CN105309245A - Method utilizing bacillus subtilis PTS-394 wettable powder for preventing and treating crop soil-borne disease - Google Patents
Method utilizing bacillus subtilis PTS-394 wettable powder for preventing and treating crop soil-borne disease Download PDFInfo
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Abstract
The invention discloses a method for utilizing bacillus subtilis PTS-394 wettable powder for preventing and treating crop soil-borne disease. The method comprises the steps that in the sowing process, the bacillus subtilis PTS-394 wettable powder and seedling culture matrix are mixed according to the mass ratio of 1:10-100, and then sowing is carried out; in the field planting process, the PTS-394 wettable powder is uniformly broadcast into field planting furrows or holes; 10-15 days after field planting, the PTS-394 wettable powder is diluted to irrigate roots and used for preventing and treating the crop soil-borne disease. The current situation that existing biological agents are used in the field disorderly is broken through, the efficient field use technology is constructed for the bacillus subtilis PTS-394 wettable powder, the crop soil-borne disease can be prevented and treated stably and efficiently, phytotoxicity can be prevented, the pesticide use cost is reduced, and ecological environment is improved.
Description
Technical field
The present invention relates to biocontrol fungicide field operation technique, particularly a kind of method utilizing bacillus subtilis PTS-394 wetting powder to prevent and treat soil-borne disease of crop.
Background technology
Facilities vegetable has consequence in China's agricultural and rural economic development, is the main path of pillar industry in modern agricultural production and increasing peasant income.Jiangsu Province in recent years facilities vegetable cultivated area is stabilized in about 4,000,000 mu, accounts for 20% of the growing vegetables gross area.But the facilities vegetable planting structure distribution of high yield, continuous cropping also provides abundant nutrition and Reproduction Conditions for various disease simultaneously, frequently broken out, the sustainable development of serious threat vegetable production in green house by facilities vegetable soil-borne disease (solanaceous vegetables bacterial wilt, fusarium wilt, capsicum damping off, Strawberry Root Rot etc.) in recent years.
The main cause that soil-borne disease occurs is that the continuous farming of facilities vegetable causes rhizosphere soil microorganism population structure to be destroyed, original inhabitants' beneficial bacterium significantly reduces, in soil, pathogen bacterium amount increases year by year, and then causes soil nutrient ratio and quantity numerous imbalances (Yang Xiurong etc., 2008).At present, in production, facility for prevention and control soil-borne diseases of vegetable mainly relies on chemical pesticide, but control efficiency is always undesirable, soil-borne disease occurs still serious, chemical control simultaneously easily causes facility ecological environmental pollution, increase the residual of vegetables agricultural product toxic chemical substance, serious harm is brought to human health.Use biological prevention to control soil-borne disease in recent years, the strategy reaching restoration of the ecosystem more and more receives the concern of domestic and international scientific research personnel, and wins initial success.Janvier etc. (2007) research shows, in soil, beneficial microorganism community distribution and suppression pathogen ability are proportionate; Along with the increase of beneficial bacterium density in soil, the breeding of banana blight bacteria Fusariumoxysporumf.sp.cubense is suppressed, and disease incidence obviously declines; Pankhurst etc. (2008), if find to there is a large amount of beneficial bacteria and fungi in rhizosphere soil, have significant inhibitory action to Rhizoctonia cereali (Rhizoctoniasolani); Wu Fengzhi (2009) etc. study discovery, and the diversity improving soil microbial community can reduce the disease index of angular leaf spot of cucumber, powdery mildew, downy mildew and fusarium wilt and the quantity of Fusarium oxysporum, improve cucumber yield.
The kind of important soil-borne disease pathogen in facilities environment has been carried out in Plant Protection Inst., Jiangsu Academy of Agriculture since 2006, the research of biological property and occurrence regularity, with several important soil-borne disease (solanaceous vegetables bacterial wilt, fusarium wilt, capsicum damping off, Strawberry Root Rot etc.) be target, set up biocontrol microorganisms resources bank, obtain plant height effect biocontrol microorganisms-bacillus subtilis (Bacillussubtilis) PTS-394 (Chinese patent 201110339098.X), and prepare corresponding biocontrol fungicide, on this basis simultaneously, develop bacillus subtilis PTS-394 wetting powder, optimize its zymotechnique (Chinese patent 201310081080.3), this bacterial strain was stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) in 2013 at present, and deposit number is CGMCCNo.8085.This bacterial strain is in laboratory, good preventive and therapeutic effect is all shown to the multiple pathogen such as fusarium wilt and bacterial wilt, but in recent years when field is applied, peasant household often uses PTS-394 wetting powder separately by rule of thumb, insecticide spraying time, application method and dosage are chaotic, not unified standard, in most cases only fill with root when field planting or mix soil or spraying process, during sowing and general not medication after field planting, mu dosage 10-40 kilogram all has, cause field, various places soil-borne disease control efficiency instability (40-85%), even also poisoning in seedling stage occurs.At present, how field effective use bacillus subtilis PTS-394 wetting powder has become the key factor restricting this microbial inoculum and apply.
Summary of the invention
Practicable field effective use technology (insecticide spraying time, application method and dosage) is lacked for current bacillus subtilis PTS-394, cause the problem of its field control soil-borne disease effect instability, there is provided a kind of method utilizing bacillus subtilis PTS-394 wetting powder to prevent and treat soil-borne disease of crop, the present invention is achieved in that
A), before sowing, after by bacillus subtilis PTS-394 wetting powder, 1:10-100 mixes in mass ratio with seedling medium, more conventionally sow;
B) before field planting, in field planting ditch or cave, PTS-394 wetting powder is evenly spread fertilizer over the fields, 4-6 gram/;
C) after field planting, 10-15 days, PTS-394 wetting powder water fills with root, 500ml/ by after the mass ratio dilution of 1:100-1000;
Wetting powder of the present invention is 20,000,000,000 bacillus subtilis PTS-394 wetting powders.
PTS-394 wetting powder field of the present invention operation technique can be used for the various crop soil-borne diseases such as control solanaceous vegetables bacterial wilt, fusarium wilt, capsicum damping off, Strawberry Root Rot.
The invention has the advantages that: between crop sowing to harvest time, build bacillus subtilis PTS-394 wetting powder and prevent and treat various crop soil-borne disease field effective use technology, reach and utilize bacillus subtilis PTS-394 stability and high efficiency to prevent and treat soil-borne disease of crop; Reduce chemical agent dosage, reduce drug cost; To improve the ecological environment simultaneously, reduce the object of chemical agent residual quantity in agricultural product.
Embodiment
Embodiment 1 bacillus subtilis PTS-394 wetting powder viable bacteria amount detects
Preparation method adopts the method disclosed in Chinese patent 201110339098.X specification embodiment 4, namely gets the raw materials ready according to following weight percents: the former powder of bacillus subtilis PTS-394 viable bacteria: 40%; NNO (sodium metnylene bis-naphthalene sulfonate): 7%; Polyethylene glycol: 3%; Neopelex: 5%; Precipitated calcium carbonate: 45%;
Preparation process: the former powder of bacillus subtilis PTS-394 viable bacteria etc. is placed in blender according to the above ratio, and after mixing, being crushed to 325 orders through airslide disintegrating mill sieves, packaging, 20,000,000,000 bacillus subtilis PTS-394 wetting powders.
Under finished product is placed in normal temperature, shady and cool place's preservation, adopted plate dilution method respectively at after the same day, 6 months, 12 months, 18 months and 24 months, detected viable bacteria content in wetting powder.Every sub-sampling 5 parts, every dilution process repeats 3 wares; Testing result is as shown in table 1:
Table 1 bacillus subtilis PTS-394 wetting powder product viable bacteria amount detects
| 10 4cfu/g | 10 6cfu/g | 10 8cfu/g | 10 9cfu/g | 10 10cfu/g |
| The same day | Many | Many | 250 | 31 | 2 |
| 6 months | Many | Many | 249 | 29 | 2 |
| 12 months | Many | Many | 213 | 28 | 2 |
| 18 months | Many | Many | 181 | 19 | 2 |
| 24 months | Many | Many | 127 | 13 | 1 |
From table 1, in 1 year storage period of bacillus subtilis PTS-394 wetting powder, clump count change is little, but more than 1 year, clump count started slow minimizing; After 2 years, clump count drops to 1.3 × 10
10cfu/g; This shows: the bacillus subtilis PTS-394 wetting powder shelf-life is 2 years, meet agriculture chemical examination institution of the People's Republic of China (PRC) to biopesticide " bin stability and temperature, storage requirement are on the bioactive impact of product " (http://www.chinapesticide.gov.cn/nydjzlgd/view.html, the Ministry of Agriculture the 10th command chapter 4: special novel pesticide registration (microbial pesticide)).
Embodiment 2 bacillus subtilis PTS-394 wetting powder field control greenhouse tomato bacterial wilt is tested
Test period: 2010-2012, twice repetition.
Test site: carry out in the greenhouse tomato booth of Shahe town, Ganyu County, Lianyungang, Jiangsu Province.
Test kind: local main breed, kind No. one, capital.
Test medicine: the bacillus subtilis PTS-394 that embodiment 1 obtains; Contrast medicament is 50% carbendazim, purchased from Huifeng Agricultural Chemistry Co., Ltd., Jiangsu.
Seedling medium: vegetable seedling substrate (Lianyun Harbour Heng Aoda fertilizer Science and Technology Ltd.).
Test method: choose 4 heavier green houses of bacterial wilt of tomato natural occurrence for test booth, test seedling operates routinely, through point disk seedling growing and two stages of field planting.During sowing, bacillus subtilis PTS-394 wetting powder and seedling medium are mixed by five different quality ratios of 1:5,1:10,1:50,1:100 and 1:200 respectively, establish clear water to contrast simultaneously, conventional cave dish planting; Dish specification 10x5 hole, cave, 0.8 thickness, footpath, end 3.2x1.7; 40 days investigation emergence rates, seedling length and seedling fresh weights after planting.
In field planting ditch, spread fertilizer over the fields bacillus subtilis PTS-394 wetting powder according to 2 grams of/seedlings, 4 grams of/seedlings, 6 grams of/seedlings, 8 grams of/seedlings and 10 grams of/seedlings, five different amounts respectively during field planting, and then transplant tomato seedling; 10-15 days after field planting, carry out filling root according to mass ratio 1:50,1:100,1:1000,1:2000 tetra-ratios with water dilution bacillus subtilis PTS-394 wetting powders respectively, every seedling fills with root 500 milliliters; About 400, each process community seedling, repeats 2 times, process community random distribution; Set up clear water to contrast simultaneously; Investigate the incidence of each process when control plot tomato seedling is fallen ill, calculate preventive effect.
The incidence of disease=investigation community morbidity strain number/investigate total strain number x100;
The incidence of disease of preventive effect=(incidence of disease of the incidence of disease-control plot of process community)/control plot;
Result of the test is in table 2 and table 3:
The emergence rate of table 2 bacillus subtilis PTS-394 wetting powder and the rear tomato seedling of seedling medium different proportion mixing and growth
Impact
| Process | Seed number | To emerge number | Emergence rate | Seedling long (cm) | Seedling fresh weight (g) |
| 1:5 | 105 | 71 | 67.62b | 18.6b | 3.9b |
| 1:10 | 105 | 90 | 85.71a | 26.8a | 5.0a |
| 1:50 | 105 | 95 | 90.48a | 27.5a | 5.4a |
| 1:100 | 105 | 94 | 89.52a | 25.1a | 5.1a |
| 1:200 | 105 | 95 | 90.48a | 20.4ab | 3.9b |
| Clear water contrasts | 105 | 95 | 90.48a | 20.5ab | 3.8b |
Note: the different lowercase of same row shows to manifest outstanding sex differernce.
After table 3 bacillus subtilis PTS-394 wetting powder field planting and field planting, different administration amount is to the control of bacterial wilt of tomato
Effect
| Process | 1:50 | 1:100 | 1:1000 | 1:2000 |
| 2 grams of/seedlings | 66.07b | 60.53b | 42.22b | 21.72b |
| 4 grams of/seedlings | 85.57a | 85.21a | 79.53a | 67.08a |
| 6 grams of/seedlings | 87.95a | 87.34a | 81.59a | 71.44a |
| 8 grams of/seedlings | 88.19a | 87.95a | 81.93a | 72.14a |
| 10 grams of/seedlings | 87.99a | 87.20a | 80.05a | 69.27a |
Note: the different lowercase of same row shows to manifest outstanding sex differernce.
From table 2, when bacillus subtilis PTS-394 wetting powder and seedling medium mix, the process of 1:5 ratio has poisoning, and emergence rate is only 67.62%, in significant difference compared with clear water control treatment.The leaf emergence of 1:200 ratio, seedling are long all suitable with clear water control treatment with seedling fresh weight; 3 leaf emergence of 1:10,1:50 and 1:100 are at 85.71-90.48%, also suitable with clear water control treatment, but seedling is long and seedling fresh weight all increases in significance, and when showing that bacillus subtilis PTS-394 wetting powder and seedling medium mix, 1:10,1:50 and 1:100 ratio has significant growth-promoting functions to tomato plant.
The result of table 3 shows: spread fertilizer over the fields bacillus subtilis PTS-394 wetting powder with the consumption of 2 grams of/seedlings during field planting, the effect of control of plant bacterial wilt is lower, is only 21.72-66.07%.Process preventive effect increase compared with the process of 4-6 gram of/seedling of amount of application 8-10 gram of/seedling is not remarkable, considers producer's drug cost burden, the consumption of preferred 4-6 gram of/seedling; After field planting, three ratios with water dilution bacillus subtilis PTS-394 wetting powders of 10-15 days, 1:50-1:1000 carry out filling root, prevent and treat bacterial wilt Be very effective; And carry out filling root with 1:2000 dilution proportion bacillus subtilis PTS-394 wetting powder, preventive effect declines a lot.Consider producer's drug cost burden, after preferably using the dilution proportion bacillus subtilis PTS-394 wetting powder of 1:100-1000, fill with root.
Therefore: before A, sowing, bacillus subtilis PTS-394 wetting powder embodiment 1 obtained mixes with the ratio uniform of seedling medium according to 1:10-100;
In the field planting ditch risen or cave, bacillus subtilis PTS-394 wetting powder is spread fertilizer over the fields according to the ratio uniform of 4-6 gram of/seedling before B, field planting, in the present embodiment, plantation density is 3000/mu, therefore bacillus subtilis PTS-394 wetting powder every mu of consumption 12-18 kilogram during field planting;
10-15 days after C, field planting, carry out filling root according to the ratios with water dilution bacillus subtilis PTS-394 wetting powder of 1:100-1000, every seedling fills with root 500 milliliters.
Embodiment 3 bacillus subtilis PTS-394 wetting powder field control greenhouse pepper bacterial wilt and tomato wilt test
Test period: 2012-2015 test in 3 years;
Test site: carry out in Hua Chong town, Shuyang County, Xiuqian City of Jiangsu Province greenhouse pepper, tomato booth;
Test kind: local main breed, Soviet Union green pepper No. 17 and kind No. one, capital;
Test medicine: the bacillus subtilis PTS-394 wetting powder that embodiment 1 obtains; Contrast medicament is 50% carbendazim, purchased from Huifeng Agricultural Chemistry Co., Ltd., Jiangsu.
Seedling medium: seeding and seedling raising organic substrate (Zhenjiang training flower bud matrix development in science and technology Co., Ltd).
Test method: choose pepper ralstonia solanacearum and heavier 2 green houses of tomato wilt natural occurrence are test booth, test seedling operates routinely, through point disk seedling growing and two stages of field planting.During sowing, bacillus subtilis PTS-394 wetting powder and seedling medium are mixed in the ratio of 1:50, conventional planting; After spreading fertilizer over the fields bacillus subtilis PTS-394 wetting powder according to the consumption of 6g/ during field planting in field planting ditch, then transplant capsicum or tomato seedling; Every 15 days after field planting, carry out filling root according to the ratios with water dilution bacillus subtilis PTS-394 wetting powder of 1:100, every seedling fills with root 500 milliliters; About 400, each process community seedling, repeats 2 times, process community random distribution; Contrast agents 50% carbendazim using method is identical, uses by product description recommended dose; Set up clear water control group simultaneously.
When control plot capsicum or tomato seedling morbidity, investigation incidence, calculates preventive effect,
The incidence of disease=investigation community morbidity strain number/investigate total strain number x100;
The incidence of disease of preventive effect=(incidence of disease of the incidence of disease-control plot of process community)/control plot;
Result of the test is as shown in table 4:
Table 4 bacillus subtilis PTS-394 wetting powder field is to the control effect testing of pepper ralstonia solanacearum and tomato wilt
Note: the different lowercase of same row shows to manifest outstanding sex differernce.
From table 4, the incidence of disease contrasting disease Tanaka pepper ralstonia solanacearum and tomato wilt is high, is respectively 62.13% and 54.08%.Bacillus subtilis PTS-394 wetting powder field operation technique is respectively 81.81% and 85.98% to pepper ralstonia solanacearum and tomato wilt field efficacy, and the preventive effect of chemical agent carbendazim is respectively 21.68% and 48.72%, the preventive effect of bacillus subtilis PTS-394 wetting powder field operation technique is apparently higher than conventional chemical medicament carbendazim.
Result of the test shows: bacillus subtilis PTS-394 wetting powder field operation technique can effectively prevent and treat pepper ralstonia solanacearum and tomato wilt.
Embodiment 4 bacillus subtilis PTS-394 wetting powder field is tested capsicum damping off control effect testing in seedling stage
Test period: in January, 2013;
Test site: Huang Ma town, Qingpu District, Huai'an, Jiangsu Province is carried out;
Test kind: local main breed, Soviet Union green pepper No. 15;
Test medicine: the bacillus subtilis PTS-394 wetting powder that embodiment 1 obtains; Contrast medicament is 50% tpn, purchased from Jiangsu lark agrochemical Co., Ltd;
Seedling medium: long agriculture organic substrate, purchased from the fertile factory of the long agriculture organic substrate in Qingpu District, Huai'an;
Test method: test seedling operates routinely, during sowing by bacillus subtilis PTS-394 wetting powder and seedling medium respectively in mass ratio the ratio of 1:1,1:10,1:100 and 1:200 mix; Set up contrast medicament group and clear water control group simultaneously; Medicament group uses 50% tpn, and using method, with PTS-394 wetting powder, mixes with matrix by product description recommended dose.
Conventional cave dish planting, dish specification 96 hole, cave, 0.6 thickness, footpath, end 4.8x2.1; 40 days Investigate incidences after planting, calculate preventive effect;
The incidence of disease=morbidity strain number/investigate total strain number x100;
Preventive effect=(the process incidence of disease-contrast incidence of disease)/contrast incidence of disease;
Result of the test is in table 5:
Table 5 bacillus subtilis PTS-394 wetting powder field is to capsicum damping off control effect testing result in seedling stage
Note: the different lowercase of same row shows to manifest outstanding sex differernce.
From table 5, PTS-394 wetting powder mixes with matrix 1:1 has poisoning to plant, although the capsicum damping off incidence of disease is very low, plant affects by poisoning, growth retardation, loses value; PTS-394 wetting powder and matrix 1:200 mixed processing to the preventive effect of capsicum damping off with to contrast medicament tpn suitable; PTS-394 wetting powder mixes with matrix 1:10-1:100, is 74.70-85.30% to the preventive effect of capsicum damping off, apparently higher than the preventive effect (54.66%) of contrast chemical agent tpn; Result of the test shows: PTS-394 wetting powder and matrix in mass ratio 1:10-100 mixed processing effectively can prevent and treat capsicum damping off in seedling stage.
The effect test of embodiment 5 bacillus subtilis PTS-394 wetting powder field control Strawberry Root Rot
Test period: in August, 2015;
Test site: Lishui, Jiangsu Province Agriculture Science Institute base of plant;
Test kind: red cheek, directly provides Strawberry Seedlings by Jiangsu Province Agriculture Science Institute Horticultural Research Institute;
Test medicine: the bacillus subtilis PTS-394 wetting powder that embodiment 1 obtains; Contrast medicament is 10% difenoconazole water dispersible granule, purchased from Xian Zhengda limited company, is the general chemical medicament preventing and treating root rot on producing.
Test method: spread fertilizer over the fields bacillus subtilis PTS-394 wetting powder according to the consumption of 4g/ and 6g/ respectively during field planting in field planting ditch, transplants Strawberry Seedlings; Every 15-20 days after field planting, then carry out filling root according to two ratios with water dilution bacillus subtilis PTS-394 wetting powders of 1:100 and 1:500 respectively, every seedling fills with root 500 milliliters; Set up contrast medicament and clear water contrast simultaneously; Contrast medicament is 10% difenoconazole water dispersible granule, and using method, with bacillus subtilis PTS-394 wetting powder, uses by product description recommended dose; About 300, each process community seedling, repeats 2 times, process community random distribution; Investigation incidence when control plot capsicum or tomato seedling morbidity, calculates preventive effect:
The incidence of disease=investigation community morbidity strain number/investigate total strain number x100;
The incidence of disease of preventive effect=(incidence of disease of the incidence of disease-control plot of process community)/control plot;
Result of the test is in table 6:
Table 6 bacillus subtilis PTS-394 wetting powder field is to the field efficacy measurement result of Strawberry Root Rot
Note: the different lowercase of same row shows to manifest outstanding sex differernce.
From table 6, during field planting, after bacillus subtilis PTS-394 wetting powder 4-6g/, field planting, the ratio root irrigation of 1:100 is 73.21-79.88% to the preventive effect of Strawberry Root Rot, apparently higher than the preventive effect of chemical agent 10% difenoconazole water dispersible granule.The ratio root irrigation of 1:500 after field planting, preventive effect drops to 67.33-76.39%, is still significant difference with contrasting chemical agent.Result of the test shows: the bacillus subtilis PTS-394 wetting powder field operation technique of structure effectively can prevent and treat Strawberry Root Rot.
Claims (2)
1. utilize bacillus subtilis PTS-394 wetting powder to prevent and treat a method for soil-borne disease of crop, it is characterized in that, concrete steps are as follows:
A) during sowing, by bacillus subtilis PTS-394 wetting powder and seedling medium in mass ratio 1:10-100 mix and sow afterwards;
B) during field planting, in field planting ditch or cave, PTS-394 wetting powder is evenly spread fertilizer over the fields, 4-6 gram/;
C) after field planting, 10-15 days, PTS-394 wetting powder water fills with root, 500ml/ by after the mass ratio dilution of 1:100-1000.
2. utilize bacillus subtilis PTS-394 wetting powder to prevent and treat the method for soil-borne disease of crop according to claim 1, it is characterized in that, described soil-borne disease of crop refers to: solanaceous vegetables bacterial wilt, fusarium wilt, capsicum damping off, Strawberry Root Rot.
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| CN106305224A (en) * | 2016-07-03 | 2017-01-11 | 安建慧 | Method for preventing or treating tomato verticillium wilt |
| CN107897179A (en) * | 2017-11-09 | 2018-04-13 | 江苏省农业科学院 | The effective use method of bacillus subtilis PTS-394 wettable powder field control Fusarium solanis |
| CN109744126A (en) * | 2019-03-28 | 2019-05-14 | 南京富一农业科技有限公司 | A kind of strawberry seedling cultivates nutrient matrix and preparation method thereof |
| CN117431195A (en) * | 2023-12-21 | 2024-01-23 | 北京绿氮生物科技有限公司 | Saline-alkali resistant growth-promoting bacterium, growth-promoting bacterium agent and application thereof |
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