DE1037752B - Nematode repellants and methods of making such an agent - Google Patents

Description

GERMAN

The invention relates to means for incorporation in the soil in order to prevent the infestation of the growing thereon Vegetation, such as B. potatoes, beets and peas, to prevent by elk or nematodes.

It is known that when grown on certain soils, the parts of these crops lying in the soil of parasitic elk, which are among the greatest pests in arable farming. These Pests are so prevalent in many areas that the cultivation of various crops, especially the mentioned above, is practically impossible on soils that used to be very productive. Until now no effective measures to control the elderly are known and it is unlikely that that parasites of this type can be combated economically by using the soil Treats chemicals that are toxic to the infesting organisms. In contrast, it is known that certain naturally occurring fungi attack free-living nematodes, which fungi are referred to below as "predatory fungi" are designated. However, very little is known about the way in which the mushrooms work, and so far Few attempts have been made to demonstrate the peculiar behavior of these mushrooms for practical purposes to take advantage of. The present invention is based on the discovery that certain species of mushrooms are capable of the to attack parasitic nematodes on plants, e.g. B. the species Heterodera, Ditylenchus, Melidogyne etc. The invention also relates to the manufacture of such agents. Part of the present invention also the statement that the predatory mushrooms are grown under the conditions of deep culture fermentation can.

According to the present invention, the agent to be incorporated into the soil is to control the infestation to combat the growing vegetation with elk or nematodes, from the hyphae or cell threads of a predatory mushroom together with a solid organic carrier material that decomposes in the soil.

The carrier substance expediently consists of manure, shredded wool or textile waste or of Bran or similar materials, such as B. humus or foliage soil. However, the hyphae can also do without them Carriers are added to the soil if such substances are already present in the soil.

The mushrooms which are found to be particularly valuable for the use according to the invention are those which belong to the class of the Moniliaceen known as predatory; among them would be the following to quote:

Dactylaria thaumasia Dreschler,
Dactylaria candida (Nees) Sacc,
Dactylella doedycoides Dreschler,
Dactylella ellipsospora (Preuss) Grove,

Nematode control agents

and method of manufacture

such a means

Applicant:
MeIvyn Lumb,

West Bridgford, Nottingham,

and Charles Lionel Duddington,

Kingston-on -Thames, Surrey

(Great Britain)

Representative: Dr. M. Eule, patent attorney,
Munich 13, Kurfürstenplatz 2

Claimed priority:
Great Britain January 27, May 31 and October 30, 1956

Melvyn Lumb, West Bridgford, Nottingham,

and Charles Lionel Duddington,

Kingston-on-Thames, Surrey (UK),

have been named as inventors

Arthrobotrys conoides Dreschler,
Trichothecium flagrans Duddington,
Arthrobotrys robusta Duddington.

Such types of mushrooms can be used individually; however, you can add two or more to the remedy Incorporate species. So one has z. B. found out that an agent that, in addition to a rapidly growing fungus, such as B. Dactylaria thaumasia, a slower growing fungus such. B. Dactylaria doedycoides, contains, gives better results than a product containing either type alone.

So far, no cultures of these fungi have been established, apart from surface cultures in a purely experimental manner Scale. The present invention relates in one aspect to methods of growing the above Class of fungi in deep cultures, with isolation of the mycelium containing the viable fungi the culture fluids. For example, it has now been found that the above-mentioned fungi can be grown in deep cultures can draw in a number of media, provided that in the latter carbon, hydrogen and nitrogen are available. Regarding the nutrients that can be contained in the appropriate media,

809 599/561

3 4

If carbohydrates, amino acids, wheat soft water, are incorporated, then the resulting mass can Yeast extract, malt extract, peptones, extracts from Vege- are added to the soil according to methods as described in the stability and trace metals in the form of their salts. Which are generally used in agriculture. So Culture medium is suitable prior to use by, for example, the mass on the to be treated

Measures made sterile, for example, spread through 5 acres and is then used during the

Treatment in an autoclave at a temperature equal to the cultivation of the field.

115 ° C for a period of 20 minutes, and is then The above procedure is useful if the treatment is vaccinated with spores of a culture of the abovementioned development of the soil for the purpose of combating the elderly in Class of Predatory Moniliaceen. Which is to be carried out on a large scale when vaccinating. But it's only one spores used are obtained by growing a small-scale treatment required in the Culture of the required organism on a solid only small amounts of the mushrooms come into question, so can Medium, for example agar-agar with corn extract and the organism used in it in a surface glucose as the essential nutrients. The culture should be grown, preferably on solid inoculated medium is stirred and placed under germ-free agar-agar, i.e. not in a deep culture. The organ relationships with a constant influx of air 15 mus comes preferably along with a verb. The temperature is carefully regulated. thinners, such as B. manure, for application, and While it is possible to cultivate in exactly the same way as in treatment Run temperatures in the range of 15 to 37 ° C on a large scale.

optimal results are achieved at a temperature in the range of 24 to 28 ° C. The 20 species supplied to the culture, such as. B. beet nematodes or pea root air volume is also an important factor and depending on älchen, a similar treatment is applied.
from the volume involved, to a certain extent also.
contains. For example, it has been found that

Using a cylindrical vessel with a 25 Example 1
Total cubic content of 18181, which is about 13641 the

Culture fluid contained an aeration with 14,161 A culture of Dactylaria thaumasia Dreschler

per minute is very satisfactory. was made according to normal methods on a solid medium

The cultivation is allowed to go on until none of the following are grown:

Increase in weight of the mycelium contained in the vessel 30 corn extract 2 percent by weight

what can be determined by taking samples from grape sugar 0 25

the vessel is checked. This condition will be normal- Aear Agar 25

wisely reached after 7 or 8 days, although some _water "/" /// ". ] [] ["[[ fill up _to I ₀₀
Procedure only 4 days, others up to 10 days

required. As soon as the cultivation is completed, the culture obtained in this way became this

the mycelia present in the liquid by using 13641 a culture medium of the following

Filtration or other suitable measures isolated; Composition made by heating for 20 minutes

If necessary, they can also be inoculated with the water that had been sterilized at 115 ° C:
withdraw, for example by drying in vacuo

at room temperature without the viability of the 40 dextrose 0.25 percent by weight

Destroy organism. Peptone 0.25 ,,

The mycelium obtained in this way becomes the control aminoacetic acid (glycine) ... 0.25

fungus of the algae parasitizing on potato roots sodium chloride 0.5 ,,

either in the dried or in the undried wheat soft water 0.5

Conditions processed. The mycelium can, for example, 45 cornmeal 0.1 "

- if desired - fill up to 100 with water

suitable diluent incorporated into a soil

are, which is contaminated with Kartoffelwurzelälchen and the _pH value of the medium has previously been no potato harvest prior to sterilization; set to 5.75 after the incorporation. The culture was carried out in a cylinder potatoes 50 drical vessel with a volume of 1818 1 on the soil treated in this way. The cultivation and an excellent harvest are obtained. Solid substances can be used as liquid during the entire cultivation by means of diluents, two turbo-paddle agitators, each 686 mm through, for example, those which agitated nutrients to the fungus at a speed of η - 115. They lead so that the same The temperature of the liquid was kept at 26 ° C and can increase in the soil. One of the best of this class of 55 with sterile air in the vessel under the surface diluent is manure, and the use of the liquid has been blown in an amount of manure, which has been banded with a mycelium, of 14.16 1 each Minute. Seven days after inoculation, the one viable fungus from the predatory class was found by examining a specimen containing the moniliaceen, which enables the farmer to fertilize not only the soil, but also the weight of the mycelium per unit volume of liquid 60 no longer increased. To handle that separates from the liquid to combat the potato root elbows. The mycelium was isolated using a drum filter. If manure is used as a diluent, then part of the mycelium has been dried at room temperature in the treated manure for a short time, such as a vacuum, and mixed with bran for one or more weeks, lying undisturbed in piles that are 1.0 percent by weight dry Mycels contained, let before it gets into the ground; this way 65

the fungus reproduces within the fertilizer heap in Example 2
Further. If necessary, the fertilizer can be used

Before using a second treatment. Well-rotted manure was treated with a lot

be subjected to viable mycelium. That is treated by 0.9 kg of vaccine per 254 kg of fertilizer, and

viable mycelium the manure in this way 70 although with a culture of Dactylaria thaumasia, the

obtained by growing on the surface of a medium of the following composition:

Liquid made by heating 10 g of corn flour in 1.2 l of water at 80 ° C ... 1,000 ml

Agar-agar 20 g

The treatment was carried out by spreading the manure heap in layers and while forming gave the vaccine on top of each shift. The treatment suffered was under after treatment with a fertilizer the designation No. 1 Lincolnshire Farmers Co-operative Fertilizer with a fertilizer dose of 711 kg to around 0.4 hectares are plowed and divided into a number of fields each 4.57 m wide and seven furrows long. After 3 months, a number of fields with two cart loads each (about 100 kg) were inoculated Treated manure, a number of fields with two cartloads each (about 100 kg) uninoculated

was repeated after 3 months using io fertilizer. A number of fields were treated

a culture of Dactylaria thaumasia produced in a similar manner, each 254 kg of fertilizer, 2.27 kg vaccine. After another month, they were in a field known as potato roots contaminated to such an extent that the cultivation of potatoes is abandoned as uneconomical had been drawn seven furrows of 27.4 m each. Two of the furrows were made with two cart loads each (about 100 kg) manure treated as above with a preparation that is 1.5 percent by weight dried Mycels from Dactylaria thaumasia 'Dreschler, prepared as in Example 1, contained in one Amount of 203 kg on 0.4 ha; the remaining fields were left untreated. Each of the fields was planted with potatoes of the Record variety, namely 10 tubers per row, i.e. 70 tubers per field. The furrows were then padded in the usual way and left to their own devices for 4 months, after which time

Dactylaria thaumasia was inoculated; another two furrows 20 each field separately harvested and the yield

were treated with an equivalent amount of fertilizer of about the same age but not inoculated with a fungus. The remaining three furrows were not treated with manure of any kind and were used as controls. Seed potatoes of the Record variety were planted in each of the seven furrows at a distance of 457 mm, ie a total of 60 tubers per row. After planting was completed, each furrow was filled in and piled up, as is customary practice, after which the field was ^{left undisturbed for 5 x} / ₂ months until the potatoes could be harvested by hand. At the time of harvest it was observed that the plants which had been treated with the inoculated fertilizer were completely healthy, whereas the plants treated with the uninoculated fertilizer showed severe damage caused by elkings, the untreated plants were completely destroyed. The foliage of the plants in the control rows, which was neither weighed with inoculation or in the usual cultivation method. the
were the following:

individual yields of potatoes

treatment

untreated ....
unvaccinated

fertilizer

inoculated

fertilizer

dry mycelium

in bran

Potato yield in kg of arable land No.

0.91
3.18
4.31

0.68 2.27 4.08

Total

1.36 0.91

0.68

5.44 7.71

2.27 10.89 16.1

2.04 4.31

It can be seen that the potato yield on the soil treated with the fungus mixed with manure was considerably higher than the yield from the untreated soil or from that with uninoculated manure treated was withered and dead, and a 40 treated soil. In a similar way was that too A lot of weeds had grown in these rows

settled. The yield of potatoes from each of the individual rows was as follows:

Row no. treatment Yield
on potatoes
in kg 1
2
3
4th
5
6th
7th inoculated fertilizer
inoculated fertilizer
uninoculated fertilizer
uninoculated fertilizer
untreated
untreated
untreated 36.0
34.5
16.3
21.0
6.8
6.0
7.7

Bran mixed mushroom, but not treated with manure, was considerably higher than the yield from the untreated soil.

45

It can be seen that the potato yield from the soil treated with the fungus is considerably higher than that the yield from the untreated soil as well as from the soil treated with non-inoculated fertilizer.

Example 3
Well-rotted manure was covered with a mycelium

Example 4

At random, an en bloc experiment was carried out using sugar beets of the genus "Sharpes British Klein" on soil contaminated with the beet nematode Heterodera Schachtii. Before sowing the sugar beets, the soil was treated with Fison's commercial fertilizer y Market Garden No. 4 « and at the same time different groups of arable land (each with an area of around 0.67 m ² ) were treated in the following way:

Treatment No. 1: Left untreated as a control.

Treatment No. 2: Treated with bran, 1.36 kg per field.

Treatment No. 3: Treated with wet mycelium from Dactylaria thaumasia (produced by deep culture fermentation, as described in Example 1), 0.45 kg per piece of land.

Treatment No. 4: Treated with bran, 1.36 kg per field, and wet mycelium from Dactylaria thaumasia

treats the viable fungus of genus 65 (produced as in 3), 0.45 kg per piece of land.
Dactylaria thaumasia Dreschler, prepared as Treatment No. 5: Treated with Bran, 1.36 kg each

described in example 1. The mycelium was mixed with the fertilizer to 254 kg of fertilizer 6.8 kg of mycelium. A field that was known to be severely affected by the potato roots 70 above.

Field, and wet mycelium from Dactylaria thaumasia (prepared as in 3), 0.45 kg per field 4 weeks a second treatment with the mycelium like

Treatment No. 6: Treated with bran, 1.36 kg per field, wet mycelium from Dactylaria thaumasia (produced as in 3), 0.45 kg per piece of land and 10 ml of a surface culture fermentation of Dactylella doedycoides.

The sugar beets were sown on April 1, 1955, the October 7, 1955 harvest. The sugar beet yield from each treatment is tabulated below specified.

Beet yield
kg / piece of land

1.70

Treatment No. 13 14

2.12 2.12

2.37

2.37

2.60

As you can see, there was a slight increase in the sugar beet yield when either bran used alone or the mycelium alone, but this increase was insignificant in either case. One a statistically significant increase in yield was obtained with the two treatments 4 and 5 with their combination of bran and mycelium, a very significant and pronounced increase in yield with treatment 6, where the combination used the mycelia of two different viable fungi.

Example 5

Peas of the Meteor variety were sown in pots; each of the latter contained 3.18 kg of potting soil, which was heavily contaminated with the pea root. These Cultures were used for an experiment in which the potting soil was pretreated has been subjected to one of the six methods listed below; each treatment was twelve times repeated. The treatment of the soil in the pots was carried out in the mass (i.e. before potting), individually for each treatment.

Treatment No. 1: Manure, 31.2 g per pot, combined with wet mycelium from D. thaumasia, 56.7 g per pot (produced by deep culture fermentation, as in Example 1

ίο described).

Treatment No. 2: Wet mycelium from D. thaumasia (prepared as in 1), 56.7 g per pot.

Treatment No. 3: As for treatment No. 1, but using A. robusta instead of D. thaumasia.

Treatment No. 4: As for treatment No. 2, but using A. robusta instead of D. thaumasia. Treatment No. 5: stable manure, 31.2 g per pot.
Treatment no. 6: Soil completely untreated, for control.

In Table I below, details of the growth in height of the plants (measured in meters) in each of the pots observed 6 weeks after sowing. It will be the total amount of each one Treatment group belonging to twelve plants indicated, whereby it can be seen that the plants in the one with the fungus potting soil treated alone showed greater height growth than the plants in the untreated one Arable soil or the arable soil treated with manure alone. Furthermore, the plants in the pots showed whose soil had been treated with manure in combination with the fungus, a greater increase in height than those in the pots whose soil had been treated with the fungus alone.

Table I

Treatment
No. Experimental plants 15th 18th 20th 20th 20th 23 20th 18th 18th 20th 18th 20th All in all
cm 1 18th 18th 18th 18th 18th 15th 18th 15th 18th 20th 18th 20th 230 2 25th 20th 20th 20th 23 23 20th 20th 23 20th 18th 25th 214 3 18th 15th 20th 18th 15th 25th 20th 20th 15th 20th 23 23 257 4th 15th 18th 18th 18th 13th 18th 18th 15th 15th 18th 15th 15th 232 5 13th 10 13th 15th 15th 15th 15th 13th 15th 15th 15th 15th 196 6th 169

Furthermore, the pods of each plant were harvested 12 weeks after sowing, the total weight of the pods produced from each plant was determined. In Table II the individual pod weights for each of the twelve plants belonging to a treatment group and the total weight of the pods each Treatment group indicated. As you can see, the plants in the with the fungus gave together with the Manure treated soil will yield higher yields than the plants in soil treated with the fungus alone, and the latter in turn gave higher yields than the plants in the control pots or in the pots, their Soil had been treated with the manure alone.

No. 4.08 7.27 12.95 3.18 Weight of the pods 4.54 per plant (kg) 7.27 2.27 9.08 10.45 8.16 All in all 1 7.72 4.08 5.67 1.77 6.81 1.81 9.53 2.72 2.72 7.27 4.54 5.44 85.54 2 7.27 3.63 4.99 5.9 4.09 7.94 3.18 7.27 7.72 13.15 2.72 14.5 51.00 3 5.22 9.52 9.08 7.04 16.55 10.9 13.15 5.44 4.08 2.49 14.5 10.9 104.79 4th 0.227 5.9 2.72 4.54 3.41 0.908 1.82 0.454 0.227 4.76 1.81 1.36 84.40 5 0 0.454 0.114 0.454 0 1.36 2.72 0.908 1.59 2.93 0.454 1.14 25.63 6th 0.227 0 9.63

Claims (7)

Patent claims: 1. Nematode control agent, characterized in that it is an active ingredient in hyphae or cell threads of a nematode-infested mold, in particular fungi of at least one species the class of the nematode-infesting Moniliaceen. 2. Nematode control agent according to claim 1 _), characterized in that it also contains a solid organic carrier which decomposes in the soil. 3. Nematode control agent according to claim 1 or 2, characterized in that it is used as a carrier manure, Contains shredded wool, textile waste or bran. 4. Nematode control agent according to one of the preceding claims, characterized in that that there is a fungus of the following species Dactylaria thaumasia Dreschler, Dactylaria Candida (Nees) Sacc, Dactylella doedycoides Dreschler, Dactylella eUipsospora (Preuss) Grove, Arthrobotrys conoides Dreschler, Trichothecium flagrans Duddington, Contains Arthrobotrys robusta Duddington. 5. A method for producing a nematode control agent according to claims 1 and 4, characterized in that the fungus is grown in a culture medium under aerobic deep culture conditions cultivates, then separates the hyphae from the culture medium and optionally the same with the Mixed carrier substance. 6. A method for producing a nematode control agent according to claims 1 and 4, characterized in that the fungus is grown in a culture medium, the hyphae from the culture medium separates and the carrier substance in a moist state with the hyphae under conditions which lead to Conduct fermentation, inoculate. 7. The method according to claim 5, characterized in that the deep culture medium from a Carbohydrate, one or more amino acids, wheat soft water and water.