CN103103136A - Effective Ustilaginoidea virens separation method - Google Patents
Effective Ustilaginoidea virens separation method Download PDFInfo
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Abstract
The invention relates to an effective Ustilaginoidea virens separation method, and belongs to the technical field of microbe. The method comprises the steps of: scraping a small amount of chlamydospore powder on surfaces of false smut balls with a pricking needle; placing the powder in a sterile petri dish; adding 0.5mL of sterile water suspension chlamydospores; then diluting the chlamydospore suspension by 2-3 gradients, with the ratio of the chlamydospore suspension to sterile water being 1:5 in a 1.8mL Eppendorf tube through a gradient dilution method; transferring 0.2mL of the diluted chlamydospore suspension to surface of a Wakimoto toceshi solid culture medium for separation (containing 50 mug / ml chloramphenicol); smearing evenly; inversely placing the petri dish into an incubator at 28 DEG C for dark culture; and effectively separating out target pathogens six days later. The method provided by the invention obviously weakens interference effect of infectious microbe in chlamydospores of rice false smut balls, has very simple operation procedure, is suitable for separation of rice false smut balls stored at normal temperature or 4 DEG C in a refrigerator within 3 months, and has the best separation effect on rice false smut ball bacteria just acquired in the field.
Description
Technical field
The present invention relates to a kind of effective ustilaginoidea virens separation method, belong to microbial technology field.
Background technology
It is sick that rice green smut is commonly called as good harvest, is a kind of important disease of Rice Panicle, and its pathogenic bacteria is that the green pyrenomycetes of Deuteromycotina belongs to green pyrenomycetes
Ustilaginoidea virens(Cook.) Tak.in recent years, along with wideling popularize of dense cluster type high yield and high quality rice, false smut generation area and the degree that causes harm increase the weight of gradually, the generation of false smut has a significant effect to yield and quality of rice, rice curve quantity on the sick fringe in field is usually at 1～10, many meetings reach 30～50, rice curve chlamydospore powder is blackish green or black, sneak into severe contamination paddy after paddy, impact rice matter and outward appearance, and mix rice curve in sick paddy and germ and contain toxin to the toxic effect of people and animals, false smut has now become the important disease on Rice Production, seriously restricting the safety in production of China paddy rice.
The material that is used at present the ustilaginoidea virens separation has rice curve and sclerotium (being produced by minority rice curve), but for sclerotium, field rice curve quantity is many, easily gathers.Aspect genetic diversity, Variation mechanism and the Study on Pathogenicity of different habitats germ, the different monospore isolates of same rice curve, adopt the rice curve to demonstrate even more important effect as the material that germ separates.
Ustilaginoidea virens belongs to the weak fungi of the stronger saprophytic property of a kind of parasitics, and the rice curve is nutritious, normal a large amount of saprophytic bacteria, fungi and other pathogenic bacterias of association, and this bacteria growing is slow in addition, easily is subject to living contaminants in sepn process, and Success rate of virus isolation is very low.Utilize at present method that the rice curve separates germ partition method, spore shake method, chlamydospore suspension method in a organized way, its semilate rice curve internal layer separate tissue fado studied person adopt.Yet in actual mechanical process, required incubation time is longer, and velocity of separation is slower, and success ratio is lower, and Yin Qichang follows the appearance of the miscellaneous bacterias such as other bacterium, fungi, affects the effect of the further separation of ustilaginoidea virens and purifying.The spore method of shaking, directly the chlamydospore on rice curve surface directly to be shaken to drop down onto on separatory substratum, because the rice curved surface is not disinfected, the a large amount of miscellaneous bacteria of rice curved surface is also easily shaken to be dropped on media surface, and Fast Growth, affect the purifying of chlamydospore sprouting, growth and bacterium colony.The chlamydospore suspension method, must sprout into mycelia and pass substratum because being mixed in chlamydospore in substratum, extend the time that chlamydospore forms single bacterium colony, easily caused the too fast miscellaneous bacteria of the speed of growth to be covered on single bacterium colony of a small amount of steamed bun shape, strengthened the difficulty of separation and purification.Liu Ming rosy clouds 2009 adopt respectively tissue isolation with the chlamydospore suspension method, rice curve germ to be separated, result shows that success ratio that two kinds of methods are separated only has respectively 8.5% and 5.3%(Liu Ming rosy clouds, 2009, ustilaginoidea virens isolation technique, culture condition research, Liaoning agricultural sciences, 2009. (6): 20-22).a kind of easy ustilaginoidea virens Rapid Isolation (ZL201010301356.0) is patent of invention of applicant, its technical characteristics is after rice curve surface disinfectant is processed, smash the rice curve to pieces with sterilized water, make and smash liquid to pieces and contain chlamydospore and internal layer tissue block, then rinse separatory side of body Ben Zheshi media surface with smashing liquid to pieces, can isolate rapidly ustilaginoidea virens, but the method has been used ethanol and mercuric chloride sterilizing agent in sepn process, due to chlamydospore to the comparatively responsive (Wang Yongqiang of sterilizing agent, 2010, the comparative studies of the green pyrenomycetes of rice (ustilaginoidea virens) separation method, fungus journal 2010, 29(1), adopt this method longer to the shelf-time, the rice curve germ that the chlamydospore germination is lower separates obvious impact.
the rice curve that the field gathers was deposited 3 months, its surperficial chlamydospore still has certain sprouting ability, though there is more miscellaneous bacteria association on rice curve surface, but population quantity is not as good as chlamydospore, therefore the objective of the invention is on patent ZL201010301356.0 technical characterictic basis, utilize the chlamydosporic Germination characteristics of rice curve and predominance thereof, by technological improvement, weaken the interference effect of miscellaneous bacteria in rice curve chlamydospore, avoid simultaneously sterilizing agent to chlamydosporic lethal effect, increase the success ratio of separating, reduce sepn process and step, and provide a kind of more easy, more efficiently ustilaginoidea virens separation method, for the false smut research worker.
Summary of the invention
The object of the present invention is to provide a kind of more easy, effective ustilaginoidea virens separation method, can isolate in a large number, rapidly ustilaginoidea virens in a short time from the rice curve, for the research of ustilaginoidea virens provides the basis.
The objective of the invention is to realize as follows:
(1) rice curve standard specimen is selected: choose the fresh yellow, yellow-green colour or the green rice curve that gather from the field, or choose with the paper bag packing and dry, and normal temperature or 4 ℃ of Refrigerator stores, the shelf time is no more than the rice curve of 3 months;
(2) preparation of chlamydospore suspension: with the aseptic lip-deep chlamydospore powder of pin scraping simple grain rice curve of choosing, be placed in sterile petri dish, add the sterilized water suspension of 0.5mL to make chlamydospore suspension;
(3) dilution of chlamydospore suspension: press after 2～3 gradients of 1:5 dilution proportion chlamydospore suspension and sterilized water standby;
(4) ustilaginoidea virens separates: inhale step (3) with liquid-transfering gun and diluted good chlamydospore suspension 0.2mL in separating with side of body Ben Zheshi solid culture primary surface, and evenly smear with aseptic glass triangle rod, the culture dish that will contain again above-mentioned substratum is inverted into 28 ℃ of dark culturing in thermostat container, observes day by day separating resulting;
(5) ustilaginoidea virens purifying: cultivate after the 6th day, choose the yellow or white small colonies of pin picking with tip and move to cultivate with purifying in the PSA solid medium and cultivated 10 days;
(6) ustilaginoidea virens is preserved: according to growth characteristics and the microscopic examination result thereof of germ at substratum, with ustilaginoidea virens move to cultivate 10 days in the PSA slant medium after, be stored in 4 ℃ of refrigerators.
Described separation with the formula of side of body Ben Zheshi substratum is: potato 300g, peptone 5g, sucrose 15g, Ca (NO
3)
24H
2O 0.5g, Na
2HPO
412H
20 2.0g, agar 16g, Jia Shui are settled to 1000mL, at 121 ℃ of lower high pressure moist heat sterilization 20～25min, add the paraxin 50 μ g/ml mixings of bacteria growing inhibiting when substratum is cooled to 45 ℃～50 ℃, pour into solidify in culture dish rear standby.
Described cultivation with the formula of PSA solid medium is: potato 200g, agar 20g, sucrose 20g, Jia Shui are settled to 1000mL, and be standby after 121 ℃ of lower high pressure moist heat sterilization 20min.
Adopt the present invention for Bemisia tabaci bacterial parasite Verticillium lecanii (
Verticillium lecanii) (from infected Bemisia tabaci Nymph body surface) separation, for Pyricularia oryzae (
Pyricularia grisea) the monospore separation of (after the moisturizing 24 of the panicle blast branch of bacteria infection stalk, can produce a large amount of conidiums) is also comparatively applicable.
The chlamydosporic Germination characteristics of rice curve and population quantity advantage thereof within 3 months are deposited in utilization of the present invention, adopt the miscellaneous bacteria quantity in thorium scraping and sterilized water gradient dilution technology minimizing chlamydospore suspension of choosing of chlamydospore powder, weaken the interference effect of miscellaneous bacteria, avoided simultaneously sterilizing agent to chlamydosporic lethal effect, ensured the original sprouting ability of chlamydospore, and provide a kind of more easy, economize material, practicality, effective ustilaginoidea virens separation method.
Description of drawings
Fig. 1 is the monospore isolate of ustilaginoidea virens; A represents the separation and Culture monospore small colonies of the 6th day, and plate is pollution-free; B represents the separation and Culture monospore small colonies of the 6th day, and plate has pollution, but does not affect the purifying of bacterium colony; C represents the separation and Culture monospore bacterium colony of the 10th day, and colonial morphology is clear.
Fig. 2 is the dull and stereotyped colonial morphology of cultivating of ustilaginoidea virens.
Fig. 3 is the form of ustilaginoidea virens liquid culture.
Fig. 4 is ustilaginoidea virens mycelia and conidium thereof.
Fig. 5 is ustilaginoidea virens conidia germination figure, and expression separates germ sprouting 12h, separation germ sprouting 24h respectively from left to right, known germ sprouts 24h.
Embodiment
Embodiment 1
(1) rice curve standard specimen source: preserve the rice curve of 3 months under normal temperature.
(2) substratum preparation
A: separatory substratum adopts side of body Ben Zheshi substratum: potato 300g, peptone 5g, sucrose 15g, Ca (NO3) 24H2O 0.5g, Na2HPO412H:0 2.0g, agar 16g, Jia Shui are settled to 1000mL.Substratum adds paraxin (the 50 μ g/ml) mixing of bacteria growing inhibiting, and pours in culture dish at 121 ℃ of lower high pressure moist heat sterilization 20min when substratum is cooled to 45 ℃～50 ℃, standby after culture medium solidifying.
B: the substratum of cultivating use adopts PSA solid medium, PS liquid nutrient medium.The PSA solid medium is that potato 200g, agar 20g, sucrose 20g, Jia Shui are settled to 1000mL.The PS liquid nutrient medium is that potato 200g, sucrose 20g, Jia Shui are settled to 1000mL.Above substratum is all standby after 121 ℃ of lower high pressure moist heat sterilization 20min.
(3) preparation of chlamydospore suspension: clamp the rice curve with aseptic nipper, with the aseptic pin of choosing with the chlamydospore powder scraping a little (approximately 0.005g) of certain, rice curve surface, make it to be placed in sterile petri dish (diameter=8.9cm), add the sterilized water of 0.5mL to suspend, make chlamydospore suspension.In the Eppendorf of 1.8mL pipe, draw 0.1ml chlamydospore suspension and 0.5ml sterilized water with liquid-transfering gun and press after 3 gradients of 1:5 dilution proportion standbyly, make the ultimate density of chlamydospore powder in diluent be about 4.6 * 10
-5G/mL.
(4) ustilaginoidea virens separates: inhale with liquid-transfering gun and diluted good chlamydospore suspension 0.2mL in separatory side of body Ben Zheshi solid culture primary surface, and evenly smear with aseptic glass curved rod.The culture dish that will contain substratum is inverted into 28 ℃ of dark culturing in thermostat container, observes day by day separating resulting.
(5) ustilaginoidea virens purifying: cultivate after the 6th day, choose the yellow or white small colonies (as shown in Figure 1) of pin picking with tip and move to cultivate with purifying in the PSA substratum and cultivated 10 days.
(6) ustilaginoidea virens is preserved: according to growth characteristics and the microscopic examination result thereof of germ at substratum, with ustilaginoidea virens move to cultivate 10 days in the PSA slant medium after, be stored in 4 ℃ of refrigerators.
(7) ustilaginoidea virens growth and microscopic examination
Get the purifying cultivation germ of 7 days and carry out the observation of flat board cultivation present situation, observe colony morphology characteristic and the speed of growth, as shown in Figure 2; The germ bacterium piece that the picking purifying is cultivated is inoculated in the PS liquid nutrient medium, and 28 ℃ of shaking culture are after 7～10 days, and form and the mycelia of observing respectively mycelium pellet produce the spore situation.
(8) the conidia germination characteristic is observed
Draw a small amount of PS liquid nutrient medium shaking culture conidium liquid of 10 days in PSA solid culture primary surface, after evenly smearing, the culture dish that will contain substratum is inverted in thermostat container 28 ℃ of dark culturing 12h or 24 h, 400 times of Microscopic observation ustilaginoidea virens Conidium germination of P.grisea situations, and make comparisons with the conidia germination feature of known ustilaginoidea virens.
As shown in Fig. 3,4,5, according to the observation of isolate cultural characteristic, spore shape microscopic examination and conidia germination characteristic observations thereof, can identify isolate is ustilaginoidea virens.Adopt false smut indoors artificial inoculation method, can cause that for the isolate that tries paddy rice produces the rice curve.(Yang Xiujuan, 2011, rice green smut indoors artificial inoculation technique. plant protection journal, 2011,38 (5): 395-400), show thus and adopt the inventive method can effectively isolate ustilaginoidea virens from the rice curve.
Several ustilaginoidea virens separation method of embodiment 2 result relatively
1, the effective partition method of ustilaginoidea virens of the present invention, its step is as follows:
1) separation of ustilaginoidea virens
With embodiment 1
2) ustilaginoidea virens growth, conidia germination characteristic and microscopic examination
With embodiment 1
2, chlamydospore suspension method
With reference to Zhou Yongli 1999 method (Zhou Yongli, 1999, the pre-test of ustilaginoidea virens isolation technique, the rice in China science, 1999.13 (3): l86-188), being the rice curve will sterilize after 30s with 75% ethanol, use again aseptic washing 3 times, then be placed in the vibration of Eppendorf pipe and be mixed with chlamydospore suspension, get 2～3 droppings in culture dish, pour in culture dish when substratum is cooled to 45 ℃～50 ℃, shake gently and make it and the spore suspension mixing, treat that culture medium solidifying is placed on 28 ℃ of dark culturing in thermostat container, observes separating resulting day by day.
3, patent ZL201010301356.0 partition method
Simple grain rice curve with 75% ethanol disinfection 1min, 0.1% mercuric chloride sterilization 2min after, then use sterile water wash 3 times.Then be placed in culture dish (d=5cm), add sterilized water 4mL, after smashing to pieces with the aseptic operation cutter, be tissue mashing liquid (include chlamydospore and organize fragment).Again with liquid-transfering gun draw smash to pieces liquid 1mL(contain organize fragment) rinse in the culture dish that contains side of body Ben Zheshi substratum (d=9cm), make media surface smash liquid to pieces and cover and organize fragment to disperse, suck subsequently the remaining liquid of smashing to pieces.And under the clean worktable aseptic condition of behaviour air-dry media surface moisture content, the culture dish after air-dry is placed in 28 ℃ of dark culturing of thermostat container, observes day by day separating resulting.
4, implementation result
1) the effective partition method of ustilaginoidea virens of the present invention, separation and Culture just yellow and white single bacterium colony of visible more polymorphic consistent, clear, cleaning-less bacteria infection after the 6th day, separation and purification is easy.These purifieds are ustilaginoidea virens through flat board cultivation, liquid culture feature, spore and mycelia microscopic examination and the equal susceptible of proof of conidia germination characteristic observation thereof.after the rice curve that the field gathers is deposited 3 months, its surperficial chlamydospore still has and sprouts preferably ability, though there is more miscellaneous bacteria association on rice curve surface, but population quantity is not as good as chlamydospore, in operating process owing to not using rice curve surface disinfectant, avoided sterilizing agent to chlamydosporic lethal effect, adopting simultaneously rice curve chlamydospore powder to choose pin scrapes to follow the example of with sterilized water gradient dilution method and reduces miscellaneous bacteria quantity in chlamydospore suspension, weakened the interference effect of miscellaneous bacteria in the chlamydospore suspension, make dull and stereotyped of reduced contamination, even pollution-free, the single colonial morphology that occurs is clear, be easy to purifying.And chlamydospore is to be uniformly dispersed in media surface, the oxygen when having guaranteed chlamydospore sprouting and mycelial growth, thereby got final product sharp separation to ustilaginoidea virens in 6 days.Adopt this method best to the rice curve separating effect that the field has just gathered, also be specially adapted to normal temperature or 4 ℃ of refrigerators and deposit and separated with interior yellow, yellow-green colour and green rice curve germ in 3 months, the sample success ratio of separation can reach more than 85%.
2) adopt the chlamydospore suspension method, because the ustilaginoidea virens growth is very slow, being mixed in chlamydospore in substratum must sprout into mycelia and pass substratum, need 10 talentes to grow the single bacterium colony (skewness of a small amount of white, bacterium colony occur to be assembled), the white that occurs simultaneously growing faster or sap green miscellaneous bacteria are covered on single bacterium colony of steamed bun shape.Chlamydospore is comparatively responsive to temperature, and with the rising of temperature, germination rate sharply descends after 28 ℃, and 45 ℃ germinate hardly, and 50 ℃ is fatal temperature.Adopt chlamydospore suspension method and substratum (45 ℃) mode of averaging, when the substratum excess Temperature can have lethal effect to spore, temperature is too low cause substratum and spore suspension method mixing insufficient, cause white single bacterium colony skewness, assembling appears in bacterium colony, when cultivating 10 days, the bacterium colony forming position the too fast miscellaneous bacteria of the speed of growth often occurs and covers, and has increased the difficulty of ustilaginoidea virens separation and purification.
3) adopt patent ZL201010301356.0 partition method, separation and Culture after the 6th day just visible more polymorphic consistent, clear, without yellow and white single bacterium colony of living contaminants, part organizes block length that the white hypha of doubtful ustilaginoidea virens is arranged, isolation and purification culture is easy.These purifieds are ustilaginoidea virens through dull and stereotyped cultural characteristic, spore and mycelia microscopic examination and the equal susceptible of proof of its conidia germination characteristic observation thereof.The liquid chlamydospore is many owing to smashing to pieces, and miscellaneous bacteria is less, even there is a small amount of miscellaneous bacteria to occur also can distinguishing fast and primary dcreening operation by the bacterium colony dominance, but the method has been used the mercuric chloride sterilizing agent of ethanol and severe toxicity.Chlamydospore is comparatively responsive to sterilizing agent, the processing of rice curve surface disinfectant can cause the part chlamydospore to sprout the forfeiture of ability, rice curve surface cleaning also can cause the minimizing that part has sprouting ability chlamydospore quantity, adopts this method to deposit the rice curve germ separating effect of 3 months to normal temperature relatively poor.The standard specimen that can isolate aimed strain from the rice curve standard specimen for 68 different sourcess of examination has 27, and the standard specimen that adopts method of the present invention can effectively isolate aimed strain has 59.Identical rice curve adopts method after separating of the present invention, adopt again patent ZL201010301356.0 partition method to separate, find that in 15 standard specimens for examination, the former has 13 standard specimens to separate successfully, and the latter only 7 standard specimens separate successfully, separate to obtain single colony number in flat board and be starkly lower than the latter.Show that method of the present invention more is applicable to the separation of rice curve germ, more easy in operation.
Method of the present invention is on patent ZL201010301356.0 technical characterictic basis, by technological improvement, reduce operation steps, and a kind of more easy, practical, the effective ustilaginoidea virens separation method that provides, have and more economize material, the advantage such as quick and easy, applicable.
Claims (3)
1. effective ustilaginoidea virens separation method is characterized in that: carry out according to following steps:
(1) rice curve standard specimen is selected: choose the fresh yellow, yellow-green colour or the green rice curve that gather from the field, or choose with the paper bag packing and dry, and normal temperature or 4 ℃ of Refrigerator stores, the shelf time is no more than the rice curve of 3 months;
(2) preparation of chlamydospore suspension: with the aseptic lip-deep chlamydospore powder of pin scraping simple grain rice curve of choosing, be placed in sterile petri dish, add the sterilized water suspension of 0.5mL to make chlamydospore suspension;
(3) dilution of chlamydospore suspension: press after 2～3 gradients of 1:5 dilution proportion chlamydospore suspension and sterilized water standby;
(4) ustilaginoidea virens separates: inhale step (3) with liquid-transfering gun and diluted good chlamydospore suspension 0.2mL in separating with side of body Ben Zheshi solid culture primary surface, and evenly smear with aseptic glass triangle rod, the culture dish that will contain again above-mentioned substratum is inverted into 28 ℃ of dark culturing in thermostat container, observes day by day separating resulting;
(5) ustilaginoidea virens purifying: cultivate after the 6th day, choose the yellow or white small colonies of pin picking with tip and move to cultivate with purifying in the PSA solid medium and cultivated 10 days;
(6) ustilaginoidea virens is preserved: according to growth characteristics and the microscopic examination result thereof of germ at substratum, with ustilaginoidea virens move to cultivate 10 days in the PSA slant medium after, be stored in 4 ℃ of refrigerators.
2. effective ustilaginoidea virens separation method according to claim 1 is characterized in that: described separation with the formula of side of body Ben Zheshi substratum is: potato 300g, peptone 5g, sucrose 15g, Ca (NO
3)
24H
2O 0.5g, Na
2HPO
412H
20 2.0g, agar 16g, Jia Shui are settled to 1000mL, at 121 ℃ of lower high pressure moist heat sterilization 20～25min, add the paraxin 50 μ g/ml mixings of bacteria growing inhibiting when substratum is cooled to 45 ℃～50 ℃, pour into solidify in culture dish rear standby.
3. effective ustilaginoidea virens separation method according to claim 1, it is characterized in that: described cultivation with the formula of PSA solid medium is: potato 200g, agar 20g, sucrose 20g, Jia Shui are settled to 1000mL, and be standby after 121 ℃ of lower high pressure moist heat sterilization 20min.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103627659A (en) * | 2013-11-27 | 2014-03-12 | 福建省农业科学院植物保护研究所 | Bacillus subtilis and application in prevention of rice ustilaginoidea virens |
| CN105154343A (en) * | 2015-10-28 | 2015-12-16 | 四川省农业科学院植物保护研究所 | Simple method for separating and preserving ustilaginoidea virens |
| CN106906147A (en) * | 2017-03-28 | 2017-06-30 | 浙江大学 | It is adapted to the culture medium of ustilaginoidea virens growth |
| CN110408550A (en) * | 2019-08-19 | 2019-11-05 | 辽宁省农业科学院 | A kind of separation method of the green pyrenomycetes of rice white and its preparation method of artificial infection liquid |
| CN112608849A (en) * | 2020-12-11 | 2021-04-06 | 华智生物技术有限公司 | Rice false smut germ separation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2090054C1 (en) * | 1993-04-27 | 1997-09-20 | Научно-исследовательский институт сельского хозяйства Центрально-Черноземной полосы им.В.В.Докучаева | Method for millet smutting |
| 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 |
-
2013
- 2013-02-25 CN CN2013100581518A patent/CN103103136A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2090054C1 (en) * | 1993-04-27 | 1997-09-20 | Научно-исследовательский институт сельского хозяйства Центрально-Черноземной полосы им.В.В.Докучаева | Method for millet smutting |
| 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 |
Non-Patent Citations (2)
| Title |
|---|
| 李卫平等: "稻曲病菌厚垣孢子的萌发特性", 《浙江农业学报》, vol. 20, no. 4, 31 December 2008 (2008-12-31), pages 278 - 281 * |
| 杨志才等: "海南蔗区甘蔗黑穗病菌的分离鉴定", 《热带作物学报》, vol. 31, no. 5, 30 May 2010 (2010-05-30), pages 828 - 833 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103627659A (en) * | 2013-11-27 | 2014-03-12 | 福建省农业科学院植物保护研究所 | Bacillus subtilis and application in prevention of rice ustilaginoidea virens |
| CN103627659B (en) * | 2013-11-27 | 2015-10-28 | 福建省农业科学院植物保护研究所 | A kind of subtilis and the application in rice green smut control thereof |
| CN105154343A (en) * | 2015-10-28 | 2015-12-16 | 四川省农业科学院植物保护研究所 | Simple method for separating and preserving ustilaginoidea virens |
| CN105154343B (en) * | 2015-10-28 | 2018-03-06 | 四川省农业科学院植物保护研究所 | A kind of easy rice aspergillus separation and store method |
| CN106906147A (en) * | 2017-03-28 | 2017-06-30 | 浙江大学 | It is adapted to the culture medium of ustilaginoidea virens growth |
| CN106906147B (en) * | 2017-03-28 | 2021-04-16 | 浙江大学 | Culture medium suitable for growth of ustilaginoidea virens |
| CN110408550A (en) * | 2019-08-19 | 2019-11-05 | 辽宁省农业科学院 | A kind of separation method of the green pyrenomycetes of rice white and its preparation method of artificial infection liquid |
| CN112608849A (en) * | 2020-12-11 | 2021-04-06 | 华智生物技术有限公司 | Rice false smut germ separation method |
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