CN103436480A - Plate culture and preparation method of ustilaginoidea virens thin-wall conidium - Google Patents
Plate culture and preparation method of ustilaginoidea virens thin-wall conidium Download PDFInfo
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Abstract
The invention discloses a plate culture and preparation method of ustilaginoidea virens thin-wall conidium. The core of the method is that the thin-wall conidium is directly utilized for culturing and breeding new thin-wall conidium through a plate culture medium. The method comprises the following steps: 1) preparing an initial thin-wall conidium solution; 3) preparing a sporulation culture medium plate; 3) performing sporulation cultivation; 4) collecting new thin-wall conidium; and 5) purifying conidium. The method has the advantages that the cultivation and preparation can be performed within a shorter time which is generally 4 to 6 days; the operation is simple, easy and convenient; oscillating culture equipment is not required; the thin-wall conidium in an obtained culture product is mainly the conidium of the same generation; the conidium solution is relatively high in quality; no purifying treatment is required; and the method is applied to a plurality of research works.
Description
Technical field
The present invention relates to agrotechnique and biotechnology.Specifically the conidial flat board of a kind of ustilaginoidea virens thin-walled is cultivated the preparation method.
Background technology
In most of fungal disease systems, the propagulum of pathogenic bacteria enlarges the effect such as self population quantity except having breeding, the more important thing is to have with the host and identify mutually, and start to infect host's function, therefore in many important research of plant disease processes, all need to use the propagulum of pathogenic bacteria to make test materials, apparent, the efficient cultivation preparation method of propagulum material easily, will be very beneficial for carrying out smoothly of research work.False smut has become a kind of great disease of China's Rice Production, and the propagulum of ustilaginoidea virens includes the thecaspore of condition, without pachypycnidium and the thin-walled conidium three types of condition.In laboratory, yet there are no so far by cultivating this germ and obtain thecasporous effective report; Cultivation is obtained pachypycnidium and is also had larger technical difficulty, and there is dormancy in pachypycnidium, uses very inconvenient; Therefore, the propagulum that relevant institute so far uses is mainly the thin-walled conidium of using this germ.
So far, the conidial cultivation preparation method of the ustilaginoidea virens thin-walled that everybody knows, mainly contain 2 kinds.A kind of is the second stage of culture method, first the germ mycelium is cultivated at solid medium, then mycelium is proceeded to liquid nutrient medium and carries out shaking culture, and whole process generally needs more than 15 days.Another kind is three phase culture methods, first the germ mycelium is cultivated at solid medium, then mycelium is proceeded to liquid nutrient medium and carry out shaking culture, then pouring the shaking culture product into solid medium again carries out biphasic cultivation, and whole process generally needs more than 20 days.These 2 kinds of methods have common step, all need to carry out the liquid oscilaltion cultivation.
These 2 kinds of methods can effectively prepare ustilaginoidea virens thin-walled conidium, but all there are some disadvantages in they in practice, as consuming time longer; Work consuming is more; Need shaking culture equipment; Thin-walled conidium in cultured products belongs to many mixtures of spore from generation to generation; Include a large amount of hypha bodies, bacterial metabolism product and nutrient media components etc. in the cultured products mixed solution, often needing to filter the subsequent processing steps such as mycelia and precipitation spore just is applicable to using, because cultured products is the thickness state, the precipitation spore often needs the high speed centrifugation machine equipment simultaneously.
Summary of the invention
The objective of the invention is according to thering is the special characteristic of growing after ustilaginoidea virens thin-walled conidia germination, provide a kind of ustilaginoidea virens thin-walled conidial dull and stereotyped preparation method of cultivation.
The technical scheme that the present invention solves the problems of the technologies described above is as follows:
The conidial dull and stereotyped preparation method that cultivates of a kind of ustilaginoidea virens thin-walled, operation steps is as follows:
1. the preparation of initial thin-walled conidium liquid:
A) the conidial source of initial thin-walled: the second stage of cultural method of knowing with everybody, obtains and contains the conidial cultured products of a large amount of thin-walleds by the shaking culture mode with mycelium; Perhaps adopt the fresh pachypycnidium of cultivating on the sick grain of rice in field, obtain and contain the conidial cultured products of a large amount of thin-walleds.
B) spore purification process: the conidial cultured products of the thin-walled of acquisition is carried out to the spore purification process, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal treating collecting precipitation spore, can remove medium component and the outer secretion matter of thalline in cultured products; Then with sterilized water, will precipitate spore suspension washing, then carry out centrifugal treating one time, obtain pure spore ball.With sterilized water, the spore ball suspended dispersed is evenly become to thin-walled conidium liquid, save backup.
2. produce the preparation of spore culture medium flat plate:
What adopt the PSA substratum partly measures composition as producing the spore substratum, produces the spore substratum and is: potato 100g; Sucrose 10g; Agar 20g; Water 1000ml, by changing into the culture dish flat board after conventional sterilizing, make and produce the product spore flat board that spore is cultivated use.
3. producing spore cultivates:
Get step 1) the thin-walled conidium drop that saves backup on the product spore culture medium flat plate face of getting ready, by level and smooth glass stick trowelling platen surface, make that spore is full and uniform to be distributed on platen surface, returning after lid and proceeding to temperature is to cultivate 5 days under 28 ℃ of conditions.
4. the conidial collection of new thin-walled:
Upper step was cultivated after 5 days, produced on the spore culture medium flat plate and grew the conidial small colonies of generation thin-walled that a large amount of diameters are about 0.5～1.5mm, now collected spore for the most in good time machine.Collection method is to get clear water and annotate on product spore platen surface, then, scrub and produce spore culture medium flat plate surface colony with common hairbrush, making spores release on bacterium colony in water, the water liquid of scrubbing spore is collected, is cultured products ustilaginoidea virens thin-walled conidium.
5. thin-walled conidium purification process:
Collected cultured products thin-walled conidium is carried out to the spore purification process, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal treating collecting precipitation spore, can remove medium component and the outer secretion matter of thalline in cultured products; Then with sterilized water, will precipitate spore suspension washing, then carry out centrifugal treating one time, and can obtain very pure thin-walled and divide spore ball.With sterilized water, this spore ball suspended dispersed is evenly become to ustilaginoidea virens thin-walled minute spore liquid.
Advantage of the present invention is:
1. cultivate preparation process time shorter, generally only need 4～6 days;
2. it is simple and easy convenient to operate, without shaking culture equipment;
3. the thin-walled conidium in the cultured products that obtained is mainly the spore of same generation;
4. the spore liquid quality is higher, without relevant purifying treatment, also is applicable to the use of many research work.
The accompanying drawing explanation
Fig. 1 is the early stage schematic diagram of thin-walled conidia germination.
In figure, A is that spore starts to sprout the generation germ tube; B is that germ tube that spore germination produces is constantly grown and formed the sporogenous hyphae of multi-branched; Black scale short-term 10 μ m.
Fig. 2 is that the sporogenous hyphae of multi-branched progressively produces thin-walled conidium figure.
In figure, black scale line 0.4mm.
Fig. 3 is that sporogenous hyphae is constantly divided branch growth and produces spore, forms the small colonies figure that a mycelia and spore are intensive.
In figure, mycelia and the spore of bacterium colony periphery are rarer, the good and imaging clear (A) of printing opacity, and mycelia and the spore at bacterium colony middle part are intensive and light tight, can't imaging (B).Black scale line: 0.6mm.
Fig. 4 is that Fig. 3 amplifies the bacterium colony figure of 2 times.
In figure, A: well reach on the camera lens focal plane more clearly spore string (group) in printing opacity, B: in printing opacity better and depart from the camera lens focal plane not clearly spore string (group); C: be in the light by thick close bacterium colony itself, the bacterium colony position that can't focus.Black scale line: 0.4mm.
Fig. 5 is the partial enlarged drawing of Fig. 3 bacterium colony.
In figure, A: spore string (group) more clearly on colony edge printing opacity good Huan district and camera lens focal plane, B: in printing opacity Cha Huan district and depart from the camera lens focal plane, not clearly intensive spore string (group) slightly; C: in mycelia and the thick close zone of spore, light tight, the imaging of can't focusing.Black scale line: 0.2mm.
Fig. 6 is that ustilaginoidea virens is producing a large amount of small colonies figure of formation on the spore culture medium flat plate.
In figure, be after the thin-walled conidium is applied in and produces on the spore substratum and cultivate 5 days, each spore germination growth, and produce spore of new generation, form the bacterium colony that diameter is about 0.5～1mm.
Fig. 7 is the mycelia figure that produces spore peak bacterium colony periphery later.
In figure, white scale line: 0.4mm.
Fig. 8 is that amplify the part of Fig. 7, shows that colony edge does not have sporulation in the mycelia zone clearly.
In figure, black scale line: 0.2mm.
Fig. 9 is that the inventive method is cultivated the thin-walled conidium liquid figure obtained.
In figure, the outward appearance of the inventive method cultured products (A): liquid is limpid evenly, without obvious hypha body; Adopt the outward appearance of shaking culture method cultured products (B): the liquid thickness has a large amount of hypha bodies.
Embodiment
Below in conjunction with accompanying drawing and embodiment, the invention will be further described.
Growth and development process uniqueness after ustilaginoidea virens thin-walled conidia germination.Contriver's discovery, a thin-walled conidium sprouts on the nutrient agar flat board, generally first produces germ tube, and germ tube produces the sporogenous hyphae (as shown in Figure 1) of multi-branched afterwards, then on sporogenous hyphae, forms spore of new generation (as shown in Figure 2); Then, sporogenous hyphae is branch and large volume production spore more fast constantly, soon, sporogenous hyphae and spore set form macroscopic minute colony, colony diameter when the product spore peaks is about 0.5～1.5mm, bacterium colony now is visible a large amount of intensive spore of new generation (as shown in Fig. 3～Fig. 5) under the microscope, and on culture medium flat plate, naked eyes directly are viewed as small particles (as shown in Figure 6).After this, sporogenous hyphae grows changes the vegetative hyphae (as Fig. 7, shown in Fig. 8) that no longer produces spore into, and step by step, mycelia is increased, and bacterium colony increases, and the spore amount descends, until can't detect spore.The conidial dull and stereotyped preparation method that cultivates of a kind of ustilaginoidea virens thin-walled of the present invention, utilize the conidial this germination and growth characteristic of thin-walled exactly, at the initial stage of spore germination grown cultures, collects in time, obtains a large amount of new thin-walled conidiums.
The second stage of cultural method that the first conidial source of thin-walled of using, 1) can adopt everybody to know,, obtain and contain the conidial cultured products of a large amount of thin-walleds by the shaking culture mode with mycelium.2) also can adopt the spore method for preparing of the present invention, cultivate the fresh pachypycnidium (chlamydospore) that field gathers, obtain and contain the conidial cultured products of a large amount of thin-walleds; This is based on after pachypycnidium is sprouted and is easy to produce the thin-walled conidium, and contriver's discovery, and the characteristic of growing after pachypycnidium is sprouted is the same with the characteristic of growing after the thin-walled conidia germination.
Two kinds of methods in the first conidial source of thin-walled of using of above-mentioned explanation can obtain and contain the conidial cultured products of thin-walled; By after the spore purification process in this cultured products, can prolonged preservation standby in sterilized water; This is based on the contriver and finds, simple thin-walled conidium is deposited in sterilized water, preserves after 4 years and still has spore energy germination and growth to produce normal offspring, therefore, can simple thin-walled conidium be made into to spore liquid with sterilized water, and prolonged preservation is standby.
Embodiment 1
Adopt a kind of ustilaginoidea virens thin-walled of the present invention conidium to cultivate the preparation method, cultivate the thin-walled conidium for preparing ustilaginoidea virens bacterial strain Uv-34, as follows operation:
1) preparation of initial thin-walled conidium liquid:
A) the conidial first source of thin-walled: the second stage of cultural method that adopts everybody to know, obtains and contains the conidial cultured products of a large amount of thin-walleds by the shaking culture mode with mycelium;
B) spore purification process: what will obtain carries out the spore purification process containing the conidial cultured products of thin-walled, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal treating collecting precipitation spore, can remove medium component and the outer secretion matter of thalline in cultured products; Then with sterilized water, will precipitate spore suspension washing, then carry out centrifugal treating one time, obtain pure spore ball.With sterilized water, that the spore ball suspended dispersed is even, and to be deployed into concentration be 10
6the spore liquid of individual spore/ml saves backup.
2) produce the preparation of spore culture medium flat plate:
What adopt the PSA substratum partly measures composition as producing the spore substratum, produces the spore medium component and is: potato 100g; Sucrose 10g; Agar 20g; Water 1000ml, after conventional sterilizing, pour the 9cm culture dish into and make the product spore flat board that produces spore cultivation use.
3) producing spore cultivates:
Get step 1) the thin-walled conidium liquid 10 μ l that save backup, drip on the product spore culture medium flat plate face of getting ready, with level and smooth glass stick trowelling platen surface, make that spore is full and uniform to be distributed on platen surface, returning after lid and proceeding to temperature is cultivation 5 days under 28 ℃ of conditions.
4) the conidial collection of new thin-walled:
Upper step was cultivated after 5 days, produced on the spore culture medium flat plate and grew the generation thin-walled conidium small colonies (as shown in Figure 6) that a large amount of diameters are about 0.5～1.5mm, now for collecting new thin-walled the most in good time conidial machine.Collection method is to get clear water and annotate on product spore platen surface, then, scrub and produce spore culture medium flat plate surface colony with common hairbrush, make spores release on bacterium colony in water, each culture plate water 15ml scrubs 2 times, collects the cultured products of common acquisition 30ml; This cultured products contains a large amount of new thin-walled conidiums, and spore concentration reaches 1.89 * 10
6individual spore/ml.
Although this cultured products also contains the outer secretion of a small amount of mycelia, nutrient media components and thalline, but the spore cultured products obtained with the shaking culture mode is compared, spore liquid product appearance character of the present invention is limpid evenly, without significantly rolling into a ball saccharoid, move liquid or packaging operation good (as shown in Figure 9), thereby the spore liquid quality is higher, without relevant purification step, also be applicable to the use of many research work, very convenient.
5) thin-walled conidium purification process:
Collected cultured products is carried out to thin-walled conidium purification process, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal treating collecting precipitation spore, can remove medium component and the outer secretion matter of thalline in cultured products; Then with sterilized water, will precipitate spore suspension washing, then carry out centrifugal treating one time, can obtain very pure thin-walled conidium group; With sterilized water, this spore ball suspended dispersed is evenly become to ustilaginoidea virens thin-walled conidium liquid.
Embodiment 2
Adopt a kind of ustilaginoidea virens thin-walled of the present invention conidium to cultivate the preparation method, cultivation prepares the thin-walled conidium of ustilaginoidea virens bacterial strain Uv-105, pressing step 1) to the step 4) operation of embodiment 1 implements, the dull and stereotyped cultured products 30ml obtained containing thin-walled conidium liquid of the every ware of result, the spore concentration of cultured products reaches 2.72 * 10
6individual spore/ml; The appearance character of this cultured products is identical with embodiment's 1;
After pressing the step 5) operation enforcement of embodiment 1, obtain the very pure thin-walled conidium of bacterial strain Uv-105.
Embodiment 3
Adopt a kind of ustilaginoidea virens thin-walled of the present invention conidium to cultivate the preparation method, cultivation prepares the thin-walled conidium of ustilaginoidea virens bacterial strain Uv-110, pressing step 1) to the step 4) operation of embodiment 1 implements, the every ware of result obtains the cultured products 30ml containing thin-walled conidium liquid, and the spore concentration of cultured products reaches 2.15 * 10
6individual spore/ml; The appearance character of this cultured products is identical with embodiment's 1;
After pressing the step 5) operation enforcement of embodiment 1, obtain the very pure thin-walled conidium of bacterial strain Uv-110.
Embodiment 4
Adopt a kind of ustilaginoidea virens thin-walled of the present invention conidium to cultivate the preparation method, cultivate the thin-walled conidium for preparing ustilaginoidea virens bacterial strain Uv-203, as follows implementation and operation:
1) preparation of initial thin-walled conidium liquid:
The conidial first source of thin-walled: the fresh pachypycnidium (chlamydospore) gathered from field, cultivate to obtain and contain the conidial cultured products of a large amount of thin-walleds; Concrete grammar is: gather the fresh rice curve in field (the sick grain of rice of false smut) and go back to laboratory; Under aseptic condition, clamp the rice curve with tweezers, knock gently tweezers, shake and abandon the pachypycnidium on rice curve top layer, then the rice curve is moved on to and produce spore substratum top, then knock tweezers, the pachypycnidium on the rice curve is shaken to fall to interspersing among and cultivate on basal plane, cover the culture dish lid, cultivate under the condition that to be placed in temperature be 28 ℃, after 5 days, cultivate on basal plane and the white small colonies that many diameters are 0.5～1mm occurs, simultaneously, some contaminants or bacterial colony also appear; By sterile razor blade, free of contamination ustilaginoidea virens small colonies is transferred in sterile petri dish together with culture medium flat plate cutting, added a small amount of sterilized water, with the sterilizing hairbrush, scrub gently small colonies, obtain and contain the conidial initial spore liquid of a large amount of thin-walleds; This spore liquid can adopt method of the present invention without purification process, directly is used for cultivating the new thin-walled conidium of preparation; Ensuing operation steps is as follows:
2) produce the preparation of spore culture medium flat plate: the step 2 of pressing embodiment 1) operation is implemented;
3) product spore cultivation: get the present embodiment step 1) cultivate the thin-walled conidium liquid 10 μ l that obtain, drip on the product spore culture medium flat plate face of getting ready, by level and smooth glass stick trowelling platen surface, make that spore is full and uniform to be distributed on platen surface, returning after lid and proceeding to temperature is to cultivate under 28 ℃ of conditions.
4) the conidial collection of new thin-walled: the step 4) operation of pressing embodiment 1 is implemented; The dull and stereotyped cultured products 30ml obtained containing thin-walled conidium liquid of the every ware of result, the spore concentration of cultured products reaches 2.52 * 10
6individual spore/ml; The appearance character of this cultured products is identical with embodiment's 1;
5) thin-walled conidium purification process: after pressing the step 5) operation enforcement of embodiment 1, obtain very pure thin-walled conidium; This bacterial strain is that directly the Pathogen culture from the sick grain of rice in field obtains, and names as Uv-203.
Embodiment 5
Adopt a kind of ustilaginoidea virens thin-walled of the present invention conidium to cultivate the preparation method, cultivate the thin-walled conidium for preparing ustilaginoidea virens bacterial strain Uv-205.Press the step 1) of embodiment 4 and implement to the step 4) operation, the every ware of result obtains the cultured products 30ml containing thin-walled conidium liquid, and the spore concentration of cultured products is 1.25 * 10
6individual spore/ml; The appearance character of this cultured products is identical with embodiment's 1;
After pressing the step 5) operation enforcement of embodiment 1, obtain very pure thin-walled conidium; This bacterial strain is that directly the Pathogen culture from the sick grain of rice in field obtains, and names as Uv-205.
Claims (1)
1. the conidial dull and stereotyped preparation method that cultivates of ustilaginoidea virens thin-walled, is characterized in that, operation steps is as follows:
1) preparation of initial thin-walled conidium liquid:
A) the conidial source of initial thin-walled: the second stage of cultural method that adopts everybody to know, obtains and contains the conidial cultured products of a large amount of thin-walleds by the shaking culture mode with mycelium; Perhaps adopt the method for cultivating the fresh pachypycnidium on the sick grain of rice in field, obtain and contain the conidial cultured products of a large amount of thin-walleds;
B) spore purification process: the conidial cultured products of the thin-walled of acquisition is carried out to the spore purification process, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal and collecting precipitation spore; Then with sterilized water, will precipitate spore and suspend and wash, then carry out once centrifugally, obtain pure spore ball, with sterilized water, the spore ball suspended dispersed evenly be become to thin-walled conidium liquid, save backup;
2) produce the preparation of spore culture medium flat plate:
What adopt the PSA substratum partly measures composition as producing the spore substratum, produces the spore substratum and is: potato 100g; Sucrose 10g; Agar 20g; Water 1000ml, by changing into the culture dish flat board after conventional sterilizing, make and produce the product spore flat board that spore is cultivated use;
3) producing spore cultivates:
Get step 1) the thin-walled conidium liquid that saves backup, drip on the product spore culture medium flat plate face of getting ready, with level and smooth glass stick trowelling platen surface, make that spore is full and uniform to be distributed on platen surface, returning after lid and proceeding to temperature is cultivation 5 days under 28 ℃ of conditions;
4) the conidial collection of new thin-walled:
Upper step was cultivated after 5 days, produced on the spore culture medium flat plate and grew the conidial small colonies of generation thin-walled that a large amount of diameters are about 0.5～1.5mm, now collected new thin-walled conidium; Collection method is, get clear water and annotate and to produce on the spore platen surface, then with common hairbrush, scrub product spore culture medium flat plate surface colony, make spores release on bacterium colony in water, the water liquid of scrubbing spore is collected, be and contain a large amount of new conidial cultured products of ustilaginoidea virens thin-walled;
5) thin-walled conidium purification process:
Collected cultured products is carried out to thin-walled conidium purification process, and purification treating method is to remove the mycelia in cultured products by 2 layers of filtered through gauze; Then filtered liquid is carried out to centrifugal treating collecting precipitation spore; Then with sterilized water, will precipitate spore suspension washing, then carry out centrifugal treating one time, can obtain very pure thin-walled conidium group, with sterilized water, this spore ball suspended dispersed evenly be become to ustilaginoidea virens thin-walled conidium liquid.
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| CN201310378599.8A CN103436480B (en) | 2013-08-27 | 2013-08-27 | Plate culture and preparation method of ustilaginoidea virens thin-wall conidium |
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| CN201310378599.8A CN103436480B (en) | 2013-08-27 | 2013-08-27 | Plate culture and preparation method of ustilaginoidea virens thin-wall conidium |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105779372A (en) * | 2016-05-24 | 2016-07-20 | 广西大学 | Potato culture medium suitable for ustilaginoidea virens spore production and application thereof |
| CN106085945A (en) * | 2016-08-24 | 2016-11-09 | 文山苗乡三七科技有限公司 | A kind of Radix Notoginseng Northern leaf spot bacterium conidium abductive approach |
| CN106635841A (en) * | 2017-01-10 | 2017-05-10 | 广西大学 | Standing form of botrytis cinerea laboratory material and preparation and application method of standing form |
| CN106635948A (en) * | 2017-01-10 | 2017-05-10 | 广西大学 | Pollution-free preparing and culturing method for rice-false-smut-case thin-wall conidia |
| CN106701653A (en) * | 2017-01-10 | 2017-05-24 | 广西大学 | Pollution-reduction preparation and culture method of ustilaginoidea virens thin-wall conidia |
| CN106867957A (en) * | 2017-01-10 | 2017-06-20 | 广西大学 | A kind of ustilaginoidea virens thin-walled of Pollution protection is conidial to prepare cultural method |
| CN113308432A (en) * | 2021-06-15 | 2021-08-27 | 广西大学 | Transfer and dispersion method of ustilaginoidea virens thin-walled conidia |
| CN113308430A (en) * | 2021-06-15 | 2021-08-27 | 广西大学 | Dispersion culture method of banana colletotrichum gloeosporioides conidia |
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| CN101875906A (en) * | 2010-08-04 | 2010-11-03 | 福建省农业科学院植物保护研究所 | Simple and convenient method for quickly separating ustilaginoidea virens |
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Cited By (8)
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| CN105779372A (en) * | 2016-05-24 | 2016-07-20 | 广西大学 | Potato culture medium suitable for ustilaginoidea virens spore production and application thereof |
| CN106085945A (en) * | 2016-08-24 | 2016-11-09 | 文山苗乡三七科技有限公司 | A kind of Radix Notoginseng Northern leaf spot bacterium conidium abductive approach |
| CN106635841A (en) * | 2017-01-10 | 2017-05-10 | 广西大学 | Standing form of botrytis cinerea laboratory material and preparation and application method of standing form |
| CN106635948A (en) * | 2017-01-10 | 2017-05-10 | 广西大学 | Pollution-free preparing and culturing method for rice-false-smut-case thin-wall conidia |
| CN106701653A (en) * | 2017-01-10 | 2017-05-24 | 广西大学 | Pollution-reduction preparation and culture method of ustilaginoidea virens thin-wall conidia |
| CN106867957A (en) * | 2017-01-10 | 2017-06-20 | 广西大学 | A kind of ustilaginoidea virens thin-walled of Pollution protection is conidial to prepare cultural method |
| CN113308432A (en) * | 2021-06-15 | 2021-08-27 | 广西大学 | Transfer and dispersion method of ustilaginoidea virens thin-walled conidia |
| CN113308430A (en) * | 2021-06-15 | 2021-08-27 | 广西大学 | Dispersion culture method of banana colletotrichum gloeosporioides conidia |
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