. Further, since the cells are maintained and preserved as plants, the occupying area of the culture vessel containing them is by no means small, and the cost such as electricity bill is increased accordingly. Furthermore, mutations during storage cannot be ignored.
【０００７】また、前述したガラス化法あるいは緩速予
備凍結法による超低温保存は操作が煩雑であるため効率
が悪いという欠点を持っている。本発明は以上のような
諸欠点を改善し、長期保存をより省力的、効率的にする
ため、ガラス化法による超低温保存法を更に発展させる
方法を提供せんとするものである。Further, the ultra-low temperature preservation by the above-mentioned vitrification method or slow-preliminary freezing method has a drawback that the operation is complicated and the efficiency is low. The present invention aims to provide a method for further developing the ultra-low temperature preservation method by vitrification method in order to improve the above-mentioned various drawbacks and to make long-term preservation more labor-saving and efficient.
【０００８】[0008]
【課題を解決するための手段】上記問題点を解決するた
めの本発明の方法は、第１に植物の茎頂をアルギン酸ビ
ーズ内に包埋した後に、超低温で急速冷却した後、超低
温域で保存することを特徴としている。Means for Solving the Problems The method of the present invention for solving the above-mentioned problems is as follows. First, after embedding the shoot apices of plants in alginate beads, the mixture is rapidly cooled at ultra-low temperature and then at ultra-low temperature range. It is characterized by saving.
【０００９】第２にアルギン酸ビーズに包埋される植物
の茎頂を脱水耐性を付与する前処理を施した後、ガラス
化液に浸漬して浸透脱水させ、超低温で冷却し、超低温
域で保存することを特徴としている。Secondly, the shoot apex of the plant embedded in alginate beads is subjected to a pretreatment for imparting dehydration resistance, then immersed in a vitrification solution for permeation dehydration, cooled at an ultralow temperature, and stored at an ultralow temperature range. It is characterized by doing.
【００１０】第３にアルギン酸ビーズに包埋した植物の
茎頂において超低温以下に急速冷却する前に、脱水耐性
を付与するため前処理をすることを特徴としている。[0010] Thirdly, the plant apex embedded in alginate beads is characterized by being subjected to a pretreatment for imparting dehydration resistance before being rapidly cooled to an ultralow temperature or lower.
【００１１】第４に植物の茎頂を前処理する前に、脱水
耐性を付与するためあらかじめ茎頂を高濃度のしょ糖液
を含む培地で前培養することを特徴としている。Fourthly, before the pretreatment of the shoot apices of plants, the shoot apices are pre-cultured in a medium containing a high-concentration sucrose solution in order to impart dehydration resistance.
【００１２】第５に超低温保存したアルギン酸を含むビ
ーズに包埋した植物の茎頂および腋芽を温水で急速加温
した後、高濃度のしょ糖液で洗浄処理することにより徐
々に吸水させ、植物体に再生させることを特徴としてい
る。Fifthly, the shoot apex and axillary buds of the plant embedded in beads containing alginic acid stored at ultra-low temperature were rapidly heated with warm water, and then washed with a high-concentration sucrose solution to gradually absorb water to give a plant body. It is characterized by being played back.
【００１３】[0013]
【発明の実施の形態】本発明においては、植物体から無
菌的に摘出した茎頂を、高濃度しょ糖を含む培地で前培
養し、いくら状に凝固したアルギン酸ビーズ中に包埋
し、同時に脱水耐性を付与するための凍害制御剤処理の
第１段階と、グリセリンを主体とした濃厚なガラス化液
に一定時間浸漬して脱水させた後に液体窒素温度に急速
冷却してガラス化させる第２段階、所定温度の温水で急
速加温した後に高濃度しょ糖液で洗浄し、固形培地上に
置床して植物体を再生させる第３段階の３つの工程から
なる。BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the shoot apex aseptically extracted from a plant is pre-cultured in a medium containing high-concentration sucrose, embedded in alginate beads which are coagulated to a degree, and simultaneously dehydrated. First step of frost damage control agent treatment for imparting resistance, and second step of vitrification by rapid cooling to liquid nitrogen temperature after immersion in a concentrated vitrification solution containing glycerin for a certain period of time for dehydration The method comprises three steps of a third stage of rapid heating with warm water of a predetermined temperature, washing with a high-concentration sucrose solution, and placing on a solid medium to regenerate plants.
【００１４】[0014]
【実施例】以下本発明の方法をわさびの茎頂について実
施した実施例につき詳述すると、まず第１段階では、無
菌的に摘出した植物（わさび）の茎頂１を、表１に示す
硝酸アンモニウムと硝酸カリウム（ＮＨ４ＮＯ３、ＫＮ
Ｏ３）を１／２に希釈した改変ムラシゲ・スクーグ培地
（１／２ＭＳ培地）に０．３Ｍしょ糖を添加した固形培
地２をシャーレ等の蓋付の容器３に収容し、その内部で
１６〜２４時間、２０℃、２，０００Ｌｘで前培養す
る。[Examples] The following is a detailed description of an example in which the method of the present invention was carried out on the shoot apices of wasabi. First, in the first step, the shoot apices 1 of plants (wasabi) aseptically extracted were treated with ammonium nitrate as shown in Table 1. And potassium nitrate (NH 4 NO 3 , KN
O 3 ) -diluted modified Murashige-Skoog medium (1/2 MS medium) into which solid medium 2 containing 0.3 M sucrose is contained in a container 3 with a lid such as a petri dish, and 16 to 16 Pre-incubate at 2,000 Lx at 20 ° C. for 24 hours.
【００１５】表１Table 1
【００１６】第２段階では、図１に示すように予めアル
ギン酸ナトリウムに第１段階で得られた植物の茎頂１を
入れたものを、ピペット４等を用いてビーカー等の容器
６中にある塩化カルシウム液に滴下することにより、各
１個の茎頂を含む多数のビーズ７を作成する。これら２
種類の溶液８ａ，８ｂには表２に示すような前処理（ｌ
ｏａｄｉｎｇ）液を含んでいるため、３０分間のビーズ
の凝固時間経過と同時に前処理も完了する。次に表２及
び図１で示す容器１１中のガラス化液（ＰＶＳ２液）９
にビーズ７を浸漬し脱水させ、ガラス化液９とともに密
閉容器１２内に収容して液体窒素１５中で−１９６℃等
の超低温で急速冷却する。このガラス化処理条件は、わ
さびの場合０℃では１００分間、２５℃では３０分間の
処理が適当であった（図２）。また、この処理中に組織
から脱水された水分でガラス化液９が希釈されるため、
途中で数回の液交換が必要である。これらの液剤処理は
ビーズガラス化法では図示するようにビーカー等の容器
６，１１を用いることができるため、液の交換を極めて
容易に行うことが可能となる。In the second step, as shown in FIG. 1, the plant apex 1 of the plant obtained in the first step is put in sodium alginate in advance and placed in a container 6 such as a beaker using a pipette 4 or the like. A large number of beads 7 each including one shoot apex are prepared by dropping the calcium chloride liquid. These two
For the types of solutions 8a and 8b, pretreatment (l
Therefore, the pretreatment is completed at the same time as the bead coagulation time of 30 minutes has elapsed. Next, the vitrification liquid (PVS2 liquid) 9 in the container 11 shown in Table 2 and FIG.
The beads 7 are immersed in and dehydrated in a closed container 12 together with the vitrification liquid 9 and rapidly cooled in liquid nitrogen 15 at an ultralow temperature such as -196 ° C. As for the vitrification treatment conditions, wasabi was suitable for treatment at 0 ° C. for 100 minutes and at 25 ° C. for 30 minutes (FIG. 2). Further, since the vitrification liquid 9 is diluted with the water dehydrated from the tissue during this treatment,
It is necessary to change the liquid several times on the way. In the bead vitrification method, as shown in the figure, containers 6 and 11 such as beakers can be used for these liquid agent treatments, so that the liquids can be exchanged very easily.
【００１７】表２Table 2
【００１８】第３段階では、超低温からの加温中におけ
るガラス化した茎頂組織内での細胞内凍結による細胞の
破壊を回避するため、例えば４０℃の温水１９中で急速
加温する。この加温によって融解した後、直ちにガラス
化液９を抜き取り、１．２Ｍしょ糖液１３に約３０時間
位浸漬し、ガラス化液の有害物を希釈するとともに急速
吸水の害を防ぐ。洗浄後のビーズ７は、わさびでは容器
１４内においてベンジルアデニン０．１ｍｇ／ｌ添加し
た固形培地１６上に置床し、翌日約２４時間後培地１６
を交換する。再培養した茎頂１は、フラスコ等の容器１
７内で２５℃、２，０００Ｌｘの条件下において７〜１
０日後にシュート１８の再生が認められる。In the third step, in order to avoid destruction of cells due to intracellular freezing in vitrified shoot apical tissue during heating from ultra-low temperature, rapid heating is performed in warm water 19 at 40 ° C., for example. Immediately after melting by virtue of this heating, the vitrification liquid 9 is drawn out and immersed in 1.2M sucrose solution 13 for about 30 hours to dilute the harmful substances of the vitrification liquid and prevent the rapid absorption of water. The beads 7 after washing were placed on a solid medium 16 containing 0.1 mg / l of benzyladenine in a container 14 with wasabi, and after 24 hours, the medium 16 was washed.
To replace. Recultured shoot apex 1 is a container 1 such as a flask
7 to 25 under conditions of 25 ° C. and 2,000 Lx
After 0 days, regeneration of shoot 18 is recognized.
【００１９】なお、上記実施例で液体窒素１５で急速冷
却完了後、第３段階の工程に移行する説明をしたが、図
１に示すようにこれを超低温フリーザー２１等により−
１３５℃以下で保存すれば半永久的に保存することがで
き、実際にはこのような保存ができる点に意義がある。In the above-mentioned embodiment, the description has been given of shifting to the step of the third stage after completion of the rapid cooling with the liquid nitrogen 15. However, as shown in FIG.
If it is stored at 135 ° C. or lower, it can be stored semipermanently, and it is significant that such storage can be carried out in practice.
【００２０】以上説明したように、本実施例において
は、植物の茎頂をそのまま、あるいは高濃度のしょ糖液
を含む培地で前培養し、アルギン酸ビーズ内に包埋す
る。次に脱水耐性を付与するため前処理を施した後、ガ
ラス化液に浸漬して浸透脱水させ、直ちに液体窒素中に
浸漬する。このことにより茎頂組織は、結晶を含まない
固体であるガラス化状態となるため危険な細胞内凍結を
回避することができ、−１３５℃以下の超低温で安定的
に保存することができる。As described above, in this example, the shoot apex of the plant is pre-cultured as it is or in a medium containing a high-concentration sucrose solution and embedded in alginate beads. Then, after performing a pretreatment for imparting dehydration resistance, it is immersed in a vitrification liquid to permeate and dehydrate, and immediately immersed in liquid nitrogen. As a result, the shoot apical tissue is in a vitrified state, which is a solid that does not contain crystals, so that dangerous intracellular freezing can be avoided and it can be stably stored at an ultralow temperature of -135 ° C or lower.
【００２１】上記方法により超低温保存した植物の茎頂
及び腋芽は、加温時においても細胞内凍結の危険がある
ため、４０℃温水で急速加温する。これらによって種子
としての保存が困難な絶滅危惧の稀少植物、育種素材と
しての野生種、栽培種等の遺伝資源、あるいは栽培種の
親株等を半永久的に安定して保存することが可能とな
る。The shoot apices and axillary buds of a plant which have been cryopreserved by the above method are rapidly heated with warm water at 40 ° C. because there is a risk of intracellular freezing even during heating. As a result, it becomes possible to semi-permanently and stably preserve endangered rare plants that are difficult to store as seeds, wild species as breeding materials, genetic resources such as cultivated species, or parent strains of cultivated species.
【００２２】そして保存に必要なスペースは、１０茎頂
当たり約２ｍｌであるため、狭い場所でも多くの品種、
系統の保存が可能となる。したがって、これまでの方法
による保存中の維持にかかる多労性、高コスト性等の欠
点を大幅に改善することができる。また、従来のガラス
化法による超低温保存法と比べ、約３ｍｍの大きさのビ
ーズ中に茎頂を包埋しているため、各処理において最も
繁雑な操作である液交換についてはピペット等を必要と
せず、ビーカー等を利用することができるため、その操
作をより容易に、且つ確実に行うことが可能となる。さ
らに、超低温下では、細胞の生体反応が停止するため、
保存期間に関わらず変異の発生が極めて低いとされてい
る。Since the space required for storage is about 2 ml per 10 shoot tips, many varieties can be stored even in a narrow space.
The strain can be preserved. Therefore, it is possible to greatly reduce the drawbacks such as the labor-intensive and high-cost performances required for the maintenance during storage by the conventional methods. In addition, compared to the conventional cryogenic preservation method by vitrification, the stem apex is embedded in beads of about 3 mm in size, so a pipette or the like is required for the most complicated operation of liquid exchange in each process. Instead, since a beaker or the like can be used, the operation can be performed more easily and reliably. Furthermore, since the biological reaction of cells is stopped under ultra-low temperature,
It is said that the occurrence of mutations is extremely low regardless of the storage period.
【００２３】特に現在、圃場あるいは試験管内で行われ
ている植物の品種、系統等の維持保存を、ガラス化法ま
たは緩速予備凍結法による超低温保存法を利用すること
により、省力的、効率的、且つ安定的となるが、本発明
にかかるビーズガラス化法により、更に容易に且つ確実
に大量の茎頂の処理を行うことが可能となる。[0023] In particular, for the preservation and preservation of plant varieties, strains, etc., which are currently conducted in the field or in a test tube, by utilizing the ultra-low temperature preservation method by the vitrification method or the slow preliminary freezing method, it is labor-saving and efficient. Although it is stable, the bead vitrification method according to the present invention makes it possible to treat a large amount of shoot tips more easily and reliably.
【００２４】[0024]
【発明の効果】以上のように構成される本発明によれ
ば、以下のような効果を奏するものである。 （１）茎頂組織は、結晶を含まない固体であるガラス化
状態となるため危険な細胞内凍結を回避することがで
き、超低温域で安定的に保存することができる。 （２）超低温保存した植物の茎頂は、温水で急速加温す
るので、加温時においても細胞内凍結の危険がなくな
る。 （３）種子としての保存が困難な絶滅危惧の稀少植物、
育種素材としての野生種、栽培種等の遺伝資源、あるい
は栽培種の親株等を半永久的に安定して保存することが
可能となり、更に容易且つ確実に大量の茎頂の処理を行
うことが可能となる。 （４）保存に必要なスペースが少なくてすむため、狭い
場所でも多くの品種、系統の保存が可能となる。したが
って、これまでの方法による保存中の維持にかかる多労
性、高コスト性等の欠点を大幅に改善することができ
る。 （５）従来のガラス化法による超低温保存法と比べ、ビ
ーズ中に茎頂を包埋しているため、各処理において最も
繁雑な操作である液交換についてはピペット等を必要と
せず、ビーカー等を利用することができる結果、その操
作をより容易に、且つ確実に行うことが可能となる。 （６）超低温下では、細胞の生体反応が停止するため、
保存期間に関わらず変異の発生が極めて低い。According to the present invention configured as described above, the following effects are exhibited. (1) Since the shoot apex tissue is in a vitrified state, which is a solid containing no crystals, dangerous intracellular freezing can be avoided and the stem apex tissue can be stably stored in an ultralow temperature range. (2) Since the shoot apex of a plant preserved at ultra-low temperature is rapidly heated with warm water, there is no danger of intracellular freezing even during heating. (3) An endangered rare plant that is difficult to store as seeds,
Genetic resources such as wild species and cultivated species as breeding materials, or parent strains of cultivated species can be stored semi-permanently and stably, and a large amount of shoot tips can be processed more easily and reliably. Becomes (4) Since the space required for storage is small, many varieties and strains can be stored even in a narrow space. Therefore, it is possible to greatly reduce the drawbacks such as the labor-intensive and high-cost performances required for the maintenance during storage by the conventional methods. (5) Compared with the conventional ultra-low temperature preservation method by vitrification, since the shoot apex is embedded in the beads, pipetting is not required for liquid exchange, which is the most complicated operation in each process, and beakers, etc. As a result, the operation can be performed more easily and reliably. (6) Since the biological reaction of cells is stopped under ultra-low temperature,
The occurrence of mutations is extremely low regardless of the storage period.
【図１】本発明方法による植物茎頂の保存及び再生方法
の工程説明図である。FIG. 1 is a process explanatory view of a method for storing and regenerating a plant apex according to the method of the present invention.
【図２】（Ａ），（Ｂ）はわさび茎頂のシュート形成率
に及ぼすガラス化処理温度と処理時間の関係を２５℃及
び０℃の場合について示す線図である。FIG. 2 (A) and (B) are diagrams showing the relationship between the vitrification treatment temperature and the treatment time that affect the shoot formation rate of wasabi shoot apices at 25 ° C. and 0 ° C.
１ 茎頂 ２ しょ糖培地 ７ ビーズ ８ａ，８ｂ 前処理及びビーズ作成用溶液 ９ ガラス化（ＰＶＳ２）液 １１ 容器 １３ 高濃度しょ糖液 １５ 液体窒素 １６ 固形培地 1 Stem tip 2 Sucrose medium 7 Beads 8a, 8b Pretreatment and bead making solution 9 Vitrification (PVS2) solution 11 Container 13 High concentration sucrose solution 15 Liquid nitrogen 16 Solid medium
【表１】 [Table 1]
【表２】 [Table 2]
Claims (5)
した後に、超低温で急速冷却した後、超低温域で保存す
る植物茎頂の保存方法。1. A method for preserving plant shoot apices, which comprises embedding a shoot apex of a plant in alginate beads, rapidly cooling it at a super low temperature, and then storing the shoot at a super low temperature range.
ーズに包埋される植物の茎頂を脱水耐性を付与する前処
理を施した後、ガラス化液に浸漬して浸透脱水させ、超
低温で冷却し、超低温域で保存する植物茎頂の保存方
法。2. The method according to claim 1, wherein a shoot apex of a plant embedded in alginate beads is subjected to a pretreatment for imparting dehydration resistance, and then immersed in a vitrification solution for permeation dehydration and cooled at an ultralow temperature. A method of preserving plant shoot apices that is stored in the ultra-low temperature range.
酸ビーズに包埋した植物の茎頂において超低温に急速冷
却する前に、脱水耐性を付与するため前処理をする植物
茎頂の保存方法。3. The method for preserving plant shoot apex according to claim 1, wherein the shoot apex of the plant embedded in alginate beads is pre-treated to impart dehydration resistance before being rapidly cooled to ultralow temperature.
頂を前処理する前に、脱水耐性を付与するためあらかじ
め茎頂を高濃度のしょ糖液を含む培地で前培養する植物
茎頂の保存方法。4. The plant shoot apex according to claim 1, wherein the shoot apex is pre-cultured in a medium containing a high-concentration sucrose solution in order to impart dehydration resistance before pretreating the shoot apex of the plant. How to save.
存したアルギン酸ビーズに包埋した植物の茎頂を温水で
急速加温した後、高濃度のしょ糖液で洗浄処理すること
により徐々に吸水させ、植物体に再生させる植物茎頂の
再生方法。5. The method according to claim 1, wherein the shoot apex of the plant embedded in the alginate beads stored at ultra-low temperature is rapidly heated with warm water, and then washed with a high-concentration sucrose solution to gradually absorb water. And a method for regenerating a plant shoot apex that allows the plant to regenerate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23931895A JPH0956285A (en) | 1995-08-24 | 1995-08-24 | Preservation and regeneration of plant shoot apex |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23931895A JPH0956285A (en) | 1995-08-24 | 1995-08-24 | Preservation and regeneration of plant shoot apex |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0956285A true JPH0956285A (en) | 1997-03-04 |
Family
ID=17042939
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP23931895A Pending JPH0956285A (en) | 1995-08-24 | 1995-08-24 | Preservation and regeneration of plant shoot apex |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0956285A (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823582A (en) * | 2012-09-18 | 2012-12-19 | 上海交通大学 | Vitrification ultralow-temperature preserving method for agapanthus embryogenic callus |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN104255709A (en) * | 2014-09-15 | 2015-01-07 | 上海交通大学 | Method for improving preservation effect of agapanthus embryonic callus |
| CN104255705A (en) * | 2014-09-04 | 2015-01-07 | 中国农业科学院作物科学研究所 | Method for preventing vitrification of regenerated seedlings of jerusalem artichoke stem tips subjected to ultralow-temperature storage |
| CN105706922A (en) * | 2016-01-29 | 2016-06-29 | 云南省农业科学院花卉研究所 | Ultralow-temperature dendrobium nobile preserving and virus removing method |
| CN111513061A (en) * | 2020-05-22 | 2020-08-11 | 上海市农业生物基因中心 | Ultralow-temperature preservation and recovery culture method for alum root clump buds |
| CN115363020A (en) * | 2022-10-24 | 2022-11-22 | 中国科学院昆明植物研究所 | Preservation method of bulbil germplasm resources |
-
1995
- 1995-08-24 JP JP23931895A patent/JPH0956285A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823582A (en) * | 2012-09-18 | 2012-12-19 | 上海交通大学 | Vitrification ultralow-temperature preserving method for agapanthus embryogenic callus |
| CN104255705A (en) * | 2014-09-04 | 2015-01-07 | 中国农业科学院作物科学研究所 | Method for preventing vitrification of regenerated seedlings of jerusalem artichoke stem tips subjected to ultralow-temperature storage |
| CN104255705B (en) * | 2014-09-04 | 2015-11-04 | 中国农业科学院作物科学研究所 | One overcomes the vitrified method of regrowth after jerusalem artichoke stem apex Excised Embryos |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN104255709A (en) * | 2014-09-15 | 2015-01-07 | 上海交通大学 | Method for improving preservation effect of agapanthus embryonic callus |
| CN104170818B (en) * | 2014-09-15 | 2015-08-26 | 上海交通大学 | A kind of method optimizing Agipanthus embryo callus vitrification ultra-low temperature preservation effect |
| CN105706922A (en) * | 2016-01-29 | 2016-06-29 | 云南省农业科学院花卉研究所 | Ultralow-temperature dendrobium nobile preserving and virus removing method |
| CN111513061A (en) * | 2020-05-22 | 2020-08-11 | 上海市农业生物基因中心 | Ultralow-temperature preservation and recovery culture method for alum root clump buds |
| CN111513061B (en) * | 2020-05-22 | 2022-05-03 | 上海市农业生物基因中心 | Ultralow-temperature preservation and recovery culture method for alum root clump buds |
| CN115363020A (en) * | 2022-10-24 | 2022-11-22 | 中国科学院昆明植物研究所 | Preservation method of bulbil germplasm resources |
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