WO1984004653A1 - Procede de culture de mycorhizes va - Google Patents

Procede de culture de mycorhizes va Download PDF

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Publication number
WO1984004653A1
WO1984004653A1 PCT/GB1984/000176 GB8400176W WO8404653A1 WO 1984004653 A1 WO1984004653 A1 WO 1984004653A1 GB 8400176 W GB8400176 W GB 8400176W WO 8404653 A1 WO8404653 A1 WO 8404653A1
Authority
WO
WIPO (PCT)
Prior art keywords
sulphur
growth
culture
agar
medium
Prior art date
Application number
PCT/GB1984/000176
Other languages
English (en)
Inventor
Christine Margaret Hepper
Original Assignee
Nat Res Dev
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority claimed from GB838314432A external-priority patent/GB8314432D0/en
Priority claimed from GB838332120A external-priority patent/GB8332120D0/en
Application filed by Nat Res Dev filed Critical Nat Res Dev
Publication of WO1984004653A1 publication Critical patent/WO1984004653A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00—Cultivation of mushrooms
    • A01G18/10—Mycorrhiza; Mycorrhizal associations

Definitions

  • VA mycorrhizal fungi are beneficial fungi which infect the feeding roots of plants and stimulate uptake of phosphorus from the soil. Hyphae of the fungi grow outwardly from the root well beyond the phosphate depletion zone, (the zone from which the available phosphate has already been consumed by the plant). The fungi have been grown well only in association with live host roots. Thus, inocula of the fungi for use in infecting plants have been produced only on a small scale from the roots of previously inoculated plants, see e.g. UK Patent 2043688B (National Research Development Corporation).
  • An important feature of the invention consists in a method of culturing a VA mycorrhizal fungus, non-biotrophically, which method comprises culturing the fungus, preferably aerobically, on a growth medium containing an inorganic sulphur compound, preferably one containing a sulphur atom of valence less than 6, most preferably of valence 4.
  • the growth medium also contains amino acid(s).
  • a carbon source can be included, but currently it seems best to avoid using a sugar.
  • the sulphur compound employed is preferably one containing an oxo-acid anion, especially
  • the sulphur compound contains two or more S-atoms, at least one of them preferably has a valence of less than 6. This is tantamount to saying that their average valence is less than 6, but it is preferred to express the preference in terms of individual atoms because the 4-valent state of an individual atom, as in disulphite, appears preferred over an average valence of 4 resulting from one atom being 6-valent and another 2-valent, as in thiosulphate.
  • the sulphur compound can also be a sulphide, for example K 2 S. Indications are that potassium salts give very much better results than sodium salts, which appear to inhibit growth in some way and are therefore less preferred.
  • the method of culture works well for Glomus caledonium and is expected to be suitable for any of the usual VA mycorrhizal fungi, for example Glomus mosseae, Glomus fasciculatus and other
  • the growth medium is conveniently agar-based, although it is contemplated that growth will also be obtained on appropriate liquid media. While growth has been produced without a carbon source in the medium, it is expected that improved results will be obtained by including one. Indications are that a non-sugar carbon source would be preferable to a sugar. Otherwise, the medium may contain any of the usual growth factors beneficial to fungal culture, for example yeast extract, peptone or thiamine and amino acid(s) such as cystine, glycine and lysine.
  • the culture conditions are normally aerobic, although subaerobic quantities of oxygen may be usable under some conditions.
  • the culture is not necessarily carried out on stationary plates, e.g. submerged shake culture would be possible. While it is preferred to pre-germinate the spores before contacting them with the growth medium, in situ germination has also been carried out successfully. Thus, using water agar and potassium sulphite, the spores germinated well and hyphal growth was stimulated.
  • the invention includes, of course, the VA mycorrhizal fungi obtained by the method of culture of the invention and inocula based on them.
  • EXAMPLE 1 Spores of Glomus caledonium were isolated from around the roots of stock plants of Nardus stricta, Allium cepa or Zea mays by wet-sieving. They were surfaced-sterilised by immersing them in a solution of 20 g/litre chloramine T containing 200 mg/litre streptomycin sulphate for 20 minutes, washed three times in sterile water and plated on to Difco Bacto agar (10 g/litre). The spores were incubated at 23o - 25 oC and transferred individually to the plates of test medium when they had just germinated (i.e. when the germ tube length was less than the spore diameter).
  • test media were prepared using Difco Bacto agar (10 g/litre) and were checked for pH after sterilisation at 121°C for 15 minutes. All pHs were between 6.6 and 7.5 and did not differ by more than 0.5 unit in any experiment.
  • the inorganic sulphur-containing compounds tested are listed in Table 1 below. Each compound was tested at the concentration given in Table 1 (equivalent to 12.3 mg S/litre) and also at one-tenth and one-hundredth this concentration.
  • EXAMPLE 2 Four sets of additional experiments were carried out in which were used (a) the agar of Example 1, (b) a different agar (Oxoid type L28) and (c) and (d) two agarose preparations. In each set of experiments the medium was used with and without potassium sulphite (60.5 mg/litre). Conditions were otherwise in accordance with Example 1. As Table 2 shows, hyphal growth from germinating spores was best when the Bacto agar of Example 1 was used.
  • Example 1 was repeated using different inorganic sulphurcontaining compounds, each compound being again tested at a concentration equivalent to 12.3 mg S/litre and also at one-tenth and one-hundredth this concentration.
  • the compounds tested and the results obtained are shown in Table 3.
  • EXAMPLE 4 If the effect of reduced inorganic sulphur compounds is due to a change in the oxidation - reduction potential of the medium, then it was thought possible that the same effect might be obtained with a compound such as ascorbic acid. This was tested at a range of concentrations by the procedure of Example 1. The results in Table 4 below show that there was no stimulation of hyphal growth at any concentration of ascorbate tested; it was in fact inhibitory to fungal growth in the concentration range 0.1 - 10 g per litre.
  • the medium did not include a carbon source for the fungus, an organic acid, oxaloacetic acid, which was found to be marginally better than other acids or sugars in earlier tests was added to the medium.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention vise à résoudre le problème de la culture des mycorhizes VA (vésiculaires-arbusculaires) suivant un procédé non-biotrophique (in vitro), en l'absence des racines des plantes) servant normalement d'hôte fongueux. Cependant, on a maintenant découvert que ces mycorhizes peuvent être cultivés sur un support de croissance contenant un composé de soufre inorganique, de préférence sous une forme réduite, contenant un atome de soufre d'une valence inférieure à 6. Du sulfite de potassium est particulièrement efficace.
PCT/GB1984/000176 1983-05-25 1984-05-23 Procede de culture de mycorhizes va WO1984004653A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB838314432A GB8314432D0 (en) 1983-05-25 1983-05-25 Culture of va mycorrhizal fungi
GB838332120A GB8332120D0 (en) 1983-12-01 1983-12-01 Culture of va mycorrhizal fungi

Publications (1)

Publication Number Publication Date
WO1984004653A1 true WO1984004653A1 (fr) 1984-12-06

Family

ID=26286226

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1984/000176 WO1984004653A1 (fr) 1983-05-25 1984-05-23 Procede de culture de mycorhizes va

Country Status (3)

Country Link
EP (1) EP0144378A1 (fr)
GB (1) GB2142933B (fr)
WO (1) WO1984004653A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991001082A1 (fr) * 1989-07-14 1991-02-07 Sean Morrison Production d'inoculum de mycorhizes par des cultures hydroponiques statiques
WO1991007868A1 (fr) * 1989-12-04 1991-06-13 Michigan State University Procede et compositions stimulant les champignons de mycorhize vesiculaire-arbusculaire
WO2007014974A1 (fr) * 2005-07-29 2007-02-08 Consejo Superior De Investigaciones Científicas Inoculant aseptique de micorhization et procedes d'application in vitro et ex vitro
US11814587B2 (en) * 2016-12-22 2023-11-14 Rainer TESCH Method for treating petroleum or natural gas

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1981003338A1 (fr) * 1980-05-15 1981-11-26 Battelle Development Corp Culture liquide de champignons ectomycorrhizes sporulants

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2043688B (en) * 1979-02-14 1983-04-27 Thompson J P Production of mycorrhizal fungi

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1981003338A1 (fr) * 1980-05-15 1981-11-26 Battelle Development Corp Culture liquide de champignons ectomycorrhizes sporulants

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991001082A1 (fr) * 1989-07-14 1991-02-07 Sean Morrison Production d'inoculum de mycorhizes par des cultures hydroponiques statiques
WO1991007868A1 (fr) * 1989-12-04 1991-06-13 Michigan State University Procede et compositions stimulant les champignons de mycorhize vesiculaire-arbusculaire
WO2007014974A1 (fr) * 2005-07-29 2007-02-08 Consejo Superior De Investigaciones Científicas Inoculant aseptique de micorhization et procedes d'application in vitro et ex vitro
ES2268984A1 (es) * 2005-07-29 2007-03-16 Consejo Superior Investig. Cientificas Inoculante aseptico de micorrizacion y procedimientos de aplicacion en condiciones in vitro y ex vitro.
US11814587B2 (en) * 2016-12-22 2023-11-14 Rainer TESCH Method for treating petroleum or natural gas

Also Published As

Publication number Publication date
GB2142933B (en) 1987-01-28
EP0144378A1 (fr) 1985-06-19
GB2142933A (en) 1985-01-30
GB8413168D0 (en) 1984-06-27

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