JPH0322930A - Formation and proliferation of crown of plant of genus asparagus and proliferation of seedling - Google Patents

Abstract

PURPOSE:To form a crown of a plant of genus Asparagus capable of proliferating large amount of plant or seedling in high efficiency by culturing the tissue or callus of the plant under aeration in a liquid medium impregnated in a supporting material. CONSTITUTION:The tissue or callus of a plant of genus Asparagus (preferably edible asparagus) is cultured under aeration in a liquid medium impregnated in a supporting material to form a crown. Furthermore, similar proliferation process comprising (slicing of crown culture of slice thickening of the crown) is repeated, as necessary.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a crown shape and a method for propagating plants of the genus Asparagus, and a method for propagating plants and seedlings. [Prior Art] Plants of the genus Asparagus have conventionally been propagated by seeding or division. Plants of the genus Asparagus are dioecious, and male plants are considered to be better in terms of quality and cultivation management. Furthermore, since it is a crop with large genetic variations, propagation by sowing leads to the contamination of female plants and causes differences in yield and quality among plants, which poses a significant problem in cultivation. In addition, propagation by division requires a lot of manpower and a long period of time, is extremely inefficient, and has a high risk of transmitting viral diseases.

In order to improve the above-mentioned problems, mass propagation of superior strain clones using tissue culture has recently attracted attention. Normally, for mass propagation of asparagus, tissue pieces are cultured, a large number of shoots are propagated, and then each is transplanted to a rooting medium, and seedlings are formed through differentiation of adventitious roots. However, since the rooting rate is generally low, there is a problem that the propagation efficiency is low. In addition, attempts have been made to culture tissue pieces to form callus, and then use somatic embryos produced from the callus to mass multiply (for example, 1985 Horticultural Society Autumn Conference Research Presentation Abstracts, page 210). ~211, 1986.ll.23-25, University of the Ryukyus, 1988 Horticultural Society Research Presentation Abstracts P254-255, 1986.1
0.7-9, Kyushu University). Although growing somatic embryos solves the problem of low rooting rate, the problem is that most isolated somatic embryos develop callus or malformation during culture.

[Problem to be solved by the invention]

The present inventors recognized that the conventional tissue culture method for plants of the genus Asparagus has the above-mentioned problems, and cultivated the seeds and seedlings of plants of the genus Asparagus by culturing tissues using a new method different from the conventional method. We investigated methods for efficient propagation. [Means for Solving the Problems] As a result, the present inventors developed a technique for tissue culture and propagation of Asparagus plants by culturing the tissue under aerated conditions using a liquid medium impregnated with a supporting material to form a crown. It was discovered that the problems of the prior art described above could be satisfactorily solved by using a plant body, and by conducting further research based on this new knowledge, the present invention was completed.

According to the method of the present invention, (1) Asparagus is cultured under aerated conditions using a liquid medium in which a supporting material is impregnated with tissue or callus of a plant of the genus Asparagus to form a crown. The crown shape of plants of the genus is grown by culturing the callus using the method described in (1) and cutting the crown. A method for propagating crowns of plants of the Asparagus genus, characterized by repeating the same propagation process of crown cutting → section culture → crown enlargement as necessary; A method for propagating plants of the genus Asparagus, characterized by growing the plants by culturing them under aerated conditions using a liquid medium impregnated with Provided is a method for propagating seeds and seedlings of plants of the genus Asparagus, characterized by:

In the present invention, any plant belonging to the genus Asparagus can be used. Specifically, the plant is Asparagus officinalis, which is an edible asparagus.
L. var. altilis L. For example, it is preferable to use this in the present invention.

In the present invention, tissues or callus (ca) of plants of the genus Asparagus are used.
Hus) is tissue cultured to form a crown. In this case, IJlwi such as embryo, shoot tip, stem, pseudoleaf, underground stem, etc. can be exemplified as the tissue used for Mi tissue culture. Regarding callus, in the present invention, callus obtained by culturing the tissue by a known culture method (for example, Horticultural Society Journal, Vol. 40, No. 4, pp. 11-17, 1971) can be used.

The medium used in the crown type of the present invention is a medium containing inorganic components and a carbon source as essential ingredients, to which are added plant hormones and vitamins, and further added with amino acids as necessary. No I of the medium! ! As a precept,
Examples include inorganic salts containing elements such as nitrogen, phosphorus, potassium, sodium, calcium, magnesium, sulfur, iron, manganese, zinc, boron, molybdenum, chlorine, iodine, and cobalt. Specifically, potassium nitrate, nitric acid Sodium, ammonium nitrate, potassium chloride, calcium chloride, potassium monohydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate, magnesium chloride, sodium sulfate}
IJum, ferrous sulfate, ferric sulfate, manganese sulfate,
copper sulfate, sodium molybdate, molybdenum trioxide,
Examples include compounds such as potassium iodide, zinc sulfate, boric acid, and cobalt chloride.

Examples of carbon sources for the medium include carbohydrates such as sucrose and glucose, derivatives thereof, organic acids such as fatty acids, and primary alcohols such as ethanol.

Examples of the plant hormones in the medium include naphthaleneacetic acid (NA^), indoleacetic acid (14A), p-chlorophenoxyacetic acid, and 2,4-dichlorophenoxyacetic acid (2.
Auxins such as 4-[)), indolebutyric acid (IB8), and their derivatives, and penzyladenine (BA
), kinetin, zeatin, and other cytokinins.

The vitamins in the medium include biotin, thiamine (vitamin B), pyridoxin (vitamin B6), pyridoxal, pyridoxamine, calcium pantothenate,
Ascorbic acid (vitamin C), inositol, nicotinic acid, nicotinamide and lipoflavin (vitamin B
6) etc. can be exemplified.

Examples of amino acids in the medium include glycine, alanine, glutamine, cysteine, phenylalanine, and lysine.

The medium of the present invention usually contains about 0.0% of the inorganic component.
1 μH to about 100 lIM, about 0.01 mg/j of the plant hormones! Preferably, the amino acids are used in an amount of 0 to about 150 mg/1 and 0 to about 1000 mg/1. Specifically, the medium used for tissue culture according to the present invention may be a conventionally known culture medium used for plant tissue culture, such as Murashige Skoog ('62
) [Murashige &Skoogl's medium,
Linsmeyer Skoog (RM-1965) [Li
nsa+aier&Skoog] medium, white
('63) [White] medium, Gamborg [Ga
mborg] B-5 medium, Mitsui Mon 9 medium, niche
An example is a medium prepared by adding the above-mentioned carbon source and plant hormone to a Nitch medium [Nitch & Nitch], and further adding the above-mentioned bitter straws and straw acids as necessary, but this In the present invention, among these, media prepared using Niche-Hetch, Linsmeyer-Skoog, and Murashige-Skoos are particularly preferred. Regarding the Mi precepts of the conventionally known culture medium mentioned above, for example, Takeuchi,
“New Plant Tissue Culture,” by Nakajima and Furuya, p3B6-p39
1, Asakura Shoten, 1979. The medium that can be used in the crown-shaped method of the present invention is a liquid medium, and includes polymers such as polyester, polyolefin, polyvinyl chloride, polyvinylidene, and polyacrylonitrile acetate alcohol, and artificial soil such as rock wool, vermiculite, and barlite. It is used by impregnating a support such as a plastic mesh or wire mesh.

In the present invention, tissue or callus of a plant of the genus Asparagus is tissue cultured using the liquid medium under aerated conditions to form a crown.

In this case, the crown as used in the present invention is a storage tissue that corresponds to the underground stem in the whole plant and is morphologically a shortened stem, and has a lump-like or crown-like shape obtained by tissue culture of tissue or callus. A cluster of cells that contains buds or bud primordia that become the basis of buds.

Aeration methods include forced ventilation using air pumps, gas cylinders, fans, etc., blowing air through culture vessels equipped with filters, and natural ventilation using air expansion and contraction due to temperature changes. As the gas to be used, a mixture of two or more types of gases such as oxygen, carbon dioxide, nitrogen, and air can be used. In particular, in the present invention, oxygen is 5 to 20 vol/χ, carbon dioxide is 4
A mixed gas containing 00 to 1000 ppm is desirable. Although the ventilation rate varies depending on the shape and size of the culture container, it is preferable that the gas phase portion within the culture container be ventilated 1 to 20 times/h.

The culture device used for crown formation is not limited in any way as long as it has a support material for impregnating it with a liquid medium and can ventilate the gas phase. Specifically, an apparatus as shown in FIG. 2 can be used.

FIG. 1 is a diagram showing the process of crown formation, proliferation, and plant growth of Asparagus plants according to the method of the present invention. The upper part of the figure shows the path through which the crown is formed by tissue culturing the cut sections obtained from the shoot stem, and the lower left part of the figure shows the route by which the crown is grown from said crown. The cycle is shown, and the bottom right side of the figure shows a plant that has been rooted and regenerated from the crown that has been propagated in this way.

In the present invention, the above-mentioned medium can be used as a medium for forming a crown, but in particular, when using a medium containing 3% or more of C.J. I can do it.

In the present invention, by employing aeration conditions during crown formation, a high-quality crown with a developed vascular system can be formed, and the shape and speed can be improved. The resulting crowns are easy to root and avoid vitrification.

In the present invention, the crown formed as described above is cut, preferably to include at least one bud, and the obtained cut pieces are further used in a liquid medium impregnated with a supporting material under aerated conditions. The crown can be enlarged and elongated by culturing it. The same liquid medium as that used for crown formation can be used for the above-mentioned crown enlargement and proliferation, and especially when using a medium containing 3% (initial concentration) or more of siam, crown proliferation rate can be improved. Ventilation conditions may be the same as described above.

In the present invention, by adopting aeration conditions during crown enlargement, a high-quality crown can be obtained, that is, a crown that is easy to root, and vitrificesidin can be prevented.

Furthermore, if necessary, crown propagation can be performed by repeating the same crown cut section tissue culture-crown enlargement propagation process.

Next, an appropriate amount of crowns are extracted from the crowns propagated as described above and cultured in a rooting medium to root, and the buds are grown into plants of an appropriate size. In this process of growing the plant, liquid aeration culture similar to that used for the crown formation described above can be used to increase the rooting rate and rooting speed of storage roots and to obtain high-quality seedlings.

Rooting is particularly promoted when an anti-gibberellin agent is added to the medium. A specific example of an anti-gibberellin agent is Ansijudol. Furthermore, in the present invention, although an acclimatization step is not necessarily required,
If necessary, the above-mentioned plants can be acclimatized to produce seedlings, and by repeating the above method, seeds and seedlings of Asparagus plants can be propagated in large quantities.

Hereinafter, the method of the present invention will be specifically illustrated by examples.

Example 1 The shoots of the asparagus variety "Hokkai 100" maintained in a sterile system were cut and the nodes adjusted to a length of 7 to 10+a+a were used as material.The medium was an MS medium containing inorganic elements and an organic element. Based on niche/niche, NH4NO3 is 1
/2 times CaCl. 2 times, glutamine 104M
Basic medium (hereinafter abbreviated as ASP medium)
And so.

The medium is based on ASP medium, shot J360g/l
, using a liquid medium with pH 5.8, and using IB as auxin.
A10-6? l, 3X1 B8 as cytokinin
0-&M, 10- and uncimidol as anti-gibberellin
hM was added. After dispensing 180 d each of this together with about 2.0 g of polyester wool into a commercially available culture box (approximately 1000 atg, with a silicone tube attached to the lid and a gas filtration filter), 100 Individually placed on the floor at 25°C.
When cultured for 4 weeks under the conditions of 30,001 ux and an aeration rate of 30 m/+win, the results shown in Table 1 were obtained.

Comparative Example 1 The same procedure as in Example 1 was performed except that air ventilation was not performed, and as a result, the results shown in Table 1 were obtained.

Example 2 The crown of the asparagus cultivar "Hokkai 100" obtained by the method of Example 1 was cut into pieces containing 2 to 3 buds as a material. The medium is basically ASP medium, and C-yl! 60g/J! The pH was adjusted to 5.7, and to this were added IBA@10-'M as auxin, 10-'M B^ as cytokinin, and 10-'M of ancymidol as anti-gibberellin. After dispensing 180 d each of this together with about 3.0 g of polyester wool into a commercially available culture box (capacity approx. 1000 rd, with a silicone tube attached to the lid and a gas filtration filter), 25 asparagus crowns/ A total of 100 boxes were placed on a bed and cultured for 4 weeks under the conditions of 25° C., 3000] tv and an air aeration rate of 30 d/+gin. The results shown in Table 2 were obtained.

Comparative Example 2 Example 2 was carried out in the same manner as in Example 2 except that air ventilation was not performed, and the results shown in Table 2 were obtained.

Example 3 The crown of the asparagus variety "Hokkai 100" maintained by the method of Example 2 was cut into pieces containing 2 to 3 buds and used as a material. The medium is basically ASP medium. F 1!60g/l, pH5. 7, IBM as auxin at 10-'M, and BA as cytokinin at 10-'? 1, IO-''H of uncimidol was added as an anti-gibberellin. This was placed in a commercially available culture box (capacity: approximately 1000 d, with a silicone tube attached to the lid and a gas filtration filter attached), and about 3.0 mm of polyester wool was added. After dispensing 180 pieces each with 0g, place the above asparagus crowns on a bed of 25 pieces/BOX for a total of 100 pieces, and heat at 25°C for 3 hours.
000]IJX, air ventilation rate 30 d/m
When cultured for 4 weeks under conditions of i.n., the results shown in Table 3 were obtained.

Comparative Example 3 Example 3 was carried out in the same manner as in Example 3 except that air ventilation was not performed, and the results shown in Table 3 were obtained.

(Margin below this page) Table 1 Example 1 Comparative Example 1 Crown shape coverage rate 85% 62% Table 3 Rooting rate Average fresh weight/rooted individual Example 3 76% 195■ Comparative Example 3 34% 180mgTable 2 Implementation Example 2 Comparative Example 2 Crown multiplication rate 4.3 times 3.5 times (blank space below this page) [Effects of the invention] According to the method of the present invention, crowns are formed from Asparagus plants by tissue culture, and , this can be efficiently propagated in large quantities, and furthermore, plants and seedlings can be efficiently propagated in large quantities via the obtained crowns.

[Brief explanation of the drawing]

Figure 1 shows the crown shape, proliferation, and growth of Asparagus plants.
FIG. 2, which is a diagram showing processes such as propagation of plants, is a culture device used for crown-shaped plants. Figure 1

Claims (1)

[Claims] 1. A method for forming a crown of a plant of the genus Asparagus, which comprises forming a crown by culturing under aerated conditions using a liquid medium in which a supporting material is impregnated with tissue or callus of a plant of the genus Asparagus. . 2. The sections obtained by cutting the crown formed by the method described in claim 1 are further cultured under aerated conditions using a liquid medium impregnated with a supporting material to enlarge the crown, and if necessary, the same A method for propagating crowns of plants of the genus Asparagus, which is characterized by repeating the propagation process of crown cutting → section culture → crown enlargement. 3. A plant of the genus Asparagus, characterized in that the plant is grown by culturing it under aerated conditions using a liquid medium in which a support material is impregnated with the crown obtained by the method according to claim 1 or claim 2. How the body multiplies. 4. The method for propagating a plant of the genus Asparagus according to claim 3, wherein the growth step of the plant includes a rooting step. 5. A method for propagating seeds and seedlings of plants of the genus Asparagus, which comprises acclimatizing the plants obtained by the method according to claim 3 or 4 to produce seedlings.