JPH067037A - Culture of field crop - Google Patents

Abstract

PURPOSE:To efficiently culture a useful field crop by directly coating a seed, etc., with a microorganism belonging to Enterobacter and exhibiting a plant growth promotion effect and sowing it into the soil or by mixing the microorganism in the soil and subsequently sowing a seed therein. CONSTITUTION:A microorganism [e.g. Enterobacter cloacae (Bikoken stipulation No. 1529)] belonging to Enterobacter and exhibiting a plant growth promotion effect is inoculated in a culture medium and cultured at 30 deg.C and 240rpm for 24hr so as to prepare a seed mother culture solution. A seed (e.g. cucumber seed) is immersed in the resultant culture solution and the treated seed is sown in the soil not treated with the microorganism for raising of seedlings. Or, the microorganism is mixed in the soil and a seed, etc., is subsequently sown in the treated soil for raising of seedlings. Or, a seedling or a field crop is cultured in a fertilizer solution for nutriculture, containing the microorganism exhibiting the plant growth promotion effect according to the nutriculture method. Thus, the objective useful field crop can efficiently be cultured.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for cultivating agricultural products,
Specifically, it relates to a method for cultivating an agricultural crop using a novel microorganism having a plant growth promoting action.

[0002]

2. Description of the Related Art A wide variety of microorganisms inhabit the rhizosphere or root surface of plants, which greatly affects the growth of organisms and the occurrence of diseases. Therefore, attempts have been made to separate industrially useful microorganisms from these rhizosphere microorganisms and utilize them for improving agricultural productivity, and there are many research reports and the like.

For example, nitrogen-fixing fungi supply nitrogen, which is one of the plant's three major nutrients, by fixing nitrogen in the air, and some mycorrhizal fungi utilize phosphorus in soil. It is known to increase efficiency and promote plant growth by supplying plants with phosphorus, which is an essential plant element.
In addition, although a wide variety of plant pathogens inhabit the soil, it is known that there are also microorganisms that have an antagonistic action against these pathogens. For example, bacteria belonging to the genus Pseudomonas as antagonistic microorganisms, etc. Has been separated and research on its use is being conducted.

However, since the mycorrhizal fungus is an active parasite, it is necessary to culture the plant itself, and it is difficult to industrially mass-produce it. for that reason,
It has not been put to practical use yet. Furthermore, even if a nitrogen-fixing bacterium can be industrially cultivated in a large amount, when the nitrogen-fixing bacterium is sprayed on the soil, the number of bacteria in the soil decreases with time. As a result, the fixed amount of nitrogen decreases, and it is said that there is a problem in comparing the economical efficiency with the case of using a commercially available nitrogen fertilizer. Further, most of the antagonistic bacteria produce an antagonistic substance (antibiotic) against the growth of plant pathogenic bacteria, and thus may exhibit a growth inhibitory effect on the growth of plants more or less.

[0005]

The object of the present invention is to provide a useful microorganism which can be easily cultured in a large amount, has good colonization to the plant rhizosphere and root surface, and has an action of promoting the growth of the plant. The purpose is to establish a method of applying the microorganism in order to improve the cultivation efficiency of agricultural crops, etc., after separating it from the root fungus and culturing the microorganism in a large amount.

[0006] In order to achieve the above object, the present inventors extensively searched for microorganisms having a plant growth promoting action from plant root fungi, and found a novel microorganism belonging to the genus Enterobacter isolated from root soil of cucumber. The present inventors have completed the present invention by discovering that Enterobacter cloacae meets the purpose of the present invention such as promoting the growth of various types of useful agricultural products including cucumber.

That is, the present invention is, firstly, that a microorganism belonging to the genus Enterobacter and having a plant growth promoting action is directly applied to seeds or the like and sown in the soil, or after the microorganism is mixed into the soil. The present invention relates to a method for cultivating agricultural crops, which comprises sowing seeds or the like on the seeds. A second aspect of the present invention is a crop characterized in that a seedling or a crop is hydroponically grown using a fertilizer solution for hydroponics to which microorganisms belonging to the genus Enterobacter and having a plant growth promoting action are added. Regarding cultivation method.

The novel microorganism, Enterobacter cloacae, used in the present invention was isolated from the rhizosphere soil of cucumber by the present inventors,
It has the following mycological properties.

1. Morphology (1) Cell shape and size: bacillus, 0.8-1.0 × 1.5
3.0 micron (2) Cell polymorphism: No (3) Motility: Yes (movement by periflagellate flagella) (4) Spores: No (5) Gram stain: Negative (6) Acidic: Negative

2. Growth state in each medium (1) Meat broth agar plate culture: Cells do not produce significant pigment,
It grows with a pale yellow cream-like appearance. The growth is vigorous. (2) Meat broth agar slope culture: Same as (1) above. (3) Liquid broth culture: The whole grows turbid and does not form pellicle. (4) Meat broth gelatin stab culture: It liquefies extremely slowly. (5) Milk culture: No noticeable changes such as liquefaction, coagulation and pH change are observed.

3. Physiological properties (1) Reduction of nitrate: Positive (2) Denitrification reaction: Negative (3) MR test: Negative (4) VP test: Positive (5) Indole formation: Negative (6) Hydrogen sulfide formation: Negative (7) Starch hydrolysis :: Negative (8) Use of citric acid: Positive (9) Inorganic nitrogen source: Nitrate or ammonium salt is used as a nitrogen source. (10) Pigment formation: No significant production of soluble and insoluble pigments is observed. (11) Urease: Positive (12) Oxidase: Negative (13) Catalase: Positive (14) Growth range: Grows in the temperature range of 15 to 45 ° C,
The optimum temperature is 28 to 37 ° C. A suitable growth pH is around neutral. (15) Attitude toward oxygen: facultative anaerobic (16) OF test: F type (17) Generation of acid and gas from sugars

[0012]

[Table 1]

4. Other properties (1) Production of DNase: negative (2) Production of tryptophan deaminase: negative (3) Production of β-galactosidase: positive (4) Arginine degradation test: positive (5) Lysine decarboxylation: negative ( 6) Ornithine decarboxylation reaction: positive (7) Esculin degradability: negative

As described above, this strain is a Gram-negative facultative anaerobic bacillus, has a morphological property that it does not make spores, and moves with periflagellates, and is oxidase-negative and nitrate-reducing-positive. Since it exhibits physiological properties, it is determined to belong to the family Enterobacteriaceae. Furthermore, due to various physiological properties, Enterobacter cloacae (Enterobacaceae)
ter cloacae) is most appropriate.
This strain has been deposited at the Institute of Microbial Science and Technology, the Agency of Industrial Science and Technology, as Microtechnology Research Institute No. 1529 (FERM BP-1529).

Next, the first and second aspects of the present invention will be described. A microorganism belonging to the genus Enterobacter such as Enterobacter cloacae and having a plant growth promoting action is applied at a rate of 10 ⁶ to 10 ¹⁰ per one seed of the plant and then directly applied to the soil, or Alternatively, the growth of plants can be promoted by mixing the microorganism at a rate of 10 ⁵ to 10 ⁸ per 1 g of soil into the soil and sowing seeds in the soil. In the case of hydroponics, the above-mentioned microorganisms are mixed in the nutrient solution at a rate of 10 ⁶ to 10 ⁸ cells / ml, and the microorganisms are added as needed during the cultivation period to grow the plants. Can be promoted.

In the cultivation of agricultural crops, the effect of inoculating the above-mentioned microorganisms firstly appears as a root growth promoting action, and then the growth of foliage is promoted.

The type of plant to which the present invention is applied is not limited, but agricultural crops are particularly preferable. Here, the agricultural products mean all agricultural products such as shells, vegetables, flowers and fruit trees, and their harvests. In addition, plants include seedlings, for example, seedlings of vegetables such as cucumber, pumpkin, eggplant, tomato, melon, and watermelon, seedlings of flowers, seedlings of cereals such as rice, and the like. be able to.
Further, in the present invention, the seeds and the like include seeds as well as seed potatoes in the case of tubers such as potatoes. Further, the hydroponics means all hydroponics such as hydroponic method, sand tillage method, gravel tillage method and rock wool method.

[0018]

EXAMPLES Next, the present invention will be described in detail with reference to Examples. Reference Example 1 (1) Enterobacter cloaca
e) Large-scale culture glucose 1.0%, peptone 0.5%, KH ₂ PO ₄ 0.1%,
400 ml of a liquid medium containing 0.05% of MgSO ₄ .7H ₂ O (hereinafter referred to as M medium) was added to a 1-liter Erlenmeyer flask, sterilized at 120 ° C. for 30 minutes, and then cooled and Enterobacter cloacae. (FERM BP-1529) Inoculate 1 platinum loop,
The seed culture medium is obtained by culturing at 240 rpm at 30 ° C for 24 hours. Charge 20 liters of M medium into a 30 liter jar fermenter, sterilize at 120 ° C for 30 minutes, and
After cooling to 0 ° C., the above seed culture solution was inoculated, and the culture temperature was 30
Culturing was carried out for 24 hours at a stirring temperature of 200 rpm and an aeration rate of 100 vvm. The number of Enterobacter cloacae bacteria in the culture was 1 × 10 ¹⁰ cells / ml.

Example 1 (1) Cucumber raising seedlings Soil for raising seedlings was added to a tray for raising seedlings (30 cm × 50 cm × 3 cm), and cucumber seeds (variety name: lady) 10
0 seeds were sown, cultivated at 20 to 30 ° C. for 1 week, then planted in a 3 inch pot, and further raised for 2 weeks. The experimental plots are as follows. Control area: The soil for raising seedlings was used as it was. Soil addition area: Enterobacter cloaca (FERM BP-15
29) 1 × 10 ⁷ pieces (obtained in the reference example, the same applies below) /
g / soil was added in the proportion of soil to raise seedlings. Seed application area: Seeds (cultivar name: lady) in the culture fluid of Enterobacter cloacae (bacteria number: 1.0 × 10 ¹⁰ cells / ml)
Was soaked and sown in soil to which the fungus was not added to raise seedlings.

The results are shown in Table 1. The control section has a ground length of 1
Soil seedlings of 0 cm or less accounted for 37%, whereas in the soil-added plot, the S-size proportion was 6%, and in the seed-applied plot, the S-size proportion was 18%. It was less than the comparison.

As described above, it was possible to grow large and healthy seedlings by treating with Enterobacter cloacae.

[0022]

[Table 2]

(2) Cucumber Cultivation The seedlings obtained as described above were planted in house soil at 80 cm intervals, and cucumber was cultivated for 3 months under the conditions of natural temperature and natural light in the house. As the fertilizer, inorganic fertilizer (KASEI No. 14) was added to the root as needed. Also, from the second week of cultivation, diisistone (trade name) was applied as needed to control oilworms. The test results are shown in Table 2.

As is clear from the table, the above-ground portion was larger in both the soil-added group and the seed-coated group than in the control group, and the amount of cucumber harvested was increased by 8 to 12% as compared with the control group.

[0025]

[Table 3]

(3) Rooting of Enterobacter cloacae on the root surface Roots were sampled over time during the cultivation period, and the number of Enterobacter cloacae growing on the root surface was measured. The root surface microorganisms were measured according to the method described in page 380 of the soil microorganism experiment method (edited by the Society for Soil Microorganism Research, published by Yokendo). In addition, Enterobacter cloaca is a Martin medium (glucose 1%, peptone 0.5%, KH ₂ PO ₄ 0.1%,
_{MgSO 4 · 7H 2 O 0.05%} , rose bengal 0.0033
%, Agar 2.0%, pH 6.8) at 30 ° C. for 24 to 48 hours, characteristic polysaccharides, color tone and shape are shown. Therefore, the number of colonies was counted using this property as an index.
The results are shown in Table 3.

[0027]

[Table 4]

Example 2 Two seeds of sunny lettuce were sowed on a 4 cm square urethane mat, and in a liquid fertilizer containing Otsuka House fertilizer No. 1 0.15% and No. 2 0.1% (both are trade names). Soaked in water, 24 ℃, 50
After cultivating for 10 days under the condition of 00 lux, it was planted in a hydroponic cultivation apparatus and cultivated under the conditions of 8000 lux and 24 ° C for 3 days.
It was cultivated for 5 days. The experimental plots are as follows.

Control group: Otsuka house fertilizer group without addition of microorganisms Addition group: Enterobacter cloacae was added to the Otsuka house fertilizer at a rate of 1 × 10 ⁵ pieces / ml to 1 × 10 ⁹ pieces / ml. One week per week for Enterobacter cloaca
A total of 4 times. Enterobacter cloacae was cultured in the same manner as in Reference Example 1 and used for the experiment.
The test results are shown in Table 4.

[0030]

[Table 5]

As is clear from the table, an increase in yield and root weight was observed by the addition of Enterobacter cloacae. In particular, the effect was remarkable at 1 × 10 ⁶ to 1 × 10 ⁸ cells / ml.

Example 3 40 g of rice (cultivar name: Akinishiki) was sown in a seedling box (50 cm × 15 cm × 20 cm) containing black soil, and the soil was covered at a thickness of 3-4 mm, and then 30-32 ° C. After sprouting for 2 days at 25 ° C., it was cultivated at 25 ° C. for 17 days. The experimental plots are as follows.

Control group: Bacteria-free soil (black soil) was used. Addition section: Black soil and cover soil to which 1.0 × 10 ⁷ Enterobacter cloacae / g soil had been added were used.

Enterobacter cloacae was prepared in the same manner as in Reference Example 1. The average foliage length of rice seedlings was 16.3 cm in the control group and 18.4 cm in the addition group, showing 113% growth compared to the control group.

Example 4 200 seeds of spinach (variety name: Jiromaru) were sown in a pot (50 cm × 15 cm × 20 cm) at 300 at 20 ° C.
It was cultivated for 40 days under the condition of 00 lux. The experimental plots are as follows.

Control group: Soil without addition of bacteria (black soil) was used. Addition section: 1.0 × 10 ⁸ pieces of Enterobacter cloacae / g soil was added.

Enterobacter cloacae was prepared in the same manner as in Reference Example 1. The results are shown in Table 5.

[0038]

[Table 6]

As is clear from the table, the addition group had an average weight of 135% as compared with the control group.

Example 5 One corn seed was sown in a 3 inch pot containing black soil and cultivated for 20 days at 25 ° C. and 5000 lux. The experimental plots are as follows.

Control section: Cultivated on black soil without addition of Enterobacter cloaca. Addition area: Enterobacter cloacae 1 per 1 g of soil
Cultivated by adding at a rate of × 10 ⁴ pieces / ml to 1 × 10 ⁹ pieces / ml.

Enterobacter cloacae was cultured in the same manner as in Reference Example 1 and used for the experiment. The test results are shown in Table 6.

[0043]

[Table 7]

As is clear from the table, Enterobacter
An increase in foliage length was observed with the addition of cloaca. Especially 1
In the area of × 10 ⁵ pieces / g soil to 1 × 10 ⁸ pieces / g soil, 114 to 137% of the growth promotion was recognized as compared with the control.

[0045]

INDUSTRIAL APPLICABILITY According to the present invention, by using a novel microorganism having a plant growth promoting action in cultivating agricultural crops and the like, useful agricultural crops can be efficiently cultivated.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Shinji Miyamichi Shinji Miyamichi, 760, Shimooka-cho, Kohoku-ku, Yokohama, Kanagawa Meiji Seika Co., Ltd.Pharmaceutical Research Institute (72) Hidemasa Hidaka 580, Horikawa-cho, Kawasaki-shi, Kanagawa Confectionery Co., Ltd. Biological Science Research Institute

Claims (4)

[Claims] 1. A microorganism belonging to the genus Enterobacter and having a plant growth promoting action is directly applied to seeds or the like and sown in the soil, or after the microorganisms are mixed in the soil, seeds or the like are added thereto. A method for cultivating agricultural products, characterized by sowing. 2. The method according to claim 1, wherein the microorganism belonging to the genus Enterobacter and having a plant growth promoting activity is Enterobacter cloacae. 3. A method for cultivating agricultural crops, which comprises hydroponically growing seedlings or agricultural crops using a fertilizer solution for hydroponic culture to which fine substances belonging to the genus Enterobacter and having a plant growth promoting action are added. 4. The method according to claim 3, which is a fine substance Enterobacter cloaca, which belongs to the genus Enterobacter and has a plant growth promoting action.