DD159507A3 - STABILIZER FOR A NEEDLE MEDIUM - Google Patents

Abstract

The invention relates to a stabilizer for a culture medium for the sterile culture in highly viscous form for the cultivation, propagation and maintenance of crops and microorganisms. It aims to provide a stabilizer by using a modified natural product of constant or by the targeted control of the production gleichzuhaltender quality, which allows to work always under the same conditions, which is universally applicable and dispersible in water and residue-free of the cultured organisms can be separated. This is achieved by a combination of agar-agar and microcrystalline cellulose.

Description

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Stabilizer for a Nährmedimn

App e ndu ngsgebiet of the invention;

The invention relates to a stabilizer for a culture medium for the sterile culture in highly viscous form for the cultivation, propagation and maintenance of crops and microorganisms.

Characteristic of the known solutions

Sterile culture is a technique for the rapid and genetically identical propagation of crops, using young plants from regenerative plant parts (such as sprouts) in a known manner under sterile conditions.

Medicinal media for this sterile culture contain salts of macronutrients, salts of certain micronutrients, various mono- or disaccharides, vitamins and possibly, natural or synthetically produced plant hormones. These components are offered in aqueous solution.

In addition, the nutrient media contain substances that solidify the medium (so-called stabilizers). By setting a certain minimum viscosity is achieved that patch small plant parts do not sink into the culture medium, where they would otherwise be cut off from the atmosphere and thus necessary gas exchange, which can lead to the death of the explants.

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At the same time, the medium must have a consistency which allows the formed roots to easily penetrate the substrate and thus deliver to the growing plant the nutrients distributed in the medium. The roots must be sufficiently well anchored in the culture medium, so that the plant receives sufficient stability even in later stages of growth, because even in this stage of development, sinking into the nutrient medium could be the cause of plants dying. Finally, after completion of the in vitro culture, the substrate must be easily, completely and easily removed from the roots with little effort. As the stabilizer, agar agar is generally used which largely satisfies the above-mentioned partially contradictory conditions.

To prepare the nutrient media, the procedure is such that components of the nutrient solution are dissolved individually or in groups in deionized water, mixed in a certain ratio and made up to a predetermined volume with deionized water. This solution is heated until complete dissolution of the agar and then metered liquid into the culture vessels. The culture media are sterilized after filling in the culture vessels in a known manner and serve as a substrate on which attracted under sterile conditions in a conventional manner from regenerative plant parts young plants or cultures of microorganisms are propagated.

However, agar-agar has some undesirable properties. Due to its chemical structure based on polysaccharide, it may be subject to certain degradation phenomena by selected strains of microorganisms. In addition, the product is not stable for the pH range present when propagating acidophilic microorganisms and explants, so that changes in the viscometric behavior may occur , The water retention capacity is of limited duration, so that it can lead to dehydration phenomena.

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The removal of the rooted plants from the agar medium is often hampered by the gel consistency of the substrate. In order to prevent the roots from breaking off in this process, the agar medium must first be liquefied by gentle heating. This process is time-consuming and must be carried out with extreme care, otherwise the plants could be thermally damaged.

To make matters worse, that as a natural product of organisms of short-lived nature it is seasonal and '

Λ, climatic fluctuations are particularly strongly subject.,

so that the quality and the offer can be significantly affected.

DE-AS 2 551 155 discloses the use of a water-adsorbing clay as a stabilizer for a nutrient medium, which eliminates the disadvantages of the agar-agar. DB-OS 2 β34 392 gives as stabilizer soluble cellulose derivatives such as eg B, carboxymethylcellulose. However, these substances have the disadvantage that they are not universally applicable due to their ion content. In addition, it is difficult to completely separate the nutrient medium from the cultured organisms after completion of the culture. Furthermore, it is disadvantageous that these substances in the

') Are difficult to solve.

The resulting materials meet the following requirements for use as substrates: they do not contain inorganic ions or only in traces; they contain only indifferent organic constituents; , .- they do not contain any components that have the osmotic values; ^to change the solutions;

they are inert to salts and pH-resistant; they are sterilizable in a conventional manner (heat sterilization).

Due to the fact that the stability of the dispersion viskosimetrically based on other factors, as

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with gels of agar-agar and no state of optimal swelling exists, it is possible to better free the crops from the adhering parts of the substrate, since the addition of water without Hachquellung a rapid drop in viscosity occurs, resulting in a simple separation of the Dispersion of the cultivated plant parts, shoots and roots.

Object of the invention ν

The object of the invention is to provide a stabilizer for a nutrient medium for the cultivation of crops and microorganisms by using a modified Haturstoffes of constant or by the targeted control of the production way a quality stabilizer, which allows always under the same conditions, the tests and propagation which can be used universally and can be separated without residue from the cultivated organisms and is dispersible in water, Mhrmedien o _o g. Composition are particularly suitable for the cultivation, propagation and conservation of the following groups of crops and microorganisms by sterile culture:

Leaf and stalk vegetables (e.g., Brassica oleracea, var. Bot-

rytis, capitata, gemmifera, etc.)

Pulses (ζ _β Β _β Phaseolus Vulgaris, Glycine

soy)

Greens (for example Lycopersicon esculentum,

Capsicum annuum) root vegetables (e.g., Daucus carota, Apium

, '_ ·.' · · · · · 'Graveolens)''-. ··· Onion vegetables (z _o B "Allium c.epa)

Medicinal plants (z, B _e Digitalis purpurea,

Datura innoxia) fiber plants (e.g., Linum usitatissimurn,

Cannabis sativa ', Gossypiiyn

herbaceum, G ₁ , hirsutum)

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forage crops

Luxury food plants

Spice plants cereals

Root crops Soft fruit

Pome fruit Stone fruit Tropical fruits Oil fruits Ornamental plants Microorganisms (e.g., Medicago sativa, Brassica oleracea var, viridis, Trifolium pratense) (e.g., Coffea arabica, Camellia

sinensis, Humulus lupulus) (e.g., Petrosllinum hortense) (e.g., Triticum aestivum, Seeal cereale, Triticale, Avena sativa, Hordeum vulgare) (e.g., Solanum tuberosum, Beta

vulgaris, Zea mays) (e.g., Ribes nigrum, Ribes and others crispa, Rubus idaeus, Fragaria

ananassa) (e.g., Malus domestica, Pyrus domestica) (e.g., Prunus doraestica, Prunus

avium, Prunus cerasifera) (e.g., Citrus aurantium, Citrus

limon) (e.g., Brassica napus var

Elaeis guinensis) (for example chrysanthemums spec. Dianthus

Caryophyllus, Petunia hybrida) e.g. Enterobacteriaceae such as Escherichia coli, Aerobacter aerogenes, - Bacillaceae such as Bacillus cerus, Bacteria such as Sarcina lutea, Actinomycetaceae such as Actinomyc'-es bövis, Mycophyta such as the oomycetes Phytophthora infestans and Peronospora spec * such as the Ascomycetes Penicillum notatum, Saccharomyces cerevisae and Aspcrgillus funiigatus.

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p; the essence of the invention

The invention has for its object to find a suitable combination of substances as a stabilizer of a nutrient medium. The object is achieved in that microcrystalline cellulose is combined with agar agar or "another suitable hydrating agent and used as a stabilizer. The intended microcrystalline cellulose is obtained by heterogeneous acid hydrolysis of a cellulosic starting material having an x-cellulose content of more than 90 % and then subjected to a colloidal wet milling with the addition of active ingredients and nutrients,

embodiments

The method according to the invention will be explained in more detail below with reference to an exemplary embodiment:

It is made using deionized water, a nutrient solution consisting of. "

Ammonium nitrate. 1,650 g

Potassium nitrate 1,900 "

Magnesium sulfate heptahydrate 0.370 "

Potassium dihydrogen phosphate. 0,170 "

Boric acid 0.0062 "

Magnesium sulfate monohydrate 0.0169

Zinc sulfate heptahydrate. 0.0086 "

Potassium iodide · 0.00083 "

Sodium molybdate dihydrate. '0,00025'

Copper sulphate pentahydrate. 0.000025 "

Cobalt chloride hexahydrate '0.000025' '

Iron II sulphate heptahydrate 0.0273 "

Ethylenediaminetetraacetic acid disodium salt O, 0373 '"

Calcium Chloride Dihydrate 0.440 "

myo-inositol. 0.100 "

Glycine 0.002 "

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O , 0005 G O , 0005 It O , 0001 Il 30 Il 1 1

nicotinic acid

Pyridoxine hydrochloride Thiamine hydrochloride Sucrose

Wassor

(Murashige T., F. Skoog, Physiol Plant 1, pp. 473-97 (1962), which serves as a basis for the production of the microcrystalline cellulose gel.) In this solution, under stirring, microcrystalline cellulose, which consists of pulp with an oC- Cellulose content of 93 % was obtained by heterogeneous acid hydrolysis to LODP, registered and homogenized.

By colloidal hate milling, a thixotropic gel is produced which, depending on the desired viscosity, has a concentration of 2 to 12% microcrystalline cellulose in the pesticide.

Agar agar is still added to the dispersion in a low-viscosity state so that its concentration is 0.6 to 0.05 %, based on the total amount of nutrient medium. After reaching the required viscosity, the dispersion is dried, preferably spray-dried. The resulting powder is portioned or compressed and then microbiologically sterilized in a conventional manner, so that when using defined amounts in the individual culture flasks always the same conditions büi addition of a defined sterile water can be maintained «The solutions in the culture vessels in a known manner to ensure the formation of the high-viscosity combination of agar-agar and microcrystalline cellulose. ·

This is followed by. Applying the crops to be propagated or grown on the substrate.

Claims (2)

221538 WP C12N / 221 598 Fit of 3 · 6β 1982 A stabilizer for a nutrient medium for sterile culture in a highly viscous form for the cultivation, propagation and maintenance of crops and microorganisms, characterized in that it consists of a mixture of 40 to 120 parts by weight of microcrystalline cellulose powder and 1 to 12 parts by weight of agar-agar. Stabilizer according to item 1, characterized in that the microcrystalline cellulose powder was obtained by heterogeneous acid hydrolysis of a cellulosic starting material with a cellulose content of more than 90 % and subsequent colloidal hate milling with the addition of Y / irk and nutrients.