CN107047620B - A kind of gray mold control method - Google Patents
A kind of gray mold control method Download PDFInfo
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- CN107047620B CN107047620B CN201710277591.0A CN201710277591A CN107047620B CN 107047620 B CN107047620 B CN 107047620B CN 201710277591 A CN201710277591 A CN 201710277591A CN 107047620 B CN107047620 B CN 107047620B
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Abstract
The invention discloses a kind of gray mold control method, photosynthetic bacteria causes pathogen spore that can not form the prevention and treatment realized to gray mold by inhibiting the mycelia growth of gray mold pathogen, carrying out teratogenesis to gray mold pathogen mycelium；Photosynthetic bacteria is seeded in solid medium when concrete application, under the intensity of illumination of 7500lx, 7 ~ 14d of constant temperature illumination cultivation in 30 DEG C of illumination boxs, red or maroon single colonie is grown in observation culture medium, it is linked into photosynthetic bacteria liquid culture medium with oese picking single colonie, expand culture in the illumination insulating box that intensity of illumination is 7500lx, temperature is 30 DEG C and obtains Fermentation by Photosynthetic Bacteria liquid to 14d or so, it is transferred in the sprayer of sterilizing, is stored in spare in 4 DEG C of refrigerator under aseptic condition.The present invention provides a kind of effective biological control method of gray mold, solves the problems, such as that present gray mold control efficiency is unstable, a large amount of noxious materials can be remained in fruits and vegetables.
Description
Technical field
The present invention relates to fruit and vegetable disease prevention techniques fields, more particularly, to a kind of gray mold control method.
Background technique
Gray mold is that open country, protecting field crop be common and a kind of fungal disease of more difficult prevention and treatment, belongs to low temperature and high relative humidity
Type disease, growth of pathogenic bacteria temperature are 20~30 DEG C, and therefore, temperature is disease when 20~25 DEG C, humidity continue 90% or more
The evil high-incidence season.Gray mold occurs in twenties kinds of crops such as strawberry, cucumber, eggplant, tomato, capsicum, grape；Gray mold is to work
Stem, leaf, flower, fruit and the carpopodium of object can cause damages；Gray mold is particularly acute the harm of fruit.
Gray mold be it is microbial by the Botrytis cinerea in Deuteromycotina hyphomycetales Botrytis fungi, which is not only
Crop is common and a kind of fungal disease of more difficult prevention and treatment, and the disease is a kind of typical aeroborne disease, can be with empty gas and water
Stream and farming operation propagate, usually have an infection center then radiate out, prevented and treated not in time after generation, sprawling compared with
Fastly；Gray mold sick seedling color is shallow, and blade, petiole morbidity are in canescence, and water stain shape, organization softening are to rotting, and surface has when high humidity
Mould ash；Young stem is mostly come into being irregular water logging spot in petiole base, softens rot quickly, hangs contracting or folding falls, last sick seedling is rotted
It is withered；When fruit is caught an illness, Chinese olive is aggrieved serious, and remaining column cap or petal are dipped more, backward fruit extension, causes the pericarp to be in
Canescence, and have the mould layer of thick grey, it is in water corruption shape；Gray mold is not only easy to infect, can also endanger from the seedling of crop
Evil arrives the fruit of crop；And the difficulty that gray mold pathogen is thoroughly eliminated is very big；Therefore, as long as once breaking out ash in crop
Mildew not only results in the death of seedling, also will affect the yield and quality of fruits and vegetables；Greatly economic damage can be caused to crop
It loses.
Therefore, now during long-term cropping, the prevention and treatment of gray mold is just particularly important.Chemical prevention at present
It is that prevention and treatment gray mold is the most universal and extensive method；Chemical prevention is to be prevented and treated by using chemical agent gray mold
Method；Currently used chemical agent is mainly that aminopyrimidine fungicide, pyroles fungicide and acid amide fungicides etc. are several
Class fungicide.The major advantage of chemical prevention can in lethality significant effect, the rapid and stabilization for gray mold pathogenic bacteria
Repeatability is strong, preventing and controlling are easy to operate, workload is small；But the use that chemical agent is long-term, so that the pathogenic bacteria of gray mold
Drug resistance constantly enhance, the strong bacterial strain of drug resistance increases, and the effect for actually resulting in chemical prevention is greatly reduced；In addition,
Above-mentioned a few class fungicide are all hypertoxic type drugs, and for chemical agent after sterilization, the residual drawback of chemical agent is more and more convex
It is aobvious；It on the one hand is that chemical agent easily largely remains in crop, remaining Toxic in the fruits and vegetables for causing people finally to eat
Matter is exceeded, and long-term consumption causes greatly to injure to the body of people；It can not meet people and yearned for safety, low toxicity effect fruit
The purchasing demand of vegetable；On the other hand it is that a large amount of chemicals can not decompose in the soil, the quality of soil is caused to change, gives ring
Border brings huge harmfulness pressure；Therefore, the advantage of the chemical prevention of gray mold is constantly reduced.
Gradually begin one's study using the means of biological control now and prevent gray mold, have studied at present trichoderma, saccharomycete,
Bacillus has the effect of the biological control of gray mold certain；But photosynthetic bacteria is in the biological control of gray mold at present
Application have not been reported.
Therefore, the photosynthetic biological control for being carefully used for gray mold is realized aobvious to gray mold pathogen lethality effect now
It writes, rapidly, effect stability and nontoxic to gray mold prevention and treatment process avoids remaining in crop during gray mold prevents and treats big
Amount noxious material is the direction that we study.
Summary of the invention
In view of the above shortcomings of the prior art, the technical problems to be solved by the present invention are: how to provide a kind of gray mold
Control method is realized to gray mold pathogen lethality significant effect, rapid, effect stability, and realizes the anti-of gray mold
It is nontoxic to control process, avoids remaining a large amount of noxious material on crop, meets the needs of people are to less toxic fruits and vegetables.
In order to solve the above-mentioned technical problem, present invention employs the following technical solutions:
A kind of gray mold control method, which is characterized in that by the way of spraying photosynthetic bacteria, inhibited by photosynthetic bacteria
The mycelia of gray mold pathogen grows and carrying out teratogenesis to gray mold pathogen mycelium causes pathogen spore can not shape
At to realize the prevention and treatment to gray mold.
Further, the photosynthetic bacteria is the one of excrement red pseudomonas, Rhodospirillum rubrum or Rhodopseudomonas palustris
Kind.
Further, the photosynthetic bacteria is Rhodopseudomonas palustris；Rhodopseudomonas palustris is not only to gray mold cause of disease
The growth of bacterium mycelia has good inhibiting effect, and has teratogenesis to gray mold pathogen mycelium, so that grey mold
Sick pathogen mycelium can not form spore；So that the 2nd day bacteriostasis rate just reaches after spraying Rhodopseudomonas palustris fermentation liquid
71.17%, and Agro-chemicals control gray mold is sprayed, the bacteriostasis rate 71.17% reached is identical within the 2nd day.After on day 4, natural pond
Damp red pseudomonas is stable 87.29% always to the bacteriostasis rate of gray mold pathogen；And chemical agent is to gray mold pathogen
Bacteriostasis rate reach up to 81.22% on day 4, since the 4th day, bacteriostasis rate dropped to 67.30%.Therefore, marsh is red
Pseudomonad is more significant to the control efficiency of gray mold, rapid and control efficiency is more stable.
Further, the mode for spraying photosynthetic bacteria, which refers to, is prepared into fermentation liquid for photosynthetic bacteria, periodically will preparation
Good light and ferment product uniformly sprays on crop, and -72h for 24 hours can be spaced on crop and periodically sprays light on crop
Ferment product is closed, it is best to the control efficiency of gray mold.
Further, the Fermentation by Photosynthetic Bacteria liquid is prepared in the following way, and photosynthetic bacteria is seeded in solid medium
In, under the intensity of illumination of 7500lx, constant temperature 7~14d of illumination cultivation in 30 DEG C of illumination boxs, observe grown in culture medium it is red
Color or maroon single colonie, are linked into photosynthetic bacteria liquid culture medium with oese picking single colonie, are in intensity of illumination
7500lx, temperature are to expand culture in 30 DEG C of illumination insulating box to obtain Fermentation by Photosynthetic Bacteria liquid to 14d or so, under aseptic condition
Fermentation by Photosynthetic Bacteria liquid is transferred in the sprayer of sterilizing, is stored in spare in 4 DEG C of refrigerator.
Further, the gray mold refers to the gray mold occurred on strawberry fruit.
Compared with prior art, the invention has the following advantages that
(1) present invention grows tool by mycelia of the experimental verification photosynthetic bacteria to the pathogen the pathogen of Botrytis cinerea of gray mold
There are inhibiting effect, the spore that there is teratogenesis to the mycelium of the pathogen of Botrytis cinerea and make the pathogen of Botrytis cinerea that can not be formed
Effect；The results show, photosynthetic bacteria have preventive and therapeutic effect to gray mold, provide a kind of effective Biological Control of Gray Mold
Method；It is nontoxic to realize gray mold prevention and treatment process, avoids remaining a large amount of noxious materials in fruits and vegetables, meets people to less toxic fruit
The demand of vegetable.
(2) present invention passes through experimental verification excrement red pseudomonas 1.2176, the red vacation of Rhodospirillum rubrum 1.5005 or marsh
Monad 1.8929 has bacteriostasis, the results show, the red false unit cell of excrement to the pathogen the pathogen of Botrytis cinerea of gray mold
The highest bacteriostasis rate of bacterium 1.2176 can achieve 70.42%, and the highest bacteriostasis rate of Rhodospirillum rubrum 1.5005 can achieve
61.36%, the highest bacteriostasis rate of Rhodopseudomonas palustris 1.8929 can achieve 87.29%；It is not only right to demonstrate photosynthetic bacteria
Gray mold pathogen the pathogen of Botrytis cinerea has bacteriostasis, and antibacterial efficiency reaches 61.36% or more, and highest bacteriostasis rate can
To reach 87.29%, illustrate that photosynthetic bacteria has good effect to the prevention and treatment of gray mold.
(3) present invention is experimentally confirmed, the Rhodopseudomonas palustris 1.8929 second days bacteriostasis rates to the pathogen of Botrytis cinerea
Just reach 71.17%, the 4th day bacteriostasis rate reaches highest, bacteriostasis rate 87.29%, the 5th day, the 6th day bacteriostasis rate it is steady
It is scheduled on 87.29%；The highest bacteriostasis rate of existing acid amide fungicides is 81.22%, but is occurred in the 4th day bacteriostasis rate bright
Aobvious decline；Therefore, it was demonstrated that Rhodopseudomonas palustris 1.8929 is to prevention and treatment efficiency of the chemical agent to the pathogen of Botrytis cinerea more significant,
It is stronger to the lethality of the pathogen of Botrytis cinerea, prevent effect more stable, and compared with chemical prevention and control method, using photosynthetic bacteria
The process prevented and treated gray mold is nontoxic, not only avoids and causes damages to worker's body, also fundamentally avoids fruits and vegetables
The case where interior residual a large amount of noxious materials, occurs, and the production of less toxic fruits and vegetables may be implemented, and meets people to the need of less toxic fruits and vegetables
It asks.
(4) present invention is experimentally confirmed, the pathogen of Botrytis cinerea bacterium after 1.2176 fermentation liquor treatment of excrement red pseudomonas
Filament is more assembled, and numbers of branches also increases, and buckling phenomena occur in a small number of mycelia, and the sporogenesis quantity of the pathogen of Botrytis cinerea subtracts
It is few；The branch of Rhodospirillum rubrum 1.5005 increases, and the sporogenesis of the pathogen of Botrytis cinerea is substantially without influence；Rhodopseudomonas palustris
The pathogen of Botrytis cinerea mycelium tubbiness, quantity after 1.8929 fermentation liquor treatments are reduced, branch increases, mycelia buckling phenomena is tight
Weight, and spore can not be formed；Therefore, the present invention can not only inhibit the normal growth of the pathogen of Botrytis cinerea, additionally it is possible to so that grey Portugal
The mycelial growthform of grape spore bacterium, has an effect on the formation of spore, realizes the prevention and treatment for fundamentally solving the problems, such as gray mold.
In conclusion the present invention can fundamentally prevent and treat gray mold, so that the control efficiency to gray mold is aobvious
It writes, control efficiency is stable, entire gray mold prevention and treatment process is nontoxic and the advantages such as control cost is low.
Detailed description of the invention
Fig. 1 is the life that the pathogen of Botrytis cinerea in the PDA culture medium for picking sterile water filter paper is placed in the embodiment of the present invention 3
Long pictorial diagram.
Fig. 2 is that the PDA culture medium for picking 1.5005 fermentation liquid filter paper of Rhodospirillum rubrum is placed in the embodiment of the present invention 3
The growth pictorial diagram of middle the pathogen of Botrytis cinerea.
Fig. 3 is that the PDA training for picking 1.8929 fermentation liquid filter paper of Rhodopseudomonas palustris is placed in the embodiment of the present invention 3
Support the growth pictorial diagram of the pathogen of Botrytis cinerea in base.
Fig. 4 is that the PDA culture for picking 1.2176 fermentation liquid filter paper of excrement red pseudomonas is placed in the embodiment of the present invention 3
The growth pictorial diagram of the pathogen of Botrytis cinerea in base.
Fig. 5 is the pathogen of Botrytis cinerea bacteria cake 4d in embodiment 4 in the mixed culture medium made of sterile water and PDA culture medium
Growth pictorial diagram.
Fig. 6 is the pathogen of Botrytis cinerea bacteria cake 4d in embodiment 4 in 1.5005 fermentation liquid of Rhodospirillum rubrum and PDA culture medium system
At mixed culture medium in growth pictorial diagram.
Fig. 7 is that the pathogen of Botrytis cinerea bacteria cake 4d is cultivated in 1.8929 fermentation liquid of Rhodopseudomonas palustris and PDA in embodiment 4
Growth pictorial diagram in mixed culture medium made of base.
Fig. 8 is the pathogen of Botrytis cinerea bacteria cake 4d in embodiment 4 in 1.2176 fermentation liquid of excrement red pseudomonas and PDA culture medium
Growth pictorial diagram in manufactured mixed culture medium.
Fig. 9 is the pathogen of Botrytis cinerea bacteria cake 4d training with PDA in the phonetic mould solution of 1000 times of dilution amides in embodiment 4
Support the growth pictorial diagram in mixed culture medium made of base.
Figure 10 is that the pathogen of Botrytis cinerea bacteria cake 4d is trained in the phonetic mould solution of 1500 times of dilution amides and PDA in embodiment 4
Support the growth pictorial diagram in mixed culture medium made of base.
Figure 11 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Fig. 5 under an electron microscope.
Figure 12 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Fig. 6 under an electron microscope.
Figure 13 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Fig. 7 under an electron microscope.
Figure 14 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Fig. 8 under an electron microscope.
Figure 15 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Fig. 9 under an electron microscope.
Figure 16 is the growth structure schematic diagram of the mycelia for the pathogen of Botrytis cinerea cultivated in Figure 10 under an electron microscope.
Figure 17 is that the pathogen of Botrytis cinerea cultivated in Fig. 5 generates the growth structure schematic diagram of spore under an electron microscope.
Figure 18 is that the pathogen of Botrytis cinerea cultivated in Fig. 6 generates the growth structure schematic diagram of spore under an electron microscope.
Figure 19 is that the pathogen of Botrytis cinerea cultivated in Fig. 7 generates the growth structure schematic diagram of spore under an electron microscope.
Figure 20 is that the pathogen of Botrytis cinerea cultivated in Fig. 8 generates the growth structure schematic diagram of spore under an electron microscope.
Figure 21 is that the pathogen of Botrytis cinerea cultivated in Fig. 9 generates the growth structure schematic diagram of spore under an electron microscope.
Figure 22 is that the pathogen of Botrytis cinerea cultivated in Figure 10 generates the growth structure schematic diagram of spore under an electron microscope.
Specific embodiment
Invention is further described in detail with Figure of description combined with specific embodiments below.The implementation case with
Implemented under premised on the technology of the present invention, provides detailed embodiment and specific operating process now to illustrate tool of the present invention
It is creative, but protection scope of the present invention embodiment not limited to the following.
1 Photosynthetic bacterium strain of embodiment and culture
Excrement red pseudomonas, number: 1.2176, it buys from China General Microbiological culture presevation administrative center.The red vacation of excrement
The formula of unit cell bacterium culture medium (being provided by China General Microbiological culture presevation administrative center) is as follows:
Excrement red pseudomonas culture medium (ingredient and final concentration): potassium dihydrogen phosphate 1.0g, calcium chloride 0.1g, sodium bicarbonate
3.0g, sodium acetate 1.0g, magnesium chloride 0.5g, ammonium chloride 1.0g, sodium chloride 1.0g, sodium succinate 1.0g, yeast extract 0.5g,
Peptone 0.5g, trace element solution 1.0ml, vitamin solution 1.0ml, distilled water are settled to 1000ml, and pH is adjusted to 6.8.
Rhodospirillum rubrum, number: 1.5005, it buys from China General Microbiological culture presevation administrative center.Dark red red spiral shell
The formula of bacterium culture medium (being provided by China General Microbiological culture presevation administrative center) is as follows:
Rhodospirillum rubrum culture medium (ingredient and final concentration): yeast extract 2.00g, trypticase peptone 2.00g, phosphorus
Acid dihydride potassium 0.30g, dipotassium hydrogen phosphate 0.30g, ammonium sulfate 0.30g, sodium chloride 0.60g, bitter salt 0.13g, seven water
Close ferrous sulfate 2.00mg, CALCIUM CHLORIDE DIHYDRATE 8.00mg, sodium acetate 2.50g, sodium formate 2.50g, sodium bicarbonate 5.00g, L-
Cysteine 0.50g, Sodium Sulphate Nine Hydroxide 0.50g, resazurin 1.00mg, Trace salts solution 10.00ml, vitamin solution
10.00ml, distilled water are settled to 1000ml, natural pH.
Rhodopseudomonas palustris, number: 1.8929, it buys from China General Microbiological culture presevation administrative center.Marsh
The formula of red pseudomonas culture medium (being provided by China General Microbiological culture presevation administrative center) is as follows:
Rhodopseudomonas palustris culture medium (ingredient and final concentration): glucose 0.5g, soluble starch 0.5g show peptone
0.5g, yeast extract 0.5g, sour hydrolyzed casein 0.5g, Sodium Pyruvate 0.3g, dipotassium hydrogen phosphate 0.3g, bitter salt
0.05g, agar 15g, distilled water are settled to 1000ml, and pH is adjusted to 7.2.
Wherein, trace element solution: Iron dichloride tetrahydrate 1.8g, cobalt chloride hexahydrate 0.25g, Nickel dichloride hexahydrate
0.01g, Copper dichloride dihydrate 0.01g, four chloride hydrate manganese 0.70g, zinc chloride 0.1g, boric acid 0.5g, two molybdic acid hydrate sodium
0.03g, five hydration sodium selenite 0.01g, distilled water are settled to 1000ml.
Vitamin solution: biotin 0.1g, niacin 0.35g, thiamine hydrochloride 0.3g, p-aminobenzoic acid 0.2g, hydrochloric acid
Pyridoxamine 0.1g, calcium pantothenate 0.1g, vitamin B12 0.05g, distilled water are settled to 1000ml.
The preparation of 2 Fermentation by Photosynthetic Bacteria liquid of embodiment
Excrement red pseudomonas 1.2176, Rhodospirillum rubrum 1.5005 and Rhodopseudomonas palustris 1.8929 are seeded in respectively
In three solid mediums (ingredient of film solid media), by three film solid medias under the intensity of illumination of 7500lx, 30 DEG C
7~14d of constant temperature illumination cultivation in illumination box observes the red or maroon single colonie grown in three culture mediums respectively,
The single colonie in three solid mediums is picked them separately with oese respectively correspond be linked into excrement red pseudomonas culture medium, dark red
In red spirillum culture medium, Rhodopseudomonas palustris culture medium, in the illumination insulating box that intensity of illumination is 7500lx, temperature is 30 DEG C
In continue to expand culture to 14d or so to respectively obtain excrement red pseudomonas fermentation liquid, Rhodospirillum rubrum fermentation liquid and marsh red
Pseudomonas fermentation liquid, aseptically respectively by excrement red pseudomonas fermentation liquid, Rhodospirillum rubrum fermentation liquid and marsh
Rhodopseudomonas fermentation liquid is transferred in the sprayer of sterilizing, is stored in spare in 4 DEG C of refrigerator.
Preliminary Determination of the 3 Fermentation by Photosynthetic Bacteria liquid of embodiment to gray mold pathogen the pathogen of Botrytis cinerea antagonistic activity
A, botrytis cinerea pathogen: the pathogen of Botrytis cinerea (separation of forestry institute, Beihua University saves).
The activation culture of the pathogen of Botrytis cinerea and the preparation of spore suspension.The Botrytis cinerea for being stored in slant medium is turned
Enter in PDA culture medium, the Botrytis cinerea of activation is seeded in cultivate 5~7 days on culture medium and covers with culture by culture 7~14d activation
After in base, under 25 DEG C of constant temperatures after lighting process 48h, sterile water is added, enables sterile water just by the table of bacterium colony
Face is covered；24~48h is continued with to a large amount of sporangiums are generated in equal conditions, discharges spore, spore suspension is made.
Wherein PDA culture medium (ingredient and final concentration): peeled potatoes 200g, boiling water bath 20min, with double gauze mistake
Filter, glucose 20.0g, agar powder 18g-20g after dissolving, are settled to 1000ml, natural pH with distilled water.
B, the preparation of the phonetic mould medicament of amide.It is phonetic mould using technology and application method guidance according to amide, it prepares respectively
The phonetic mould solution of the amide of 1000 times and 1500 times dilutions.
C, Preliminary Determination of the photosynthetic bacteria to the pathogen of Botrytis cinerea antagonistic activity.Using filter paper enzyme, to be picked on filter paper
It is blank control group CK in the PDA culture medium for being coated with Botrytis cinerea bacterium suspension that sterile water, which is placed on, will pick class depth respectively
The circle of red 1.5005 fermentation liquid of spirillum, 1.2176 fermentation liquid of 1.8929 fermentation liquid of Rhodopseudomonas palustris and excrement red pseudomonas
Hole filter paper is individually positioned in No. 1 PDA culture medium for being coated with Botrytis cinerea bacterium suspension, No. 2 PDA culture mediums and No. 3 PDA trainings
It supports on base, carries out four repetitions and test.
It is observed within every 24 hours after inoculation, obtains Fig. 1-4, Fig. 1-4 is the 4th day photo clapped after inoculation, from Fig. 1-4
As can be seen that the pathogen of Botrytis cinerea normal growth in PDA culture medium, bacterium colony growth is intensive, and bacterium colony is raw in discovery blank control group CK
Long color is deeper；In No. 1 PDA culture medium, the pathogen of Botrytis cinerea growth is more normal, and the concentration of bacterium colony growth compares blank control
The concentration of group CK is low, and colony colour is shallower；In No. 2 PDA culture mediums, in the filter paper of inoculation Rhodopseudomonas palustris fermentation liquid
Around piece, a transparent inhibition zone without any strain growth is formed, inhibition zone boundary is obvious, and inhibition zone is wide and transparent；No. 3
In PDA culture medium, around the filter paper of inoculation excrement red pseudomonas fermentation liquid, minority forms inhibition zone, and majority does not form suppression
Bacterium circle, and the inhibition zone formed is more relatively narrow than the inhibition zone formed in No. 2 PDA culture mediums, and inhibition zone boundary is unobvious.
The influence that 4 Fermentation by Photosynthetic Bacteria liquid of embodiment grows gray mold pathogen the pathogen of Botrytis cinerea mycelia is by dark red red spiral shell
1.5005 fermentation liquid of bacterium, 1.8929 fermentation liquid of Rhodopseudomonas palustris and 1.2176 fermentation liquid of excrement red pseudomonas respectively with dissolve
PDA culture medium afterwards is mixed and made into No. 1, No. 2, No. 3 mixing PDA culture mediums according to the ratio of volume 1:1, mixes PDA culture medium
Total volume be 40ml；The sterile water and PDA culture medium that equivalent is added are mixed and made into negative control group CK, equivalent 1000 is added
Again, the phonetic mould solution of amide of 1500 times of dilutions is mixed and made into positive controls CK with PDA culture medium respectively1And CK2。
The bacteria cake that diameter is 6mm is made in the pathogen of Botrytis cinerea for cultivating 6d, in No. 1, No. 2, No. 3 mixing PDA culture mediums, yin
Property control group CK, positive controls CK1And CK2The middle the pathogen of Botrytis cinerea bacteria cake for being inoculated with 3 production respectively, three bacteria cakes are in triangle
Shape is inoculated in the medium；Then whole culture mediums are placed in dark culturing in 25 DEG C of constant incubator, carry out four repetitions
Experiment.4d observation the pathogen of Botrytis cinerea hypha growth condition obtains Fig. 5-10, and records colony diameter, and calculate Mycelial growth
Rate:
Inhibiting rate (%)=(control colony diameter-processing colony diameter)/control colony diameter × 100
No. 1, No. 2, No. 3 PDA culture mediums, the grey grape in positive controls CK1 and CK2 are obtained by above-mentioned calculation formula
The percentage mycelial inhibition of spore bacterium obtains table 1.
By observing above-mentioned experiment: the pathogen of Botrytis cinerea normal growth in negative control group CK, second day starts
Existing white hypha, at the 4th day, mycelia is covered with entire culture medium；Rhodopseudomonas palustris 1.8929 has obviously the pathogen of Botrytis cinerea
Inhibiting effect, from after generating inhibition zone, the pathogen of Botrytis cinerea mycelia does not continue to spread, and bacteriostasis rate constantly rises, and 4d starts to reach
Up to 87.29%；Inhibition zone is generated in the feeding base of excrement red pseudomonas 1.2176, the pathogen of Botrytis cinerea mycelia does not continue to expand
It dissipates, and is not generated in the culture dish of inhibition zone for most of, the pathogen of Botrytis cinerea mycelia then continues to spread, it is seen that the red false unit cell of excrement
Bacterium 1.2176 is not sufficiently stable the pathogen of Botrytis cinerea inhibiting effect, and it is 70.72% that inhibiting rate, which reaches peak in 3d,；It is first in inoculation
Phase, Rhodospirillum rubrum 1.5005 have inhibiting effect to the pathogen of Botrytis cinerea, and it is 61.36% that inhibiting rate, which reaches peak in 3d, is connect
The kind later period, the growth of the pathogen of Botrytis cinerea mycelia was almost similar with blank control group without obvious inhibiting effect；1000 times of equivalent of addition is dilute
The PDA culture medium of the phonetic mould solution of degree of releasing amide has an obvious inhibiting effect to the pathogen of Botrytis cinerea, and when 3d starts grey grape occur
Spore bacterium mycelia, and slow growth, inhibiting rate reach 81.22%, start the pathogen of Botrytis cinerea mycelia in 5d and no longer grow；Be added etc.
Measuring the PDA culture medium of the phonetic mould solution of 1500 times of dilution amides has a good inhibiting effect to the pathogen of Botrytis cinerea, and when 3d opens
There is the pathogen of Botrytis cinerea mycelia, and slow growth in beginning, and inhibiting rate peak can reach 74.72%.
5 Fermentation by Photosynthetic Bacteria liquid of embodiment is to the mycelial teratogenesis of gray mold pathogen the pathogen of Botrytis cinerea and spore shape
At influence
By No. 1 in embodiment 4, No. 2, in No. 3 mixing PDA culture mediums, negative control group CK, positive controls CK1And CK2
The pathogen of Botrytis cinerea mycelia of culture is observed and is compared by electron microscope to obtain Figure 11-16 and Figure 17-22.
From in Figure 11-16 and Figure 17-22 as can be seen that negative control group CK in the pathogen of Botrytis cinerea mycelia robust growth,
Normal growth, plasm is uniform, diaphragm is obvious, the spore quantity of sprouting is more；The pathogen of Botrytis cinerea in No. 2 mixing PDA culture mediums
The branch of mycelia increases, and the spore quantity of sprouting is more；Botrytis mycelium tubbiness, quantity subtract in No. 3 mixing PDA culture mediums
Less, branch increase, mycelia buckling phenomena it is serious, without sporogenesis；Botrytis is more assembled in No. 4 conjunction PDA culture mediums, point
Number increases, and buckling phenomena occur in a small number of mycelia, and spore germination quantity is more；Positive controls CK1And CK2In Botrytis cinerea
The mycelial branch of bacterium, which increased significantly, be broken serious, mycelium attenuates, and has a certain number of spore germinations.
It can analyze and obtain from the data in above-mentioned figure and in table 1: (1) excrement red pseudomonas 1.2176, dark red red spiral shell
Bacterium 1.5005, these three photosynthetic bacterias have certain bacteriostasis to the pathogen the pathogen of Botrytis cinerea of gray mold；Therefore, explanation
Photosynthetic bacteria has certain preventive and therapeutic effect to the gray mold occurred in fruits and vegetables；(2) as can be seen that due to marsh from figure and table
Red pseudomonas 1.8929 has teratogenesis to the mycelium of the pathogen of Botrytis cinerea and spore can not be formed；So that marsh is red
Pseudomonad 1.8929 is still stable effective at six days or more to the fungistatic effect of gray mold pathogen the pathogen of Botrytis cinerea, and the
The bacteriostasis rate of two days Rhodopseudomonas palustris 1.8929 can achieve 71.17%, the fungistatic effect with first day of chemical agent
It is identical；In addition, the bacteriostasis rate of the 4th day Rhodopseudomonas palustris 1.8929 reaches maximum 87.29%, and bacteriostasis rate later
It is always held at 87.29%；This just illustrates that Rhodopseudomonas palustris 1.8929 is strong to the lethality of the pathogen of Botrytis cinerea, fungistatic effect
Rapidly, and fungistatic effect is stablized, and antibacterial effective time is long；(3) it can analyze to obtain from table, chemical agent is for antibacterial
In the process, in the bacteriostasis rate highest in third day, but the highest bacteriostasis rate of chemical agent is again below Rhodopseudomonas palustris
1.8929 to the bacteriostasis rate of the pathogen of Botrytis cinerea；Illustrate that Rhodopseudomonas palustris 1.8929 compares chemical drugs to the control efficiency of gray mold
Agent is good to the control efficiency of gray mold；And at the 4th day, CK1And CK2In bacteriostasis rate the case where occurring as soon as decline, finally
CK1And CK2Bacteriostasis rate stablize in 67.30% and 55.72%, and the bacteriostasis rate of Rhodopseudomonas palustris 1.8929 is from third day
It is afterwards always 87.29%；Illustrate the more longlasting stabilization of the fungistatic effect of Rhodopseudomonas palustris 1.8929, and gray mold is prevented
It controls more efficient.
6 Rhodopseudomonas palustris 1.8929 of embodiment evaluates the field control effect of grey mould fruit rot of strawberry
(1) test method
Perpetual strawberry is by sowing, temporary planting, after field planting 30 days, as the test field of field efficacy measurement.By experimental plot
Be divided into 15 sample prescriptions, each sample prescription is 4 meters long × wide 1.5 meters of rectangle, the 48h before test process works, each sample prescription according to
Equidistant sampling method takes 10 points, and every is investigated three plants, the hair the number of sick fruits of all strawberry fruits of investigation records and corresponding tight
Weight degree, the disease incidence and disease index of each sample prescription are calculated according to the grade scale of grey mould fruit rot of strawberry.Each sample prescription later
The pathogen of Botrytis cinerea is sprayed, then does following five groups of processing:
The processing of A group is that spray-on process sprays 1.8929 bacterial strain fermentation liquor 10ml/m of Rhodopseudomonas palustris2；
The processing of B group is that spray-on process sprays 1.8929 bacterial strain fermentation liquor 7.5ml/m of Rhodopseudomonas palustris2；
The processing of C group is that spray-on process sprays 1.8929 bacterial strain fermentation liquor 5ml/m of Rhodopseudomonas palustris2；
The phonetic mould wettable dry powder doses of amide that D group sprays 1000 times of dilution are positive controls CK+；
The processing of E group clear water is negative control group CK-.
Each group treatment fluid is sprayed application on strawberry fruit with sprayer, three sample prescriptions are randomly selected in every group of processing, as three
It is secondary to repeat to test.
(2) result is recorded and calculated
Each group treatment fluid, and investigation records the number of sick fruits and its severity are sprayed every 48h, investigates four (each samples altogether
Side takes 10 points, three plants of strawberry fruits of every statistics according to equidistant sampling method), calculate the disease incidence recorded each time, the state of an illness refers to
Several and control efficiency.During entire test in addition to not spraying any fungicide, all progress as usual of other field management.
The grade scale (as unit of fruit) of grey mould fruit rot of strawberry:
0 grade: fruit disease-free spot；
1 grade: there is fragmentary small scab in fruit；
2 grades: lesion area account for the gross area 15% and it is following；
3 grades: lesion area accounts for the 16%-30% of total area；
4 grades: lesion area accounts for the 30%-50% of total area；
5 grades: lesion area accounts for 50% of total area or more；
Disease incidence, disease index and control efficiency calculation formula are as follows:
Disease incidence: referring in investigated data, and morbidity strawberry fruit number accounts for the percentage of investigation strawberry fruit sum.
Disease incidence (%)=(investigation the number of sick fruits)/(investigation the total number of fruits) × 100
Disease index: referring to the weight occurred first according to disease, carries out classification numeration investigation, then according to being recorded
The disease fruit numerical value of each disease grade is calculated according to the following formula.By the calculating of disease index, the generality of disease can be taken into account
And severity.
Disease index (%)=∑ [sick grade fruit number × grade represents numerical value]/(investigation the total number of fruits × morbidity is highest level
Represent numerical value) × 100
In formula, CK0For the disease index of control prevention and treatment front vane, CKnFor the disease index of control prevention and treatment rear blade；Pt0For
Chemicals treatment prevents and treats the disease index of front vane, and Pt is the disease index that chemicals treatment prevents and treats rear blade.
Inoculation processing is carried out to Field strawberries according to testing program, investigate and record grey mould fruit rot of strawberry diseased plant morbidity grade and
Corresponding strain number calculates disease incidence obtained by each processing, disease index and control efficiency according to formula and obtains table 2.
Table 2 is control efficiency of the Rhodopseudomonas palustris to grey mould fruit rot of strawberry
(spss variance analysis, p=0.05)
It can analyze and obtain from upper table: (1) compared with blank control group, using 1.8929 bacterial strain of Rhodopseudomonas palustris
Fermentation liquid, the disease incidence and disease index of strawberry reduce.(2) Rhodopseudomonas palustris 1.8929 is to occurring on strawberry fruit
Gray mold has good control efficiency, can achieve identical as using the control efficiency of chemical agent；It is proved by realizing, natural pond
The control efficiency of damp red pseudomonas 1.8929 is up to 41.93%.(4) 1.8929 bacterial strain fermentation liquor of Rhodopseudomonas palustris
7.5ml/m2When, it is best to the gray mold control efficiency occurred on strawberry fruit.
Claims (4)
1. a kind of gray mold control method, which is characterized in that by the way of spraying photosynthetic bacteria, inhibit ash by photosynthetic bacteria
The mycelia of mildew pathogen grows and/or carrying out teratogenesis to gray mold pathogen mycelium causes pathogen spore can not shape
At to realize the prevention and treatment to gray mold；
The photosynthetic bacteria is one kind of excrement red pseudomonas or Rhodopseudomonas palustris.
2. gray mold control method according to claim 1, which is characterized in that the mode for spraying photosynthetic bacteria refers to
Photosynthetic bacteria is prepared into fermentation liquid, periodically uniformly sprays the Fermentation by Photosynthetic Bacteria liquid prepared on crop.
3. gray mold control method according to claim 2, which is characterized in that the Fermentation by Photosynthetic Bacteria liquid is according to following
Prepared by mode, photosynthetic bacteria is seeded in solid medium, under the intensity of illumination of 7500lx, constant temperature in 30 DEG C of illumination boxs
7 ~ 14d of illumination cultivation is observed and is grown red or maroon single colonie in culture medium, is linked into oese picking single colonie photosynthetic
In bacterial liquid culture medium, expand culture in the illumination insulating box that intensity of illumination is 7500lx, temperature is 30 DEG C to 14d or so
Fermentation by Photosynthetic Bacteria liquid is obtained, Fermentation by Photosynthetic Bacteria liquid is transferred in the sprayer of sterilizing under aseptic condition, is stored in 4 DEG C of ice
It is spare in case.
4. gray mold control method according to claim 1, which is characterized in that the gray mold refers to generation in strawberry fruit
Gray mold in reality.
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