JP2001186814A - Method for preparing new rooted cutting - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for preparing new rooted cutting, applicable even to a plant whose growth by cuttage regarded as impossible or difficult, having high productivity. SOLUTION: A rooting bed wetted with a liquid medium containing nitrogen, phosphorus and potassium as essential elements but not a carbon source is prepared in a culture container, a cutting ear is inserted into the rooting bed, cultured and rooting is carried out from the cutting ear while controlling the concentration of carbon dioxide gas in the culture container.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a technique for commercial mass production of cloned seedlings having excellent traits by a cutting method.

[0002]

2. Description of the Related Art Cutting is a tradition in which a cut portion of an artificially cut plant tissue (cutting) is inserted into a rooting bed and roots are formed in the rooting bed to produce an independent plant. This is a popular vegetative propagation method, and is widely used as a method for producing and growing a large number of individuals having the same genetic properties as the parent plant, that is, clone seedlings, from herbaceous plants to woody plants.

This method is simple and suitable for obtaining a large number of seedlings at one time, and is capable of mass-producing individuals with excellent traits at low cost. Plants cannot be propagated this way.
Even if this method is used, the cuttings do not root at all,
There are many species and varieties whose rooting rate is extremely low and cannot be propagated.

Cutting is performed outdoors or in a greenhouse or frame. On the other hand, in this cutting method, the environment must be maintained at a relatively high humidity until the cuttings start rooting and form healthy seedlings. When the humidity is low,
This is because the cuttings are withered and weakened due to the transpiration from the leaves. However, even when cutting in a greenhouse or frame, as well as outdoors, it takes a lot of time to maintain a high humidity environment.

[0005] Under such circumstances, there are still many plants that have excellent traits but cannot reproduce the individual at a commercially acceptable level.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for producing a cutting seedling which can be applied to a plant which has heretofore been impossible or difficult to propagate by cutting.

[0007] Further, the present invention can maintain the environment from cutting to the formation of a seedling at high humidity,
It is another object of the present invention to provide a method for producing cuttings that can improve the productivity of seedlings.

Another object of the present invention is to provide a commercially advantageous method for mass-producing cloned seedlings that can be applied to many plants.

[0009]

Means for Solving the Problems As a result of intensive studies, the inventors of the present invention have found that the above object can be achieved by cultivating cuttings by providing nutrients necessary for plant growth in a culture vessel. Heading, the present invention has been completed.

That is, the present invention provides a rooting bed which contains nitrogen, phosphorus and potassium as essential elements and is moistened with a liquid medium containing no carbon source, in a culture vessel, into which cuttings are inserted. In addition, the present invention provides a method for producing a cutting seedling, wherein rooting is performed from cuttings while controlling the concentration of carbon dioxide in a culture vessel.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail.

There is no particular limitation on the types of plants to which the present invention can be applied. The present invention can be applied to herbaceous plants such as chrysanthemums and carnations in addition to woody plants such as eucalyptus, acacia, bayberry, oak, grape, apple, cherry, rose, camellia, and plum. In addition, the present invention can be applied to plants that have conventionally been difficult to reproduce by the cutting method and those that do not.

As cuttings, in addition to green branches (current branches) and mature branches (branches that have grown before the previous year), shoots and leaves can also be used. In the case of woody plants, it is common to use green branches and mature branches, and in the case of herbaceous plants, leaves and buds are used. In the conventional method for producing cuttings, when using a branch as a cutting, it was necessary to remove a part of the leaf in order to suppress the transpiration of the leaf attached to the branch. In the invention,
Since the cuttings are placed under high humidity, the transpiration is suppressed even without such treatment. Therefore, such pre-processing is not required.

The method for producing cuttings of the present invention uses a rooting bed moistened with a liquid medium and is carried out in a culture vessel, so that it is easy to maintain the humidity of the environment surrounding the cuttings and the seedlings formed therefrom. I do. For example, if a wide-mouthed flask or the like with an opening higher than the height of the cuttings produced in this culture vessel is used, the culture environment can be compared until healthy seedlings are formed from the cuttings. Naturally kept at high humidity. By devising not only such a container but also the size and position of the container opening, the environment in the culture container can be maintained at a high humidity in various types of containers. Therefore, in the present invention, various types of containers can be selected as the culture container in consideration of workability and the like. From the viewpoint of maintaining the environment in the culture vessel at a high humidity, the use of a closed vessel is the most preferable, but even in this case, carbon dioxide must be supplied into the culture vessel. For example, a method of covering the container opening with a carbon dioxide gas permeable membrane can be employed for such a purpose.

The rooting bed is one that is substantially uniformly wetted by the liquid medium and that can hold the cuttings inserted thereinto in the inserted state. For example, sand, natural soil such as akadama, artificial soil such as vermiculite, perlite, glass beads, or a porous molded product such as foamed phenolic resin and rock wool are placed in a culture vessel and used as a rooting bed. can do.

In the present invention, the liquid medium contains nitrogen, phosphorus and potassium as essential elements. The concentration of each of these elements is adjusted according to the type of plant to which the element is applied. As such a liquid medium, a commercially available complex fertilizer for home gardening or a known liquid medium for plant tissue culture can be used as it is or appropriately diluted. For example, as a compound fertilizer for home gardening, "Hyponex liquid 5-10-5 (registered trademark)" containing nitrogen, phosphorus, and potassium as main components is used.
(Manufactured by Hyponex Japan Co., Ltd.)
A 0-fold diluted solution is used as a liquid medium for plant tissue culture as Murashige and Skoog (19).
62), hereinafter abbreviated as MS medium. ) Can be used as a highly versatile liquid medium in the present invention.

In addition to the above-mentioned MS medium, known mediums for culturing plant tissues include nitrogen, phosphorus, potassium, hydrogen, carbon, oxygen, sulfur, calcium, and magnesium as major elements and iron, manganese as trace elements. , Copper, zinc, molybdenum, boron, chlorine, inorganic salts, or thiamine,
Contains as vitamins such as pyridoxine and nicotinic acid. Therefore, as the liquid medium of the present invention, those containing these elements as inorganic salts or vitamins in addition to nitrogen, phosphorus and potassium can also be used.

The liquid medium used in the present invention may further contain a plant growth regulator. For example, auxins, such as indoleacetic acid, indolebutyric acid (IBA), and naphthaleneacetic acid, which promote adventitious root development from plant tissues, alone or in combination of two or more,
By adding 0.1 to 10 mg / l to the liquid medium of the present invention, rooting from cuttings, that is, formation of cuttings, can be promoted.

On the other hand, the liquid medium of the present invention does not contain a carbon source such as sucrose. The carbon source is an energy source common to many organisms. However, in the present invention, as in the ordinary method for producing cuttings and seedlings, operation under aseptic conditions is not a particularly essential condition. When a medium containing is used, various bacteria attached to the cuttings and various bacteria in the culture environment also propagate using the carbon source in the medium as a nutrient source, causing the cuttings and seedlings formed therefrom to die. .

However, not only the seedlings formed from the cuttings but also the cuttings themselves have the ability to perform photosynthesis by themselves. Therefore, by giving light of an appropriate intensity to the growth of plants, carbon dioxide can be removed. Into an energy source. Therefore, even in the cuttings of the present invention, rooting and the like should be able to be performed using carbon dioxide gas in the atmosphere without providing a carbon source as a nutrient. But,
In the present invention, cuttings to which nutrients such as nitrogen, phosphorus and potassium are given in a liquid medium actively perform photosynthesis, so that it is necessary to artificially control the concentration of this carbon dioxide gas. That is, the active photosynthesis of the cuttings lowers the concentration of carbon dioxide in the culture vessel, and it is necessary to artificially compensate for this. Otherwise, even if nutrients other than carbon dioxide are sufficiently supplied to the cuttings, their photosynthetic ability will eventually decrease and rooting from the cuttings,
That is, seedling formation is inhibited.

[0021] The carbon dioxide concentration required for vigorously photosynthesizing the cuttings in the culture vessel and improving the rooting rate varies depending on the type of plant used as the cuttings. However, generally, the concentration of carbon dioxide in the culture vessel is set to 300.
It is preferable to control to ５００1500 ppm. If the concentration of carbon dioxide in the culture vessel is lower than 300 ppm, neither photosynthetic ability nor rooting rate of cuttings can be expected to be significantly improved. Also,
Even if the concentration of carbon dioxide in the culture vessel is increased to more than 1500 ppm, the photosynthetic ability and rooting rate of the cuttings will not show an improvement corresponding to the carbon dioxide concentration. Control of carbon dioxide concentration
Although it may be carried out for each culture vessel, it is simpler and more cost effective to control the inside of the culture vessel to a predetermined carbon dioxide concentration by controlling the carbon dioxide concentration of the environment in which the culture vessel is placed. It is advantageous. At this time, the culture vessel may be one having the opening portion opened as it is or one having the opening portion covered with a carbon dioxide permeable membrane as described above.

In the method for producing cuttings of the present invention,
There is no particular limitation on other conditions, that is, the conditions of temperature and light intensity for culturing the cuttings. Conditions suitable for the plant from which the cuttings are derived to perform photosynthesis may be appropriately adopted. Generally, a temperature of 20 to 30 ° C. and a light intensity of 40 to 1
Conditions on the order of 00 μmol / m ² / sec are employed for this purpose. In the present invention, a light period in which the culture is performed by irradiating light and a dark period in which the culture is performed in the dark may be set, and the culture may be performed by alternately repeating the light period and the dark period. In this case, since photosynthesis is performed only during the light period, the control of carbon dioxide in the culture vessel may be performed only during the light period.

As described above, in the present invention, it is not necessary to perform the operation under aseptic conditions. However, in order to ensure a healthy seedling, the culture container, liquid medium, and rooting bed must be subjected to heat sterilization, autoclave sterilization, etc. before inserting cuttings. Is preferred.

The cuttings produced in the present invention are:
Immediately after rooting, it can be taken out of the culture container, transplanted into a seedling raising container, and grown. Conditions such as soil used for transplanting to the seedling raising container and temperature and light intensity for growing seedlings may be appropriately set so as to be suitable for the plant. Through such a breeding process, a seedling that can be used for a predetermined purpose such as tree planting can be obtained.

[0025]

In the past, it has been contraindicated to provide nutrients before rooting of cuttings when producing and growing seedlings with cuttings. This is because the pre-root cuttings do not have the normal ability to absorb nutrients, and the application of nutrients at this stage was thought to only cause rot of the cuttings (eg, "grafting, cuttings, The Reality of Toriki ”(Taikodo Co., Ltd., issued on July 1, 1986, p. 34).

However, the present inventors can utilize elements such as nitrogen, phosphorus, and potassium even in cuttings before rooting, and if these can prevent the propagation of various bacteria, they can nourish these elements. It has been found that the rooting rate is drastically increased by giving the root. Here, it is the carbon source that causes the propagation of various bacteria. Therefore, in the present invention,
The cuttings are cultured using a liquid medium containing nitrogen, phosphorus, and potassium as essential elements and no carbon source.

On the other hand, in plants, carbon dioxide present in the atmosphere is required as an energy source instead of a carbon source. In the case of the present invention, the seedlings are produced by artificially controlling the concentration in the culture environment so that the cuttings cultivated in the above medium are rooted. This is because the use of carbon dioxide in the atmosphere alone is not sufficient for rooting from cuttings.

When seedlings are to be produced by the cutting method, it is desirable that cuttings be kept at high humidity until healthy seedlings are formed. However, cutting has conventionally been performed outdoors or in a greenhouse or frame. Therefore, in order to maintain the culture environment of the cuttings and the seedlings formed from the cuttings at a high humidity, considerable effort must be taken. However, in the present invention, cuttings are produced in a culture vessel using a rooting bed moistened with a liquid medium. As a result, the humidity in the culture environment such as cuttings can be kept at a high level without any effort, and healthy cuttings can be easily produced.

[0029]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below based on embodiments.

Example 1 From the current year branch of a 40-year-old Yoshino cherry tree, a branch serving as a cutting was collected. The branches were cut to a length of 3 cm, leaving only one bud, and one third of the leaves were cut off to adjust the cuttings.

On the other hand, as a culture vessel, holes having a diameter of 1 cm were formed in two upper portions of a 10 cm square polycarbonate container, and the opening was formed with a carbon dioxide permeable membrane ("Milliseal" manufactured by Nippon Millipore Co., Ltd.). A covered one was prepared.
As a rooting bed, 100 ml of 8-fold diluted MS medium or 100 ml of 500-fold diluted "Hyponex solution 5-10-5 (registered trademark)" added with 2 mg / l of IBA, respectively.
A foamed phenolic resin molded product (“Oasis” manufactured by Nippon Soda Co., Ltd.) moistened by the above method was used.

For the production of cuttings, cuttings prepared as described above were inserted into the rooting bed at a rate of 25 per culture vessel, and the concentration of carbon dioxide in each culture vessel was reduced.
350, 500, 1000 or 1500 ppm only during the light period
At a temperature of 22 to 24 ° C. and a light intensity of 50 μm.
It was performed by culturing at mol / m ² / sec for 16 hours in the light period and 8 hours in the dark period. The concentration of carbon dioxide in the culture vessel was controlled by controlling the concentration of carbon dioxide in the environment where the culture vessel was placed.

Three weeks later, Table 1 shows the results of investigation on the presence or absence of rooting for 100 cuttings per test plot.

[Comparative Example 1] Culture of cuttings collected and adjusted from Yoshino cherry was carried out in the same manner as in Example 1 except that water containing 2 mg / l of IBA was used as a liquid medium for moistening the rooting bed. We tried to produce cuttings.
In this experiment, the concentration of carbon dioxide in the culture vessel was set to 1000.
This was performed only when the control was performed at ppm (only during the light period).

Three weeks after the cuttings were inserted into the rooting bed, 100 cuttings were checked for rooting, and the results are shown in Table 1.

[Comparative Example 2] In the same manner as in Example 1, except that the carbon dioxide concentration in the culture vessel was not controlled at all, the cuttings collected and adjusted from Yoshino cherry were cultured to produce cuttings. Tried. Note that this experiment was performed only when an 8-fold diluted MS medium containing 2 mg / l of IBA was used as a liquid medium for moistening the rooting bed.

Three weeks after the cuttings were inserted into the rooting bed, 100 cuttings were examined for the presence or absence of rooting. Table 1 shows the results. At this time, the concentration of carbon dioxide in the culture vessel had already been 100 ppm or less by the end of the light period immediately after the start of the culture.

Comparative Example 3 Production of cuttings was attempted by a conventional method.

A talc powder mixed with 1% by weight of IBA was applied to the base of a cutting of Yoshino cherry which was collected and adjusted in the same manner as in Example 1, and this was wetted with tap water. It was inserted into a phenolic resin molded product.
The phenolic resin molded product was left as it was in a greenhouse shaded by cold gauze 70%, and the cuttings were cultured.

Three weeks after the cuttings were inserted into the rooting bed, the results of an investigation on the presence or absence of rooting for 100 cuttings are shown in Table 1.

[0041]

[Table 1] As is clear from Table 1, in the test plot in which the concentration of carbon dioxide in the culture vessel was controlled to 350 ppm or more, and a medium for plant tissue culture or a fertilizer for plant was applied as a liquid medium,
In all cases, the rooting rate from the cuttings was extremely high as compared with those obtained by adding water instead of the liquid medium, those without controlling the concentration of carbon dioxide, and those obtained by the conventional cutting method.

The highest rooting rate is obtained in a test plot in which the concentration of carbon dioxide in the culture vessel is controlled to 500 ppm when a medium for plant tissue culture is used as the liquid medium, while the fertilizer for plants is used as the liquid medium. Was achieved in a test plot in which the concentration of carbon dioxide in the culture vessel was controlled at 1000 ppm.

Example 2 Branches are extended from axillary buds by excision of apical buds of an annual Eucalyptus globulus (hereinafter abbreviated as E. globulus), and after 3 weeks, apical ends of the branches From the first to second joints were cut into cuttings.

The cuttings were cultured in the same manner as in Example 1 to produce cuttings. However, at this time, in the test plot in which the carbon dioxide concentration in the culture vessel was controlled to 350 ppm or 500 ppm, in addition to the 8-fold diluted MS medium, a 4-fold diluted MS medium was also used as a plant tissue culture medium. Cutting seedlings were produced (all were IBA2
Add mg / l. ).

Three weeks after the cuttings were inserted into the rooting bed, the results of an investigation on the presence or absence of rooting of 100 cuttings are shown in Table 2.

Comparative Example 4 E. coli was used in the same manner as in Example 2 except that water containing 2 mg / l of IBA was used as a liquid medium for moistening the rooting bed. The cuttings collected from Globras were cultured to try to produce cuttings. In this experiment, the concentration of carbon dioxide in the culture vessel was set to 500 pp.
m (only during the light period).

Three weeks after the cuttings were inserted into the rooting bed, Table 2 shows the results of investigation on the presence or absence of rooting for 100 cuttings.

Comparative Example 5 E. coli was prepared in the same manner as in Example 2 except that the concentration of carbon dioxide in the culture vessel was not controlled at all. Culture the cuttings collected from Globras,
We tried to create cuttings. Note that this experiment was performed only when an 8-fold diluted MS medium containing 2 mg / l of IBA was used as a liquid medium for moistening the rooting bed.

Three weeks after the cuttings were inserted into the rooting bed, the results of investigation on the presence or absence of rooting of 100 cuttings are shown in Table 2.

[Comparative Example 6] Production of cuttings was attempted by a conventional method.

E. coli collected in the same manner as in Example 2. Example 2 A talc powder mixed with 1% by weight of IBA was applied to the base of a Globras cutting and wetted with tap water.
And inserted into the same foamed phenolic resin molded product as in The phenolic resin molded product was left as it was in a greenhouse shaded by cold gauze 70%, and the cuttings were cultured.

Three weeks after the cuttings were inserted into the rooting bed, the results of investigation on the presence or absence of rooting of 100 cuttings are shown in Table 2.

[0053]

[Table 2] As is clear from Table 2, in the test plot in which the concentration of carbon dioxide in the culture vessel was controlled to 350 ppm or more, and a plant tissue culture medium or a plant fertilizer was applied as a liquid medium,
In all cases, the rooting rate from the cuttings was extremely high as compared with those obtained by adding water instead of the liquid medium, those without controlling the concentration of carbon dioxide, and those obtained by the conventional cutting method. That is, no matter whether a plant tissue culture medium or a plant fertilizer is used as the liquid medium, rooting from cuttings of almost 90% or more is obtained when the concentration of carbon dioxide in the culture vessel is controlled to 350 ppm or more. Was observed.

[0054]

According to the present invention, the rooting rate of a plant that has been difficult to root from cuttings by the conventional cutting method is greatly improved.

Further, according to the present invention, the environment from cutting to the formation of seedlings can be easily maintained at high humidity.

Therefore, according to the present invention, it is possible to produce a cutting seedling from a plant which has been difficult to produce a cutting seedling by the conventional method. In addition, the productivity of a plant from which cuttings and seedlings can be produced relatively easily by the conventional method can be improved. In addition, the operation for this is simple and easy because it inherits the advantages of the conventional cutting method.

In addition, in the present invention, since cuttings are formed in a culture vessel, not only humidity but also temperature,
Control of conditions such as light is easier than the conventional method for producing cuttings.

Therefore, according to the invention of the present application, it is possible to easily carry out year-round production of cuttings.

That is, the present invention makes it possible to mass-produce cloned seedlings which are commercially advantageous.

 ────────────────────────────────────────────────── ─── Continuing from the front page (72) Inventor Kunimitsu Murakami 2-8-1 Iida-cho, Iwakuni-shi, Yamaguchi Japan F-term in Iwakuni Technical Research Laboratory (Reference) 2B022 AA01 AB20 DA15 EA01 2B030 AA03 AB03 CD03 CD06 CD09 CD14 CD28

Claims (7)

[Claims] 1. A rooting bed containing nitrogen, phosphorus, and potassium as essential elements and moistened with a liquid medium containing no carbon source is prepared in a culture vessel, into which cuttings are inserted and cultured. A method for producing a cutting seedling, wherein rooting is performed from a cutting while controlling the concentration of carbon dioxide in a culture vessel. 2. The method for producing cuttings according to claim 1, wherein a known plant fertilizer is used as it is or diluted as a liquid medium for moistening the rooting bed. 3. The method for producing cuttings according to claim 1, wherein a known culture medium for plant tissue culture is used as it is or diluted as a liquid medium for moistening the rooting bed. 4. A liquid medium to which a plant growth regulator is added is used as a liquid medium for moistening a rooting bed.
Or the method for producing cuttings according to 3 above. 5. The method for producing cuttings according to claim 4, wherein auxin is used as a plant growth regulator. 6. The concentration of carbon dioxide in the culture vessel is 300 to 1
5. The method according to claim 1, wherein the control is performed at 500 ppm.
Method for producing cuttings described in 1. 7. The method for producing cuttings according to claim 1, 2, 3, 5, or 6, wherein the cuttings are derived from a woody plant.