CN108103001A - A kind of method that phytophthora infestans is promoted to generate a large amount of sporangiums - Google Patents
A kind of method that phytophthora infestans is promoted to generate a large amount of sporangiums Download PDFInfo
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- CN108103001A CN108103001A CN201810127449.2A CN201810127449A CN108103001A CN 108103001 A CN108103001 A CN 108103001A CN 201810127449 A CN201810127449 A CN 201810127449A CN 108103001 A CN108103001 A CN 108103001A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
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Abstract
The present invention provides a kind of methods that phytophthora infestans is promoted to generate a large amount of sporangiums, belong to oomycetes field, and this method specifically includes：The selection of bacterial strain, the selection of culture vessel, the selection of culture medium and inoculation and culture, wherein, the volume of blake bottle is 240~500mL, and blake bottle has ventilative bottle cap.The present invention is on the basis of using medium culture late blight bacterial strain, conventional culture dish is used instead and is used as culture vessel for space bigger and the blake bottle that has certain air permeability effect, can effectively solve the problems, such as to produce sporangium quantity in previous phytophthora infestans culture with the method few.Bacterial strain generation sporangium quantity is cultivated in the opposite conventional culture dish of the present invention to be obviously improved, it provides safeguard for the research of the late blight of potato, and this method is at low cost, easy to operate, reduce the risk of suspension cell line pollution, it is a kind of efficient, quick new method, also provides a platform for the correlative study by the use of phytophthora infestans as experiment material.
Description
Technical field
The present invention relates to a kind of methods that phytophthora infestans is promoted to generate a large amount of sporangiums, belong to oomycetes field.
Technical background
The late blight of potato is drawn by Oomycete phytophthora infestans (Phytophthora infestans (Mont.) de Bary)
It rises, is one of destructive disease of potato, the general time can cause the underproduction 10%~20%, and the time for weight of falling ill can the underproduction
50%~70% in addition total crop failure.
To prevent late blight of potato disease, scientists have carried out substantial amounts of research, and research shows the late blight of potato
Bacterial strain generate sporangium be late blight of potato Disease Cycle key point, therefore will be directed to the disease carry out deeper into
Research, extraction phytophthora sporangium is one of necessary work.However the production spore quantity of late disease bacteria is artificially cultivated in an experiment
It is unsatisfactory, especially it is much more long-term be commissioned to train educate after, the sporangium quantity for cultivating generation is difficult meets the needs of experiment, to studying
Form bottleneck.To ensure being normally carried out for experiment, it is top priority that improvement production spore method, which increases sporulation quantity,.
Generally using culture dish, culture dish is covered with ware lid and prevented with sealed membrane sealing for common for the culture of all kinds of strains
Pollution operation, but due to the reason of structure, air can not circulate, therefore be caused centainly for the culture of aerobic microorganism
Obstruction.And blake bottle is generally used for the in vitro culture of cell and tissue, it is advantageous that space is big in bottle, bottle cap is ventilative and
Anti-pollution ability is strong.
The content of the invention
Spore amount deficiency is generated for existing culture dish culture phytophthora infestans, the present invention proposes a kind of promotion
The method that phytophthora infestans generate a large amount of sporangiums, the air containment of this method combination blake bottle is big and air is negotiable
Advantage and culture dish are used to cultivate the purposes of strain, dramatically increase phytophthora infestans production spore quantity, are potato late blight
The research of disease provides safeguard.
Technical scheme is as follows：A kind of method that phytophthora infestans is promoted to generate a large amount of sporangiums, tool
The method of body includes the following steps：
A. the selection of bacterial strain：Phytophthora infestans strain is selected, number XH05-5-4 cultivates 10 in 20 DEG C of incubators
~15 days spare；
B. the selection of culture vessel：Bottle cap breathable blake bottle is selected, wherein, culture bottle cap band filter membrane, filter membrane can
It is ventilative；
C. the selection of culture medium：Select rye-tomato juice culture medium；
D. it is inoculated with and cultivates：Aseptically, the culture medium described in step c is added in the blake bottle described in b step,
Target fungus block is laid using card punch and is inoculated in culture medium central；It covers culture bottle cap and is put into dark culturing in 20 DEG C of incubators
12~15 days to get substantial amounts of phytophthora infestans sporangium.
Further, the blake bottle volume is 240~500mL.
Further, the preparation method of the culture medium is：Weigh 50~60 grams of ryes, with distilled water immersion 24 it is small when,
It shows money or valuables one carries unintentionally until most of rye all germinates, heating is boiled, until wheat capsule ruptures, filtering measures 900 milliliters, adds in tomato juice 100
15 grams of milliliter and agar shake up, and are put into high pressure sterilization and sterilize 30 minutes, that is, complete the preparation of rye-tomato juice culture medium.
The features of the present invention is：It is covered during general culture dish culture strain with culture dish lid and seals to subtract with sealed membrane again
The pollution of few air, by the present invention in that replacing culture dish with blake bottle to cultivate phytophthora infestans, the blake bottle
Bottle cap band filter membrane, filter membrane is ventilative, this kind of bottle cap can increase air in blake bottle and exchange, while the filter membrane of bottle cap institute band is ventilative
While effectively block influence of the pollutant in air to bacterial strain, promote the big volume production spore of strain.
Compared with prior art, beneficial effects of the present invention are：The present invention uses the blake bottle conduct different from culture dish
To bacterial strain with relatively open space and air circulation, potato evening can be effectively solved using the cultural method for culture vessel
The problem of epidemic disease bacterium production spore amount is insufficient is obviously improved with respect to bacterial strain generation sporangium quantity is cultivated in conventional culture dish.This
It invents and provides safeguard for the research of the late blight of potato, and this method is at low cost, easy to operate, reduces the wind of bacterial strain pollution
Danger is a kind of efficient, quick new method, is also provided for the correlative study by the use of phytophthora infestans as experiment material
One platform.
Description of the drawings
Fig. 1 is the pattern schematic diagram of blake bottle in the present invention；
Fig. 2 is the mycelium growth vigor schematic diagram of blake bottle culture 12 days in the embodiment of the present invention 1；
Fig. 3 is the mycelium growth vigor schematic diagram of culture dish culture 12 days in the embodiment of the present invention 1；
Fig. 4 is the schematic diagram that fungus block is inoculated into blake bottle and culture dish respectively in the embodiment of the present invention 2；
Fig. 5 is that fungus block is inoculated into the schematic diagram of culture medium central in blake bottle in the embodiment of the present invention 2；
Fig. 6 is blake bottle and the mycelium growth vigor schematic diagram of culture dish culture 15 days in the embodiment of the present invention 2；
Fig. 7 is that the schematic diagram that sporangium is made to fall into sterile water is gently scraped using small spatula in embodiment 2；
Fig. 8 is the schematic diagram of the bacterium solution extracted in embodiment 1 after culture dish culture under the microscope；
Fig. 9 is the schematic diagram of the bacterium solution extracted in embodiment 1 after blake bottle culture under the microscope；
Figure 10 is the schematic diagram of the bacterium solution extracted in embodiment 2 after culture dish culture under the microscope；
Figure 11 is the schematic diagram of the bacterium solution extracted under the microscope after 2 medium and small sizes blake bottle culture of embodiment；
Figure 12 is the schematic diagram of the bacterium solution extracted in embodiment 2 after medium size blake bottle culture under the microscope；
Figure 13 is the schematic diagram of the bacterium solution extracted in embodiment 2 after large size blake bottle culture under the microscope.
Specific embodiment
Technical scheme is further described in detail with reference to the accompanying drawings and embodiments.
Embodiment 1 (strain of blake bottle culture phytophthora infestans)
The present embodiment is using phytophthora infestans strain XH05-5-4 as experiment material, using rye-tomato juice culture medium
It is cultivated, spore variation is produced in the case that air exchanges to have probed into.
It is carried out at the same time, concretely comprises the following steps under experimental group and control group the same terms in the present embodiment：
(1) experimental group：Make rye-tomato juice culture medium after poured into blake bottle 30ml (blake bottle pattern is shown in Fig. 1,
Specification is diameter 68mm, bore 62mm, bottle height 90mm), card punch (diameter 6mm) is taken to punch after high pressure sterilization solidification to be cooled
Method lay fungus block from the bacterial strain that original preserves, be inoculated into culture medium central, culture bottle cap is inverted that (lid is downward after covering
Lower placement) it is put into 20 DEG C of incubators, dark culturing 12 days (such as Fig. 2).Blake bottle is taken out, adds in 10ml sterile waters to culture base table
Face and gently scraping hyphal surface with small spatula makes sporangium fall into the water；Gained liquid is transferred to and has wrapped 2 layers of Mira-cloth
Small beaker in, filtering gained liquid be spore suspension；Suspension is taken to be counted on blood counting chamber (25 × 16 type)
Number.
(2) control group (control check, CK)：Make after rye-tomato juice culture medium that (culture dish is in culture dish
The Conventional glass culture dish of a diameter of 90mm) in pour into 30ml, take card punch (diameter 6mm) after high pressure sterilization solidification to be cooled
The method of punching lays fungus block from the bacterial strain that original preserves, and is inoculated into culture medium central, culture dish is sealed to hinder with sealed membrane
Disconnected air exchange, is put into 20 DEG C of incubators, dark culturing 12 days (such as Fig. 3).Culture dish is taken out, adds in 10ml sterile waters to culture
Primary surface and gently scraping hyphal surface with small spatula makes sporangium fall into the water；Gained liquid is transferred to and has wrapped 2 layers of Mira-
In the small beaker of cloth, filtering gained liquid is spore suspension；Take suspension enterprising in blood counting chamber (25 × 16 type)
Row counts.
More than experimental group and control group are respectively repeated once, and the average value counted twice in two groups is taken to compare.
Wherein, spore quantity result is produced according to blood counting chamber Counting Formula：
Sporangium number/1mL=80 lattice sporangium sum/80*400*10000
It is calculated in blake bottle culture：First phialosporae capsule quantity is 350000/mL, is 250000 in second bottle
A/mL, the average value counted twice in blake bottle are 300000/mL.
It is calculated in culture dish culture：First ware sporangium quantity is 50000/mL, is 25000 in the second ware
A/mL, the average value counted twice in culture dish are 37500/mL.
Result statistics and comparison see the table below 1 in embodiment 1.
Table 1
Upper table 1 the experimental results showed that：Spore quantity average out to is produced using the phytophthora infestans of the method for the present invention culture
Using 8 times of cellar culture ware culture production spore quantity.
Embodiment 2 (strain of different size blake bottle culture phytophthora infestans)
The present embodiment is using phytophthora infestans strain XH05-5-4 as experiment material, using rye-tomato juice culture medium
It is cultivated, production spore is influenced with probing into space size in bottle.
It is carried out at the same time, concretely comprises the following steps under experimental group and control group the same terms in the present embodiment：
(1) experimental group：First make rye-tomato juice culture medium, after autoclave sterilization respectively toward it is 3 small, in, it is different
30ml culture mediums are added in the blake bottle of specification, and (three kinds of specifications of blake bottle are divided into 240ml, 350ml, 500ml, and detail parameters are shown in Table
2)；The method that card punch (diameter 6mm) punches is taken to lay fungus block from the bacterial strain that original preserves after condensation, is inoculated into culture medium
It entreats (such as Fig. 4 and Fig. 5)；Culture bottle cap is inverted (lid is placed downwards) after covering and is put into 20 DEG C of incubators, and dark culturing 15 days is (such as
Fig. 6).Blake bottle is taken out, adding in 10ml sterile waters, gently scraping hyphal surface makes spore to media surface and with small spatula (such as Fig. 7)
Capsule falls into the water；Gained liquid is transferred in superfine metal strainer and filters off mycelia, filtering gained liquid is spore suspension；
Suspension is taken to be counted on blood counting chamber (25 × 16 type).
2 blake bottle of table is small, in, big three kinds of specifications
| Blake bottle specification | Volume (mL) | High (mm) | Diameter (mm) | Bore (mm) |
| It is small | 240 | 90 | 68 | 62 |
| In | 350 | 108 | 75 | 69 |
| Greatly | 500 | 140 | 95 | 60 |
(2) control group (control check, CK)：Rye-tomato juice culture medium is first made, it is past after autoclave sterilization
30ml culture mediums are added in culture dish (culture dish is the Conventional glass culture dish of a diameter of 90mm)；Card punch is taken after condensation
The method of (diameter 6mm) punching lays fungus block from the bacterial strain that original preserves, and is inoculated into culture medium central (such as Fig. 4)；By culture dish
(lid is placed downwards) is inverted after being sealed with sealed membrane and is put into 20 DEG C of incubators, dark culturing 15 days (such as Fig. 6).Take out culture
Ware adds in 10ml sterile waters to media surface, and gently scraping hyphal surface with small spatula makes sporangium fall into the water；By gained liquid
Body, which is transferred in superfine metal strainer, filters off mycelia, and filtering gained liquid is spore suspension；Suspension is taken in blood count
It is counted on plate (25 × 16 type).
More than experimental group and control group difference in triplicate, take the average value counted three times in three groups to compare.
As a result：According to blood counting chamber Counting Formula：
Sporangium number/1mL=80 lattice sporangium sum/80*400*10000
It is calculated：1,2,3 miospore capsule number of bottle is respectively 100000/mL, 150000/mL, 75000/
ML, 108333/mL of average out to；
It is calculated：Middle 1,2,3 miospore capsule number of bottle is respectively 50000/mL, 75000/mL, 75000/mL,
66667/mL of average out to；
It is calculated：Big 1,2,3 miospore capsule number of bottle is respectively 100000/mL, 100000/mL, 75000/
ML, 91666/mL of average out to；
It is calculated：1,2,3 miospore capsule number of culture dish is respectively 25000/mL, 0/mL, 25000/mL, is put down
Equal 16667/mL.
Wherein, the situation that cultivation results production spore number is 0/mL in culture dish 2 is not that the culture dish does not produce spore completely, and
It is since overall sporulation quantity is less, is counted on blood counting chamber (25 × 16 type), by diagonal orientation, several upper lefts,
Lower-left, center, upper right, bottom right five grids in sporangium number when being counted, spore be distributed in five grids it is outer and
Its sporulation quantity is given tacit consent to for 0.
Result statistics and comparison see the table below 3 in embodiment 2.
Table 3
From the test result analysis of upper table 3：It is not as notable as the reason for embodiment 1 for Comparative result in this experiment
Strainer selection preciosity is speculated as so that part sporangium fails under filter, but compared to control (culture dish), blake bottle is trained
It is respectively 650%, 400%, the 550% of control group that the sporulation quantity for the mode of supporting, which remains unchanged, average out to 533%.Trained using blake bottle
The mode sporulation quantity for supporting late blight bacterial strain is significantly more than the mode of culture dish culture.
Meanwhile by count results in tri- kinds of blake bottles of 240ml, 350ml, 500ml from the point of view of, to maximum volume 500ml when trains
Production spore quantity in bottle is supported to have no and dramatically increases, the volume for illustrating blake bottle culture phytophthora infestans strain be not it is more big more
It is good, also have the disadvantages such as pollution rate increases, culture medium dosage increases and inoculation difficulty is big on the contrary, being cultivated in bigger bottle
End, therefore blake bottle volume is advisable no more than 500mL in the present invention.
The foregoing is merely presently preferred embodiments of the present invention, and the equivalent change that all scopes under this invention are done should all belong to
The covering scope of the present invention.
Claims (3)
- A kind of 1. method that phytophthora infestans is promoted to generate a large amount of sporangiums, it is characterised in that the method includes as follows Step：A. the selection of bacterial strain：Phytophthora infestans strain is selected, number XH05-5-4 cultivates 10~15 in 20 DEG C of incubators It is spare；B. the selection of culture vessel：Bottle cap breathable blake bottle is selected, wherein, bottle cap band filter membrane is cultivated, filter membrane is ventilative；C. the selection of culture medium：Select rye-tomato juice culture medium；D. it is inoculated with and cultivates：Aseptically, the culture medium described in step c is added in the blake bottle described in b step, is used Card punch lays target fungus block and is inoculated in culture medium central；Cover culture bottle cap be put into dark culturing 12 in 20 DEG C of incubators~ 15 days, bacterium is scraped and is used after two layers of filtered through gauze up to phytophthora infestans sporangium.
- 2. a kind of method that phytophthora infestans is promoted to generate a large amount of sporangiums as described in claim 1, it is characterised in that： The blake bottle volume is 240~500mL.
- 3. a kind of method that phytophthora infestans is promoted to generate a large amount of sporangiums as described in claim 1, it is characterised in that： The preparation method of the culture medium is：Weigh 50~60 grams of ryes, with distilled water immersion 24 it is small when, until most of rye all sends out Bud shows money or valuables one carries unintentionally, and heating is boiled, until wheat capsule ruptures, filtering measures 900 milliliters, adds in 100 milliliters of tomato juice and 15 grams of agar shakes It is even, it is put into high pressure sterilization and sterilizes 30 minutes, that is, complete the preparation of rye-tomato juice culture medium.
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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 |
| CN110643517A (en) * | 2019-10-17 | 2020-01-03 | 河南科技学院 | Phytophthora infestans culture medium MYEB and method for detecting phytophthora infestans pathogenicity to plants |
| CN110819583A (en) * | 2018-08-10 | 2020-02-21 | 云南农业大学 | Spore-producing culture method of phytophthora sojae |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003059936A2 (en) * | 2002-01-10 | 2003-07-24 | Syngenta Participations Ag | Phytophthora Infestans CDC 14 Phosphatase Gene, Protein and Methods of Use |
| CN101117639A (en) * | 2007-06-28 | 2008-02-06 | 贵州省生物技术研究所 | Method for acquiring disease-resistance expression of agrobacterium-mediated potato transgenic hrap |
| CN101250573A (en) * | 2008-04-11 | 2008-08-27 | 云南省农业科学院生物技术与种质资源研究所 | Method for inducing phytophthora infestans to produce pathogenic secretory protein |
| CN105567675A (en) * | 2016-02-01 | 2016-05-11 | 福建农林大学 | Method for rapidly extracting DNA of single sporangium of phytophythora infestans |
| CN106834135A (en) * | 2016-12-26 | 2017-06-13 | 西北农林科技大学 | A kind of method for inducing grape Elsinochrome to produce spore |
| CN107603891A (en) * | 2017-11-03 | 2018-01-19 | 云南省农业科学院生物技术与种质资源研究所 | A kind of simple store method of phytophthora infestans |
-
2018
- 2018-02-08 CN CN201810127449.2A patent/CN108103001A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003059936A2 (en) * | 2002-01-10 | 2003-07-24 | Syngenta Participations Ag | Phytophthora Infestans CDC 14 Phosphatase Gene, Protein and Methods of Use |
| CN101117639A (en) * | 2007-06-28 | 2008-02-06 | 贵州省生物技术研究所 | Method for acquiring disease-resistance expression of agrobacterium-mediated potato transgenic hrap |
| CN101250573A (en) * | 2008-04-11 | 2008-08-27 | 云南省农业科学院生物技术与种质资源研究所 | Method for inducing phytophthora infestans to produce pathogenic secretory protein |
| CN105567675A (en) * | 2016-02-01 | 2016-05-11 | 福建农林大学 | Method for rapidly extracting DNA of single sporangium of phytophythora infestans |
| CN106834135A (en) * | 2016-12-26 | 2017-06-13 | 西北农林科技大学 | A kind of method for inducing grape Elsinochrome to produce spore |
| CN107603891A (en) * | 2017-11-03 | 2018-01-19 | 云南省农业科学院生物技术与种质资源研究所 | A kind of simple store method of phytophthora infestans |
Non-Patent Citations (1)
| Title |
|---|
| 岳海梅: "《植物病理学实验及研究技术》", 31 December 2015 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110819583A (en) * | 2018-08-10 | 2020-02-21 | 云南农业大学 | Spore-producing culture method of phytophthora sojae |
| 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 |
| CN110643517A (en) * | 2019-10-17 | 2020-01-03 | 河南科技学院 | Phytophthora infestans culture medium MYEB and method for detecting phytophthora infestans pathogenicity to plants |
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