CN101514325A - Isaria javanicanicus mould strain and application thereof in preventing and controlling prodenia litura - Google Patents
Isaria javanicanicus mould strain and application thereof in preventing and controlling prodenia litura Download PDFInfo
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- CN101514325A CN101514325A CNA200810219403XA CN200810219403A CN101514325A CN 101514325 A CN101514325 A CN 101514325A CN A200810219403X A CNA200810219403X A CN A200810219403XA CN 200810219403 A CN200810219403 A CN 200810219403A CN 101514325 A CN101514325 A CN 101514325A
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Abstract
The invention discloses an Isaria javanicanicus mould strain and application thereof. The high-efficiency Isaria javanicanicus (Friedrichs & Bally) strain Ij01 is classified to belong to Corynespora, Hyphomycetaceae, Hyphomycetales, Hyphomycetes, Mitosporic Fungi, Eumycota. The strain has been preserved in China Center for Type Culture Collection (CCTCC), the preservation date is Oct 27, 2008, and the preservation number is CCTCC M 208171. The high-efficiency Isaria javanicanicus mould is entomopathogenic fungi to have high efficiency for preventing and controlling prodenia litura, has the characteristics of brand-new action mechanism different from the prior chemical insecticides, and no environmental pollution and residue when taken as a living biological pesticide, and meets the requirement of organic food production.
Description
Technical field
The invention belongs to biological pesticide technical field, be specifically related to efficient Isaria javanicanicus mould strain and the application aspect pest control thereof.
Background technology
Prodenia litura Spodoptera litura (Fabricius) be a kind of concern all very high worldwide insect.Its feeding habits are extremely assorted, 99 sections, 290 multiple kinds of crops such as desirable food Cruciferae, Solanaceae, Curcurbitaceae.Prodenia litura is originally the intermittent insect of taking place, about 3～4 years big generations 1 time, after the nineties in 20th century, because the adjustment of agricultural planting structure, cash crop area such as vegetables and facility cultivation area rapidly increase expansion, has increased prodenia litura and got food, breeding place and hibernaculum, and summer high temperature is short of rain and the pesticide abuse killed natural enemies, make the big year after year generation of prodenia litura calamity, bring very big loss to agriculture production.
At present to the control of prodenia litura still based on chemical prevention.Because a large amount of uses of chemical pesticide, add the standard that is difficult in operational lack of standardization or peasant's actually operating, make prodenia litura generally produce resistance, and brought problem of environmental pollution thereupon.Since the sixties in 20th century, areas such as the Taiwan of ground such as India, Egypt, Japan, Turkey and China, Shanghai have reported that in succession prodenia litura has all produced resistance to multiple sterilant, comprising organochlorine, organophosphorus, carbamate, pyrethroid and Bacillus thuringiensis (Bt) etc.Problems such as resistance of being brought in view of a large amount of use chemical pesticide control prodenia lituras and environmental pollution are necessary to seek the prophylactico-therapeutic measures of continuous and effective more, and especially bio-control method is to overcome the negative effect that chemical pesticide brings.
The natural enemy of prodenia litura is more, that has write down has 169 kinds, wherein great majority are natural enemy insect, the fungi of infecting prodenia litura has only 3 kinds, is respectively Aspergillus ochraceus (Aspergillusochraceus), beauveria bassiana (Beauveria bassiana) and Nomuraea rileyi (Nomuraea rileyi).The former paecilomyces (Paecilomyces) that belongs to of the mould genus of Isaria (Isaria), be that the important insect cause of disease of a class is very former, widely distributed, can be used at present prevent and treat insects such as nematode, aphid, carmine spider mite, Bemisia tabaci, small cabbage moth and fall army worm, but do not see the report that utilizes the mould control of Isaria bacterium prodenia litura.
Summary of the invention
An object of the present invention is to provide a kind of efficient Isaria javanicanicus mould strain.
Another object of the present invention provides the application of described efficient Isaria javanicanicus mould strain control prodenia litura.
Purpose of the present invention realizes by following technical proposals:
The invention provides a kind of mould Isariajavanicus of efficient Java Isaria (Friedrichs ﹠amp that prevents and treats prodenia litura; Bally) bacterial strain Ij01.Described strain classification status belongs to Mycophyta (Eumycota), mitospore Mycophyta (Mitosporic fungi), hyphomycetes (Hyphomycetes), hyphomycetales (Hyphomycetales), silk spore section (Hyphomycetaceae), the mould genus of excellent spore (Isaria).Described bacterial strain is in China typical culture collection center (CCTCC) preservation, and preservation date is on October 27th, 2008, preserving number CCTCC M 208171, and particular content is: the mould Ij01 Isaria of Java Isaria javanicus (Friedrichs; Bally) Ij01.
The present invention uses the nature sieve method, by gathering natural susceptible insect, conventional separation and purification bacterial strain, pathogenic mensuration, biological characteristics and production traits comparison, producing bacterial strain and determine etc. that step obtains Java Isaria Isariajavanicus (the Friedrichs ﹠amp of a strain and good stability efficient to prodenia litura; Bally) bacterial strain Ij01.Described strain classification status: Mycophyta (Eumycota), mitospore Mycophyta (Mitosporic fungi), hyphomycetes (Hyphomycetes), hyphomycetales (Hyphomycetales), silk spore section (Hyphomycetaceae), the mould genus of excellent spore (Isaria).Find no sexual generation.
The morphological feature of Ij01 bacterial strain: on the PDA flat board, bacterium colony circle, white mycelium, spore layer gray purple, no coremium.On the Czapek-Dox flat board, colony growth is slower, subcircular, and white mycelium, spore layer color is light slightly.
The microscopically mycelia is transparent, and is colourless, width 2.66 ± 0.18 μ m; Conidiophore is similar to mycelia, does not become coremium, long 10.38 ± 3.26 μ m, wide 2.73 ± 0.25 μ m; Conidiogenous cell bottle stalk cucurbit shape, the widest part diameter 2.75 ± 0.35 μ m, long 7.92 ± 1.11 μ m, colyliform direct be born on the mycelia or on the conidiophore; The bottle stalk; Conidium is colourless, oblong, and two ends are point, long 5.47 ± 1.23 μ m, wide 2.09 ± 0.36 μ m.
The present invention provides the application of described efficient Isaria javanicanicus mould strain aspect the control prodenia litura simultaneously.The Ij01 bacterial strain is to the virulence of prodenia litura: under different spore concentrations, the average cumulative mortality ratio of prodenia litura 2 instar larvaes significantly increases along with the rising of concentration of treatment and the prolongation of time, when concentration is 50000 conidiums/mL, after processing the 6th day and the 8th day, the mortality ratio of prodenia litura promptly reached 48.33% and 51.26% respectively.Observed data calculates medial lethal dose through logarithm-probability value analytical method, and along with the prolongation of handling the back time, toxic limit medium dose progressively diminishes, LC
50Value is 10
5Conidium number/mL order of magnitude fluctuates up and down.Between the median lethal time, when concentration is 1 * 10
5During individual spore/mL, the median lethal time is 5.0 days, and 95% fiducial interval is 4.1～6.3 days.
The present invention compared with prior art has the following advantages and beneficial effect:
Efficient Java of the present invention Isaria mould is efficient to the control prodenia litura; The Isaria mould is a kind of insect pathogenic fungus in efficient Java of the present invention, as a kind of living body biological agricultural chemicals, have the feature of the brand-new mechanism of action that is different from existing chemical insecticide, environmentally safe, noresidue, adapt to the requirement that Organic food is produced.
Description of drawings
The bacterium colony of Fig. 1 Ij01 bacterial strain on the PDA plate culture medium
The bacterium colony of Fig. 2 Ij01 bacterial strain on the Czapek-Dox plate culture medium
The spore structure of Fig. 3 Ij01 bacterial strain
The conidial fructification of Fig. 4 Ij01 bacterial strain
Embodiment
Further elaborate the present invention below in conjunction with specific embodiment.Following embodiment is a preferred implementation of the present invention, the present invention mainly sets forth the invention thought of described bacterial strain, embodiment is not related one by one in an embodiment, but therefore do not limit the present invention, other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify, should be regarded as the substitute mode of equivalence, be included in the present invention.
Embodiment 1
By gathering a kind of susceptible prodenia litura worm corpse in Guangzhou, adopt conventional current techique separation and purification bacterial strain, pathogenic mensuration, biological characteristics and production traits comparison, producing bacterial strain and determine etc. that step obtains Java Isaria Isariajavanicus (the Friedrichs ﹠amp of a strain and good stability efficient to prodenia litura; Bally) bacterial strain Ij01.
Spore suspension preparation: picking spore on the slant strains, insert in 0.05% the tween-80 solution of having sterilized, measure spore concentration with blood counting chamber, and adjust to 10
7Spore/mL.
Seed culture: the 2mL spore suspension is inoculated in the 98mL seed culture medium, under 180r/min and 26 ℃ of conditions shake-flask culture 2～3 days seed liquor.
Seed culture based component: the Czapek-Dox of 0.5% peptone: peptone 5g, NaNO
33g, K
2HPO
41g, MgSO
47H
2O 0.5g, KCl 0.5g, FeSO
47H
2O 0.01g, sucrose 30g, agar 15～20g, water 1000mL.
Fermentation culture: earlier rice is cooked, in the freshness protection package of packing into, treat that it is cooled to normal temperature after, inoculate above-mentioned seed liquor, inoculative proportion is 5～15% (V/W), is cultivating about 2 weeks the results conidium under 25～28 ℃ of conditions.
The morphological feature of Ij01 bacterial strain: as shown in Figure 1, on the PDA flat board, bacterium colony circle, white mycelium, spore layer gray purple, no coremium.As shown in Figure 2, on the Czapek-Dox flat board, colony growth is slower, subcircular, and white mycelium, spore layer color is light slightly.
Shown in accompanying drawing 3 and accompanying drawing 4, the microscopically mycelia is transparent, and is colourless, width 2.66 ± 0.18 μ m; Conidiophore is similar to mycelia, does not become coremium, long 10.38 ± 3.26 μ m, wide 2.73 ± 0.25 μ m; Conidiogenous cell bottle stalk cucurbit shape, the widest part diameter 2.75 ± 0.35 μ m, long 7.92 ± 1.11 μ m, upright then being born on the mycelia of bottle, verticillate, conidium is born on the bottle stalk, bottle stalk Pear-Shaped; Perhaps bottle is obstructing and is being born on the conidiophore, and conidiophore is born on the mycelia; The bottle stalk; Conidium is colourless, oblong, and two ends are point, long 5.47 ± 1.23 μ m, wide 2.09 ± 0.36 μ m.
Embodiment 2 utilizes this bacterial strain control prodenia litura
Being mixed with concentration with reference to the conidium of the described method results of embodiment 1 with tween-80 solution is 10
6～10
7The suspension of conidium/mL adopts the foliar spray method that this suspension is applied to the field, uses at the second instar larvae occurrence in peak period of prodenia litura.
The Ij01 bacterial strain is to the virulence of prodenia litura: under different spore concentrations, the average cumulative mortality ratio of prodenia litura 2 instar larvaes significantly increases along with the rising of concentration of treatment and the prolongation of time, see Table 1, when concentration is 50000 conidiums/mL, after processing the 6th day and the 8th day, the mortality ratio of prodenia litura promptly reached 48.33% and 51.26% respectively.
Observed data calculates medial lethal dose through logarithm-probability value analytical method, sees Table 2, and along with the prolongation of handling the back time, toxic limit medium dose progressively diminishes, LC
50Value is 10
5Conidium number/mL order of magnitude fluctuates up and down.
Between the median lethal time, when concentration is 1 * 10
5During individual spore/mL, the median lethal time is 5.0 days, and 95% fiducial interval is 4.1～6.3 days.
Table 1 Ij01 bacterial strain is to the insecticidal activity of prodenia litura 2 instar larvaes
Table 2Ij01 bacterial strain is to the LC of prodenia litura 2 instar larvaes
50And 95% fiducial interval (10
5Individual conidium/mL)
| Handled back 3 days | Handled back 5 days | Handled back 7 days | Handled back 9 days | Handled back 11 days |
| 4.39 (3.12～6.19) | 1.24 (0.95～1.61) | 1.05 (0.82～1.35) | 0.92 (0.72～1.19) | 0.82 (0.64～1.05) |
Claims (6)
1, a kind of Isaria javanicanicus mould strain, described bacterial strain Ij01 classification position belong to Mycophyta, mitospore Mycophyta, hyphomycetes, hyphomycetales, silk spore section, the mould genus of excellent spore.Be preserved in Chinese typical culture collection center (CCTCC) preservation, preservation date is on October 27th, 2008, preserving number CCTCC M 208171.
2, the application of the described Isaria javanicanicus mould strain of a kind of claim 1 is characterized in that being applied to prevent and treat prodenia litura.
3, according to the application of the described Isaria javanicanicus mould strain of claim 2, it is characterized in that described control prodenia litura is a results Isaria javanicanicus mould strain conidium, being mixed with concentration with tween 80 solution is 10
6～10
7The suspension of conidium/mL adopts the foliar spray method that suspension is applied to the field in the second instar larvae occurrence in peak period of prodenia litura.
4,, it is characterized in that described results Isaria javanicanicus mould strain conidium may further comprise the steps according to the application of the described Isaria javanicanicus mould strain of claim 3:
(1) spore suspension preparation: picking spore on the slant strains, insert in 0.05% the tween-80 solution of having sterilized, measure spore concentration with blood counting chamber, and adjust to 10
7Spore/mL;
(2) seed culture: spore suspension is inoculated in the seed culture medium, under 180r/min and 26 ℃ of conditions shake-flask culture 2～3 days seed liquor;
(3) fermentation culture: rice is cooked, in the freshness protection package of packing into, treat that it is cooled to normal temperature after, the inoculation seed liquor was cultivated for 2 weeks under 25～28 ℃ of conditions, the results conidium.
5,, it is characterized in that the Czapek-Dox that the described seed culture based component of step (2) is 0.5% peptone: peptone 5g, NaNO according to the application of the described Isaria javanicanicus mould strain of claim 4
33g, K
2HPO
41g, MgSO
47H
2O 0.5g, KCl 0.5g, FeSO
47H
2O 0.01g, sucrose 30g, agar 15～20g and water 1000mL.
6, according to the application of the described Isaria javanicanicus mould strain of claim 4, the ratio that it is characterized in that the described inoculation seed liquor of step (3) is 5～15% (V/W).
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| CN106479943A (en) * | 2017-01-03 | 2017-03-08 | 南京林业大学 | The Java Isaria BE01 of one plant height effect preventing and treating fall webworms and its application |
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| CN109479903B (en) * | 2018-11-28 | 2021-03-19 | 华南农业大学 | Isaria javanicus conidium oil suspending agent and application thereof |
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| CN110720471A (en) * | 2019-11-15 | 2020-01-24 | 韶关市华实现代农业创新研究院 | Compound pesticide of Isaria javanicus mutagenic strain IJUV-6 and thiamethoxam and application thereof |
| CN110720471B (en) * | 2019-11-15 | 2021-07-27 | 韶关市华实现代农业创新研究院 | Compound pesticide of Isaria javanicus mutagenic strain IJUV-6 and thiamethoxam and application thereof |
| CN111961598A (en) * | 2020-09-18 | 2020-11-20 | 华南农业大学 | Isaria javanicus strain and application thereof in aspect of preventing and treating phyllotreta striolata |
| CN111961598B (en) * | 2020-09-18 | 2021-11-02 | 华南农业大学 | Isaria javanicus strain and application thereof in aspect of preventing and treating phyllotreta striolata |
| CN114456949A (en) * | 2022-01-13 | 2022-05-10 | 贵州民族大学 | Beauveria bassiana JSHA-MD912 and application thereof |
| CN114456949B (en) * | 2022-01-13 | 2023-06-16 | 贵州民族大学 | Beauveria bassiana JSHA-MD912 and application thereof |
| CN114847304A (en) * | 2022-06-07 | 2022-08-05 | 江苏省农业科学院 | Method for improving drought resistance of plants by using insecticidal fungi |
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| CN101514325B (en) | 2011-09-14 |
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