CN101617691A - Biological preparation capable of preventing and treating rice streak disease and promoting growth and application thereof - Google Patents
Biological preparation capable of preventing and treating rice streak disease and promoting growth and application thereof Download PDFInfo
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Abstract
The invention relates to a biological preparation capable of preventing and treating rice streak disease and promoting growth and application thereof, belonging to the technical field of biological pesticide. The biological preparation is prepared by a conventional method, and a producing strain is YFY02 (Lysobacter ginsengisoli). The strain of the biological preparation is characterized in that (1) the colony is round and in a opaque dilute liquid shape, and is dark yellow at initial stage of culture and chocolate brown or dark brown at later stage of culture, and the thallus is in the shape of a long rod; (2) the strain has strong bacteriostatic ability and wide antibacterial spectrum, can keep value on leaf surfaces of crops such as rice and the like and roots of crops such as konjak, calla and the like, can generate substances similar to antibiotics to promote the growth of crops of rice and the like, and has obvious and stable effect on preventing and treating rice streak disease. In the invention, the strain is cultured in liquid or solid and is prepared to liquid preparation or wettable powder. The invention has obvious and stable effect on preventing and treating bacterial diseases of rice streak disease, and good comprehensive properties, and is easy for industrial production.
Description
Technical field:
The present invention relates to a kind of biologic product and application thereof that prevents and treats the bacterial leaf streak of rice evil, belong to biological pesticide technical field.
Background technology:
Bacterial leaf streak of rice (Xanthomonoas oryzae pv.oryzicola) is called for short cecospora spot, be distributed widely in the torrid zone and the subtropical zone in Asia, the domestic areas such as south China, Central China, southwest that mainly are distributed in, being the fourth-largest rice disease of China after rice blast, banded sclerotial blight and bacterial leaf-blight, also is quarantine venereal disease evil important on China paddy rice.Because the susceptible in recent years rice varieties particularly large tracts of land of hybrid rice is promoted, Nan Fan and seed allocation and transportation etc. cause progressively its spread in china of this disease, and the lesion constantly enlarges, and have now become one of the main disease in rice district, China south.When this disease takes place, spread rapidly, cause that the flavescence of rice leaf is withered, unfilled gtains increases, output descends, general underproduction 15%-25% can reach 40%-60% in the time of seriously, and this disease is serious is threatening Rice Production.
At present, also do not control the special efficacy chemical agent of paddy rice cecospora spot in the production.The main medicament of preventing and treating bacterial diseases such as paddy rice cecospora spot as China at present is organic copper series bactericidal agent (dragon gram bacterium, the gloomy copper of thiophene etc.) and antibiotic class agricultural chemicals (middle living rhzomorph etc.), and they are all satisfactory inadequately aspect efficient, economy, safety; Long-term simultaneously extensive such chemical agent that uses makes pathogen produce pesticide resistance to such medicament.In addition, because the abuse chemical pesticide has caused chemical pesticide residual at natural world and food chain midium or long term, serious consequences such as environmental pollution, ecological balance destruction, social effects of pollution have been caused.Raising along with science and technology development and living standards of the people, the drug injury problems such as two big subject matters, particularly residue of pesticide that the production problems of quality of agricultural product problems such as residue of pesticide and agricultural product has become current China's agricultural development have caused almost commonwealthn's the awareness of unexpected development.
And that microbial pesticide has is little to non-target organism safety, toxic and side effect, to characteristics such as environment compatibility are good, gaining great popularity aspect the controlling plant damage by disease and insect.In today that human environmental consciousness strengthens, adopt efficient, nontoxic, safe noresidue, microbial pesticide easy to use to substitute the common recognition that the big chemical pesticide of present side effect progressively becomes countries in the world especially.
Summary of the invention:
The objective of the invention is to overcome the deficiency of prior art, a kind of efficient, nontoxic, safe noresidue, control paddy rice cecospora spot easy to use are provided and the biologic product of growth-promoting functions is arranged.
The biocontrol bacteria that the present invention adopts is the molten bacillus YFY02 of antibiotic (Lysobacter ginsengisoli), depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. Beijing. the Zhong Guan-cun; Preservation date: on March 17th, 2008; Preserving number CGMCC NO:2403.
The objective of the invention is to research, develop the biologic product of a kind of efficient, nontoxic, safe noresidue, control paddy rice cecospora spot easy to use by the molten bacillus YFY02 of antibiotic (Lysobacter ginsengisoli).
The molten bacillus YFY02 of antibiotic (Lysobacter ginsengisoli) is domestic not the appearing in the newspapers that the inventor found in 2006, can be used for the novel species of paddy rice cecospora spot biological control.
The molten bacillus YFY02 of production bacterial strain antibiotic of the present invention (Lysobacter ginsengisoli), separation is from the corn rhizosphere in bamboo plantation town, Fuyuan County, Yunnan Province, measure through morphology, cultivation proterties, conventional Physiology and biochemistry and the full-automatic identification systems of Biolog, this biocontrol bacteria is the new bacterial strain of the molten bacillus of antibiotic (Lysobacter ginsengisoli).
This bacterial strain has following feature:
(1) bacterium colony circle, opaque thin liquid shape, the cultivation initial stage is a dark yellow, the later stage transfers sepia or pitchy bacterium colony, thalline elongated rod shape to.
(2) this bacterial strain antimicrobial spectrum is wide, bacteriostasis is strong, can be at the rhizosphere colonization of crops such as rice leaf and potato, konjaku.Some bacterial diseases there is fungistatic effect preferably, wherein xanthomonas oryzae pv. oryzicola (Xanthomonoasoryzae pv.oryzicola) is had significant inhibitory effect, antibacterial circle diameter average out to 14.0-26.0mm.In addition, this bacterial strain is better except the viable bacteria fungistatic effect, and the bacteriostatic activity of its metabolite crude extract is also very strong, and some bacteriosises are all had good fungistatic effect.
(3) have following physiological and biochemical property: Gram-negative, oxidase and hydrogen peroxide enzyme positive, nitrate does not reduce, gelatin liquefaction, starch and not hydrolysis of Tween 80, indoles and arginine dihydrolase and urease negative.Can utilize glucose to produce acid, hydrolyzable casein; Routine and Biolog physiological and biochemical property: can utilize N-acetyl-D-galactose, N-acetyl-D-aminoglucose, D-mannose, a-D-glucose, D-trehalose, citric acid, 3-glycolic acid, a-batanone acid, a-ketoglutaric acid, propionic acid, succinic acid, bromine succinic acid, alanimamides, L-proline, L-threonine; Can not utilize L-serine, L-histidine, arabinose, maltose, close disaccharides, histidine, cellobiose etc.
(4) the molten bacillus YFY02 of antibiotic of the present invention (Lysobacter ginsengisoli) bacterial strain belongs to the short living bacterium of plant rhizosphere (Plant growth-promoting rhizobacteria is called for short PGPR), is the important factor of biological control.This bacterial strain is a kind of plant growing is had facilitation and pathogen had the beneficial bacteria of antagonism.The main mechanism of action of PGPR is: the A.PGPR bacterium can secrete the plant growth-promoting material, as plant hormone, vitamin, amino acid, other active organic molecule and derivative etc., thereby can promote the growth of crop.The B.PGPR bacterium has the effect that alleviates, reduces many crop pests generations.The biological disease regulatory mechanism of PGPR mainly is to produce antibiotic, the outer lyase of born of the same parents, cyanide and siderophore, also might be to change root circle microenvironment balance, promotes plant growth.After also finding some crop inoculation PGPR, the compound level of relevant crop pest defence rises, so this bacterial strain has control action to pathogen.
Biologic product of the present invention is the biologic product of a kind of efficient, nontoxic, safe noresidue, bacterial diseases such as control paddy rice cecospora spot easy to use.This bacterial strain can wait by generation antibiotics material stimulates paddy growth, and bacterial leaf streak of rice is had the sick effect of control efficiently, can be used for the crop pest biological control, has possessed the basic proterties of suitability for industrialized production simultaneously.
The present invention prepares according to a conventional method.Be about to biocontrol bacterial strain YFY02, cultivate, obtain to comprise bacterium thalline culture by liquid or solid, by liquid fermentation production, add wetting agent, spreader-sticker, or with after absorption carrier mixes by a certain percentage, after filtration, concentrate, pulverize and dry, be prepared into liquor or wetting powder.
The concrete preparation process of biologic product of the present invention is as follows:
One, the liquid fermentation preparation is produced
1. obtain the molten bacillus YFY02 of antibiotic (Lysobacter ginsengisoli).
2. the test tube kind is cultivated (following all be weight percentage):
On the test tube culture medium slant, culture medium prescription is 1000 milliliters of NA medium with the molten bacillus YFY02 of antibiotic inoculation: peptone 5g, sucrose 10 grams, dusty yeast 1g, beef extract 3g, agar 17-20g (medium is used), distilled water (supplying 1000ml).Cultivate 24-48h down, obtain the test tube kind for 22-26 ℃.
3. bacterial classification shaking table enlarged culture
On inclined-plane seed inoculation eggplant bottle slant medium, medium is the NA medium, and its prescription is identical with aforementioned test tube kind cultivation (2).Cultivate 24-48h down for 24-28 ℃, obtain eggplant bottle seed, use for fermenting and producing.
4. the pilot scale liquid fermentation is produced
Technological process is as follows:
Test tube strains → eggplant bottle seed → spore suspension → fermentation tank (filtrated air, culture fluid) → zymotic fluid → filling (precipitated calcium carbonate) → plate and frame filter press (discarding filtrate) → filter cake → wetting agent, the spreader-sticker etc. → 60 ℃ of oven dry → pulverizing → bacterium powder → quality examinations of sizing mixing → add.
Concrete fermentation production process is as follows:
1. the preparation of eggplant bottle inclined-plane seed liquor
Lawn with on the cultured eggplant bottle inclined-plane washes with aqua sterilisa, the preparation bacterial suspension.
2. inoculation
Adopt eggplant bottle seed, by the inoculation mouth, utilize pressure reduction to insert fermentation tank seed suspension, operation is strict, avoids polluting.Tank pressure can not be above 1.5/ square centimeter when boosting.
3. liquid fermentation production
(1) fermentation tank culture medium: analysis for soybean powder 2%, glucose 0.3%, peptone 0.2%, starch 3%, calcium carbonate 0.2%, fish meal 0.5%, peanut oil 0.1%, ammonium sulfate 0.05%, magnesium sulfate 0.03%, potassium dihydrogen phosphate 0.03%, dipotassium hydrogen phosphate 0.03%, defoamer 0.01%, NaOH600g.PH8.5 before the sterilization.
(2) sterilization: total vapour pressure of boiler should be in the 4-5 kg/cm, to guarantee the steam pressure of each pipeline.The sterilization of pipeline, air cleaner, fermentation tank and medium is all undertaken by the production technology standard.
(3) control of fermentation condition
The jar temperature: the jar Wen Jun of fermentation tank is controlled at 25-32 ℃, by inserting the thermometer measure of medium, regulates with the method that feeds cooling water or hot water in the interlayer.
Tank pressure: the tank pressure of fermentation tank is controlled at 0.5/ square centimeter, regulates by filtrated air inlet and Waste gas outlet.
Stir: the mixing speed of fermentation tank is 200 rev/mins.(air mass flow: the air flow control of seeding tank is 1: 0.5-0.8, favourable to growing of seed, enter fermentation tank after, throughput is increased to 1: 1.)
Sampling check: measured pH from probe tube sampling 1 time in per 2 hours, and smear is with violet staining microscopy thalli morphology, microscopy counting when having or not the pollution of assorted bacterium etc., fermentation tank to put jar, and carries out the counting of viable bacteria with the culture medium flat plate method.
Cultivation cycle: the cultivation cycle of fermentation tank is about 36-48 hour, can stop to cultivate when the bacterium number in the medium no longer increases, and puts jar immediately.
The liquid fermentation preparation: measure bacteria containing amount through plate count, bacteria containing amount reaches 18,000,000,000 CFU/ml, uses for the field control disease.
Two, the production of solid biologic preparation
At first carry out the liquid fermentation production of biocontrol bacteria.Through adding filler, plate-frame filtering, making beating, drying, pulverizing, make wetting powder then, use for experiment.
Detailed process is as follows:
1. carry out the liquid fermentation production (identical) of biocontrol bacteria with aforementioned one.
2. specifically add filler: zymotic fluid is pressed into basin, adds diatomite (precipitated calcium carbonate) as filler, add the back and stirred 30 minutes according to following computing formula.
3. the remnant (kilogram) in the addition (kilogram) of filler diatomite (precipitated calcium carbonate)=" zymotic fluid bacterium number (hundred million/milliliter) * put tank volume (liter) * yield "/finished product bacterium number-zymotic fluid.
4. plate-frame filtering: the pressure with 2 kg/cm, material is pressed into sheet frame by basin, filter by No. 7 filter clothes, pressure should be regulated at any time, makes the bacteria containing amount in the filtrate be no more than 0.2 hundred million/milliliter.
5. making beating: filter cake is discharged into the making beating jar, presses 8% of adding calcium carbonate quantity and add dense newborn No. 100, or, add the even stirring of an amount of filtrate 30 minutes by 3% adding SDS.
6. dry: aeration-drying is to water content 5%-8% in the baking room of temperature below 60 ℃.
7. pulverize: drop temperature can not surpass 60 ℃ during pulverizing, to prevent the thalline inactivation.
8. end product quality index determining: the mensuration of bacteria containing amount, water content, suspensibility, fineness is all carried out with reference to national company standard (Q/KWL02-2003), bacteria containing amount 1,800,000,000/gram, and all the other every indexs all meet standard.
Embodiment:
The main controlling object of biologic product of the present invention is bacterial diseases such as paddy rice cecospora spot.This preparation can be used by modes such as seed soaking, sprayings.Biologic product of the present invention in suburbs, Kunming, Yunnan, the greenhouse and the sub-district, field on ground such as state, Chuxiong Yuanmou County, Fuyuan County, Qujing and Mengzi County, state, Red River carried out efficiency test, proved that this biologic product preventive effect is remarkable, stable, can control the paddy rice cecospora spot effectively, the control efficiency of paddy rice cecospora spot is reached 60.24%-67.70%.
This biological prevention and control agent can with other kill bacterial pesticide, plant growth regulator mix use, but must guarantee mixed medicament the molten bacillus bacterium of the antibiotic YFY02 (Lysobacter ginsengisoli) of the present invention's employing is had no side effect.
Biocontrol bacteria preparation of the present invention is generally used in crop sowing or the disease phase of starting.Before method is sowing, adopts fermentation preparation and plant ball or seed and soak after (mixing) plant, dry and be manured into soil, or this preparation is passed through spray application in the disease phase of starting.Serious area is taken place the bacterial leaf streak of rice evil can impose in process of crop growth 1 to 2 time.
The present invention has obviously stable protection effect to bacteriosises such as paddy rice cecospora spots, and crops such as paddy rice are had good growth-promoting functions.Not only sick, the short fruit of coming into force of viable bacteria control is better for this bacterial strain YFY02, and the stripped fungistatic effect of its metabolite crude extract is also more outstanding, and the comprehensive proterties of this bacterial strain is better, is easy to suitability for industrialized production.
Be described in further detail the present invention with embodiment below, but content of the present invention is not limited thereto.
Embodiment one: biologic product (YFY02) is to the test of pesticide effectiveness of bacterial leaf streak of rice
(1) the stripped bacteriostatic test of biocontrol bacterial strain
1. for the examination material:
1.1 the molten bacillus strain YFY02 of strains tested antibiotic (Lysobacter ginsengisoli).
1.2 for the examination pathogen
Bacterium: xanthomonas oryzae pv. oryzicola (Xathomonas oryzae pv.oryzicola), Xanthomonas oryzae (Xanthomonas oryzae pv.oryzae), wildfire bacterium (P.syringae pv.tabaci) and anthurium bacterial eqpidemic disease (Xanthomonas axonopodis pv.dieffenbachiae), above pathogenetic bacteria is provided by this laboratory.
1.3 for having a try agent and medium
Ethyl acetate, acetone, methyl alcohol, benzinum etc., it is pure to be analysis.R
2A medium (yeast extract 0.50g; Matter peptone 0.50g between Ruan; Acid hydrolyzed casein 0.50g; Glucose 0.50g; Soluble starch 0.50g; Sodium Pyruvate 0.30g; Potassium hydrogen phosphate 0.30g; Epsom salt 0.05g; Distilled water 1000.00mL, pH7.2), PDA medium (potato 200g, agar 15g, glucose 15g, water 1000mL), NA medium (peptone 5g, beef extract 3g, yeast extract 1g, sucrose 10g, agar 17g, water 1000mL, pH7.0).
2. test method
2.1 the preparation of metabolite crude extract
At the sterilized R of dress 400mL
2The molten bacillus YFY02 of inoculation antibiotic in the 800mL conical flask of A liquid nutrient medium cultivates under 25 ℃ of conditions and stopped fermentation in 12 days.(12,000rpm/min) the collection supernatant adds the equal-volume ethyl acetate extraction 3 times in supernatant with the centrifugal 20min of zymotic fluid.Organic solvent layer 5%NaHCO
3And washing, use anhydrous NA again
2SO
4Dehydration, concentrating under reduced pressure obtains the metabolite crude extract.
2.2 the metabolite crude extract is measured the stripped antibacterial power of pathogenetic bacteria
Adopt cylinder-plate method, ethyl acetate extract is diluted to 5000 μ g/mL with acetone.Each concentration repeats 3 times, measures the inhibition zone size.
Table 1 variable concentrations crude extract is to the average diameter of inhibition zone of Different Kinds of Pathogens bacterium
Annotate: average diameter of inhibition zone is inoculation back 2d observed result.
The molten bacillus YFY02 of antibiotic metabolite crude extract sees Table 1 to the inhibition situation of 4 kinds of pathogenetic bacterias, table 1 shows, the metabolite crude extract all has the obvious suppression effect to 4 kinds of bacteriums, fungistatic effect to bacterial leaf streak of rice is the strongest, when concentration is 2mg/mL, antibacterial circle diameter reaches 3.5cm, and when concentration was 0.25mg/mL, bacteriostatic diameter was still greater than 1.5cm; Inhibitory action to Xanthomonas oryzae is taken second place, and when concentration was 2mg/mL, antibacterial circle diameter was 3.1cm, and when concentration was 0.25mg/mL, bacteriostatic diameter was still greater than 1.0cm; Inhibitory action to the anthurium bacterial epidemic disease bacterium is medium, and when concentration was 2mg/mL, antibacterial circle diameter was 2.6cm, and when concentration was 0.25mg/mL, bacteriostatic diameter was still greater than 0.7cm; Inhibitory action to the wildfire bacterium is relatively poor, and when concentration was 2mg/mL, antibacterial circle diameter was 2.0cm, when concentration is 0.25mg/mL, did not just have bacteriostasis.
(2) biologic product is to the sick test of the greenhouse pot culture control of paddy rice cecospora spot
1, experiment material and method
1.1, for the examination microbial inoculum: the molten bacillus YFY02 of biocontrol microorganisms antibiotic liquor
1.2, for studying thing: paddy rice
1.3, controlling object: bacterial leaf streak of rice
1.4. experiment place: plant protection institute of Yunnan Province agriculture university experiment booth, the yellow shell of the high sense of potted plant growth paddy rice cecospora spot kind is glutinous, and 3 repetitions are established in every processing.
1.5 the preparation of pathogen and biocontrol fungicide:
The preparation of paddy rice streak germ bacteria suspension: strains tested is the 3d activation before inoculation, paddy rice cecospora spot bacteria strain thin 10 is transplanted to cultivates 36～48h (28 ℃) on the NA flat board, pathogen washed with sterile water from the solid culture medium make bacteria suspension, on shaking table, shake up, in 600nm wavelength downward modulation OD value to 0.5, bacteria suspension concentration is equivalent to 3 * 10 with spectrophotometer
8CFU/mL uses for inoculation.
The preparation of biocontrol fungicide: molten bacillus YFY02 is inoculated in the triangular flask that 300mL NB liquid nutrient medium is housed with antibiotic, (180rpm in shaking table, 25 ℃) concussion cultivation 3d, in 600nm wavelength downward modulation OD value to 0.5, bacteria suspension concentration is equivalent to 3 * 10 with spectrophotometer
8CFU/mL.
1.6 inoculation method:
Earlier with the molten bacillus YFY02 of biocontrol microorganisms antibiotic liquor spray inoculation (booting stage) on rice leaf, inoculum concentration is the 100ml/ basin, establishes pure bacterium CK (only connecing xanthomonas oryzae pv. oryzicola), and 3 repetition are established in each processing, early, each spray in evening heat and moisture preserving 2 days also once.After 2 days, with thin 10 spray inoculations of paddy rice streak germ that prepare to the paddy rice of spraying biocontrol microorganisms YFY02, early, each spray in evening once and heat and moisture preserving 2 days.21 days " Invest, Then Investigate "s are investigated the disease number of sheets, and sick leaf-size class value, total number of sheets are calculated the incidence of disease, disease index.
Investigation record and state of an illness grade scale are as follows:
0 grade: no scab.
1 grade: lesion area accounts for below 10% of leaf area.
3 grades: lesion area accounts for 11%～25% of leaf area.
5 grades: lesion area accounts for 26%～45% of leaf area.
7 grades: lesion area accounts for 46%～65% of leaf area.
9 grades: lesion area accounts for more than 65% of leaf area.
Disease index=∑ [diseased plants at different levels (leaf) number * this disease level value] * 100/ investigation total strain number of sheets * the highest sick value of series
Total number of sheets * 100% of the sick leaf rate=investigation morbidity number of sheets/investigation
Relative control effect=(contrast disease index-processing disease index) * 100/ contrast disease index
2. test results and analysis
By the potted plant efficiency test of booth, as can be seen from Table 2: it is obvious to paddy rice cecospora spot control efficiency to use biocontrol microorganisms YFY02 biologic product, and control efficiency reaches 66.23%.Compare with the antibiotic class agricultural chemicals (middle living rhzomorph) of the control bacterial disease of using always in contrast and the production, the incidence of disease and the disease index of using each processing of biologic product YFY02 all have remarkable reduction, the preventive effect of this biologic product is remarkable, has shown the application and development prospect that this bacterial strain is potential.
The potted plant test result of table 2 booth biocontrol microorganisms YFY02 (21d)
(3) biologic product is to the sick test of field control of paddy rice cecospora spot
1, experiment material and method
1.1, reagent agent: the molten bacillus YFY02 of biocontrol microorganisms antibiotic liquor
1.2, for studying thing: paddy rice
1.3, controlling object: bacterial leaf streak of rice
1.4. experiment place: Yuanmou County rice test field, state, Chuxiong, Yunnan Province; middle fertility; rich No. seven of plantation sense paddy rice cecospora spot kind river; the sub-district is the single file district, 30 square metres of areas, and 3 repetitions are established in every processing; randomized arrangement; totally 9 sub-districts are established guard row all around, press the standardized management of high-grade rice cultivation step.
1.5 the preparation of pathogen and biocontrol fungicide:
The preparation of paddy rice streak germ bacteria suspension: strains tested is the 3d activation before inoculation, paddy rice cecospora spot bacteria strain thin 10 is transplanted to cultivates 36～48h (28 ℃) on the NA flat board, pathogen washed with sterile water from the solid culture medium make bacteria suspension, on shaking table, shake up, in 600nm wavelength downward modulation OD value to 0.5, bacteria suspension concentration is equivalent to 3 * 10 with spectrophotometer
8CFU/mL uses for inoculation.
The preparation of biocontrol fungicide: molten bacillus YFY02 is inoculated in the triangular flask that 300mL NB liquid nutrient medium is housed with antibiotic, (180rpm in shaking table, 25 ℃) concussion cultivation 3d, in 600nm wavelength downward modulation OD value to 0.5, bacteria suspension concentration is equivalent to 3 * 10 with spectrophotometer
8CFU/mL.
1.6 inoculation method:
Earlier with the molten bacillus YFY02 of biocontrol microorganisms antibiotic liquor spray inoculation (booting stage) on rice leaf, establish pure bacterium CK (only connecing xanthomonas oryzae pv. oryzicola), 3 repetition are established in each processing, early, each spray in evening heat and moisture preserving 2 days also once.After 2 days, with thin 10 spray inoculations of paddy rice streak germ that prepare to the paddy rice of spraying biocontrol microorganisms YFY02, early, each spray in evening once and heat and moisture preserving 2 days.21 days " Invest, Then Investigate "s are investigated the disease number of sheets, and sick leaf-size class value, total number of sheets are calculated the incidence of disease, disease index.
State of an illness grade scale:
0 grade: no scab.
1 grade: lesion area accounts for below 10% of leaf area.
3 grades: lesion area accounts for 11%～25% of leaf area.
5 grades: lesion area accounts for 26%～45% of leaf area.
7 grades: lesion area accounts for 46%～65% of leaf area.
9 grades: lesion area accounts for more than 65% of leaf area.
Disease index=∑ [diseased plants at different levels (leaf) number * this disease level value] * 100/ investigation total strain number of sheets * the highest sick value of series
Total number of sheets * 100% of the sick leaf rate=investigation morbidity number of sheets/investigation
Relative control effect=(contrast disease index-processing disease index) * 100/ contrast disease index
2. test results and analysis
By the field trial of carrying out at Yuanmou County, as can be seen from Table 3: it is obvious to paddy rice cecospora spot control efficiency to use biocontrol microorganisms YFY02 biologic product, and control efficiency reaches 62.40%.Compare with the antibiotic class agricultural chemicals (middle living rhzomorph) of the control bacterial disease of using always in contrast and the production, the incidence of disease and the disease index of using each processing of biologic product YFY02 all alleviate than agricultural chemicals and contrast morbidity.With twice efficiency test combination, illustrate that all biologic product YFY02 has good control efficiency to the paddy rice cecospora spot, and this efficiency test has repeatability.Above field trial further specifies this biologic product YFY02 the paddy rice cecospora spot is had stable, better prevention effect, has also possessed to produce the primary condition that goes up large-scale popularization.
Table 3 Yuanmou County experimental field biocontrol microorganisms YFY02 test result (21d)
Embodiment two: biocontrol bacterial strain YFY02 handles the growth-promoting functions to rice seedling
1, handle the preparation and the processing method of bacterium liquid:
Earlier the biocontrol microorganisms switching is activated at the NA medium, the enlarged culture of ruling then 36-48h (28 ℃) washes biocontrol microorganisms from the solid culture medium and to make bacteria suspension with sterile water, shake up on shaking table, in 600nm wavelength downward modulation 0D value to 0.5, bacteria suspension concentration is equivalent to (3 * 10 with spectrophotometer
8CFU/ml), be that 0.5 YFY02 biocontrol microorganisms liquid is by 10 times (3 * 10 with the OD value
7Cfu/mL), 100 times (3 * 10
6Cfu/mL), 200 (1.5 * 10
6Cfu/mL) doubly, 500 (6 * 10
5Cfu/mL) doubly, 1000 (3 * 10
5Cfu/mL) doing seed soaking respectively after the concentration gradient dilution doubly handles.Seed soaking 48h, every 12h changes treatment fluid 1 time.Rice material adopts yellow shell glutinous.Clear water is contrast.Each is handled 3 times and repeats, and each repeats 30 seeds.Paddy rice is cultivated and adopts conventional method: behind 30 ℃ of soak seed 48h and vernalization 12h, seed is put in the culture dish (diameter is 100mm, the paper water-soaked) that is lined with two-layer germination paper, 28 ℃ of illumination cultivation, the short fruit of coming into force of 10d investigation.
2, testing index and method:
Height of seedling and root are long: measure the length respectively handle seedling and root with ruler, wherein root is a measuring object with the longest one, with the mean value of all seedling of participating in the experiment of each processing as height of seedling and root long (cm).
Radical: the statistics all length is got the mean value of all seedling of participating in the experiment of each processing at last equally greater than the bar number of the root of 1cm.
Dry weight: seedling separated with blotting paper with root wrap, complete in 105 ℃ of baking ovens, 80 ℃ of baking 24h down weigh with the 0.001g balance after being cooled to room temperature in the drier, and its weight is dry weight (mg) divided by the seedling number.
3, bacterial strain is handled the facilitation to young rice seedlings growth
Result of the test shows (table 4): the growth that biocontrol microorganisms YFY02 handles rice seedling has good facilitation.In the processing of soaking seed with bacterium liquid, find that bacterial concentration is 200 (1.5 * 10
6Cfu/mL) times liquid is optimum the processing, and it is the most obvious to the facilitation of rice seedling growth, and five testing indexs such as height of seedling all are significantly higher than contrast and other processing.But 10 times, 50 times dilution seed soaking have suppressed growth of seedlings, and every index all significantly is lower than contrast.This test has been adopted 10 times, 50 times, 200 times, 500 times and 1000 times five concentration of bacterium liquid dilution to do seed soaking and has been handled, and the result shows that the short fruit of coming into force of 200 times of dilutions generally is better than other concentration.
Table 4 bacterial strain is handled the growth-promoting effect to rice seedling
Claims (4)
1, a kind of biologic product with control paddy rice streak disease and growth-promoting functions, it is characterized in that: the production bacterial strain of this biologic product is the molten bacillus of antibiotic (Lysobacter ginsengisoli) YFY02, preserving number CGMCC N0:2403.
2, a kind of biologic product with control paddy rice streak disease and growth-promoting functions, the step of its production process is:
(1) solid fermentation: comprise that the cultivation of test tube kind, bacterial classification shaking table enlarged culture, solid fermentation prepare biologic product;
(2) liquid fermentation: comprise that the cultivation of test tube kind, bacterial classification shaking table enlarged culture, liquid fermentation prepare biologic product;
Cultivate by liquid or solid, obtain to comprise bacterium thalline culture,, add wetting agent, spreader-sticker, or with after absorption carrier mixes by a certain percentage, after filtration, concentrate, pulverize and dry, be prepared into liquor or wetting powder by liquid fermentation production.
3, a kind of application of biologic product in control bacterial leaf streak of rice evil with control paddy rice streak disease and growth-promoting functions.
4, a kind of application of biologic product in paddy rice urgees to give birth to control paddy rice streak disease and growth-promoting functions.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100947636A CN101617691B (en) | 2009-07-24 | 2009-07-24 | Biological preparation capable of preventing and treating rice streak disease and promoting growth and application thereof |
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| CN2009100947636A CN101617691B (en) | 2009-07-24 | 2009-07-24 | Biological preparation capable of preventing and treating rice streak disease and promoting growth and application thereof |
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| CN101617691B CN101617691B (en) | 2012-03-07 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421724A (en) * | 2013-09-09 | 2013-12-04 | 云南农业大学 | Lysobacter antibioticus HY strain and application thereof |
| CN103667129A (en) * | 2013-12-06 | 2014-03-26 | 云南农业大学 | Lysobacter antibioticus 06-4 and application thereof |
| CN107400639A (en) * | 2017-06-22 | 2017-11-28 | 江苏省农业科学院 | A kind of molten bacillus of antagonism rice Xanthomonas and its separation method and application |
| CN110352958A (en) * | 2019-08-23 | 2019-10-22 | 云南农业大学 | A kind of Bio-seed Coating agent and preparation method thereof |
-
2009
- 2009-07-24 CN CN2009100947636A patent/CN101617691B/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421724A (en) * | 2013-09-09 | 2013-12-04 | 云南农业大学 | Lysobacter antibioticus HY strain and application thereof |
| CN103667129A (en) * | 2013-12-06 | 2014-03-26 | 云南农业大学 | Lysobacter antibioticus 06-4 and application thereof |
| CN107400639A (en) * | 2017-06-22 | 2017-11-28 | 江苏省农业科学院 | A kind of molten bacillus of antagonism rice Xanthomonas and its separation method and application |
| CN107400639B (en) * | 2017-06-22 | 2020-03-17 | 江苏省农业科学院 | Lysobacter for antagonizing xanthomonas oryzae as well as separation method and application thereof |
| CN110352958A (en) * | 2019-08-23 | 2019-10-22 | 云南农业大学 | A kind of Bio-seed Coating agent and preparation method thereof |
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| Publication number | Publication date |
|---|---|
| CN101617691B (en) | 2012-03-07 |
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