CN1188040C - Recompounded microbe germicide for preventing and controlling banded sclerotial blight and green smut of rice - Google Patents
Recompounded microbe germicide for preventing and controlling banded sclerotial blight and green smut of rice Download PDFInfo
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- CN1188040C CN1188040C CNB021383790A CN02138379A CN1188040C CN 1188040 C CN1188040 C CN 1188040C CN B021383790 A CNB021383790 A CN B021383790A CN 02138379 A CN02138379 A CN 02138379A CN 1188040 C CN1188040 C CN 1188040C
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Abstract
The present invention relates to a microbe composite agricultural chemical used for preventing and controlling the sheath blight and the green smut of rice. Bacillus subtilis is fermented and cultivated, and then fermentation liquid with certain bacterium content and validamycin are compounded to obtain the microbe composite agricultural chemical. The advantages of the bacillus subtilis and the validamycin can complement each other. The present invention used for preventing and controlling the sheath blight and the green smut of the rice not only has the efficient and quick bacteriostatic function of antibiotic, but also has the regulating function of the bacillus subtilis on the fair distribution of microbial flora after field colonization and mass propagation. Therefore, the present invention has synergistic effect, can play the role of optimal chemical effect, and can reduce the use quantity and the use frequency of the antibiotic validamycin to maintain the natural quality of the rice at maximum. The product has the characteristics of stable chemical effect and long preservation period (which can reach two years).
Description
(1) technical field under
The present invention relates to the microorganism Recompounded pesticide, particularly a kind of microorganism Recompounded pesticide that rice sheath blight disease and false smut is had antagonism.
(2) background technology
Rice sheath blight disease and rice green smut are the main diseases on the Rice Production.Rice sheath blight disease all has generation in the main rice workspace of countries such as the U.S., South America, Europe And Africa.The south China of China, Central China and East China morbidity are more serious, and North China, northeast and Yunnan Dao Qu also have generation, and some areas are heavier, the general field piece underproduction 5-10% that falls ill, and the serious field piece underproduction reaches 50-70%.False smut all has generation in each rice workspace of China, and false smut can cause production loss, more seriously influences the paddy rice quality.It is reported that the false smut infected seed has stronger toxicity to the people, raise livestock and poultry, can cause growth of animals or poultry to slow down with infected seed, the internal organ pathology, fecundity descends, even dead.
For a long time, the control of rice sheath blight disease mainly depends on jinggangmeisu on the Rice Production.Jinggangmeisu is longer service life, and dosage increases year by year, and the occurrence frequency and the severity of rice sheath blight disease show ascendant trend.The number of times that now is used to prevent and treat banded sclerotial blight 2 times/year before 20 years is increased to present 4-5 time/year, usage amount also from before 1.5 kilograms/hectare be increased to present 4.5-6.0 kilogram/hectare, continue to prevent and treat rice sheath blight disease and exist huge risk with jinggangmeisu.The control of rice green smut mainly depends on the medicament of organic copper, organic tin, and this type of heavy metal medicament easily produces poisoning to paddy rice and be easily rich long-pending residual in rice, influences the quality of rice.Along with the raising of rapid economy development and living standards of the people, people are more and more higher to the requirement of paddy rice quality and output.Green rice production presses for when reducing the production loss that is caused by damage by disease and insect as far as possible, the consumption of minimizing chemical pesticide and residual, the natural quality of maintenance rice, therefore, develop new control medicament, especially biopesticide seems very important and urgent.
According to the knowledge of the applicant, a kind of microbial bacterial bacillus subtilis (Bacillussubtilis) is arranged, have the bacterial strain that the pathogen of multiple fungal disease is had antagonism in its flora.Prevent and treat crop pest with bacillus subtilis with antagonism, research is all arranged both at home and abroad, abroad report mainly to cotton, fusarium wilt (the Mahaffee of tomato, W.Rosales A.M.Effects of seed factors on Spermoshpere and rhizoshpere colonization ofcotton by Bacillus subtilis GB03, Phtopathology83,1120-1125), research report (the Kong Jian etc. that domestic researcher has delivered the bacillus subtilis antibacterial material and its antibacterial protein is separated, 1992 " research of bacillus subtilis B-903 bacterial strain antibacterial material " " microorganism journal " 32 (6)), carry out in the laboratory but above-mentioned research is many, large tracts of land is not used for the field control crop pest.
At present, do not see the microorganism formulation that efficient bacillus subtilis is arranged, also not seeing has large-area applications to prevent and treat the microbial bacteria preparation of rice disease and microbial bacteria preparation and chemical pesticide composite dose in the farmland.
(3) summary of the invention
Technical problem to be solved by this invention is: overcome the deficiency of preventing and treating the prior art of rice sheath blight disease and false smut in the present production, screening and separating goes out the bacterial strain that a kind of bacillus subtilis has antagonism, itself and conventional pesticide is composite, obtain a kind of microorganism Recompounded pesticide that can effectively prevent and treat rice sheath blight disease and false smut.Simultaneously, develop the industrial process of this microorganism Recompounded pesticide.
The applicant finds that by a large amount of soil antagonistic microbes of screening bacillus subtilis (Bacillus subtilis) (the common micro-organisms center C GMCC.No.0808 of China Committee for Culture Collection of Microorganisms on September 30th, 2002) has had strong inhibitory effects to the growth of Rhizoctonia solani Kuhn and rice false smut.
The biological character of antagonism bacterium bacillus subtilis CGMCC0808:
Colonial morphology: unicellular formation bacterium colony circle, smooth all around, YPGA (yeast extract 5.0g, tryptone 5.0g; Glucose 5.0g, agar 17.0g; Water 1000ml) the bacterium colony 3-5mm of growth 24h on the culture medium flat plate, cream-coloured, there is gauffer on the surface.Diagnostic character: the Gram reaction negative, strict good gas, 2%NaCl can not grow, and lactulose, L-arabinose, L-lactic acid, melezitose and D-malic acid utilization reaction are all negative, and glycerine, sucrose, maltose and cellobiose utilization reaction are all positive.
The main points of the technology of the present invention solution are: bacillus subtilis CGMCC0808 is carried out fermented and cultured, will have the zymotic fluid and the composite jinggangmycin of certain bacterial content again, obtain a kind of microorganism Recompounded pesticide that can be used to prevent and treat rice sheath blight disease and false smut.
The industrial process of microorganism Recompounded pesticide of the present invention comprises the steps: (1), according to conventional method, carry out the bacillus subtilis strain activation in the laboratory, (2), according to conventional method, carry out strain culturing in the laboratory, (3), press inoculum concentration 1: 150-1: 300, cultured bacterial strain is moved into seeding tank carry out fermented and cultured, regularly detect thalline gemma content and bacterial content, fermentation time is 18-48 hour (4), press inoculum concentration 1: 150-1: 300, zymotic fluid in the seeding tank is moved into large fermentation tank to ferment, regularly detect thalline gemma content and bacterial content, fermentation time is 20-40 hour, (5), zymotic fluid goes out large fermentation tank, will go out zymotic fluid and composite jinggangmycin behind the jar, stir, promptly get microorganism Recompounded pesticide product.
The beneficial effect of microorganism Recompounded pesticide of the present invention is as follows:
1, according to the microecology principle, by introducing microbial bacterial agent, handle management rice root leaf encloses the reasonable distribution of microbiologic population in the ecotope, worsen the living environment of pathogen, in conjunction with cultivation management, agricultural and chemical control measure, form stable, balance, a bio-diversity paddy rice ecosystem, thereby control the popular of rice disease effectively, more enduringly.
2, microbial pesticide has the not available advantage of chemical agent, it is controlled the disease mechanism of action and is mainly: the 1. competition effect of dominant population---bacillus subtilis CGMCC0808 has the advantages that colonization ability is strong on rice plant, breeding is fast, can infect the site with the pathogen effective competition, reduce banded sclerotial blight morbidity risk rate (referring to table 1).2. the plain antibiosis of antagonism---bacillus subtilis CGMCC0808 can secrete generation antagonism element, its can suppress effectively the withered bacterium of line growth, infect and spread (referring to table 2).3. the induction of resistance effect of antagonism bacterium---bacillus subtilis CGMCC0808 can produce the enzyme relevant with disease resistance in the inducing paddy rice plant in a large number behind foliage-spray, resist the ability that pathogen is invaded, spreads thereby strengthened rice plant.
3, composite dose synergistic effect, behind bacillus subtilis CGMCC0808 and the composite jinggangmycin, form the effect of having complementary advantages, can bring into play antibiotic efficiently, bacteriostasis fast, can bring into play bacillus subtilis CGMCC0808 again and grow surely in the field, breed the adjustment function that back microbiologic population rationally distributes in a large number, thereby reach the purpose of sustainable Control disease.
4, can carry out composite with arbitrary proportion bacillus subtilis CGMCC0808 zymotic fluid and jinggangmeisu, as long as choose appropriate compound proportion, just can make the application of agricultural chemicals of the present invention reach the optimum medicine efficacy effect of can bringing into play, again can reduce antibiotic jinggangmeisu usage amount and access times, keep the purpose of rice natural quality as far as possible.
Show after tested that 5, the jinggangmeisu in the Recompounded pesticide product of the present invention forms noiseless effect to the clump count of bacillus subtilis CGMCC0808, product has efficacy stability, the characteristic of retention time long (can reach 2 years).Our experiments show that product of the present invention is environmentally safe, to the people, animal no pathogenicity, to the diseases and pests of agronomic crop natural enemies security.
(4) description of drawings
Biomaterial preservation situation: on September 30 2002 date, the common micro-organisms center C GMCC of the committee is managed in unit Chinese microorganism strain keeping, No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, address, numbering No.0808.
Fig. 1 is the flow chart of microorganism Recompounded pesticide industrial process of the present invention.
(5) embodiment
Embodiment 1, bacillus subtilis CGMCC0808 zymotic fluid becomes product of the present invention with composite jinggangmycin: with gemma content more than or equal to 90% and bacterial content of thalline more than or equal to 2 * 10
10The bacillus subtilis CGMCC0808 zymotic fluid of cfu/ml and 5% jinggangmeisu carry out composite according to 1: 1 ratio of volume, stir, and promptly get microorganism Recompounded pesticide product of the present invention (line Qu Ning).Can in the mentioned microorganism Recompounded pesticide, add the benzoic acid preservative that accounts for cumulative volume 2 ‰, prevent that medicament is putrid and deteriorated.After adding the benzoic acid preservative, stir, carry out packing and get final product.
Embodiment 2, the industrial process of microorganism Recompounded pesticide product of the present invention (line Qu Ning): at first according to conventional method, in the laboratory bacillus subtilis strain CGMCC0808 is activated, carry out strain culturing in the laboratory according to conventional method then, bacterial strain after the activation is moved into 500mlYPG (tryptone 5g, yeast extract 5g, glucose 5g, water 1000ml) in the culture fluid, at 120 commentaries on classics/min, cultivate after 48 hours down for 28 ℃, press inoculum concentration 1: 150-1: 300, cultured bacterial strain is moved into (temperature 28-32 ℃ of seeding tank, acidity-basicity ph value 7.0-7.4,) carry out fermented and cultured, fermentation time is 18-48 hour, every 2 hours seeding tank is carried out one-time detection therebetween, when bacterial content wherein more than or equal to 10
10During cfu/ml, press inoculum concentration 1: 150-1: 300, change the zymotic fluid in the seeding tank over to large fermentation tank fermentation (temperature 28-32 ℃, acidity-basicity ph value 7.0-7.4, foam can add defoamer more for a long time in right amount), fermentation time is 20-40 hour in large fermentation tank, carried out one-time detection therebetween every 2 hours, observe bacterial content, measure the acid-base value of zymotic fluid, after the gemma content of thalline in the large fermentation tank is more than or equal to 90%, detect bacterial content wherein, when bacterial content more than or equal to 2 * 10
10During cfu/ml zymotic fluid is gone out jar, zymotic fluid and 5% jinggangmeisu are carried out composite according to 1: 1 ratio of volume, stir, last, in Recompounded pesticide, add the benzoic acid preservative that accounts for cumulative volume 2 ‰, stir, carry out packing.
The content's index of jinggangmeisu and bacillus subtilis CGMCC0808 is the gross index of product.Jinggangmeisu content adopts liquid chromatography to measure (A 〉=2.0%), and the bacterium number of bacillus subtilis CGMCC0808 adopts the dull and stereotyped dilution method of counting to measure (〉=10
10Cfu/ml).Show that after tested the jinggangmeisu in the Recompounded pesticide product forms noiseless effect to the clump count of bacillus subtilis CGMCC0808.
The using method of microorganism Recompounded pesticide of the present invention is as follows: 1, prevent and treat rice sheath blight disease, at the rice sheath blight disease their early stage, every mu is used the bent peaceful 200-250ml of this product line, converts water 65-80Kg and slightly spray; Can advance control for the second time every 15-20 days; 2, the control rice green smut is prevented and treated about the last week at the paddy rice cut, and every mu is used the bent peaceful 250-300ml of this product line, converts the water 65-80Kg control of slightly spraying.
Microorganism Recompounded pesticide of the present invention is applied to prevent and treat rice sheath blight disease, as shown in table 3, showing has good control efficiency, when the bent peaceful consumption of agricultural chemicals line of the present invention is 200-300ml/ mu, 15 days control efficiency reaches 65.2-77.5%, and the lasting period increases to some extent than jinggangmeisu.Wherein single also have certain preventive effect with bacillus subtilis CGMCC0808 zymotic fluid to rice sheath blight disease.Table 3 be in Anhui, Jiangsu and Jiangxi San Sheng be with the test of pesticide effectiveness result of pesticide control rice sheath blight disease of the present invention.
Microorganism Recompounded pesticide of the present invention is applied to prevent and treat rice green smut, and is as shown in table 4, and showing has good control efficiency, and when the bent peaceful consumption of agricultural chemicals line of the present invention was 300ml/ mu, control efficiency reached 75.9-79.1%.Wherein single also have certain control efficiency with bacillus subtilis CGMCC0808 zymotic fluid to false smut.Table 4 be in Anhui, Jiangsu and Jiangxi San Sheng be with the test of pesticide effectiveness result of pesticide control rice green smut of the present invention.
Table 1, bacillus subtilis CGMCC0808 zymotic fluid are to the inhibitory action of Rhizoctonia solani Kuhn and rice false smut growth
Pathogen contrast bacterium colony stoste 1: 100 1: 500
Diameter (mm)
Colony diameter inhibiting rate colony diameter inhibiting rate colony diameter inhibiting rate
(mm) (％) (mm) (％) (mm) (％)
Rhizoctonia solani Kuhn 90 11.8 95.4 17.1 88.9 17.8 88.1
Rice green smut bacterium 90 90 100 90 100 90 100
Table 2, bacillus subtilis CGMCC0808 secretion are to the inhibitory action of Rhizoctonia solani Kuhn and rice false smut growth
Pathogen contrast bacterium colony stoste 1: 100 1: 500
Diameter (mm) colony diameter inhibiting rate colony diameter inhibiting rate colony diameter inhibiting rate
(mm) (％) (mm) (％) (mm) (％)
Rhizoctonia solani Kuhn 90 70.7 23.5 72.2 21.7 73.7 19.9
Rice green smut bacterium 90 90 100 90 100 90 100
Table 3, microorganism Recompounded pesticide of the present invention are prevented and treated the rice sheath blight disease result of the test
| Handle | Anhui (1999) | Jiangsu (2000) | Jiangxi (1999) | |||
| 7 days preventive effects | 15 days preventive effects | 7 days preventive effects | 15 days preventive effects | 7 days preventive effects | 15 days preventive effects | |
| The bent peaceful 150ml/ mu of line | 73.5b | 64.2c | 73.5b | 68.4c | 65.4c | 60.21bc |
| The bent peaceful aqua 200ml/ mu of line | 79.8a | 70.5b | 88.9b | 77.9c | 69.99bc | 65.20b |
| The bent peaceful aqua 300ml/ mu of line | 81.3a | 77.5a | 95.5c | 85.2c | 80.70c | 76.55c |
| Single bacterium aqua 250ml/ mu | 58.2c | 54.8d | 62.4bc | 38.4b | 60.05b | 56.63b |
| 5% jinggangmeisu 250ml/ mu | 80.1a | 70.6b | 85.5c | 74.4c | 74.46c | 71.32c |
| Blank (ck) | ---- | ---- | ----a | ----a | --a | --a |
Table 4, microorganism Recompounded pesticide of the present invention control rice green smut result of the test
| Handle | Anhui (1999) | Jiangsu (2000) | Jiangxi (1999) |
| The bent peaceful 150ml/ mu of line | 65.2b | 66.7b | 68.24c |
| The bent peaceful aqua 250ml/ mu of line | 73.5a | 81.9c | 79.10d |
| The bent peaceful aqua 300ml/ mu of line | 77.2a | 84.78c | 83.59d |
| Single bacterium aqua 250ml/ mu | 45.6c | 68.97b | 60.25b |
| 5% jinggangmeisu 250ml/ mu | 74.6a | 78.15c | 76.28cd |
| Blank (ck) | ---- | ----a | ----a |
Claims (9)
1, a kind of microorganism Recompounded pesticide that is used to prevent and treat rice sheath blight disease and false smut is characterized in that by bacillus subtilis (Bacillus subtilis) CGMCC.No.0808 bacterial strain bacterial content more than or equal to 2 * 10
10Cfu/ml, gemma content form more than or equal to the zymotic fluid and the composite jinggangmycin of bacterium amount 90%.
2, a kind of microorganism Recompounded pesticide that is used to prevent and treat rice sheath blight disease and false smut according to claim 1 is characterized in that by described fermentation of bacillus subtilis liquid and 5% jinggangmeisu according to composite the forming of 1: 1 ratio of volume.
3, a kind of microorganism Recompounded pesticide that is used to prevent and treat rice sheath blight disease and false smut according to claim 2 is characterized in that having the benzoic acid preservative that accounts for cumulative volume 2 ‰ in the described microorganism Recompounded pesticide.
4, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut is characterized in that comprising the steps: (1), according to conventional method, carries out bacillus subtilis CGMCC0808 bacterial strain B-916 activation culture in the laboratory; (2), according to conventional method, carry out bacterial strain breeding in the laboratory; (3), in inoculum concentration 1: 150-1: 300 ratio moves into seeding tank with cultured bacterial strain and carries out fermented and cultured, and fermentation time is 18-48 hour, regularly detects the bacterial content in the seeding tank; (4), in seeding tank bacterial content more than or equal to 10
10During cfu/ml, in inoculum concentration 1: 15-1: 30 ratio moves into large fermentation tank with the zymotic fluid in the seeding tank and ferments, and fermentation time is 20-40 hour, regularly detects thalline gemma content and bacterial content in the large fermentation tank; (5), when the zymotic fluid bacterial content of large fermentation tank more than or equal to 2 * 10
10The gemma content of cfu/ml, thalline went out jar more than or equal to 90% o'clock of the bacterium amount with zymotic fluid; (6), will go out zymotic fluid and composite jinggangmycin behind the jar, stir, promptly get microorganism Recompounded pesticide product.
5, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut according to claim 4, the controlling of production process condition that it is characterized in that seeding tank and large fermentation tank is: 28 ℃-32 ℃ of temperature, acidity-basicity ph value 7.0-7.4.
6, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut according to claim 4 is characterized in that every 2 hours seeding tank being carried out a bacterial content detects, and bacterial content is more than or equal to 10 in seeding tank
10During cfu/ml, change the zymotic fluid in the seeding tank over to large fermentation tank.
7, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut according to claim 4 is characterized in that every 2 hours large fermentation tank being carried out one-time detection, when the zymotic fluid bacterial content more than or equal to 2 * 10
10The gemma content of cfu/ml, thalline was discharged large fermentation tank more than or equal to 90% o'clock of the bacterium amount with zymotic fluid
8, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut according to claim 7, it is characterized in that the zymotic fluid and 5% jinggangmeisu that will go out large fermentation tank carry out composite according to 1: 1 ratio of volume, stir, promptly get microorganism Recompounded pesticide product.
9, a kind of industrial process that is used to prevent and treat the microorganism Recompounded pesticide of rice sheath blight disease and false smut according to claim 8 is characterized in that adding the benzoic acid preservative that accounts for cumulative volume 2 ‰ in described microorganism Recompounded pesticide.
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