JP4154458B2 - Strawberry culture method - Google Patents

Description

尚、エチレンジアミン四酢酸鉄を含有させておく場合には、硫酸鉄７水和物、及び、エチレンジアミン４酢酸２ナトリウムは省略する。
【００１７】

【００１８】
ＭＳ培地のビタミン類要素：
ニコチン酸 …………………… ０．５
チアミン塩酸塩 …………………… ０．１
ピリドキシン塩酸塩 …………………… ０．５
ｍ−イノシト−ル …………………… １００
グリシン …………………… ２
【００１９】
【実施例】
以下に本発明の実施例を図面に基づいて説明する。
常法に従って生長点を培養して得たイチゴの苗１を３本づつ、培地Ａ１、Ａ２，Ｄ１〜Ｄ４の５０ｍｌずつ入った密封容器３それぞれに密封し、前記振とう台を用いて液体培地４を常時攪拌し続ける条件下、３５日間イチゴ苗の培養を行って苗の増殖数を調べた。
尚、各培地の組成は、表１に示す通りである。
その結果表２のようになった。
【００２０】
【表１】

【００２１】
【表２】

【００２２】
つまり、先の液体培地組成物Ａ１でイチゴ苗１を培養して増殖を行うと、実験誤差によるばらつきがあるものの、１３０倍程度増殖することが分かり、糖としてブドウ糖を用いた場合にはショ糖を用いた場合に比べ高い増殖性を示すことがわかった。
また、１／４ＭＳに、カイネチンを添加した液体培地４（培地Ｄ２〜Ｄ５）を用いた場合には、ＢＡを用いた場合に比べ、増殖速度が全体的に低いものの、ＢＡを用いた場合に匹敵する増殖率を示した。つまり、１／２ＭＳ〜１／１０ＭＳにカイネチンを添加した液体培地４を用いれば、高い増殖率を得られ、この培地にＢＡを添加すれば、試験例１以上の増殖率の向上を期待することが出来る。尚、カイネチンは、１ｍｇ／リットル以下の範囲で用いていればＢＡと同様の効果を奏することが経験的に知られている。
【００２３】
上述の方法によって増殖したイチゴの苗１を３本づつ、培地Ｂ１、Ｂ２，Ｄ６，Ｄ７の５０ｍｌずつ入った密封容器３それぞれに密封し、前記振とう台を用いて液体培地４を常時攪拌し続ける条件下、培養を行ってイチゴ苗の１５ｍｍ以上のものの数を調べた。
尚、各培地は液体培地Ａ１の基本培地の種類や濃度を様々に変更したものであって、表３に示す通りである。
【００２４】
その結果、平均増殖本数は表４のようになった。
尚、培養期間は９週間である。
【００２５】
【表３】

【００２６】
【表４】

【００２７】
〔試験例２〕
表３に示す液体培地Ｄ及び液体培地Ｅを用いて、試験例１と同様にイチゴ苗を６週間培養して、１５ｍｍ以上の苗の数を調べた。その結果表４のようになった。
【００２８】
表４より、ＨＹ培地を用いた場合には、２５ｍｍ〜３５ｍｍに伸長した比較的長い芽の割合が高く、かつ、取り扱い容易な長い芽数も多く培養されていることがわかる。また同様に、表５に示す液体培地Ｂ２及びＢ３を用いて、イチゴ苗を６週間培養して１５ｍｍ以上の苗の数を調べた。その結果表６のようになった。
【００２９】
【表５】

【００３０】
【表６】

【００３１】
その結果、伸長期に用いる液体培地組成物に用いる糖としては、ショ糖に比べてブドウ糖が好ましいことがわかった。
【００３２】
次に、先の培養によって伸長したイチゴの苗１を、培地Ｃ１〜Ｃ５のそれぞれ５０ｍｌとともに各別に密封容器３に密封し、前記振とう台を用いて液体培地４を常時攪拌し続ける条件下、培養を行ってイチゴ苗の草丈が１５ｍｍ以上で、かつ、赤みがかった太いもの肥大したものの数を調べた。尚、各培地の組成は、表３に示す通りである。
【００３３】
その結果、平均増殖本数は図のようになった。
【００３４】
【表７】

【００３５】
図２より、ショ糖濃度の高い１／４ＭＳ培地を用いた場合には、２５ｍｍ〜３５ｍｍで肥大した芽の割合が高く培養されていることがわかる。
また、各培地Ｃ６〜Ｃ８のそれぞれ５０ｍｌとともに各別に密封容器に封入し、同様に培養したところ、図３のようになった。図３より、培地に含有させる糖分はショ糖よりもブドウ糖が好ましく、糖濃度としては３〜５％程度が好ましいということがわかる。
【図面の簡単な説明】
【図１】イチゴ培養方法を示す工程図
【図２】イチゴ肥大度のショ糖濃度依存性を示すグラフ
【図３】イチゴ肥大度の糖度依存性を示すグラフ[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a technology for mass-culturing strawberry seedlings.
[0002]
[Prior art]
Conventionally, liquid medium compositions for strawberries are known in which MS medium contains sucrose, and the liquid medium composition for strawberries contains benzyladenine (hereinafter abbreviated as BA). The number of strawberry buds is increased (growth), the plant height is increased (elongation), and it is enlarged to a reddish and thicker one using only the liquid medium composition for strawberry. Formation of a structure that can easily withstand the drying of the environment has been performed.
[0003]
[Problems to be solved by the invention]
According to the above-mentioned conventional strawberry cultivation method, the strawberry seedling grows and grows, but there is an insufficient aspect in the degree of growth and the degree of growth, the propagation efficiency is low, and the plant height is sufficient. There were situations where only seedlings that were difficult to handle were obtained. In addition, it has been pointed out that it is difficult to form a tissue that can withstand the drying of the natural environment, and even if it is adapted to the natural environment, it is difficult to grow into a sufficiently strong seedling. A technique for obtaining easy-to-cultivate strawberry seedlings has been desired.
[0004]
Therefore, the object of the present invention has been made in view of the above circumstances.
[0005]
[Means for Solving the Problems]
The present inventors considered that by changing the composition of the liquid medium, it would be possible to obtain strawberry seedlings that can be easily cultivated, and examined various liquid medium compositions. Various new findings were obtained regarding the number of growth, plant height elongation and seedling hypertrophy. The present invention has been made based on the above new findings,
In order to achieve this object, the characteristic means of the present invention includes 0.5% or more and less than 2% glucose in a 2 to 10-fold diluted MS liquid medium in the growth phase to increase the number of strawberry seedling buds, and 0 Cultivating the strawberry seedling in a medium containing not more than 5 mg / liter of benzyladenine (BA),
In the extension period for extending the grown strawberry seedlings, fertilizers with a ratio of nitrogen, phosphorus, and potassium (N: P: K) of 6.5: 6: 19 were added to the trace elements of the MS medium and the MS medium. In a liquid medium containing a vitamin component, 30 mg / liter of iron ethylenediaminetetraacetate, 0.5% to 3% glucose, and 0.5 mg / liter or less of benzyladenine (BA) Cultivate strawberry seedlings,
Thereafter, in the hypertrophy stage where the cultured tissue of the stretched strawberry is grown so that it can be planted in the ground, 2% to 5% glucose or sucrose, and 0.5 mg / liter or less to a 10-fold diluted MS medium The strawberry seedling is cultured in a liquid medium containing benzyladenine (BA), and its actions and effects are as follows.
[0006]
[Function and effect]
First, the present inventors have obtained the knowledge that when a strawberry seedling is grown in a liquid medium, the above-described MS medium can be used by diluting it to increase the growth efficiency of the strawberry seedling ( (See Japanese Patent Application No. 5-158429). Based on this, when the composition of the liquid medium was changed variously, the new finding that the growth rate of the strawberry seedling can be increased if the sugar content is suppressed to a relatively low concentration. Obtained. Furthermore, the present inventors considered that monosaccharides are better absorbed by strawberry seedlings than polysaccharides, and examined various liquid media. As sugar, glucose was used rather than sucrose. In some cases, new findings have been obtained that a higher proliferation efficiency can be obtained. Furthermore, the present inventors have examined the glucose content, and have found that it is desirable to set the content to 0.5% or more and less than 2%.
In addition, a fertilizer having a ratio of nitrogen, phosphorus and potassium (N: P: K) of 6.5: 6: 19, a trace element of MS medium, a vitamin element of MS medium, and ethylenediamine of 30 mg / liter When using a medium containing iron tetraacetate, a new finding was obtained that the plant height of strawberries grew extremely efficiently. Moreover, it is desirable that the liquid medium composition for strawberries contains BA and sugar.
In this case, there is a new finding that it is preferable to use glucose as the sugar rather than sucrose.
Furthermore, when 2% or more and 5% or less of sugar is added to a 4-fold dilution of MS medium, especially strawberry seedlings tend to become reddish and thick (hypertrophies), and the natural environment We obtained new knowledge that it is easy to form a tissue that can withstand the drying of the skin.
In this case, it is also known that if BA is added to the medium composition for growing the plant, the plant is efficiently propagated.
In other words, when mass-cultivating strawberry seedlings, it is possible to efficiently provide strawberry seedlings that are easy to cultivate reliably in the natural environment when the three elements of growth, elongation and hypertrophy are available. In order to satisfy these three elements, culture using a medium composition suitable for satisfying each of these elements is considered to have been impossible with conventional culture methods in that it is intended to satisfy only one kind of medium composition. In other words, it can be said that strawberry seedlings that satisfy all the above-mentioned three elements and can be easily cultivated in a natural environment can be provided.
Therefore, from the various new findings described above, in the growth phase for growing strawberry seedlings, 0.5 to 2% glucose and 0.5 mg / liter or less in a 2 to 10-fold diluted MS liquid medium The strawberry seedlings were grown in a medium containing benzyladenine (BA), and fertilizers having a ratio of nitrogen, phosphorus and potassium (N: P: K) of 6.5: 6: 19 , MS medium medium element and MS medium vitamin element, 30 mg / liter iron ethylenediaminetetraacetate, 0.5% to 3% sugar, and 0.5 mg / liter benzyladenine ( Strawberry seedlings grown in a medium containing BA) are elongated, and 2 to 4 times dilution of MS medium with 2% to 5% sugar and 0.5 mg / liter or less benzyl With adenine (BA) If grown in a certain medium, the strawberry seedling grows and grows into a strong and strong seedling (hereinafter referred to as “healthy seedling”). Strawberry seedlings can be efficiently provided.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below with reference to the drawings.
First, a growth point at the tip of a strawberry stalk is obtained according to a conventional method, and cultured according to a conventional method to form one plant body 1 (hereinafter, the plant body at this stage is referred to as a seedling). The seedling is supported on a support base 2 made of a stainless steel net 2 a and placed on a placement portion 3 a provided in the sealed container 3. In addition, a medium A (described later) is placed as a liquid medium 4 in the sealed container 3 so as to be in contact with the lower part of the plant body. The sealed container is fixed on a shaking table and continuously shaken during the culture period. The medium A is constantly stirred to continuously supply nutrients to the crown portion. Cultivate and grow for a week. (See Figure 1)
[0008]
When strawberry seedlings were grown in this way, the average increased about 130 times. Further, the proliferated strawberry seedlings are separated and cut at 2 cm above and below the crown part, and after removing the root part 1a, the leaf part 1b and the stem part 1c not including the crown part 1d, the strawberry seedlings are cut including the crown part. It is further cultured and propagated as a plant body 1 in which the strawberry seedling is regenerated.
[0009]
Three of the grown seedlings were cut at 2 cm above and below the crown part, and after removing the root part 1a, the leaf part 1b, and the stem part 1c not including the crown part 1d, the liquid medium was similarly used. 4 is put in a sealed container 3 and fixed on a shaking table, and while being continuously shaken during the culture period, the accommodated seedlings are cultured for 9 weeks to proliferate and extend. At this time, the medium B was used as the liquid medium 4.
[0010]
When strawberry seedlings were cultured in this manner, the number of plants having a plant height of 15 mm or more (healthy seedlings) increased to an average of about 130 per container, and extended to an easily handleable length of 2 cm or more in general.
[0011]
Next, the expanded seedling is fixed in a sealed container 3 together with the liquid medium 4 and is continuously shaken during the culture period. The liquid medium 4 is constantly stirred to supply nutrients to the crown portion. Then, the accommodated plant body is cultured and grown for 5 weeks. At this time, the medium C was used as the liquid medium 4.
[0012]
It was found that when the strawberry seedlings were grown in this way, the seedlings were strong reddish. Further, the enlarged strawberry seedlings are stocked and transplanted to a culture soil, then acclimatized to grow strawberry fruits and transplanted in a natural environment.
[0013]
The shaking table used here can rotate the sealed container 3 fixed on the shaking table around a vertical axis so as to draw a circle on a horizontal plane, and stir the culture medium 4 in the sealed container 3. It is. Moreover, the sealed container used here means a container that allows gas to enter and exit inside and outside and blocks invasion of germs and the like.
[0014]
As liquid medium A, MS medium (described later) is diluted 4-fold (hereinafter referred to as 1 / 4MS) (hereinafter referred to as 1 / nMS refers to MS medium diluted n-fold). It is a medium in which glucose is added at 1% by weight and BA is added at 0.03 mg / liter. In the liquid medium B, as a liquid fertilizer having a ratio of nitrogen, phosphorus and potassium (N: P: K) of 6.5: 6: 19, a 0.1% aqueous solution of fine powder Hyponex (manufactured by Hyponex) is used. A medium containing a trace element of MS medium (described later) and a vitamin element of MS medium (described later) as a basic medium (hereinafter referred to as HY medium), 0.5% to 3% glucose, and What contained 30 mg / liter of iron diaminetetraacetate and 0.5 mg / liter or less of benzyladenine (BA) was used. In addition, as the liquid medium C, a medium in which 4% by weight of glucose and BA were added to a 1/4 MS medium so as to be 0.03 mg / liter was used.
[0015]
The composition of the MS medium, and the composition of the trace elements and vitamin elements of the MS medium are shown below in terms of weight per liter (unit: mg).
[0016]

In addition, when including ethylenediaminetetraacetic acid iron, iron sulfate heptahydrate and ethylenediaminetetraacetic acid disodium are omitted.
[0017]

[0018]
Vitamin elements of MS medium:
Nicotinic acid …………………… 0.5
Thiamine hydrochloride …………………… 0.1
Pyridoxine hydrochloride …………………… 0.5
m-Inositol …………………… 100
Glycine …………………… 2
[0019]
【Example】
Embodiments of the present invention will be described below with reference to the drawings.
Three strawberry seedlings 1 obtained by cultivating the growing point according to a conventional method are sealed in sealed containers 3 each containing 50 ml of medium A1, A2, D1 to D4, and liquid medium using the shaking table. Strawberry seedlings were cultured for 35 days under the condition of constantly stirring No. 4 to examine the number of seedlings grown.
The composition of each medium is as shown in Table 1.
As a result, it became as shown in Table 2.
[0020]
[Table 1]

[0021]
[Table 2]

[0022]
That is, when the strawberry seedling 1 is cultured and proliferated with the liquid medium composition A1 described above, it is found that the strawberry seedling 1 grows about 130 times although there is variation due to experimental error. It was found that the growth property was higher than that when using.
In addition, when the liquid medium 4 (mediums D2 to D5) supplemented with kinetin is used for 1/4 MS, the growth rate is generally lower than when BA is used, but when BA is used. The growth rate was comparable. In other words, a high growth rate can be obtained by using liquid medium 4 with kinetin added to 1/2 MS to 1/10 MS, and if BA is added to this medium, an improvement in growth rate over Test Example 1 is expected. I can do it. It is empirically known that kinetin has the same effect as BA when used in a range of 1 mg / liter or less.
[0023]
Each of the three strawberry seedlings 1 grown by the above method is sealed in a sealed container 3 containing 50 ml of each of medium B1, B2, D6, and D7, and liquid medium 4 is constantly stirred using the shaking table. Under the continued conditions, culture was performed to examine the number of strawberry seedlings of 15 mm or more.
Each medium is obtained by changing the type and concentration of the basic medium of the liquid medium A1 as shown in Table 3.
[0024]
As a result, the average number of growth was as shown in Table 4.
The culture period is 9 weeks.
[0025]
[Table 3]

[0026]
[Table 4]

[0027]
[Test Example 2]
Strawberry seedlings were cultured for 6 weeks in the same manner as in Test Example 1 using the liquid medium D and liquid medium E shown in Table 3, and the number of seedlings of 15 mm or more was examined. As a result, it became as shown in Table 4.
[0028]
From Table 4, it can be seen that when the HY medium is used, the proportion of relatively long shoots extending to 25 mm to 35 mm is high, and many long shoots that are easy to handle are also cultured. Similarly, strawberry seedlings were cultured for 6 weeks using the liquid media B2 and B3 shown in Table 5, and the number of seedlings of 15 mm or more was examined. As a result, it became as shown in Table 6.
[0029]
[Table 5]

[0030]
[Table 6]

[0031]
As a result, it was found that glucose is preferable as sucrose as the sugar used in the liquid medium composition used in the extension phase.
[0032]
Next, the strawberry seedling 1 elongated by the previous culture is sealed in a sealed container 3 with 50 ml of each of the media C1 to C5, and the liquid medium 4 is constantly stirred using the shaking table. After culturing, the number of strawberry seedlings with a plant height of 15 mm or more and thick reddish and thickened was examined. The composition of each medium is as shown in Table 3.
[0033]
As a result, the average number of growth was as shown in the figure.
[0034]
[Table 7]

[0035]
FIG. 2 shows that when a ¼ MS medium with a high sucrose concentration is used, the proportion of buds enlarged at 25 mm to 35 mm is high.
Moreover, when each 50 ml of each culture medium C6-C8 was enclosed with each sealed container separately and culture | cultivated similarly, it became like FIG. As can be seen from FIG. 3, the sugar contained in the medium is preferably glucose rather than sucrose, and the sugar concentration is preferably about 3 to 5%.
[Brief description of the drawings]
FIG. 1 is a process chart showing a strawberry culture method. FIG. 2 is a graph showing the dependency of strawberry enlargement on sucrose concentration. FIG. 3 is a graph showing the dependency of strawberry enlargement on sugar content.

Claims (1)

A method for cultivating strawberries,
In the growth phase to increase the number of shoots of strawberry seedlings, 0.5 to 2% glucose and 0.5 mg / liter or less of benzyladenine (BA) are added to the MS liquid medium diluted 2 to 10 times. Cultivate the strawberry seedling in a certain medium,
In the extension period for extending the grown strawberry seedlings, fertilizers with a ratio of nitrogen, phosphorus, and potassium (N: P: K) of 6.5: 6: 19 were added to the trace elements of the MS medium and the MS medium. A liquid medium containing vitamin elements, 30 mg / liter of iron diaminetetraacetate, 0.5% to 3% of sugar, and 0.5 mg / liter or less of benzyladenine (BA) Cultivating the strawberry seedling,
Then, in the hypertrophy stage where the seedlings of the elongated strawberries are grown so that they can be planted in the ground, 2% to 5% diluted MS medium with 2% to 5% glucose or sucrose, and 0.5 mg / liter or less A strawberry culturing method, wherein the strawberry seedling is cultured in a liquid medium containing benzyladenine (BA).

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