CN110521736A - Application of the Metarhizium anisopliae on prevention and treatment sclerotinia sclerotiorum - Google Patents
Application of the Metarhizium anisopliae on prevention and treatment sclerotinia sclerotiorum Download PDFInfo
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- CN110521736A CN110521736A CN201910659178.XA CN201910659178A CN110521736A CN 110521736 A CN110521736 A CN 110521736A CN 201910659178 A CN201910659178 A CN 201910659178A CN 110521736 A CN110521736 A CN 110521736A
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- metarhizium anisopliae
- rape
- sclerotinia sclerotiorum
- prevention
- treatment
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Abstract
The invention belongs to technical field of plant disease biological control, specifically disclose application of the Metarhizium anisopliae IBCCM321.93 on prevention and treatment sclerotinia sclerotiorum, it is found by the applicant that Metarhizium anisopliae IBCCM321.93 has antagonism to rape endophytic bacterial, the bacterium is on PDA plate to the inhibiting rate of sclerotinite up to 45.43%, on in vitro rape leaf, playing fermentation liquid is 40.21% to the inhibiting rate of sclerotiniose, in field efficiency test, the bacterium can significantly inhibit the generation of sclerotinia sclerotiorum, control efficiency is 40.72%, and rapeseed mass of 1000 kernel can be dramatically increased, single plant rapeseed yield, it is not significant with rape difference of the procymidone after processed.
Description
Technical field
The invention belongs to the invention belongs to technical field of plant disease biological control, and in particular to Metarhizium anisopliae
Application of the IBCCM321.93 on prevention and treatment sclerotinia sclerotiorum.
Background technique
Rape is the important oil crops in China, the sclerotium as caused by sclerotinite (Sclerotinia sclerotiorum)
Disease is one of the main disease on Rape-seed production.Sclerotinite is distributed in the main growing area of rape, is typically considered economically
One of most destructive rape pathogen.Host range is wide, can infect soybean, cucumber, tomato, potato and rape etc. 500
Various plants lead to yield and quality degradation.It is reported that causing the loss of yield of rape reachable by sclerotiniose in China
10%-80%.
The prevention and treatment of sclerotiniose is the significant challenge faced in current production.Since host range is wide for sclerotinite, sclerotium survival
Time is long, causes prevention and treatment very difficult.The various Eco-friendly measures of integrated use are the optimal paths for preventing and treating sclerotiniose.
The breeding of resistant variety is the prevention and treatment most economical most effective control measure of sclerotinia sclerotiorum, but since sclerotinite can be with
It is parasitic in various plants, the rape variety that there is complete resistance to sclerotiniose is cultivated not yet in production at present.
Cultural control mainly passes through the sclerotium quantity reduced in soil or creates the environment item for being unfavorable for pathogen morbidity
Part carries out the prevention and treatment of disease, and main method has crop rotation, deep ploughing and Rational Irrigation etc..
The use of chemical bactericide is the major measure of current prevention and treatment sclerotinia sclerotiorum, but long-term use not only makes disease
Opportunistic pathogen brings very big harm to environmental condition and human health to which create resistances.
Biological control is more promising control method in current production, the antagonistic bacterium for sclerotinia sclerotiorum prevention and treatment
Mainly there are Bacillus subtillis, Pseudomonas alba, wax printing fabric and bacillus pumilus etc., biocontrol fungi mainly has
Trichoderma, shield shell are mould etc..
Metarhizium anisopliae (Metarhizium anisopliae) is a kind of insect pathogenic fungus, belongs to Ascomycota
(Ascomycota), Hypocreales (Hypocreales), Clavicipitaceae (Clavicipitaceae), Metarhizium
(Metarhizium), other than playing a significant role in control of insect, moreover it can be used to the prevention and treatment of plant disease, it is withered to cotton
Wither germ (Fusarium oxysporium), rhizoctonia cerealis (Rhizoctonia cerealis), fusarium graminearum
(F.graminearum) etc. a variety of pathogens antagonism with higher, there is no Metarhizium anisopliae can antagonism nuclear disk at present
The report of bacterium.
Summary of the invention
The object of the present invention is to provide Metarhizium anisopliae IBCCM321.93 answering in prevention and treatment sclerotinia sclerotiorum
With providing a kind of new means for the prevention and treatment of sclerotinia sclerotiorum.
In order to achieve the above object, the present invention uses following technical measures:
Application of the Metarhizium anisopliae IBCCM321.93 in prevention and treatment sclerotinia sclerotiorum, is prepared into including the use of the bacterial strain
Nuclear disk bacteria inhibitor；Or it is prepared into the preparation of prevention and treatment sclerotinia sclerotiorum；Above-described inhibitor or preparation can be with cockchafers
Sub- green muscardine fungus IBCCM321.93 is sole active ingredient, or is prepared together with other effective components.
Compared with prior art, the invention has the following advantages that
1. bacterial strain uses therefor Metarhizium anisopliae IBCCM321.93 of the present invention is a kind of important entomogenous fungi, answered extensively
For in the biological control of pest, but in control of plant disease using less, the present invention reports that it can be used for preventing and treating for the first time
Sclerotinia sclerotiorum.
2. the bacterial strain has preferable control efficiency to sclerotiniose.Have on PDA plate to nuclear fungal hyphae growth bright
Aobvious inhibiting effect, inhibiting rate 45.43%, and cause mycelia tip to attenuate, become close.In in vitro rape leaf, to sclerotium
The inhibiting rate of disease is 40.21%.Crop field efficiency test the result shows that, which can significantly inhibit the generation of sclerotiniose, control efficiency
It is 40.72%, and applied the rapeseed mass of 1000 kernel of the bacterium, single plant rapeseed yield, is all significantly increased compared with the control.
3. bacterial strain no pollution to the environment used in the present invention, be harmful to human health, production cost is low, simply easily makes
With.
Detailed description of the invention
Fig. 1 is the shadow that Metarhizium anisopliae IBCCM321.93 of the present invention grows nuclear fungal hyphae in PDA culture medium
It rings；
Figure 1A is blank control nuclear fungal hyphae growing state；
Figure 1B is the influence that Metarhizium anisopliae IBCCM321.93 grows nuclear fungal hyphae.
Fig. 2 is for Metarhizium anisopliae IBCCM321.93 of the present invention to the shadow of nuclear fungal hyphae form in PDA culture medium
It rings；
Fig. 2A is blank control nuclear fungal hyphae form；
Fig. 2 B is influence of the Metarhizium anisopliae IBCCM321.93 to nuclear fungal hyphae form.
Fig. 3 is for Metarhizium anisopliae IBCCM321.93 fermentation liquid of the present invention to the suppression of sclerotiniose in vitro rape leaf
Production is used；
Fig. 3 A is that a situation arises for disease on the in vitro rape leaf of control group；
A situation arises for disease on rape leaf in vitro after Metarhizium anisopliae IBCCM321.93 fermentation liquor treatment by Fig. 3 B.
Fig. 4 is Metarhizium anisopliae IBCCM321.93 fermentation liquor treatment of the present invention to rape mass of 1000 kernel and single plant rapeseed
The influence of weight；
Fig. 4 A is influence of each processing to rape mass of 1000 kernel；
Fig. 4 B is influence of each processing to single plant yield of rape.
Specific embodiment
Technical solution of the present invention is if not otherwise specified the ordinary skill in the art.Reagent of the present invention or
Material derives from commercial channel if not otherwise specified.
Embodiment 1:
The plate dual test of Metarhizium anisopliae and sclerotinite
It is beaten with the punch that internal diameter is 6mm and activated Metarhizium anisopliae fungus block is taken to connect in the side of PDA plate, 28
After cultivating 2d in DEG C dark culturing case, in the plate other side away from meeting activated sclerotinite A367 at plate edge same distance,
It is fungi-proofing as control, 4 repetitions of each processing not deliver a child.It Observe and measure hypha growth condition and is calculated after 20 DEG C of culture 2d
Bacteriostasis rate, test result such as Fig. 1, microscopically observation nuclear fungal hyphae metamorphosis, as a result such as Fig. 2.
The inhibitory effect (48h, 20 DEG C) that 1 Metarhizium anisopliae of table grows nuclear disk mycelia
Plate dual test the result shows that, Metarhizium anisopliae IBCCM321.93 has suppression to the growth of sclerotinite A367
Production is used, and bacteriostasis rate has reached 45.43% (table 1), the variation of microscopically observation nuclear fungal hyphae tip morphology, it is seen that its point
End attenuates, becomes close.
Embodiment 2:
Excised leaf measures Metarhizium anisopliae to the inhibiting effect of sclerotiniose
The preparation of fermenting green muscardine fungus liquid: green muscardine fungus bacteria cake (diameter 6mm) is inoculated in PDB culture medium (every bottle 100
ML), the shake culture 5d on 28 DEG C, the shaking table of 160r/min, falls bacterium with sterilized filtered through gauze under sterile conditions
Fermentation liquid is made in silk.
Fresh and healthy, the consistent rape leaf of size growth are acquired, is rinsed well with the tap water of flowing, after natural drying
It is dipped in fermentation mixed liquor (adding a drop Tween 80) about 30s, impregnated in sterile water as control, each processing is set 4 times
It repeats.It is put into the blotting paper of wetting in porcelain dish to keep certain humidity, and cover petiole with the blotting paper of wetting, covers sealing
Film is placed in 28 DEG C of constant temperature illumination boxs handle for 24 hours after, activated nuclear disk is connect at the same position of each blade
Bacterium Tip Splitting block, fungus block size are 6mm, are put into 20 DEG C of illumination boxs and cultivate, incidence are observed after 48h, and measure
Its lesion diameter calculates bacteriostasis rate, test result such as Fig. 3.
Inhibiting rate (48h, 22 DEG C) of the Metarhizium anisopliae to sclerotiniose on 2 excised leaf of table
After the in vitro rape leaf of Metarhizium anisopliae fermentation liquor treatment, leaf spot lesion diameter can be observed less than control group
Lesion diameter, and in 5% level, and has compareed significant difference, reaches 40.21% (table 2) to the inhibiting rate of sclerotiniose.
Embodiment 3:
The test of Metarhizium anisopliae control in field sclerotinia sclerotiorum
Being inoculated with diameter is the Metarhizium anisopliae fungus block of 6mm in 100mL PDB culture solution, 160r/min, 28 DEG C of cultures
Metarhizium anisopliae fermentation liquid is made with two layers of filtered through gauze mycelia in 5d, adjusts spore concentration to 107CFU/mL, dilution 10
Times, every processing sprays 2L, plot area 10m2, the another procymidone that sets of test is as processing, 500 times of dilutions, dosage 30g/
Mu, as control, each processing is repeated 4 times for clear water processing.
Spraying treatment is carried out in rape flowering, activated sclerotinite A367 fungus block is inoculated with afterwards for 24 hours, is twined with sealed membrane
Around holding humidity.Scab length is measured in stable disease phase (before rape harvest), and carries out significance difference analysis.In rape
After harvest, each processing rape mass of 1000 kernel and single plant rapeseed weight are measured, as a result as (321.93 be chafer to Fig. 4 in Fig. 4
Green muscardine fungus IBCCM321.93).
Field control effect (stable disease phase) of 3 different disposal of table to sclerotinia sclerotiorum
Different disposal shows the field trial of sclerotinia sclerotiorum: rotten in stable disease phase (before rape harvest) investigation
The preventive effect of mould benefit is relatively high, is 48.05% (table 3), and Metarhizium anisopliae IBCCM321.93 processing is 40.72%, and two
Person with compare with significant difference, but therebetween difference it is not significant.Mass of 1000 kernel and single plant rapeseed produce after investigation harvest
Amount, as shown in Figure 4, the rape mass of 1000 kernel of procymidone processing and Weight per plant highest, Metarhizium anisopliae processing are only second to procymidone
Processing, with compareed significant difference.
Claims (3)
1. application of the Metarhizium anisopliae IBCCM321.93 in preparation prevention and treatment sclerotinia sclerotiorum preparation.
2. Metarhizium anisopliae IBCCM321.93 is preparing the application in nuclear disk bacteria inhibitor.
3. Metarhizium anisopliae IBCCM321.93 is promoting the application in growth of rape.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112425609A (en) * | 2020-12-18 | 2021-03-02 | 重庆谷百奥生物研究院有限公司 | Metarhizium anisopliae preparation for promoting growth of banana plants and preventing and treating banana wilt |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112425609A (en) * | 2020-12-18 | 2021-03-02 | 重庆谷百奥生物研究院有限公司 | Metarhizium anisopliae preparation for promoting growth of banana plants and preventing and treating banana wilt |
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