CN105441375A - Method for generating sporangia and releasing zoospore by inducing phytophthora capsici - Google Patents
Method for generating sporangia and releasing zoospore by inducing phytophthora capsici Download PDFInfo
- Publication number
- CN105441375A CN105441375A CN201510899825.6A CN201510899825A CN105441375A CN 105441375 A CN105441375 A CN 105441375A CN 201510899825 A CN201510899825 A CN 201510899825A CN 105441375 A CN105441375 A CN 105441375A
- Authority
- CN
- China
- Prior art keywords
- pepper
- zoospore
- phytophthora blight
- induction
- sporocyst
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for generating sporangia and releasing zoospore by inducing phytophthora capsici. The method includes the steps that phytophthora capsici is activated, an inducing solution for generating sporangia is prepared, sporangia are induced to be generated, and zoospore is induced to be released. The inducing solution used in the method is originally imported V8 vegetable juice with the volume concentration being 10-15% and a CaCO3 solution with the volume concentration being 0.2-0.3 g/L. When phytophthora capsici is induced to generate the sporangia, continuous light culture is carried out under a white fluorescent lamp. The method is simple, convenient to use, quick, free of pollution and capable of being used in phytophthora capsici pathogenicity measurement and biological activity measurement, achieved through an antibacterial agent, of phytophthora capsici.
Description
Technical field
The invention belongs to field of microbial culture technology, be specifically related to induce phytophthora blight of pepper to produce sporocyst and discharge the method for zoospore.
Background technology
Capsicum epidemic disease is the crushing fungal disease generally occurred in a kind of world wide, first it found in 1918 in New Mexico, cause the phytophthora blight of pepper PhytophthoracapsiciLeonian of capsicum epidemic disease to be that LeonLeonian named in nineteen twenty-two, it belongs to pathogenic oomycetes.Phytophthora capsici is survived the winter in soil invalid body with oospore or chlamydospore, and can survive even longer time several months, host range is wide, mainly infects the multiple important vegetable such as capsicum, tomato, eggplant, cucumber, pumpkin.Capsicum epidemic disease open country and protecting field all can occur, and the popular of this disease causes capsicum to lose seriously, and general diseased plant rate is 15 – 30%, reaches more than 80% time serious.Since nineteen eighty, capsicum epidemic disease increases the weight of increasingly at China's growing vegetables region occurring degree, has had a strong impact on the economic benefit of growing vegetables industry, produces cause huge financial loss to the vegetable safety of China.Therefore, phytophthora blight of pepper is subject to the height extensive concern of various countries phytopathologist and geneticist.
Under natural condition, Phytophthora capsici infects from the root of plant or basal part of stem mainly through zoospore, during artificial culture, can be passed through mycelium culture, induction produces sporocyst, subzero treatment promotes Zoospore liberation, thus obtain more zoospore.Tradition cultivates revulsion: mycelia is placed in oat medium or rye substratum or V8 juice nutrient solution, after activation culture longer for some time, add soil extract or Pi Shi liquid again, under illumination condition, induction produces sporocyst, after subzero treatment, produce zoospore.The method induction phytophthora produces sporocyst needs more than 10 days, and the time is long; Meanwhile, soil extract, without stringent sterilization, Pi Shi nutrient solution moist heat sterilization, is induced the effect instability of generation zoosporangium, is also easily polluted.Also have other induction method also can produce a large amount of zoospores: rye culture medium culturing mycelia covers with flat board, after smearing with aseptic L-type glass rod the culture medium flat plate covering with mycelia, be placed in 28 DEG C of light irradiation biochemical incubators and cultivate 24h with the generation of inducing spore capsule, 1h in 4 DEG C of refrigerators is moved to again after producing sporangial culture medium flat plate and adding the sterilized water of 18mL, Phytophthora capsici can be induced to produce zoospore, but aforesaid method is more loaded down with trivial details, also available zoospore to could be obtained through filtering.In addition, fresh host plant material revulsion also can induce generation sporocyst, produce zoospore, but the method easily suffers the pollution of bacterium after subzero treatment, and the more difficult acquisition of vegetable material.
Summary of the invention
The shortcoming that the object of the invention is to overcome prior art, with not enough, provides induction phytophthora blight of pepper produce sporocyst and discharge the method for zoospore, for phytophthora blight of pepper Pathogenic Tests and sterilant to researchs such as the biological activity determinations of phytophthora blight of pepper.
Object of the present invention is achieved through the following technical solutions: induction phytophthora blight of pepper produces sporocyst and discharges the method for zoospore, comprises the steps:
(1) phytophthora blight of pepper activation: phytophthora blight of pepper is inoculated on Radix Dauci Sativae substratum and carries out activation culture.
(2) preparation produces sporangial induced liquid: market will be bought imported with original packaging V8 vegetables juice and be mixed with the juice of volumetric concentration 10%-15% with deionized water, for subsequent use after sterilizing; Preparation 0.2-0.3g/LCaCO
3solution, for subsequent use after sterilizing.
(3) induction produces sporocyst: get the phytophthora blight of pepper that step (1) activated and be inoculated in the V8 vegetables juice of 10%-15% prepared by step (2), 25 DEG C of dark culturing 2d; Then to the greatest extent V8 vegetables juice, with sterilizing washed with de-ionized water once, change 0.2-0.3g/LCaCO prepared by step (2)
3solution, 2d is cultivated in the lower 25 DEG C of continuous illuminations of white fluorescence in intensity of illumination 700-800Lux, can induce generation sporocyst.
(4) induction release zoospore: step (3) is placed 15min through the phytophthora blight of pepper 4 DEG C of induction, is then placed in 25 DEG C, a large amount of zoospores can be discharged after 30min.
Activation culture described in step (1) is 25 DEG C of dark culturing 2d.
Water described in step (2) is sterilizing deionized water.
The condition optimization of the sterilizing described in step (2) is 121 DEG C of moist heat sterilization 30min.
Induction described in step (3) produces sporangial method and is preferably: get the phytophthora blight of pepper edge mycelia block that step (1) activated and be placed on aseptic culture dish, mycelia block be 1cm × 1cm, 20 pieces, add the V8 vegetables juice of volumetric concentration 15% prepared by step (2), make juice just dipped mycelia block, 25 DEG C of dark culturing 2d; Then confide all V8 vegetables juice, with the washed with de-ionized water of sterilizing once, then add 0.3g/LCaCO
3solution makes just dipped mycelia block, and 2d is cultivated in the white fluorescence illumination box 25 DEG C of continuous illuminations being placed in intensity of illumination 800Lux, can induce generation sporocyst.
Described Radix Dauci Sativae substratum is that silk is cut in the peeling of 200g fresh carrot, and the 800ml that adds water boils 1h, and 4 layers of filtered through gauze remove residue, agar 15g, and heating makes agar melt, constant volume 1000ml, pH=6.0.
The present invention has following advantage and effect relative to prior art:
(1) quick: method of the present invention can obtain sporocyst and the zoospore of a large amount of phytophthora blight of pepper in 4.5d, and more traditional cultivation induction method is compared, induction duration shortens.
(2) pollution-free: the present invention the adding of pollution-free source in operation, the sporocyst induced and zoospore all do not pollute.
(3) induced liquid can save backup for a long time: the 0.2-0.3g/LCaCO that the present invention is used
3through sterilising treatment, room temperature preservation can be put.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Specifications of raw materials
V8 vegetables juice: Campbell Soup Company, the U.S..
The V8 vegetables juice of 15%: mix with deionized water 850mL and V8 juice beverage 150mL, for subsequent use after 121 DEG C of moist heat sterilization 30min.
CaCO
3: sigma company, the U.S..
0.3g/LCaCO
3: the CaCO taking 0.3g
3, be dissolved in 1000mL deionized water, for subsequent use after 121 DEG C of moist heat sterilization 30min.
Culture dish required for other experimental implementation, Radix Dauci Sativae substratum, pipettor, knife blade, Autoclave, refrigerator, deionized water, illumination box etc. are laboratory conventional tool or material.
Embodiment 1 phytophthora blight of pepper standard order-checking bacterial strain produces sporocyst and zoospore
(1) on Bechtop, phytophthora blight of pepper (strain number: LT1534, order-checking reference culture) is inoculated on Radix Dauci Sativae substratum, 25 DEG C of dark culturing 2d.
(2) on Bechtop, extract the mycelia block of 0.3 × 0.3cm at colony edge place knife blade, get 20 ferfas silk blocks and be placed in the sterile petri dish that diameter is 11cm, add the V8 vegetables juice 30mL that volumetric concentration is 15%, 25 DEG C of dark culturing 2d; Then to the greatest extent V8 vegetables juice, with sterilizing washed with de-ionized water once, then change aseptic 0.3g/LCaCO
315mL, is placed in the illumination box of the white fluorescence of intensity of illumination 800Lux, and 2d is cultivated in 25 DEG C of continuous illuminations.
(3) by the phytophthora blight of pepper through above-mentioned inducing culture in 4 DEG C of refrigerators, place 15min; Then be placed in 25 DEG C of incubators, after 30min, collect liquid.
(4) be that 10 μ L liquid-transfering guns draw the liquid of 5 μ L as on blood counting slide glass with range, under 10 times of mirror visuals field, calculate the quantity of zoospore.Sample 3 times, calculate the mean number of zoospore in 5 μ L liquid, be then converted into the concentration of every milliliter of zoospore suspension.The zoospore number discharged after measured reaches 7.32 × 10
5individual spore/mL, release rate more than 90%.
Embodiment 2 induces the Phytophthora capsici bacteria strain be separated in the sick sample of fresh chilli epidemic disease to produce sporocyst and zoospore
(1) get the sick sample sample of capsicum epidemic disease of fresh morbidity, be good in disease the tissue block that intersection clip size is about 5mm × 5mm with scissors.
(2) on Bechtop, tissue block is placed in 75% alcohol and soaks 30s, then soak rinsing lmin in 5% chlorine bleach liquor; Then sterilizing washed with de-ionized water is used 3 times.After sterilizing filter paper blots water, transferred on Radix Dauci Sativae culture medium flat plate, 25 DEG C of dark culturing 3d.
(3) after growing bacterium colony, on Bechtop, picking colony color is the edge mycelia block of white, carries out mycelia purifying, can obtain phytophthora blight of pepper to it.
(4) on Bechtop, extract the mycelia block of 1cm × 1cm at colony edge place knife blade, get 20 ferfas silk blocks and be placed in the V8 vegetables juice that volumetric concentration is 10%, make fruit juice liquid just dipped mycelia block, 25 DEG C of dark culturing 2d; Then confide all liquid, with sterilizing washed with de-ionized water once, change to sterilized 0.2g/LCaCO
3just dipped mycelia block, is placed in the illumination box of the white fluorescence of intensity of illumination 700Lux, and 2d is cultivated in 25 DEG C of continuous illuminations.
(5) the above-mentioned bacterial strain through inducing culture is placed in 4 DEG C of refrigerator 15min, is then positioned in 25 DEG C of incubators, after 30min, collects liquid.Be that 10 μ L liquid-transfering guns draw the liquid of 5 μ L as on blood counting slide glass with range, under 10 times of mirror visuals field, calculate the quantity of zoospore.Sample 3 times, calculate the mean number of zoospore in 5 μ L liquid, be then converted into the concentration of every milliliter of zoospore suspension.After measured, the zoospore number 5.31 × 10 of release
5individual spore/mL, release rate more than 90%.
Claims (3)
1. induce phytophthora blight of pepper produce sporocyst and discharge the method for zoospore, it is characterized in that:
(1) phytophthora blight of pepper activation: phytophthora blight of pepper is inoculated on Radix Dauci Sativae substratum and carries out activation culture;
(2) preparation produces sporangial induced liquid: the juice with deionized water, V8 vegetables juice being mixed with volumetric concentration 10%-15%, for subsequent use after sterilizing;
(3) induction produces sporocyst: get the phytophthora blight of pepper that step (1) activated and be inoculated in the V8 vegetables juice of volumetric concentration 10%-15% prepared by step (2), 25 DEG C of dark culturing 2d, mycelia block be 0.3cm × 0.3cm-1cm × 1cm, 20 pieces; Then confide all V8 vegetables juice, with sterilizing washed with de-ionized water once, change the 0.2-0.3g/LCaCO of sterilizing
3solution, 2d is cultivated in the lower 25 DEG C of continuous illuminations of white fluorescence in intensity of illumination 700Lux-800Lux, can induce and produce a large amount of sporocysts;
(4) induction release zoospore: step (3) is placed 15min through the phytophthora blight of pepper 4 DEG C of induction, is then placed in 25 DEG C, a large amount of zoospores can be discharged after 30min.
2. induction phytophthora blight of pepper according to claim 1 produces sporocyst and discharges the method for zoospore, it is characterized in that: the induction described in step (3) produces sporangial method and is: get the phytophthora blight of pepper edge mycelia block that step (1) activated and be placed on aseptic culture dish, mycelia block be 1cm × 1cm, 20 pieces, add the V8 vegetables juice of volumetric concentration 15% prepared by step (2), make juice just dipped mycelia block, 25 DEG C of dark culturing 2d; Then confide all V8 vegetables juice, with the washed with de-ionized water of sterilizing once, then add 0.3g/LCaCO
3solution makes just dipped mycelia block, and 2d is cultivated in the white fluorescence illumination box 25 DEG C of continuous illuminations being placed in intensity of illumination 800Lux, can induce generation sporocyst.
3. induction phytophthora blight of pepper according to claim 1 produces sporocyst and discharges the method for zoospore, it is characterized in that: described Radix Dauci Sativae substratum is that silk is cut in the peeling of 200g fresh carrot, the 800ml that adds water boils 1h, 4 layers of filtered through gauze remove residue, agar 15g, heating makes agar melt, constant volume 1000ml, pH=6.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510899825.6A CN105441375B (en) | 2015-12-09 | 2015-12-09 | Induction phytophthora blight of pepper generates sporangium and discharges the method for zoospore |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510899825.6A CN105441375B (en) | 2015-12-09 | 2015-12-09 | Induction phytophthora blight of pepper generates sporangium and discharges the method for zoospore |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105441375A true CN105441375A (en) | 2016-03-30 |
| CN105441375B CN105441375B (en) | 2018-10-02 |
Family
ID=55552016
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510899825.6A Active CN105441375B (en) | 2015-12-09 | 2015-12-09 | Induction phytophthora blight of pepper generates sporangium and discharges the method for zoospore |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105441375B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886451A (en) * | 2016-04-26 | 2016-08-24 | 东北农业大学 | Single spore isolation method of phytophthora capsici zoospores |
| CN106636302A (en) * | 2016-12-27 | 2017-05-10 | 福建省农业科学院植物保护研究所 | Method for evaluating pepper phytophthora blight resistance of crops |
| CN106613555A (en) * | 2016-12-29 | 2017-05-10 | 福建省农业科学院植物保护研究所 | Method for inducing pepper against pestilence by using benzothiadiazole as exciton |
| CN108034629A (en) * | 2018-01-19 | 2018-05-15 | 北京农学院 | CA culture mediums are for cultivating the application in strawberry brown leaf spot fungi production spore |
| CN108410793A (en) * | 2018-04-27 | 2018-08-17 | 江西省农业科学院植物保护研究所 | A kind of induction phytophthora blight of pepper production culture medium of spore, preparation method and application |
| CN109825443A (en) * | 2019-02-12 | 2019-05-31 | 安徽农业大学 | It is a kind of induce Phytophthora capsici Germination of Oospores culture medium and its application |
| CN110564667A (en) * | 2019-09-25 | 2019-12-13 | 云南省烟草农业科学研究院 | method for efficiently suspending and concentrating zoospores of plant pathogenic oomycetes and application of zoospores |
| CN110819583A (en) * | 2018-08-10 | 2020-02-21 | 云南农业大学 | Spore-producing culture method of phytophthora sojae |
| CN111172182A (en) * | 2019-12-06 | 2020-05-19 | 长江大学 | Phytophthora capsici PcMPK12 gene and vector and application thereof |
| CN113373064A (en) * | 2021-07-14 | 2021-09-10 | 海南大学 | Method for inducing phytophthora litchi sporangium to release zoospores |
| CN115044528A (en) * | 2022-06-10 | 2022-09-13 | 华南农业大学 | Method for inducing oomycetes saprophyta to generate zoospores |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154193A (en) * | 2011-01-20 | 2011-08-17 | 天津市植物保护研究所 | Method for inducing Phytophthora capsici to produce large amount of sporangia |
| CN102391980A (en) * | 2011-11-25 | 2012-03-28 | 福建省农业科学院植物保护研究所 | Simple and effective method for producing zoospores through inducing Phytophthora capsici |
| CN102807957A (en) * | 2012-08-06 | 2012-12-05 | 中国农业大学 | Method for increasing phytophthora capsici oospore output and promoting oospore germination |
| CN103045529A (en) * | 2012-12-31 | 2013-04-17 | 广东省农业科学院植物保护研究所 | Method of inducing phytophthora nicotianae breda de haan bacteria to generate sporangiums and release zoospores |
| CN104726390A (en) * | 2015-03-17 | 2015-06-24 | 内蒙古农业大学 | Culture medium for prompting Phytophthora capsici Leonian to generate zoosporangium |
-
2015
- 2015-12-09 CN CN201510899825.6A patent/CN105441375B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154193A (en) * | 2011-01-20 | 2011-08-17 | 天津市植物保护研究所 | Method for inducing Phytophthora capsici to produce large amount of sporangia |
| CN102391980A (en) * | 2011-11-25 | 2012-03-28 | 福建省农业科学院植物保护研究所 | Simple and effective method for producing zoospores through inducing Phytophthora capsici |
| CN102807957A (en) * | 2012-08-06 | 2012-12-05 | 中国农业大学 | Method for increasing phytophthora capsici oospore output and promoting oospore germination |
| CN103045529A (en) * | 2012-12-31 | 2013-04-17 | 广东省农业科学院植物保护研究所 | Method of inducing phytophthora nicotianae breda de haan bacteria to generate sporangiums and release zoospores |
| CN104726390A (en) * | 2015-03-17 | 2015-06-24 | 内蒙古农业大学 | Culture medium for prompting Phytophthora capsici Leonian to generate zoosporangium |
Non-Patent Citations (4)
| Title |
|---|
| 兰成忠等: "辣椒疫霉菌产孢培养基及诱导方法筛选", 《热带作物学报》 * |
| 孙广宇等: "《植物病理学实验技术》", 31 August 2002, 中国农业出版社 * |
| 王晓敏等: "辣椒疫霉菌孢子诱导技术研究", 《西北农业学报》 * |
| 许亚池等: "一种便捷高产的辣椒疫霉菌游动孢子产生方法", 《江苏农业科学》 * |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886451B (en) * | 2016-04-26 | 2020-07-10 | 东北农业大学 | Single spore separation method of phytophthora capsici zoospores |
| CN105886451A (en) * | 2016-04-26 | 2016-08-24 | 东北农业大学 | Single spore isolation method of phytophthora capsici zoospores |
| CN106636302A (en) * | 2016-12-27 | 2017-05-10 | 福建省农业科学院植物保护研究所 | Method for evaluating pepper phytophthora blight resistance of crops |
| CN106613555A (en) * | 2016-12-29 | 2017-05-10 | 福建省农业科学院植物保护研究所 | Method for inducing pepper against pestilence by using benzothiadiazole as exciton |
| CN108034629A (en) * | 2018-01-19 | 2018-05-15 | 北京农学院 | CA culture mediums are for cultivating the application in strawberry brown leaf spot fungi production spore |
| CN108034629B (en) * | 2018-01-19 | 2021-05-25 | 北京农学院 | Application of CA culture medium in culturing strawberry brown leaf spot pathogen sporulation |
| CN108410793A (en) * | 2018-04-27 | 2018-08-17 | 江西省农业科学院植物保护研究所 | A kind of induction phytophthora blight of pepper production culture medium of spore, preparation method and application |
| CN108410793B (en) * | 2018-04-27 | 2020-11-06 | 江西省农业科学院植物保护研究所 | Culture medium for inducing phytophthora capsici to produce spores, preparation method and application |
| CN110819583B (en) * | 2018-08-10 | 2021-07-06 | 云南农业大学 | Spore-producing culture method of phytophthora sojae |
| CN110819583A (en) * | 2018-08-10 | 2020-02-21 | 云南农业大学 | Spore-producing culture method of phytophthora sojae |
| CN109825443A (en) * | 2019-02-12 | 2019-05-31 | 安徽农业大学 | It is a kind of induce Phytophthora capsici Germination of Oospores culture medium and its application |
| CN110564667A (en) * | 2019-09-25 | 2019-12-13 | 云南省烟草农业科学研究院 | method for efficiently suspending and concentrating zoospores of plant pathogenic oomycetes and application of zoospores |
| CN110564667B (en) * | 2019-09-25 | 2023-02-28 | 云南省烟草农业科学研究院 | Method for efficiently suspending and concentrating zoospores of plant pathogenic oomycetes and application of zoospores |
| CN111172182A (en) * | 2019-12-06 | 2020-05-19 | 长江大学 | Phytophthora capsici PcMPK12 gene and vector and application thereof |
| CN111172182B (en) * | 2019-12-06 | 2021-09-24 | 长江大学 | Phytophthora capsici PcMPK12 gene and vector and application thereof |
| CN113373064A (en) * | 2021-07-14 | 2021-09-10 | 海南大学 | Method for inducing phytophthora litchi sporangium to release zoospores |
| CN113373064B (en) * | 2021-07-14 | 2023-04-14 | 海南大学 | Method for inducing phytophthora litchi sporocyst to release zoospores |
| CN115044528A (en) * | 2022-06-10 | 2022-09-13 | 华南农业大学 | Method for inducing oomycetes saprophyta to generate zoospores |
| CN115044528B (en) * | 2022-06-10 | 2023-06-02 | 华南农业大学 | Method for inducing oomycetes saphenous pythium to generate zoospores |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105441375B (en) | 2018-10-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105441375A (en) | Method for generating sporangia and releasing zoospore by inducing phytophthora capsici | |
| CN103045529B (en) | Method of inducing phytophthora nicotianae breda de haan bacteria to generate sporangiums and release zoospores | |
| CN101724569B (en) | Method for inducing nematode-trapping fungi to synchronously produce capturing organs | |
| CN105200019B (en) | The preparation method of environmental protection ferment and algal tufa inhibitor | |
| CN105052498B (en) | A kind of liquid cultivating method for setting up DSE and corn syntaxial system | |
| CN105145676B (en) | Algal tufa inhibitor | |
| CN102771394A (en) | Method for cloning and culturing seaweed gametophytes | |
| CN106212281A (en) | A kind of method for tissue culture improving Fructus Musae survival rate | |
| CN109722390B (en) | Separation and purification of rhododendron erinaceus mycorrhizal fungi strain DPS-A and inoculation application method thereof | |
| CN107446842A (en) | One bacillus subtilis and its application in purifying water | |
| CN110257316A (en) | A kind of plant anthrax bacteria conidium rapid separation and purification method | |
| CN102835310A (en) | Double-tray combined type plant tissue culture container | |
| CN105255746B (en) | One plant of Paecilonyces variotii strain and its application for having High pathogenicity to diaphorina citri | |
| CN101984042B (en) | Preservation method of pathogenic bacteria of potato late blight and special medium for the same | |
| CN105217801B (en) | Application of the environmental protection ferment in terms of lake algal bloom harm is prevented | |
| CN108410793B (en) | Culture medium for inducing phytophthora capsici to produce spores, preparation method and application | |
| CN104726349A (en) | Tobacco phytophthora culture medium and preparation method thereof | |
| CN103146597A (en) | Method for preparing photosynthetic bacteria liquid | |
| CN101380333A (en) | Method for extracting cellular fluid from human homogeneous variant cell | |
| KR101779857B1 (en) | Manufacturing method of cordyceps sinensis activated rice and cordyceps sinensis activated rice by the manufacturing method | |
| CN107372117A (en) | Taxus chinensis var. mairei Callus abductive approach | |
| CN103555649B (en) | Solid-phase preservation method for kelp gametophytes | |
| CN106613358A (en) | White fungus spawn extracting method | |
| CN102643776B (en) | Culture medium appropriate for phytophthotacactorum to produce sporangiums by taking radish as raw material | |
| CN104920064B (en) | A kind of preparation method of the special parent species of Cordyceps militaris production |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |