CN107630064A - A kind of phytophthora infestans antagonistic effect culture medium and its preparation method and application - Google Patents
A kind of phytophthora infestans antagonistic effect culture medium and its preparation method and application Download PDFInfo
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- CN107630064A CN107630064A CN201711055271.7A CN201711055271A CN107630064A CN 107630064 A CN107630064 A CN 107630064A CN 201711055271 A CN201711055271 A CN 201711055271A CN 107630064 A CN107630064 A CN 107630064A
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Abstract
The invention discloses a kind of preparation method of phytophthora infestans antagonistic effect culture medium, comprise the following steps：1）It is prepared by PDA culture medium；2）It is prepared by HX culture mediums；3）Prepared by mixed culture medium, you can obtain the phytophthora infestans antagonistic effect culture medium；And provide the culture medium and application.Beneficial effects of the present invention are：The present invention has taken into full account influence of the culture medium to microorganism growth conditions, pass through the determination of Phytophthora infestans healthy growth on culture medium, the determination of secretory substance can be produced on culture medium to antagonistic strain HC5, a kind of culture medium of suitable phytophthora infestans antagonistic effect is obtained, the screening operation for running into such antagonistic strain again for phytophthora infestans provides a kind of new practical matrix.
Description
Technical field
The present invention relates to microbiological culture media technical field, and in particular to a kind of phytophthora infestans antagonistic effect training
Support base and its preparation method and application.
Background technology
The late blight of potato is by phytophthora infestans（Phytophthora infestans(Mont.) de Bary）Cause
, it is a kind of destructive disease in potato produces, drastically influence the yield and quality of potato.At present, potato
Late blight preventing and treating mainly includes chemical prevention, breeding for disease resistance etc..Chemical prevention is quick, but it is dirty residues of pesticides, environment to be present
The problems such as dye；Application of the disease-resistant seed selection on potato is extremely limited, because the cultigen of potato is homologous four times
Body asexually propagated crop, there is the characteristics of height heterozygosis, hereditary form complexity, hybridize with most wild diploid species not affine.
With development in science and technology progress, biological interspecies relation, intraspecific relationship, the biology of regulation harmful organism population density are utilized
Means of prevention is increasingly valued by people, this bio-control method have the characteristics that safely, effectively with it is pollution-free, with guarantor
The requirement of shield ecological environment and socio-intangible asset development matches.
At present, more scholar Filippov, Jindal K, Yang Huaiwen, Xia Liqin etc. were carried out to potato both at home and abroad
The biological control research of late disease bacteria, achieve preferable prevention effect.These researchs are prevented for the biology of the late blight of potato
Control and established theoretical and application foundation, illustrate the good application prospect of biological prevention.In the research process of applicant,
Because the culture matrix that the growth of phytophthora infestans needs is more special, it is necessary to can health life on rye culture medium
It is long, and the preferable bacterial strain HC5 of one plant of Antagonism that applicant filters out from soil, it can not be produced on rye culture medium short of money
Anti- secretion, research finds preferably produce antagonism secretion in culture matrix PDA culture medium, but potato late blight
Germ can not but produce spore in PDA culture medium.Based on this particularity, applicant is attempted two kinds of culture medium groups
Close, in this combination culture medium, phytophthora infestans can healthy growth, while bacterial strain HC5 also can be produced preferably point
Secretion so that this antagonistic experiment is effectively implemented, and the screening operation for running into such antagonistic strain again for phytophthora infestans carries
A kind of new practical matrix is supplied.
The content of the invention
The purpose of the present invention aiming at it is above-mentioned in the prior art the defects of, there is provided a kind of phytophthora infestans antagonism
Experiment culture medium and its preparation method and application, such a culture medium, both met antagonistic strain HC5 and produced antagonism secretion,
The healthy growth of phytophthora infestans is met again, so that antagonistic experiment is effectively implemented.The culture medium is true first
Secretion can normally produced thereon by having determined antagonistic strain HC5, and also determining phytophthora infestans can be thereon
Healthy growth；Then carry out antagonistic experiment thereon, make experimental result distortion this avoid the single satisfaction of condition, simultaneously
Also avoid the single satisfaction of condition and increase screening operation or miss some effective results, be that the antagonism of phytophthora infestans sieves
Work is selected to provide a kind of new practical matrix.
To achieve these goals, technical scheme provided by the invention is：A kind of phytophthora infestans antagonistic effect is used
The preparation method of culture medium, comprises the following steps：
1）It is prepared by PDA culture medium：
Potato dextrose medium PDA preparation method is to take potato 200g, glucose 20g, agar 18g, is added water to
1L；2）It is prepared by HX culture mediums：
Rye-tomato culture medium HX preparation method is rye 60g, Tomato juice 100ml, calcium carbonate 0.4g, agar 15g,
Add water to 1L；
3）It is prepared by mixed culture medium：
By step 1）With step 2）Obtained PDA culture medium and HX culture mediums, conventional culture medium HTHP is carried out respectively and is gone out
Bacterium, then in an aseptic environment, according to volume ratio 1:The two is well mixed by 1 ratio, you can obtains the late blight of potato
Bacterium antagonistic effect culture medium.
Second object of the present invention there is provided a kind of system of above-mentioned phytophthora infestans antagonistic effect culture medium
The culture medium that Preparation Method is prepared.
Third object of the present invention there is provided application of the above-mentioned culture medium in phytophthora infestans antagonistic effect,
Antagonistic effect comprises the following steps：
a）Antagonism is inoculated with：
A diameter of 5-7mm phytophthora infestans are accessed with culture medium flat plate center in phytophthora infestans antagonistic effect
Cake, 20 DEG C or room temperature dark-grown are being soaked with activation culture 24h antagonistic strain HC5 away from access at central 2-2.5cm after one week
A diameter of 5-7mm of bacterium solution filter paper dick, 20 DEG C or room temperature dark continue to cultivate, and the processing for setting not inoculating strain HC5 is
Control；
b）Antagonistic results count：
The colony radius of pathogen are measured after flat board opposite culture 7-10d, calculate bacteriostasis rate；Calculation formula is：Bacteriostasis rate=[（It is right
According to colony radius-processing colony radius）/ control colony radius] × 100%..
Culture medium is the key of microculture, and different culture mediums, the state difference that microorganism grows above is very big,
It is directly connected to the development of later stage work.Therefore, the growth conditions of antagonistic strain in different culture media have been repeated in the present invention
With the growth conditions of phytophthora infestans, to solve in antagonistic experiment, can antagonistic strain HC5 antagonistic substances preferably divide
Secrete, at the same phytophthora infestans can healthy growth the problems such as.Basal culture medium is finally determined, antagonistic strain HC5 can be met
Antagonistic substance can preferably be secreted, while phytophthora infestans also being capable of healthy growth.This not only avoids the single of condition
Meet and make experimental result distortion, while it also avoid the single satisfaction of condition and increase screening operation or miss some effectively knots
Fruit.
Beneficial effects of the present invention are：
The present invention has taken into full account influence of the culture medium to microorganism growth conditions, by Phytophthora infestans in culture medium
The determination of upper healthy growth, the determination of secretory substance can be produced on culture medium to antagonistic strain HC5, obtain a kind of be adapted to
The culture medium of phytophthora infestans antagonistic effect, the screening operation for running into such antagonistic strain again for phytophthora infestans carry
A kind of new practical matrix is supplied.
Brief description of the drawings
Fig. 1 is shown as the antagonistic experiment result on PDA+HX culture mediums.
Fig. 2 is shown as the antagonistic experiment result on HX culture mediums.
Embodiment
Above-described potato, rye, tomato are purchased from market；Other medicines are analytical grade reagent；It is used
Water is distilled water.
Embodiment 1：
（1）Material to be tested：Phytophthora infestans, antagonistic strain HC5.
（2）Flat board face-off 1：It is a diameter of in a diameter of 9cm PDA+HX flat boards center access using flat board face-off growth method
5mm phytophthora infestans cake, 20 DEG C（Or room temperature）After dark-grown one week, at away from central 2.5cm access be soaked with activation
A diameter of 5mm of culture 24h bacterial strain HC5 bacterium solutions filter paper dick, 20 DEG C（Or room temperature）Dark continues to cultivate, if not being inoculated with point
Processing from bacterial strain is control, and each bacterial strain processing is repeated 3 times.The colony radius of pathogen are measured after 10d, calculate bacteriostasis rate.
Bacteriostasis rate=[（Compare colony radius-processing colony radius）/ control colony radius] × 100%.
（3）Flat board face-off 2：Using flat board face-off growth method, a diameter of 5mm is accessed in a diameter of 9cm HX flat boards center
Phytophthora infestans cake, 20 DEG C（Or room temperature）After dark-grown one week, at away from central 2.5cm access be soaked with activation culture
A diameter of 5mm of 24h bacterial strain HC5 bacterium solutions filter paper dick, 20 DEG C（Or room temperature）Dark continues to cultivate, if not being inoculated with separation bacterium
The processing of strain is control, and each bacterial strain processing is repeated 3 times.The colony radius of pathogen are measured after 10d, calculate bacteriostasis rate.It is antibacterial
Rate=[（Compare colony radius-processing colony radius）/ control colony radius] × 100%.
Test result indicates that as depicted in figs. 1 and 2, the face-off experiment carried out in PDA+HX flat boards, bacterial strain HC5 states
For blueness, the antagonistic substance of yellow green is secreted out of, Phytophthora infestans has very strong inhibition, inhibiting rate 90%；In HX
The face-off experiment carried out in flat board, bacterial strain HC5 states are lime color, and no Antagonism material secrets out of, phytophthora infestans
Bacterial strain HC5 has been surrounded in growth, does not show Antagonism.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used
To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic.
Within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc., it should be included in the present invention's
Within protection domain.
Claims (3)
1. a kind of preparation method of phytophthora infestans antagonistic effect culture medium, it is characterised in that comprise the following steps：
1）It is prepared by PDA culture medium：
Potato dextrose medium PDA preparation method is to take potato 200g, glucose 20g, agar 18g, is added water to
1L；
2）It is prepared by HX culture mediums：
Rye-tomato culture medium HX preparation method is rye 60g, Tomato juice 100ml, calcium carbonate 0.4g, agar 15g,
Add water to 1L；
3）It is prepared by mixed culture medium：
By step 1）With step 2）Obtained PDA culture medium and HX culture mediums, conventional culture medium HTHP is carried out respectively and is gone out
Bacterium, then in an aseptic environment, according to volume ratio 1:The two is well mixed by 1 ratio, you can obtains the late blight of potato
Bacterium antagonistic effect culture medium.
2. a kind of phytophthora infestans antagonistic effect according to claim 1 is prepared with the preparation method of culture medium
Culture medium.
3. application of the culture medium according to claim 2 in phytophthora infestans antagonistic effect, it is characterised in that short of money
Anti- experiment comprises the following steps：
a）Antagonism is inoculated with：
A diameter of 5-7mm phytophthora infestans are accessed with culture medium flat plate center in phytophthora infestans antagonistic effect
Cake, 20 DEG C or room temperature dark-grown are being soaked with activation culture 24h antagonistic strain HC5 away from access at central 2-2.5cm after one week
A diameter of 5-7mm of bacterium solution filter paper dick, 20 DEG C or room temperature dark continue to cultivate, and the processing for setting not inoculating strain HC5 is
Control；
b）Antagonistic results count：
The colony radius of pathogen are measured after flat board opposite culture 7-10d, calculate bacteriostasis rate；Calculation formula is：Bacteriostasis rate=[（It is right
According to colony radius-processing colony radius）/ control colony radius] × 100%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110042062A (en) * | 2019-04-22 | 2019-07-23 | 云南农业大学 | A kind of phytophthora infestans culture medium and preparation method thereof |
| CN111378602A (en) * | 2020-02-27 | 2020-07-07 | 宜春学院 | Bacterial strain capable of inhibiting phytophthora infestans of potato late blight and separation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101984042A (en) * | 2010-10-11 | 2011-03-09 | 中国农业大学 | Preservation method of pathogenic bacteria of potato late blight and special medium for the same |
| CN107136122A (en) * | 2017-04-19 | 2017-09-08 | 福建省农业科学院植物保护研究所 | A kind of biocontrol agent for preventing and treating the late blight of potato |
-
2017
- 2017-11-01 CN CN201711055271.7A patent/CN107630064A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101984042A (en) * | 2010-10-11 | 2011-03-09 | 中国农业大学 | Preservation method of pathogenic bacteria of potato late blight and special medium for the same |
| CN107136122A (en) * | 2017-04-19 | 2017-09-08 | 福建省农业科学院植物保护研究所 | A kind of biocontrol agent for preventing and treating the late blight of potato |
Non-Patent Citations (3)
| Title |
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| 李海鹰等: "不同成分培养基影响致病疫霉生长的比较研究", 《安徽农学通报》 * |
| 杨永梅等: "一株抗马铃薯晚疫病的放线菌的分离鉴定", 《内蒙古农业大学学报》 * |
| 毕朝位等: "致病疫霉（Phytophthora infestans)的分离与培养方法", 《植物保护科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110042062A (en) * | 2019-04-22 | 2019-07-23 | 云南农业大学 | A kind of phytophthora infestans culture medium and preparation method thereof |
| CN111378602A (en) * | 2020-02-27 | 2020-07-07 | 宜春学院 | Bacterial strain capable of inhibiting phytophthora infestans of potato late blight and separation method and application thereof |
| CN111378602B (en) * | 2020-02-27 | 2023-05-09 | 宜春学院 | Bacterial strain capable of inhibiting phytophthora infestans and separation method and application thereof |
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Application publication date: 20180126 |