CN109628329A - A kind of modified PDA culture medium and preparation method thereof - Google Patents
A kind of modified PDA culture medium and preparation method thereof Download PDFInfo
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- CN109628329A CN109628329A CN201910079892.1A CN201910079892A CN109628329A CN 109628329 A CN109628329 A CN 109628329A CN 201910079892 A CN201910079892 A CN 201910079892A CN 109628329 A CN109628329 A CN 109628329A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The present invention relates to a kind of modified PDA culture mediums and preparation method thereof, obtain after then manufacturing process addition tomato, the carrot of traditional PDA culture medium are spray-dried culture medium.Such culture medium once can be manufactured in high volume, easy to use for being used for multiple times.The present invention can preferably overcome the problems, such as and improve that PDA culture medium encounters in above-mentioned two Bacteria culturing, enhance its special function.
Description
Technical field
The invention belongs to technical field of plant protection, and in particular to one kind is used for Alternaria alternate (Alternaria
Alternata) and alternaria leaf spot of apple (Alternaria alternaria f.sp mali) separates and what culture used changes
Into type PDA culture medium and preparation method thereof.
Background technique
PDA culture medium is abbreviation of the people to potato dextrose agar, i.e. Potato Dextrose Agar.
PDA culture medium is plant protection field for one of fungal diseases of plants pathogenicbacteria separation, the most common culture medium of culture.More anti-mildews
Largely make commonly using Alternaria alternate and alternaria leaf spot of apple pathogen as indicator bacteria in plain product minor biological cycling
With PDA culture medium, ready-to-use usually more wasteful time, and having differences property between batch usually influence detection effect, and meeting
There is cultivation cycle to have arrived, but expected the case where requiring is not achieved in growth.
The production method that Chinese patent application 201610081117.6 discloses a kind of high grade of transparency PDA culture medium belongs to PDA training
Support the production method field of base, comprising the following steps: A, potato cutting；B, it boils block: being first cooked by slow fire again with high fire, make potato
Fourth is thoroughly well cooked but not mushy；C, it filters；D, it feeds: by glucose 20g, potassium dihydrogen phosphate 0.8-1.4g, in magnesium sulfate 1.2g-1.7g addition
Filtrate is stated, is stirred evenly, is settled to 1000mL with water；E, adjust pH value: using hydrochloric acid solution and sodium hydroxide solution adjust pH value to
6.5-7 obtaining fluid nutrient medium；First high-purity agar powder 19-23g is dispensed into several allocates containers, then trains liquid
Feeding base is dispensed into several allocates containers, is mixed；F, inverted plate；G, it dries: by culture dish ware lid on alcolhol burner flame
Steam is dried；H, it seals, Sterility testing.This method can precisely, rapidly carry out the proportion of each raw material, avoid systematic error and
Operating error；And moisture in culture dish can be controlled well, avoid miscellaneous bacteria from breeding.
Chinese patent application 201610740833.0 discloses application of the potato full-powder in preparation PDA culture medium, belongs to
In microbiological culture media field；The invention also discloses a kind of potato full-powder culture medium, constituent and its ratio are as follows: horse
Bell potato complete 10~20g of powder, 2~10g of glucose, 12~20g of agar powder, water 1000ml.Also disclose potato full-powder culture medium
Application in fungal culture.By starch, the proteins,vitamins,and minerals etc. in potato in potato full-powder culture medium
Nutriment all saves, and nutrition is comprehensive, and edible mushroom mycelial growth rate on potato full-powder culture medium is fast, and grows prosperous
It contains；Secondly, potato full-powder culture medium preparation method is simple, the processes such as potato juice are not produced, preparation time greatly shortens；
In addition, potato full-powder culture medium production cost is low；Potato full-powder medium component is all dry powder, and preservation, transport are all very square
Just.
But the PDA culture medium of research in the prior art not can be good at overcoming and improve PDA culture medium and exist
The problem of being encountered in above-mentioned two Bacteria culturing.
Summary of the invention
In order to solve the above-mentioned problems in the prior art, the present invention provides a kind of modified PDA culture medium and its system
Preparation Method, can preferably overcome the problems, such as and improve PDA culture medium to encounter in above-mentioned two Bacteria culturing, enhance it specially
Use function.
The present invention adopts the following technical scheme: a kind of preparation method of modified PDA culture medium, in traditional PDA culture medium
Manufacturing process addition tomato, carrot then culture medium is spray-dried after obtain.Such culture medium once can be with
High-volume manufactures, easy to use for being used for multiple times.
As preferred embodiment of the invention, the preparation method the following steps are included:
(1) peeled potatoes, carrot, tomato are cleaned, cut into serving pieces, wherein in terms of mass parts, peeled potatoes 1000-
2000 parts, 300-500 parts of carrot, 80-100 parts of tomato；
(2) potato and carrot slice are put into heat in 10000 parts of cold drinking liquids and be extracted, etc. water temperatures start to count when rising to 95 DEG C
When, tomato piece is added, boils extraction 20 minutes, does not stir in the process, is filtered using 4 layers of medical sterile gauze；
(3) filter residue is discarded, and filtrate is spray-dried under the conditions of at 75-85 DEG C, obtains water content not higher than 5% dry powder, 0-15 DEG C dry
It is dry to be sealed；
(4) 8-10 mass parts dry powder is weighed when using, is added in 1000 mass parts water, and agar is added according to culture medium hardness requirement
Afterwards, steam sterilizing can be used.
As preferred embodiment of the invention, in step (1), peeled potatoes, carrot, tomato are cut into respectively
The piece of 0.5cm, 0.5cm and 1cm thickness is spare.
As preferred embodiment of the invention, in step (2), etc. water temperatures start timing when rising to 95 DEG C, the 15th point
Tomato piece, the heating of stopping in the 20th minute is added in clock；Extractive juice is equal using glucose dissolution is added after 4 layers of medical sterile gauze filtering
It is even.
As preferred embodiment of the invention, in step (3), the moisture determination method is 105 DEG C of conditions
Lower drying 2h calculates loss of weight amount and accounts for dry powder percentage before drying.
As preferred embodiment of the invention, in step (4), the steam sterilizing is that 121 DEG C of saturated vapors go out
Bacterium.
As preferred embodiment of the invention, the preparation method the following steps are included:
(1) peeled potatoes 2000g is weighed, 500 grams of carrot, 100 grams of tomato, wash clean, potato cuts along short stem direction
At the disk of 0.5cm thickness, carrot is cut into the disk of 0.5cm thickness, and the piece that tomato cuts 1 cm thick is spare；
(2) potato and carrot slice are put into heat in 10Kg cold drinking liquid and are extracted, leaching is boiled in timing when water temperature reaches 95 DEG C
It mentions 20 minutes, does not stir in the process；Extractive juice is uniformly dissolved using 200 grams of glucose of addition after 4 layers of medical sterile gauze filtering；
(3) filter residue is discarded, and filtrate is spray-dried under the conditions of 75-85 DEG C, controls moisture dry powder below 5%, and 0-15 DEG C of sealing is protected
It deposits；
(4) 8-10g dry powder is weighed when using, adds water 1000g, 121 DEG C of saturated vapors go out after agar powder is added according to hardness requirement
Bacterium can be used.
The modified PDA culture medium that the present invention also protects above-mentioned preparation method to be prepared.
Compared with prior art, the present invention adds tomato, carrot and right by the manufacturing process in traditional PDA culture medium
The improvement of culture medium preparation process, by above-mentioned improvement, the present invention achieve it is below the utility model has the advantages that
1) pass through improvement, it will be apparent that improve training of the PDA culture medium to pathogens such as Alternaria alternate and alternaria leaf spot of apple
Nourish one's nature can, mycelium morphology factor dramatically increases.
2) after improving, which can produce once, and be used for multiple times, and significantly improve the confecting efficiency of PDA culture medium,
Avoid the influence of difference between batch.
3) after improving, culture medium raw material is easier to store, and effectively extends the shelf-life of culture medium raw material.
Specific embodiment
The present invention is described in further details below with reference to embodiment, but the present invention is not limited only to following implementations
Example.
Embodiment 1
4000g peeled potatoes, 1000g carrot and 200g tomato are taken, is cut into respectively with a thickness of 0.5cm, 0.5cm and 1cm thickness
The piece of degree.First potato and carrot slice are put into 20000g tap water and begun to warm up, temperature reaches 95 DEG C of beginning timing,
Tomato piece is added after 15min, then is filtered after boiling 5min with 4 layers of medical sterile gauze, obtains filtrate 19100g.It is spraying with water adjustment
Drying tower temperature is that 82 DEG C of laggard filtrates start to be spray-dried, and obtains 783 grams of dry powder.Moisture content is 4.1% after detection.Take 8 grams
Above-mentioned dry powder is put into 1000g water, and agar powder 18g is added, obtains modified PDA culture medium 1026g.
Embodiment 2
2000g peeled potatoes, 500g carrot and 100g tomato are taken, is cut into respectively with a thickness of 0.5cm, 0.5cm and 1cm thickness
Piece.First potato and carrot slice are put into 10000g tap water and begun to warm up, temperature reaches 95 DEG C of beginning timing,
Tomato piece is added after 15min, then is filtered after boiling 5min with 4 layers of medical sterile gauze, obtains filtrate 9133g.It is spraying with water adjustment
Drying tower temperature is that 75 DEG C of laggard filtrates start to be spray-dried, and obtains 361 grams of dry powder, and moisture content is 4.7% after detection.Take 10
Gram above-mentioned dry powder is put into 1000g water, and agar powder 18g is added, obtains modified PDA culture medium 1028g.
Embodiment 3
2000g peeled potatoes, 500g carrot and 100g tomato are taken, is cut into respectively with a thickness of 0.5cm, 0.5cm and 1cm thickness
Piece.First potato and carrot slice are put into 10000g tap water and begun to warm up, temperature reaches 95 DEG C of beginning timing,
Tomato piece is added after 15min, then is filtered after boiling 5min with 4 layers of medical sterile gauze, obtains filtrate 9133g.It is spraying with water adjustment
Drying tower temperature is that 85 DEG C of laggard filtrates start to be spray-dried, and obtains 343 grams of dry powder, and moisture content is 3.7% after detection.Take 9 grams
Above-mentioned dry powder is put into 1000g water, and agar powder 20g is added, obtains modified PDA culture medium 1029g.
Embodiment 4
After 1,2,3 culture medium inoculated Alternaria alternate of common PDA culture medium and embodiment, alternaria leaf spot of apple after 3 days and 4 days
Bacterium colony growing state comparative situation is as shown in table 1 below.
The bacterium colony growing state of 1,2,3 culture medium of the common PDA culture medium of table 1 and embodiment
From table 1 it follows that improved PDA culture medium significantly improves the increment of bacterium colony, but spore is produced to unit area
Amount has little effect.As it can be seen that improved PDA culture medium prepared by the present invention significantly improves Alternaria alternate and apple spot
The culture performance of the pathogens such as defoliation, mycelium morphology factor dramatically increase.
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Claims (8)
1. a kind of preparation method of modified PDA culture medium, which is characterized in that added in the manufacturing process of traditional PDA culture medium
Tomato, carrot obtain after being then spray-dried to culture medium.
2. preparation method according to claim 1, which comprises the following steps:
(1) peeled potatoes, carrot, tomato are cleaned, cut into serving pieces, wherein in terms of mass parts, peeled potatoes 1000-
2000 parts, 300-500 parts of carrot, 80-100 parts of tomato；
(2) potato and carrot slice are put into heat in 10000 parts of cold drinking liquids and be extracted, etc. water temperatures start to count when rising to 95 DEG C
When, tomato piece is added, boils extraction 20 minutes, does not stir in the process, is filtered using 4 layers of medical sterile gauze；
(3) filter residue is discarded, and filtrate is spray-dried under the conditions of at 75-85 DEG C, obtains water content not higher than 5% dry powder, 0-15 DEG C dry
It is dry to be sealed；
(4) 8-10 mass parts dry powder is weighed when using, is added in 1000 mass parts water, and agar is added according to culture medium hardness requirement
Afterwards, steam sterilizing can be used.
3. preparation method according to claim 2, which is characterized in that in step (1), by peeled potatoes, carrot, kind
The piece that eggplant is cut into 0.5cm, 0.5cm and 1cm thickness respectively is spare.
4. preparation method according to claim 2, which is characterized in that in step (2), etc. water temperatures start to count when rising to 95 DEG C
When, the 15th minute addition tomato piece, the heating of stopping in the 20th minute；Portugal is added after filtering using 4 layers of medical sterile gauze in extractive juice
Grape sugar is uniformly dissolved.
5. preparation method according to claim 2, which is characterized in that in step (3), the moisture determination method is
2h is dried under the conditions of 105 DEG C, is calculated loss of weight amount and is accounted for dry powder percentage before drying.
6. preparation method according to claim 2, which is characterized in that in step (4), the steam sterilizing is 121 DEG C
Saturated vapor sterilizing.
7. preparation method according to claim 2, which is characterized in that the preparation method the following steps are included:
(1) peeled potatoes 2000g is weighed, 500 grams of carrot, 100 grams of tomato, wash clean, potato cuts along short stem direction
At the disk of 0.5cm thickness, carrot is cut into the disk of 0.5cm thickness, and the piece that tomato cuts 1 cm thick is spare；
(2) potato and carrot slice are put into heat in 10Kg cold drinking liquid and are extracted, leaching is boiled in timing when water temperature reaches 95 DEG C
It mentions 20 minutes, does not stir in the process；Extractive juice is uniformly dissolved using 200 grams of glucose of addition after 4 layers of medical sterile gauze filtering；
(3) filter residue is discarded, and filtrate is spray-dried under the conditions of 75-85 DEG C, controls moisture dry powder below 5%, and 0-15 DEG C of sealing is protected
It deposits；
(4) 8-10g dry powder is weighed when using, adds water 1000g, 121 DEG C of saturated vapors go out after agar powder is added according to hardness requirement
Bacterium can be used.
8. the modified PDA culture medium that preparation method described in any one of -7 is prepared according to claim 1.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910079892.1A CN109628329A (en) | 2019-01-28 | 2019-01-28 | A kind of modified PDA culture medium and preparation method thereof |
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| CN201910079892.1A CN109628329A (en) | 2019-01-28 | 2019-01-28 | A kind of modified PDA culture medium and preparation method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1077493A (en) * | 1993-05-10 | 1993-10-20 | 北京大学 | The manufacture method of potato dextrose agar dehydrated medium |
| EP1203811A2 (en) * | 2000-11-03 | 2002-05-08 | L'oreal | Production of metabolites of interest by co-culture of plant cells and non-plant cells |
| CN107043710A (en) * | 2017-03-04 | 2017-08-15 | 安徽莱姆佳生物科技股份有限公司 | Improved culture medium and its compound method for screening, purifying the pathogen of Botrytis cinerea |
-
2019
- 2019-01-28 CN CN201910079892.1A patent/CN109628329A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1077493A (en) * | 1993-05-10 | 1993-10-20 | 北京大学 | The manufacture method of potato dextrose agar dehydrated medium |
| EP1203811A2 (en) * | 2000-11-03 | 2002-05-08 | L'oreal | Production of metabolites of interest by co-culture of plant cells and non-plant cells |
| CN107043710A (en) * | 2017-03-04 | 2017-08-15 | 安徽莱姆佳生物科技股份有限公司 | Improved culture medium and its compound method for screening, purifying the pathogen of Botrytis cinerea |
Non-Patent Citations (1)
| Title |
|---|
| 田文等: "直投式泡菜乳酸菌发酵剂培养基的优化及离心条件的探索", 《食品工业》 * |
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Application publication date: 20190416 |