JPH0591869A - Plant pathogenic fungus-inhibiting microorganism and its utilization - Google Patents

Abstract

PURPOSE:To obtain the subject microorganism effective for the ecological control of plant pathogenic fungi by separating and screening microorganismic strain from Bacillus which inhibits the growth of the plant pathogenic fungi from soil. CONSTITUTION:Microorganismic strain belonging to the genus Bacillus having strong antagonistic properties against the Fusarium fungi, the Phytopathola fungi, etc., are separated and screened to obtain the objective Bacillus SP BS-0001 and Bacillus SP BS-0001-SMCP III. The obtained microorganism is added to or mixed with seeds, roots and leaves of plants or with soil.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microorganism which is effective in controlling plant pathogens and a method of using the microorganism for ecological control.

[0002]

2. Description of the Related Art Soil-borne diseases are important problems in agriculture and horticulture, and among them, wilt disease, creeping vine disease, half-blight disease, yellow rot disease, and dry rot disease caused by Fusarium bacteria such as vegetables, flowers and fruit trees. , Etc., root rot, plague, seedling blight caused by Phytopasora, Pythium, Rhizoctonia, etc. Fruit tree, vegetable crotch disease, and bacterial wilt of tomato and eggplant are major problems.

Also in airborne diseases, diseases such as powdery mildew and Botrytis cinerea are important problems. In agriculture, avoiding these diseases is the most important issue for stable production of crops. As means for avoiding these diseases, use of resistant varieties and chemical disinfection are currently the main means.

However, it is difficult to avoid diseases by these methods alone. On the other hand, environmental, pollution, and health issues have led to a stricter view on the safety of products, and environmental issues are being highlighted worldwide, and there is a need for environmental conservation and ecologically harmonized agriculture. From such a viewpoint, many attempts have been made to control diseases using antagonistic microorganisms.

Rhizoctonia on cotton
(solani) Seed treatment of cotton with Pseudomonas fluorescens in contaminated soil has been shown to increase survival rates [see
Howell et al .: Fight Pathology (CRHOWELL et
c .: Phytopathology) Vol.69, No.5, 1979]. Fusarium oxysporum f. S of carnation
against the wilt disease caused by p. dianthi, we showed the possibility of controlling the wilt disease by soaking carnation seedlings in a bacterial suspension of Alcaligenes sp. [G.Y.Yuyuan et al .: Fight Pathology (G.
Y.YUEN etc .: Phytopathology) Vol.76, No.2, 1986
Year].

Rhizoctoni of chrysanthemum
a solani) for the control of chrysanthemum stem disease caused by the antagonist Gliocladium delique essence (Gliocladium del
iqu-escens), Trichoderma spp (Trichode
rma spp.), Bacillus spp.
Etc. are used to test [Chen Sheng Ah et al .: Report of the Mycological Society of Japan, 29, 1988].

Rhizoc of the Japanese radish
The effect of seed bacteriostasis of Pseudomonas cepacia on the inhibition of bacterial wilt disease caused by P. tonia solani has been confirmed [Yoshihisa Honma et al., Report of the Japanese Society for Plant Pathology, 55, 1989]. Enterobacter cloacé (Pythium ultimum) against lettuce
ae) has reported a biocontrol attempt [D.R. Fravel et al .: Plant & Soil]
(DRFRAVEL etc .: Plant and Soil), 125, 1990
Year].

[Sclerothum Lolfushi]
tiumrolfsii) diseased cotton Rhizoctonia solani
Diseases caused by (Rhizoctonia solani) have a chitin-degrading activity (Serratia marcescen)
s) are conducting tests to control disease [I.CHET etc .: Plan
t and Soil), 129 1990] Ecological disease control technology using such antagonistic microorganisms is expected to become more important in the future.

[0009]

The present invention is an effective method for controlling plant diseases, which is an important problem in agriculture, and provides a situation of application to eco-friendly agriculture.

[0010]

[Means for Solving the Problems] The present inventors have found that wilt fungus of tomato, cucumber creeping fungus, melon creeping fungus,
Fusarium such as radish yellow rot, strawberry yellow rot, etc., seedling blight, plague, root rot phytopasora genus, Pythium genus, Rhizoctonia genus, tomato half wilt, Chinese cabbage yellow disease and other verticillium We searched for fungi that have antagonism against plant pathogens such as genus fungi, Rosella genus that causes white rot, and Helicobacter bacillus that causes purple rot, and studied how to use them.

As a result, Bacillus sp. Having a strong antagonistic activity against various pathogens was isolated and selected from the soil, and Bacillus sp. BS-0001 (Microorganisms Research Institute 12534) was selected.
No.) The identification results of this strain are as follows. 1) Morphology (1) Cell shape and size 0.8 × 2-3 μm bacilli (2) Gram staining method Negative (3) Presence or absence of spores, shape, spores formed in the center of cells and sporangia Is swollen (4) With or without motility 2) Physiological properties (1) Anaerobic properties No growth (2) VP reaction- (3) Egg-Yolk reaction + (4) Maximum growth temperature 45 ° C (5) Growth in pH 5.7 medium + (6) Growth in nutritive broth + (7) Growth in NaCl (5-10%) medium- (8) Acid production from sugars a. Glucose + b. Arabinose- c. Xylose + d. Mannitol + (9) Hydrolysis of starch + (10) Decomposition of casein + 3) G + C content in DNA This strain is classified into the genus Bacillus based on the mycological properties of 53.2 or higher.

Bacillus macerans (Ba
cillus macerans), but Bacillus macerans differs from this strain in that it has spore formation at the cell edge, undergoes anaerobic growth, produces acid from arabinose, and does not decompose casein. Is a new strain belonging to the genus Bacillus. Also, Bacillus SP
(Bacillus sp.) BS-0001 (Microtechnology Research Institute, No. 12534) was treated with nitrosoguanidine to give chloramphenicol 5 pp.
A new strain of Bacillus sp.
sp.) BS-0001-SMCPI III (Microtechnology Research Institute, No. 12516). This strain is Bacillus sp. BS-0001 except that it is resistant to chloramphenicol.
It had similar bacteriological properties to. The present invention also relates to a method of using the novel microorganism for ecological control.

In the method of using the present invention, Bacillus sp. BS-0001 (Microtechnological Research Institute No. 12534) or BS-0001-SMCPI III (Microtechnical Research Prof. No. 1
No. 2516) can be applied to each crop seed, added to the nutrient solution in the hydroponic culture, or irrigated to the base of the plant in the rockwool mat or soil culture. In addition, this strain is adsorbed on a porous material such as vermiculite, zeolite, charcoal, cultivated on a substrate such as rapeseed meal, steamed bone meal, fish meal, or a mixture of a substrate and a porous material, and the culture Can be used as a seed bed, mixed with seedling cultivation soil,
It can also be applied to the field. The substrate and the porous material are not particularly limited as long as they do not inhibit the growth of this strain.

In addition, the culture of this strain can be applied to the foliar surface against airborne infectious fungal diseases such as powdery mildew, mildew and gray mold. Further, this strain can be mixed and used in combination with other effective bacteria.

[0015]

The present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited to these examples.

[0016]

Example 1 Microorganism according to the present invention BS-0001-SMCP II
Using a liquid medium containing 0.5 g of peptone, 0.3 g of yeast extract, 0.3 g of meat extract, 1.0 g of glucose, 100 ml of distilled water and pH 7.0, I was shake-cultured at 30 ° C. for 72 hours. Soak the seeds of cucumber (southern: Takii seedling) in this culture for 24 hours at 27 ℃.
Germination, 1/2 unit hydroponic solution (NO ₃ -N: 9me / l, NH ₄ -N: 0.7me
/ l, Ur-N: 0.3me / l, P: 2.5me / l, K: 4.15me / l, Mg:
1.9me / l, S: 2.8me / l, Ca: 4.65me / l, Mn: 0.6ppm, B:
0.225ppm, Fe: 1.85ppm) was sowed on a rockwool sowing mat impregnated with the same and grown in a greenhouse for 9 days until two-leaf development,
It was transplanted to a 9 cm × 9 cm rock wool seedling mat.

One week after transplantation, the cucumber creeping vine fungus (Fu
sariumoxysporum f.sp. cucumerinumIFO 6384 with a spore suspension of) was 10 ⁸ / 10ml inoculation in nursery mat. The cucumber vine cracking fungus was prepared by culturing in a potato dextrose liquid medium at 27 ° C. for 7 days and filtering with triple gauze to obtain spore fluid. During the nursery period, the above 1/2 unit hydroponic solution was supplied at appropriate times. As a control, seeds were soaked in distilled water at 27 ° C. for 24 hours and germinated, and then treated in the same manner. The test was conducted in 10 stations in each section, and the growth and disease on the 22nd day after transplantation were investigated, and the ecological control effect against cucumber vine cracking disease was compared and examined. The results are shown in Table 1.

[0018]

[Table 1]

[0019] The seed treatment of BS-0001-SMCP III bacterium tended to suppress the development of cucumber vine cracking disease, the above-ground weight was heavy, and the growth was excellent.

[0020]

[Example 2] 500 g of raw bone powder and 500 g of vermiculite were sterilized by autoclave, and the microorganism BS-
0001-SMCP III was liquid-cultured in the same manner as in Example 1 and 10 ml of a culture and 300 ml of sterilized water were added and mixed, and the mixture was cultured for 5 days and dried under ventilation at 40 ° C or lower to obtain a test sample having a water content of 15%. It was

2% of this test sample was added to nursery soil (200 mg of nitrogen, 1500 mg of phosphoric acid, 200 mg of potassium, 4000 mg / l of magnesia lime) and filled in a 9.5 cm polypot. Cucumber seedlings that had been sown on a vermiculite floor and expanded into two leaves (Southward: Takii seedling) were transplanted to this. Fusarium oxysporum f.sp. cucumerinum IFO 6384, which had been pre-cultured in potato dextrose liquid medium at the same time as transplantation
Spore suspension was inoculated 10 ^8/10 ml in 1 pot.

As a control, using BS-0001-SMCP III culture material-free seedling-raising soil, a test was conducted in triplicate. In the test, 21 days after sowing, 15 days after transplantation and inoculation of pathogenic bacteria, disease severity, aboveground weight, and microbial properties of cultivated soil were investigated. The results are shown in Table 2.
It was shown to.

[0023]

[Table 2]

By adding the BS-0001-SMCP III culture material to the nursery soil, BS-0001-SMCP III bacteria grow in the soil and the number of F. oxysporum bacteria is decreased. As a result, cucumber vine Cracking diseases were suppressed and growth tended to be excellent.

[0025]

Example 3 Tomato wilt soil was filled in a Wagner pot of 1 / 5000a, and an organic chemical compound (8-8-8) was added so that the amount of fertilizer was N and 15kg / 10a. BS-0001 was used for the test section, and a test sample obtained in the same manner as in Example 2 was used for 500 kg / 10a.
Was added. Tomato seeds (Large Fukuju:
Sakata seeds) were sown and the disease was investigated two months later. The test was performed in triplicate. The results are shown in Table 3.

[0026]

[Table 3]

The addition of the BS-0001 culture material to the soil tended to suppress the degree of tomato wilt disease onset.

[0028]

[Example 4] Using BS-0001, a test sample obtained in the same manner as in Example 2 was used to raise seedlings (200 mg of nitrogen, 1500 mg of phosphoric acid,
2% was added to Kari 200 mg and magnesia lime 4000 mg / l), and the mixture was filled in a 9.5 cm polypot. The seeds were sown on a vermiculite floor and expanded into two leaves (Prince: Sakata seeds).
Transplanted seedlings. Melon vine cracking fungus (Fusarium) pre-cultured in potato dextrose liquid medium at the same time as transplantation
oxysporum f.sp.melonis IFO 6385) 1 spore suspension
It was inoculated with 10 ⁸ / 10ml in the pot.

As a control, using BS-0001 culture material-free seedling-raising soil, tests were conducted in triplicate. 41 days after sowing, the severity and aboveground weight were investigated. The results are shown in Table 4.

[0030]

[Table 4]

By adding the BS-0001 culture material to the nursery soil, melon cracking disease was suppressed and melon growth tended to be excellent.

[0032]

[Example 5] Tomato seedlings raised using a rockwool sowing mat were transplanted to rockwool seedling cubes when two leaves were developed. BS-0001 grown in a YPMG liquid medium at the time of transplantation was inoculated with 10 ⁸ cells / 5 ml per cube. Two days after transplantation, Pseudomonas solanacearu
m MAFF 03-01843). The test was carried out in quadruplicate using the BS-0001 non-inoculated section as a control.

Cultivation was carried out for 31 days after sowing, and the degree of disease and growth were investigated. The results are shown in Table 5.

[0034]

[Table 5]

With BS-0001, the occurrence of bacterial wilt of tomato was suppressed and the growth of tomato tended to be excellent.

[0036]

EFFECTS OF THE INVENTION According to the present invention, diseases caused by phytopathogenic fungi can be suppressed and healthy crops can be grown.

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Claims (3)

[Claims] 1. A novel microorganism Bacillus sp. BS-0001 which suppresses the growth of plant pathogens. 2. A novel microorganism Bacillus sp. BS-0001-SMCP I that suppresses the growth of plant pathogens.
II. 3. A novel microorganism Bacillus sp.
us sp.) A method of using the microorganism for ecological control, which comprises treating BS-0001 or BS-0001-SMCP III on seeds, roots or leaves of plants or soil.