CN109929792A - A kind of rice straw powder rice blast product spore culture medium and preparation method - Google Patents
A kind of rice straw powder rice blast product spore culture medium and preparation method Download PDFInfo
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- CN109929792A CN109929792A CN201910155529.3A CN201910155529A CN109929792A CN 109929792 A CN109929792 A CN 109929792A CN 201910155529 A CN201910155529 A CN 201910155529A CN 109929792 A CN109929792 A CN 109929792A
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Abstract
The invention belongs to culture medium field, discloses a kind of rice straw powder rice blast product spore culture medium and preparation method, rice straw powder rice blast product spore culture medium are made of corn flour, rice straw powder and agar；The preparation method of rice straw powder rice blast product spore culture medium includes: to provide the bacterium using rice straw powder as carbon source and nitrogen source and produce carbon and nitrogen required for spore；Vitamin and glycogen in corn flour is added.The vitamin and glycogen that the present invention is added in corn flour are then necessary to adjusting microorganism eubolism, to meet the nutritional need of Pyricularia oryzae；Sporogenous Medium For Pyricularia Oryzae of the invention, cultivates 20d, and sporulation quantity is 5 × 103A spore/milliliter culture medium.
Description
Technical field
The invention belongs to culture medium field more particularly to a kind of rice straw powder rice blast product spore culture mediums and preparation method.
Background technique
Currently, the prior art commonly used in the trade is such that
The rice blast as caused by Pyricularia oryzae [Magnaporthe grisea (Hebert) Barr] is rice three in the world
One of major disease causes serious influence to the yield and quality of rice.Long-term production practices prove that breeding and plantation are anti-
Rice blast kind is that rice blast is most economical, measure effectively and safely for prevention and treatment, meets requirement of the mankind to green food.Breeding is anti-
Rice blast kind needs to treat material selection seedling stage, tillering regularity and booting initial stage progress artificial infection idenfication in field, selects
Disease-resistant and crop field shows preferable rice material, wherein a crucial link is exactly the inoculum for obtaining sufficient amount --- rice
Seasonal febrile diseases bacterium conidium.This just needs the monospore of isolated difference rice blast bacterial strain from the rice standard specimen of morbidity and is protected
It is numerous to deposit expansion.Past generallys use the agar medium of sorghum seed, naked barley grain, rice plant (stalk, blade etc.) or oatmeal
Spore is produced Deng culture rice blast.But some cultural method operating process are cumbersome, and incubation time is long, and production spore effect is not satisfactory, and
It is easy pollution microbes during the cultivation process；Some culture sills are difficult to obtain.
In conclusion problem of the existing technology is:
In the prior art, culture medium is not able to satisfy the nutritional need of Pyricularia oryzae.Product spore culture medium in the prior art does not have
Have using rice straw powder as carbon source and nitrogen source, causes not to be able to satisfy carbon and nitrogen required for Involved in Sporulation in Magnaporthe grisea.
Summary of the invention
For technical problem of the existing technology, the present invention provides a kind of rice straw powder rice blast product spore culture medium and systems
Preparation Method.
The invention is realized in this way a kind of rice straw powder rice blast product spore culture medium, the rice straw powder rice blast produces spore training
It supports base to be made of corn flour, rice straw powder and agar, 4g corn flour 5g rice straw powder, 2g agar is matched in every 100ml sterile water water.
A kind of preparation method another object of the present invention is to provide rice straw powder rice blast product spore culture medium includes:
The first step, with the conical flask of 500ml, first measurement 300ml sterile water when configuring culture medium, is used for convenience of sterilizing
Essence amount balance weighs 12g corn flour, 15g rice straw powder, 6g agar respectively, is added in the container equipped with sterile water, is put into microwave
Heating in furnace, is heated one time 10 seconds several times, in order to by rice straw powder and in corn flour by thermogenetic gas as far as possible
It drains, liquid overflows when pressure cooker being avoided to sterilize.
Second step, the culture medium for being heated to configure take out after holding fluidized state, wrap one-way ventilating film and be put into high pressure sterilization
121 DEG C of sterilizings of pot, take out after high pressure pot temperature is down to 40 DEG C, and 0.2mg/1000ml antibiotic is added, and (rifampin or chlorine are mould
Element) prevention and treatment germ contamination.To reduce germ contamination probability.
Further, it after second step, also needs to carry out after inoculation: be cultivated 18~23 days in 25 DEG C of incubators of constant temperature.
In conclusion advantages of the present invention and good effect are as follows:
The vitamin and glycogen that the present invention is added in corn flour are then necessary to adjusting microorganism eubolism, to meet
The nutritional need of Pyricularia oryzae.
By the present invention in that data analysis shows, produce the normal bacterial strain of spore ability cultivated in 25 DEG C of incubators of constant temperature
After 20 days, media surface is scrubbed with 60ml sterile water, 80% bacterial strain can obtain 100, every visual field under 100 power microscopes
The aaerosol solution of spore.
The results showed that
Ningxia Pyricularia oryzae was carried out using basal culture medium to 2017 within 2014 expanding numerous identification, be concluded that
51 parts of Pyricularia oryzaes in 2013 are detected from 3 batches in October in May-point within 2014, bacterial strain is resisted according to single-gene differential host
From the point of view of resistant frequency, resistant gene Piz, Pizt, Pi9, PiB are higher to bacterial strain fastness frequency, are greater than 40%, see Table 1 for details.Obtain with
Upper resistant gene is larger in disease resistance contribution rate of the Ningxia to rice varieties, should reinforce utilizing.
Table 1: the fastness frequency of rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2013
4 batch Pyricularia oryzaes are detected altogether from October in May-within 2015, detect rice blast single-ascospore strain 103 in 2014 altogether
Part, according to single-gene differential host to the fastness frequency of bacterial strain from the point of view of, resistant gene Pikh, Piz, Piz5 Pizt, Pi9,
It is Pita2, higher to bacterial strain fastness frequency, it is greater than 44%；See Table 2 for details.Show that the above resistant gene resists Rice Production in Ningxia kind
Characteristic of disease contribution rate is relatively large, should reinforce utilizing.
Table 2: the fastness frequency of rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2014
6 batch, 98 parts of Pyricularia oryzaes are identified altogether from October in May-within 2016, bacterial strain is resisted according to single-gene differential host
From the point of view of resistant frequency, resistant gene Pikh, Piz, Piz5 Pizt, Pi1, Pi5, Pi9, Pita2, pi25 were to 2015 years rice blast
Bacterial strain fastness frequency is higher, is greater than 60%, see Table 3 for details.Show that the above resistant gene contributes the disease resistance of Rice Production in Ningxia kind
Rate is larger, should reinforce utilizing；Especially disease-resistant gene pi25 fastness frequency is up to 90.82, it should reinforce introduction of use.
The fastness frequency of 3 rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2015 of table
5 batches are detected altogether from October in May-within 2017 and identify 2016 altogether 120 parts of single-ascospore strain, by anti-to qualification result
The analysis of resistant frequency, Pizt, Pikh, Piz, Pib, Pi9, Pi20, Pita2, gene pairs strains expressed are resistance, and Pia
The effect of Piks Pish Pit Pi3Pi19 several resistant genes Ningxia rice blast is acted on it is then little, wherein Pi5 fastness frequency by
67.02% in 2016 drops to 21.67%, and fastness frequency is greatly reduced, and needs to continue to identify that this gene pairs of observation is peaceful from now on
The effect of summer rice blast.Fastness frequency is shown in Table 4.
Table 4: fastness frequency of the resistant gene to Ningxia rice blast bacterial strain in 2016
The present invention provides the bacterium and produces carbon and nitrogen required for spore using rice straw powder as carbon source and nitrogen source.
And the vitamin in corn flour is added and glycogen guarantees Involved in Sporulation in Magnaporthe grisea nutritional need.
Sporogenous Medium For Pyricularia Oryzae of the invention, after cultivating 20d, sporulation quantity is 5 × 103A spore/milliliter culture medium.
Detailed description of the invention
Fig. 1 is the preparation method flow chart of rice straw powder rice blast product spore culture medium provided in an embodiment of the present invention.
Fig. 2 is provided in an embodiment of the present invention from 3 batches detection in October in May, 2014-point, 51 parts of rice blast in 2013
Bacterium single-ascospore strain, and analyze and obtain the monogenic fastness frequency of single-ascospore strain antagonism.
Fig. 3 is provided in an embodiment of the present invention to detect 4 batch, 98 parts of Pyricularia oryzaes in 2014 altogether from October in May, 2015-
Single-ascospore strain, logical more analyses obtain the monogenic fastness frequency of single-ascospore strain antagonism.
Fig. 4 is provided in an embodiment of the present invention to detect 6 batch, 103 parts of rice blast in 2015 altogether from October in May, 2016-
Bacterium single-ascospore strain, logical more analyses obtain the monogenic fastness frequency of single-ascospore strain antagonism.
Fig. 5 is provided in an embodiment of the present invention to detect 5 batch, 120 parts of rice blast in 2016 altogether from October in May, 2017-
Bacterium single-ascospore strain, logical more analyses obtain the monogenic fastness frequency of single-ascospore strain antagonism.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
In the prior art, culture medium is not able to satisfy the nutritional need of Pyricularia oryzae.
In order to solve the above technical problems, being described in detail with reference to the accompanying drawing to the present invention.
Rice straw powder rice blast product spore culture medium provided in an embodiment of the present invention, is made of corn flour, rice straw powder and agar.
Sporogenous Medium For Pyricularia Oryzae of the invention, cultivates 20d, and sporulation quantity is 5 × 103A spore/milliliter culture medium.
As shown in Figure 1, the preparation method of rice straw powder rice blast product spore culture medium provided in an embodiment of the present invention includes:
S101: measuring 300ml sterile water, weighs 12g corn flour 15g rice straw powder, 6g agar respectively；It is added to equipped with sterile
In the container of water, it is put into heating in micro-wave oven.
S102: taking out after being heated to the culture medium fluidized state of configuration, wrap one-way ventilating film, sterilizes, and after cooling, is added
Antibiotic.
It in step S101, heats, one time 10 seconds, will be arranged in rice straw powder and by thermogenetic hot gas in corn flour several times
Out.
In step S102,121 DEG C of high-pressure sterilizing pot sterilizings are put into, are taken out after high pressure pot temperature is down to 40 DEG C, is added
0.2mg/1000ml antibiotic.
It after step S102, also needs to carry out after inoculation: be cultivated 18~23 days in 25 DEG C of incubators of constant temperature.
Below with reference to specific experiment, the invention will be further described.
It is special from starting autonomous region breeding in 2013 in Ningxia Academy of Agri-Forestry Sciences's crops research institute plant pathology laboratory
So far, the method and formula are constantly improved and is verified, the present invention successfully expands 898 parts of numerous rice blast single-ascospore strain.And it is right
Wherein 517 parts of single-ascospore strains are identified.Including 2014 51 parts, 2015 103 parts, 2016 98 parts, 2017
120 parts, 145 parts of single-ascospore strains in 2018 can be found in Rice Production in Ningxia breeding special project acceptance report in 2017 2014-
Experimental data in 2017.
It is specific as follows:
1) rice blast monospore qualification result in 2014:
As shown in Fig. 2, 51 parts of physiological races of rice blast fungus in 2013 are detected from 3 batches in October in May-point, by inspection
Survey result fastness frequency analysis, according to single-gene differential host to the fastness frequency of bacterial strain from the point of view of, resistant gene Piz, Pizt,
Pi9, PiB are higher to bacterial strain fastness frequency, are greater than 40%, see Table 1 for details.Obtain the above resistant gene to the disease resistance of rice varieties
Contribution rate is larger, should reinforce utilizing.
Table 1: the fastness frequency of rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2013
2) rice blast monospore qualification result in 2015:
As shown in figure 3, detecting 4 batch physiological races of rice blast fungus altogether from October in May-, rice blast in 2014 is detected altogether
103 parts of single-ascospore strain, according to single-gene differential host to the fastness frequency of bacterial strain from the point of view of, resistant gene Pikh, Piz,
It is Piz5Pizt, Pi9, Pita2, higher to bacterial strain fastness frequency, it is greater than 44%；See Table 2 for details.Show the above resistant gene to my area
It wants the disease resistance contribution rate of rice varieties relatively large, should reinforce utilizing.
Table 2: the fastness frequency of rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2014
3) rice blast monospore qualification result in 2016:
As shown in figure 4, detecting 6 batch physiological races of rice blast fungus altogether from October in May-, identifies 98 parts, reflected according to single-gene
From the point of view of other host is to the fastness frequency of bacterial strain, resistant gene Pikh, Piz, Piz5
Pizt, Pi1, Pi5, Pi9, Pita2, pi25 are higher to rice blast bacterial strain fastness frequency in 2015, are greater than 60%,
See Table 3 for details.It show that the above resistant gene wants the disease resistance contribution rate of rice varieties larger in my area, should reinforce utilizing；It is especially anti-
Ospc gene pi25 fastness frequency is up to 90.82, it should reinforce introduction of use.
The fastness frequency of 3 rice blast key resistance gene pairs Ningxia rice blast bacterial strain in 2015 of table
4) rice blast monospore qualification result in 2017:
As shown in figure 5, detecting 5 batch physiological races of rice blast fungus altogether from October in May-, single-ascospore strain in 2016 is detected altogether
120 parts, pass through the analysis to testing result fastness frequency, Pizt, Pikh, Piz, Pib, Pi9, Pi20, Pita2, gene pairs bacterium
Strain be demonstrated by it is resistance, and the effect of Pia Piks Pish Pit Pi3 Pi19 several resistant genes to the effect of Ningxia rice blast then
Less, wherein Pi5 fastness frequency by 2016 67.02% dropped to 21.67%, and fastness frequency is greatly reduced, and needs from now on
Continue the effect that this gene pairs Ningxia rice blast is observed in identification.Fastness frequency is shown in Table 4.
Table 4: fastness frequency of the resistant gene to Ningxia rice blast bacterial strain in 2016
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (5)
1. a kind of rice straw powder rice blast product spore culture medium, which is characterized in that the rice straw powder rice blast product spore culture medium is by corn
Powder, rice straw powder and agar composition；4g corn flour, 5g rice straw powder and 2g agar are formulated in every 100ml sterile water.
2. a kind of preparation method of rice straw powder rice blast product spore culture medium as described in claim 1, which is characterized in that the rice straw
The preparation method of powder rice blast product spore culture medium includes:
The first step measures 300ml sterile water, weighs 12g corn flour, 15g rice straw powder, 6g agar respectively；It is added to equipped with sterile
In the container of water, it is put into heating in micro-wave oven；
Second step takes out after being heated to the culture medium fluidized state of configuration, wraps one-way ventilating film, sterilizes, and after cooling, is added anti-
Raw element.
3. the preparation method of rice straw powder rice blast product spore culture medium as claimed in claim 2, which is characterized in that
It in the first step, heats, one time 10 seconds, will be drained in rice straw powder and in corn flour by thermogenetic gas several times.
4. the preparation method of rice straw powder rice blast product spore culture medium as claimed in claim 2, which is characterized in that
In second step, 121 DEG C of high-pressure sterilizing pot sterilizings are put into, are taken out after high pressure pot temperature is down to 40 DEG C, 0.2mg/ is added
1000ml antibiotic.
5. the preparation method of rice straw powder rice blast product spore culture medium as claimed in claim 2, which is characterized in that after second step,
It also needs to carry out: be cultivated 18-23 days in 25 DEG C of incubators of constant temperature after inoculation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300978A (en) * | 2020-11-04 | 2021-02-02 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Rice blast pathogen spore separation and preservation intelligent system and method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104355815A (en) * | 2014-10-30 | 2015-02-18 | 湖南原生生物科技股份有限公司 | Water-soluble medicinal fertilizer containing strong-antagonism anti-rice blast bacterial strain, preparation method and application of medicinal fertilizer |
| CN108753937A (en) * | 2018-06-28 | 2018-11-06 | 扬州大学 | A kind of detection method of blast resistant gene expression and its application |
| CN113788883A (en) * | 2021-10-20 | 2021-12-14 | 福建农林大学 | Magnaporthe grisea MoSpc2 gene and application thereof |
-
2019
- 2019-03-01 CN CN201910155529.3A patent/CN109929792A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104355815A (en) * | 2014-10-30 | 2015-02-18 | 湖南原生生物科技股份有限公司 | Water-soluble medicinal fertilizer containing strong-antagonism anti-rice blast bacterial strain, preparation method and application of medicinal fertilizer |
| CN108753937A (en) * | 2018-06-28 | 2018-11-06 | 扬州大学 | A kind of detection method of blast resistant gene expression and its application |
| CN113788883A (en) * | 2021-10-20 | 2021-12-14 | 福建农林大学 | Magnaporthe grisea MoSpc2 gene and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 吴斯骏等: "一种稻瘟病菌培养产孢培养基的配制与效比分析", 《安徽农业科学》 * |
| 孔秀英: "稻瘟病菌分生孢子的诱发及保存方法", 《微生物学通报》 * |
| 路金才: "《全国高等医药院校药学类第四轮规划教材 药用植物学 第3版》", 31 January 2016, 北京：中国医药科技出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300978A (en) * | 2020-11-04 | 2021-02-02 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Rice blast pathogen spore separation and preservation intelligent system and method |
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Inventor after: Shi Yanli Inventor after: Li Wenqiang Inventor after: Chen Dongsheng Inventor after: Liu Wei Inventor after: Huang Xinling Inventor after: Sha Rong Inventor after: Pu Lili Inventor after: Yang Shenglong Inventor after: Wang Jian Inventor before: Shi Yanli Inventor before: Chen Dongsheng Inventor before: Liu Wei Inventor before: Li Wenqiang Inventor before: Huang Xinling Inventor before: Sha Rong Inventor before: Pu Lili Inventor before: Yang Shenglong Inventor before: Wang Jian |