JPS59187706A - Microorganism coated seed - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to seeds coated with microorganisms. Conventionally, seed bags made of water-soluble resin or a mixture of water-soluble resin and fungicides or insecticides have been known for Seedlings to prevent seed deterioration or insect damage, mainly from the standpoint of seed preservation. However, little is known about the 4p effect of promoting seed germination and growth.

In general, early seed germination is extremely desirable because it eliminates the waste of sown seeds, saves seeds, makes germinated seedlings uniform, and reduces variation in growth. has been done.

From this point of view, the present inventors have conducted extensive research on yuzu coating, which promotes seed germination, and have found that seeds coated with a combination of a specific microorganism kept dormant and a coating agent. The present inventors have discovered that this has an extremely favorable germination and growth promoting effect, leading to the present invention.

That is, the present invention utilizes a plurality of microorganisms selected from the genera Pseudomonas, Bacillus, Arthrobacterium, Curthia, Aspergillus, Bacteroides, Brevibacterium, Nocardia, and Azotobacter, which are kept dormant and The present invention is a microorganism-coated seed characterized by being coated with a mixture of a bacterial cell preparation consisting of a mixture with an active carrier and a coating agent.

The microorganisms used in the present invention are classified into four groups: inorganic or organic nitrogen compound-degrading bacteria, cellulose-degrading bacteria, fat-degrading bacteria, and aerial nitrogen-fixing bacteria. The bacteria belonging to each group are as follows. be.

Inorganic or organic nitrogen compound degrading bacteria (1) Pseudomonas denitrificans (Ps
eudomonas denitritrif'1ca
ns ) (FEI Research Institute No. 6121) (2) Bacillus subtilis (FEI Research Institute No. 6114) (3i Bacillus thuringiensis
(4) Arthrobacter agilis (February 6113) (5) Kurtia zopf'ii (February 2011) No. 6119) Cellulose-degrading eggplant +6) Aspargillus tarreus (Feikoken Bacteria No. 6112) (7) Bacteroides succinoges
Bacteroides succinogenes (Feikoku Senboku Yori No. 6117) Fat N bacteria (8) Bacteroides lipolyticum (Bacteroides succinogenes)
(9) Brevibacterium lipolyticum (Br.
evibacterium lypolyticum
) (Norcardia corallina) (Norcardia corallina) (Number 6120) Airborne nitrogen-fixing bacteria αυ Azotobacter paspali (Number 7000) 0 Ka Azotobacter chroococcum
) (Feikoken Bacteria No. 6999) P&denitrificans is a bacterium isolated from soil, and is an aerobic and facultative aerobic bacterium.

B. subtilis and B. thuringiensis are both aerobic bacteria isolated from soil and have spore-forming ability. N. Coralina is an aerobic bacterium and has the ability to form chlamydospores. Many hydrocarbons, fats and oils, and phenols can be used as carbon sources. Agilis is a polymorphic aerobic bacterium and has a wide temperature range. In addition to organic nitrogen compounds, a large number of hydrocarbons can be used as carbon sources. Bacteroides succinoges is an anaerobic bacterium isolated from cow rumen. Bacteroides ribolyticum is a halophilic aerobic bacterium. Brev + IJ politicum is a bacterium isolated from a whey digestion plant, is an aerobic and facultative aerobe, and has a wide temperature range. and,
It can quickly break down fat. K. Sophie is
It was isolated from the decomposition product of chicken manure, and is an aerobic and facultative aerobic bacterium. A! ! P. tereus is an aerobic filamentous fungus that has spore-forming ability.

The bacteria II) to (4) and Ql described above can be cultured in a large-capacity fermentation tank made of stainless steel. Medium composition: 3% soybean meal, 1.0% starch, 0 ammonium sulfate.
．． 2% and 0.3% calcium carbonate, and the pH is adjusted to 70 before the culture medium is sterilized. Bacteria are grown by aeration and agitation culture at 30°C. The above-mentioned bacteria (71, +91) can be similarly cultured in a stainless steel large-capacity fermentation tank. The pH of the medium is adjusted to 7.0 before sterilization.Cultivation is carried out at 30°C, but under slow and mild conditions without aeration or excessive stirring. The above bacteria (6) (i.e.
Aspergillus s can also be cultured in the same stainless steel fermentation tank. Regarding the culture medium, the culture medium used for the bacteria in (1) to (4) is also used, but
However, the pH is adjusted to 6.0. Above (1) - (
Perform aeration agitation culture in the same manner as for the bacteria in 4) to allow the bacteria to grow.

The bacterium in (8), Bacteroides ribolyticum, is produced by adding 3% salt to nutrient broth (Difco, USA).
Inoculate into a medium adjusted to pH 6.5 to 7.0 before sterilization,
Grow at 35°C by aerated shaking or aerated stirring culture.

Bacteria 0υ and 02 are both aerial nitrogen-fixing bacteria extracted and isolated from agricultural land, and unlike the 10 types of bacteria mentioned above, they are grown in culture tubes on Chiten medium. The composition of the medium is HP
o, 1. (l f/1, MgSO4'7H)
00.2 Riri, NaC40,2VA, Fe5040.
OQ 55'/11 soil extract 100ml1. distilled water9
00 ml, 7 for mannide Q,Q? 7ZtSQ
Heaven 15. OVl, and the pH was adjusted before sterilizing the medium.
Adjust it to 7.5. Next, aerate culture at 25℃! Rather than being busy, let things multiply. In addition, as mentioned above (1i~0″
For the culture medium of A (D), other ingredients like the above-mentioned medium, such as minerals, vitamins, yeast extract, fish meal, plain heart infusion, NZ
-Additional additions such as amines and other commonly used ingredients are also possible.

In the present invention, each bacterial cell belonging to the four groups may be used alone, but inorganic or organic nitrogen compounds M111! one or more types of l and cellulose content M
It is most effective to use a combination of bacteria, fat-degrading bacteria, and air nitrogen-fixing bacteria, or to use a combination of all of the bacteria in (]) to (I2) above.

When mixing these microbial cells, the amount is usually increased or decreased as appropriate by slowing down the mixing of equal amounts. Moreover, instead of mixing these microbial cells, those that have been individually pure cultured by the above method are mixed together. That is, the culture was continued for 12 to 48 hours in the culture medium, and the bacterial cell paste was collected by centrifugation, and then freeze-dried in an acetone-dry ice bath to make it dormant, and then mixed with each other. do.

Furthermore, it is also possible to purely separate only the bacterial cells from the bacterial cell paste and freeze-dry it, or to leave it in a wet state without freeze-drying.

However, if it remains in a wet state such as a bacterial cell paste, it is not only inconvenient to handle and store, but also the bacteria may become inactivated or become excessively active. Hence these points? To solve this problem, it is preferable to mix the bacterial paste with an inert carrier. As the inert carrier, any material can be used as long as it is inert to these fungi, for example clays or clay minerals such as diatomite, montmorillonite, kaolin, vermiculite, bentonite, alumina, Silica, calcium carbonate, aluminum carbonate, apatite, attapulgite, etc. can be widely used. These inert carriers and bacterial cells are mixed in the most suitable ratio according to the carrier used and the moisture content of the bacterial cells to prepare a bacterial preparation.

The inert carrier acts not only as a drying agent for the bacterial cells but also as a retention agent for the bacterial cells. Therefore, the ratio of the bacterial cells and the inert carrier to the inert carrier is, for example, 1 to 9:9 to 1, more preferably. 1:
If mixed at a ratio of 1 to 1:5, the lifespan of the bacteria will be extended, and germination and proliferation of the bacteria will also be suppressed, so the mixture of bacteria and inert carrier should be packaged in special packaging. There will no longer be a need for packaging materials or packaging methods.

There are no special precautions to be taken when storing it, just store it in a cool, dry place.

Although microbial products usually require very delicate storage and preservation precautions, the simple operation of simply mixing the wet cake with the carrier without lion-aising the product can last a long time. Can be stored and preserved for a period of time.

The bacterial cell preparation thus obtained is applied to seed coats in the following manner. That is, first, seeds with a good germination rate (preferably a germination rate of 98% or more) are carefully selected and an appropriate amount of these seeds are delivered to the neck to be dressed.

The drying device has an inlet at one end and an outlet at the other end, and is equipped with a rotating screw or a rotating brush inside, as well as drying air outlets provided at appropriate intervals on the rotating shaft. A coating machine or a pelletizing machine can be used.

Next, while gradually rotating the screw or brush of the coating device, a water-soluble adhesive with relatively low water content, such as sodium or carboxymethyl cellulose (Na-C), is applied.
MC), polyvinyl alcohol (Pv), etc. are added in small quantities, and a mixture of the bacterial cell preparation and the M& agent is further added. The nucleating agents used here include clay minerals such as diatomaceous earth and montmorillonite, inorganic compounds such as calcium carbonate and magnesium carbonate, and 5orb (US Materials Co., Ltd.) consisting of potassium salts of starch polyacrylate bonded polymers.
It can also be called Bib-8orb, a product name manufactured by Science.

) It is possible to revitalize the town. In addition, 5Orb is an odorless and harmless powder exhibiting a light yellowish brown color, and has the property of adsorbing a large amount of water or aqueous solution. Further, the mixing ratio of the bacterial cell preparation and the tonic agent is desirably 25:5 to 5:95 (weight ratio).

When this range is exceeded, no significant improvement in seed germination rate is observed.

Next, compressed air L2 is supplied into the rotating shaft to dry the generated pellet. Also, depending on the size of the seeds, the operations after adding the adhesive are repeated several times to adjust the amount of phase preparation and compounding agent to obtain the desired particle size. In the final drilling process, warm air is blown to dry the adhesive and the pellet is forced to dry.

The microorganism of the present invention produced in this way r + m j
bt oak seeds have remarkable germination and growth promoting effects and disease control effects. ffl, when the microorganisms/nucleophiles of the present invention are seeded with Th and run out of water, the drug to be treated first absorbs water and swells, and furthermore, the dried fine powder of the keratin preparation also absorbs water and becomes dormant. Activates useful microorganisms, approximately 3 times after the first watering.
Microbial activity begins after 0 minutes to 1 hour. This state becomes an ideal environment for microbial activity, and the activated microorganisms absorb nutrients and minerals from the soil at the same time as absorbing water from the lubricant, and decompose them with enzymes while cell division. repeat. This division period lasts about 24 hours, and microorganisms change generations in this short period of time.

The protoplasm of dead microorganisms contains soluble organic or inorganic substances, enzymes, vitamins, growth hormones, etc. that have been broken down into easily absorbable forms, and these substances are ideal for plant germination and growth. It becomes a source of nutrients for the body. In addition, a slight increase in temperature is observed due to microbial activity, which leads to seed germination and
This provides the most ideal environment for rooting and growth. in this way,
Seeds kept in pellets are encased in an ideal environment with appropriate moisture, temperature, and nutrition, and grow straight, without bending and elongating like normal roots.

Therefore, less energy is required for the germination and growth of the plant, which is thought to contribute to a marked improvement in the germination rate and to the healthy growth of each seedling.

In addition, the microorganisms used in the seed compounding machine of the present invention are useful microorganisms obtained by pure culture, such as Pysium
m), Rhizophus, Trichoderma, etc., and prevents plant diseases. -According to theory, of the soil organisms that normally exist in soil, 2 to 3% are useful microorganisms and 5 to 12% are pathogenic microorganisms, and these beneficial microorganisms and pathogenic bacteria are Parents and offspring survive in search of a common nutrient source for reproduction, and when pathogenic bacteria win and begin reproduction, they cause disease to the plants. Therefore, in order to prevent the growth of pathogenic bacteria, disinfectants, a-drugs, etc. are usually used. The effects cannot be expected. Therefore, like the present invention, h! Is it pure culture of Dai 11 in child covering? ] When useful microorganisms that are not contaminated with Ryobe are used, the useful microorganisms, which are significantly superior in number, size, activity, etc., suppress the proliferation of pathogenic bacteria and do not cause diseases to A&1. In this way, the effect of vocal mimics is that, unlike furnace chemicals or fungicides, they do not kill pathogenic bacteria, but rather suppress their reproduction and keep them in a state where they are no longer active enough to cause disease to plants. it is conceivable that.

As falsely explained above, when using the rice plant 83 of the present invention, the germination and growth of seeds is promoted due to the synergistic effect of the specific useful microorganisms and the reagent. There is no wastage of sown seeds and it is possible to save seeds,
The germinated seedlings will be uniform, with no variation in growth, and will be healthy seedlings, and you can expect increased yields, better absorption of nutrients, and improved quality, and improved fruit aroma and flavor. ! It has an i-point effect.

Next, the effects of the present invention will be explained using test examples.

First, the following microorganisms were cultured according to their respective methods.

(1) Ps, Denitrificans (2)
B, subtilis (31B, thuringensis (41A,
Agilis (5) K., Zophii (61Asp, Tereus (71Bact), Succinogenes (81Bact)
, Lipolyticum (91Brev, Libolyticum αQ N, Coralina (Ill Company 0. Paspari Q3 Azo, Crocoticum soybean meal 3%)
A medium with a composition of 1% starch, 0.3% ammonium sulfate, and 0.3% charcoal was adjusted to pH 7, sterilized, and inoculated with cultures (1) to (4) and 0α, respectively. . Each culture was carried out in a stainless steel fermentor, maintained at a temperature of 30° C., and continued for 24 hours with aeration and stirring. +51, (7),
For f9+ sono, culture is carried out in a stainless steel fermenter in the same way as above, but the culture is carried out in a medium with a composition of 6% soybean meal, 2.0% starch, and 0.5% charcoal. Each culture was cultured at 30° C. for 24 hours with slow stirring and no aeration. Regarding the shield in (6), (1) to (
Cultivation was carried out in the same manner as in case 4), but the pH of the medium was
γ was adjusted to 6.017C instead of 0.

The corner of (8) is nutritional broth (manufactured by Difco, USA) K salt 3
The cells were inoculated into a medium whose pH was adjusted to 6.5 to 70 before sterilization, and grown at 35°C by aerated shaking or aerated agitation culture.

(The bacteria of Itl. 03 were cultured under the same conditions as above. After the culture was completed, the culture was centrifuged to obtain a bacterial cell paste. This paste was freeze-dried in an acetone-dry ice bath.

Cultures of 12 types of bacteria thus produced were mixed with diatomaceous earth at a ratio of 10:90 (weight ratio) to obtain a mixture of bacteria and carrier.

Next, the mixtures of these bacteria and carriers were combined to prepare bacterial cell preparations having the following blending ratios. In addition,
The numbers shown on the right are the number of bacterial cells per y of preparation (unit:
billion).

Among the bacterial cell preparations below, N is a combination of all 12 types of bacteria listed above.
The powder formulation of No. 0, 9 is a pale pink to brown color and is a fluid powder with a slight Kl odor.
It is abbreviated as 0-40ON.

No, IPg, Denitrificans 2.5N
0.2 B, Subtilis 2.5A
sp, Tereus 0,5 Brema,
Lipolyticum 05No, 3A, Agilis 2.5 Willow, Tereus
0.5Bact, Succinogenes 0
．． 5No, 4 Ps,, denitrifying force y
, r, j, QB, Thuringensis 0.4, Zoffii 0.4
8rev, Revoliticam 04N,
Coralina 04No, 5B, Subtilis 1.0m, Agilis
0.5, Sophie 05
Brev, Livoliticam 05N1
Coralina 0.5 companies 0. Crocotscum 0.5N'o, 5 Ps, Denitrificans 1.0m, Agilis 0
．． 5Bait, Zakushino Kenneth 05Bac
t, Revoliticam 0.5K/Zoffii
゛ 0.5Brev, Livolitycum 05No, γ Bact, Livolitycum 0.5Pg, Denitrificans 0.5
B, S. subtilis 02B, S. thurinkensis 05N, Coralina j. 0. Passbari 0.5 excitation, Kurokotsukamu 0.5No, 8Ps, Denitrificans 0.5B, Subtilis
05B, Thuringiensis 02 people, Agilis 05 Bact, Libolyticum 0.5N, Coralina 0.5mu Sp, Tereus
0,2, Sophie
02Azo, Pass Bali 05N
o, 9A, Agilis 0.5B,
subtilis 1. OB, Thuringiensis 0.5, Sophie
0, 1h, Denitrificans 1.0mu Sp, Tereus Brev, Rivoliticum N, Coralina 01AZO, Pasvari 05 Beni 0. Crocodile cam
0.5 Test Example 1゜ U, S, G-4 (using ION,
The water-binding polymer 5arb (product name of American Materials Science Co., Ltd.) was used as a stupefying agent, and the mixture ratios shown in Table 1 were prepared. BL manufactured by Gustafson, USA
A comparative test of effectiveness was conducted based on the difference in the blending ratio of the bacterial preparation and the coating agent.

The seed covering operation was carried out in accordance with the procedure prescribed for the above-mentioned beretized machine.

First, after thoroughly degassing the black soil (field soil) that had been delighted with chloropicrin two weeks before the test, each 1.5 m
X 1.5 m Xo, 3004 each for 6 2 m seedling boxes
The surface was leveled. Nql in each of these glaze boxes
Six types of test plots corresponding to the mixing ratios shown in Table 1 were set up, labeled from No. to 6i. For each test area, 100 fC'PIi child pellets were prepared using the mixing ratio shown in Table 1.
The seeds were sown in cr++ squares at intervals of one grain, and the soil that had passed through a 2-diameter sieve was spread over the soil to a thickness of 5 m.

Irrigation was carried out once a day from the time of sowing, using a watering can to uniformly water each plot. Furthermore, germination was determined at 10 a.m. every day when cotyledons appeared on the ground surface. The test results are shown in Table 2.

Chapter 1 Table The results in Table 2 can be summarized as follows.

1) The number of days to reach 80%, which is the minimum standard for the germination rate of sold seeds, is sown in No, 3, No, 4 areas FE] Color 4
The germination rate was 80% even on the 8th day in the No. 1 plot where the pollinating agent was not changed. did not reach
The results were lower than that of bare seeds (control). This is thought to be because there are too many microorganisms around the seeds, and nutrients such as nitrogen components are wasted on the multiplication of microorganisms, causing malnutrition in the seeds themselves.

2) No. 3, No. 3, and No. 004 plots are about 1 on the 8th day of sowing.
A germination rate of 0.00% was obtained, and it was found that good results were obtained when the mixing ratio of the dusting agent and the bacterial cell preparation was 75:25 to 95:5 (by weight ratio).

Test Example 2: Using the bacterial preparation with a mixing ratio of N081 to NO.9 as the bacterial preparation, and using the same 5o as in Test Example 1 as the dusting agent.
rb'%- was used, and the self-combination ratio of each bacterial cell preparation and 5orb was adjusted to 5:95 (if ratio), and eggplant (product 8i
: Senryo) and cabbage (variety: Early Akiho) 1 seed each
Seeds were coated on 00 grains using the same pelletized machine as in Test Example 1, and a comparison test was conducted between each m-form preparation.

First, field soil that had been sterilized in the same manner as in Test Example 1 was placed in two sets of planters (6 in each) labeled with numbers from A1 to A10.
0m x 25 crn x 25 crn) and sow 100 seeds in each section (4 rows x 25 if! I)
L-: Field soil passed through a sieve with a diameter of 2 mm was sieved to a thickness of 5 mm. Irrigation was carried out three times a day from the time of sowing, using a watering can to uniformly water each plot. Further, germination was determined at 10 am every day when cotyledons appeared from the ground surface. The test results are shown in Table 3.

The results in Table 3 are summarized as follows.

1) For both eggplant and cabbage seeds, germination was accelerated compared to 1-year-old control seeds (A10) of pine in each district, and this tendency was observed in many mixed types of fungi. flat 7
The effect was particularly remarkable in ~S9, and the U, 8.0-40ON formulation (A9), which was a mixture of all 12 types of bacterial cells, showed the highest effect.

2) After the test was over, we pulled out the young canopy and observed the underground part.
As the number of mixed types of fungi increased, the degree of fullness of the roots also increased, and in particular, in plot 49, there were many fine roots, confirming good early growth from germination.

Test Example 3 USG-40ON was used as a bacterial preparation, and various glazing agents were used to conduct comparative trial sales using different dusting agents to be mixed into the bacterial preparation. However, the mixing ratio of the coating agent and the bacterial cell preparation was 95:5 (weight ratio), which showed the best results in Test Example 1.
And so.

The Kumung roe that was used on the branch was lettuce (variety: Great Lakes 54), tomato (product 8I: Fukuju No. 2), and carrot (variety:
There are 3 types of pellets, 3 cm in bright red), and all pellet sizes are 3 in diameter, (
I got 1maK.

Three types of nuclide agents were used: water-binding polymer 5orb, calcium carbonate, and montmorillonite.

Regarding the above conditions and types, according to the classification shown in Table 4, seed nuclide was conducted on 100 seeds in each group using a pelletized machine in the same manner as in Test Example 1, and a comparative test was conducted based on the differences in the mulch agent. Example 4 Table First, field soil treated with anti-killing in the same manner as in Test Example 1 was placed in seedling raising boxes labeled with the symbols of the six categories M to F above (18 in total) for each of lettuce, tomato, and carrot (1.5 in each). m x
1.5 m x 0.2 m) packed with 300,
After leveling the surface, 100 seeds were added to each section.
The seeds were sown at a spacing of 1 seed per 0m square, and the soil was cultivated so that there were 5 grains of soil that passed through a sieve with 2 diameters.

Water was sprinkled uniformly once a day from the time of sowing to each plot using a watering can. Germination was determined every day at 10:00 a.m. when cotyledons emerged from the ground surface. The test results are shown in Table 5. In addition, on the final day of the trial, 20 seedlings were randomly removed from each plot, and their lj'n weights were measured. Table 6 shows the results.

The results in Tables 5 and 6 are summarized as follows.

1) The germination rate of the flannel lees agent also improved when the bacterial cell preparation was added to the mixture compared to when the flannel lees agent was used alone.

Furthermore, among the active agents, the water-binding polymer 5orb was the most effective.

2) New seedlings after the test! Even if you look at I ¥ weight, it is 5
In the case of the combination nuclide of orb and Use-40ON, the fresh type was the heaviest, confirming the healthy growth of the seedlings.

Patent applicant Ni Nis C International Company 1 31-

Claims (1)

[Scope of Claims] (11) A plurality of species selected from the genus Pseudomonas, Bacillus, Arthrobacterium, Kurtia, Aspergillus, Bacteroides, Brevibacterium, Nocardia, and Azotobacter and kept dormant. A microorganism-covered seed characterized by being coated with a mixture of a bacterial preparation consisting of a mixture of microorganisms and an inert carrier and a seeding agent. (2) The blending ratio of the bacterial preparation and the seeding agent is 25. Near 5
The microorganism-coated seed according to claim 1, which has a weight ratio of 5:95.