CN106119120A - A kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof - Google Patents
A kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof, this plasmodiophora brassicae inoculation nutrient matrix component according to volume ratio is: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2, the present invention discloses the preparation method of a kind of plasmodiophora brassicae inoculation nutrient matrix, the method includes uniformly mixing peat soil, Vermiculitum, perlite according to volume ratio, being distributed in round plastic nutritive cube after the sterilized process of nutrient matrix, present invention simultaneously discloses a kind of nutrient matrix component applied in plasmodiophora brassicae expanding propagation is 100% quartz sand.Host's Seedling sickness rate of the nutrient matrix of the present invention reaches 100%, and disease index reaches 69.6, is the nutrient matrix of preferable plasmodiophora brassicae inoculation；Its sickness rate is high, morbidity stably can be utilized for differentiating and the evaluation and screening of anti-clubroot breeding material of plasmodiophora brassicae biological strain.
Description
Technical field
The invention belongs to microbial technology field, particularly relate to a kind of plasmodiophora brassicae inoculation nutrient matrix and preparation side thereof
Method.
Background technology
Clubroot is the Cruciferae vegetables caused by Caulis et Folium Brassicae campestris plasmodiophora brassicae (Plasmodiophora brassicae Woron.)
Dish disease.It is generally thought to comprise two eposides at present: resting spore, protoplasm group, and three phases: rank of surviving in soil
Section, Root hair infection stage, cortex infect the stage (Kageyama&Asano, 2009).These resting spores can be promoted to sprout
Condition is currently not it is clear that but they easily become under the conditions of ambient temperature 6 DEG C to 27 DEG C, acid ph value (less than 6.0)
Merit infects host (Karling, 1968).Occurring with the moisture in soil close to time saturated susceptible host, resting spore is sprouted
Discharge nascent zoospore, there is a pair preposition flagellum of different length (Brown, 1997).When nascent zoospore arrives
During the epidermis cell of root hair or root, he loses flagellum and becomes amebicide apperance (Roberts&Boothroyd, 1972).Travelling
The inclusions of spore is injected into host cell, and the protoplasm of multinuclear is rolled into a ball at intracellular germinate (Rush, 2003).Although root
The effect that hair infects also is not very clear, it is generally understood that Root hair infection can increase plasmodiophora brassicae colony's number in susceptible host
Amount, and cause the serious morbidity (Mitani et al., 2003) of host.In the First aggression stage, sporangial protoplasm is rolled into a ball at table
Develop into a thin-walled zoosporangium in chrotoplast, include 4 to 8 secondary zoospores (Agrios, 2005).But second
The cortical cell of host's root and the middle pillar cells (Dixon&Page, 1998) are being infected in the deep generation of infecting in stage.Infect
Result cause the abnormal increment of host plant tissue, produce spore protoplasm group and grow and also form knee (Rush, 2003).Work as quilt
Plasmodiophora brassicae is infected the knee cracking substantial amounts of hypnocyst of rotten generation later of parasitism and discharges into soil, and can be long-term
In soil, survival reaches more than 7 years.
Inoculation method aspect, owing to plasmodiophora brassicae is strict obligate parasite, the artificial propagation of plasmodiophora brassicae and inoculated identification pair
Used inoculation method and test material require strict, and this work is also the key link carrying out clubroot research.Root at present
The inoculation identification method of swollen bacterium dips in root method, drop method and bacterium local method；Inoculum density is mostly 1 × 107～1 × 108spores/
ml；The more complicated requirement of operation sequence is strict, and during easily form the cross-contamination between different bacterium source.Use different
After nutrient matrix inoculation plasmodiophora brassicae, the morbidity effect of host's Seedling is different, plasmodiophora brassicae inoculation morbidity de-stabilising effect test for identification
Result.
Summary of the invention
It is an object of the invention to provide a kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof, it is intended to solve at present
The more complicated requirement of inoculation identification method operation sequence of plasmodiophora brassicae is strict, easily forms the cross-contamination between different bacterium source and connects
The problem that after bacterium, plant morbidity is unstable.
The present invention is achieved in that
A kind of plasmodiophora brassicae inoculation nutrient matrix, this plasmodiophora brassicae inoculation nutrient matrix component according to volume ratio is: peat
Soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2.
Another object of the present invention is to provide the preparation method of a kind of plasmodiophora brassicae inoculation nutrient matrix, described plasmodiophora brassicae
The preparation method of inoculation nutrient matrix comprises the following steps:
Peat soil, Vermiculitum, perlite are mixed according to volume ratio 1:0.8～1.2:0.8～1.2 ratio uniform, and will system
Standby nutrient matrix is distributed in sterilizing bag；
The sterilizing bag that will be equipped with nutrient matrix is put in autoclave, sterilizing；
Nutrient matrix after sterilized process is distributed in round plastic nutritive cube and is compacted；
Configure acid water with concentrated hydrochloric acid, the pallet bottom nutritive cube pours into the acid water with concentrated hydrochloric acid configuration, makes battalion
Foster matrix soil is impregnated with acid moisture stand-by.
Further, sterilizing methods is: 121 DEG C of moist heat sterilizations 30 minutes.
Further, the acid water pH value of described concentrated hydrochloric acid configuration is 6.0.
Another object of the present invention is to provide a kind of nutrient matrix of application in plasmodiophora brassicae expanding propagation, should expand plasmodiophora brassicae
The nutrient matrix component of numerous middle application is 100% quartz sand.
Further, quartz sand particle size is 1mm～2mm.
Another object of the present invention is to provide a kind of plasmodiophora brassicae inoculation nutrient matrix to breed at plasmodiophora brassicae inoculated identification
In application.
Plasmodiophora brassicae inoculation nutrient matrix that the present invention provides and preparation method thereof, by the comparison to 6 kinds of culture matrixes,
The formula of No. 6 culture matrixes found i.e. includes according to volume ratio: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～
1.2, the sickness rate susceptible host reaches 100%, disease index reaches 69.6 (more than the threshold values of disease index 50), is that one can
Good culture matrix for plasmodiophora brassicae inoculated identification；In plasmodiophora brassicae inoculation test, susceptible host falls ill to show and stablizes, energy
Enough Screening and Identification carrying out plasmodiophora brassicae Race Identification and the anti-clubroot of kind；Plasmodiophora brassicae inoculation nutrient matrix according to
Volume ratio includes: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2, uses the nutrient matrix of this proportioning formula to connect
Planting plasmodiophora brassicae, the sickness rate of susceptible variety reaches 100%, and morbidity is stable；
The sickness rate of host's Seedling of No. 2 culture matrix formula that is 100% quartz sands is relatively low, the rear batallion but its host's Seedling is fallen ill
The well-grown supporting body easily forms big single tumor, and No. 2 culture medium can be used to the expanding propagation as plasmodiophora brassicae, easy
Use to the biggest lump.
Accompanying drawing explanation
Fig. 1 is the preparation method flow chart of the plasmodiophora brassicae inoculation nutrient matrix that the embodiment of the present invention provides.
Detailed description of the invention
In order to make the purpose of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, to the present invention
It is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not used to
Limit the present invention.
A kind of method that the invention provides simple and effective plasmodiophora brassicae inoculated identification breeding, uses drop inocalation method to compare
The inoculation efficiency of plasmodiophora brassicae under the nutrient matrix of different ratio and different inoculum densities, provides for carrying out the clubroot present invention
Unified inoculated identification standard.
Below in conjunction with the accompanying drawings the application principle of the present invention is explained in detail.
A kind of plasmodiophora brassicae inoculation nutrient matrix, this plasmodiophora brassicae inoculation nutrient matrix component according to volume ratio is: peat
Soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2.
As shown in Figure 1: another object of the present invention is to provide the preparation method of a kind of plasmodiophora brassicae inoculation nutrient matrix,
The preparation method of described plasmodiophora brassicae inoculation nutrient matrix comprises the following steps:
S101: peat soil, Vermiculitum, perlite are mixed according to volume ratio 1:0.8～1.2:0.8～1.2 ratio uniform, and
The nutrient matrix of preparation is distributed in sterilizing bag；
S102: the sterilizing bag that will be equipped with nutrient matrix is put in autoclave, sterilizing；
S103: the nutrient matrix after sterilized process is distributed in round plastic nutritive cube and is compacted；
S104: configure acid water with concentrated hydrochloric acid, pours into the acid water with concentrated hydrochloric acid configuration the pallet bottom nutritive cube,
Make that nutrient matrix soil is impregnated with acid moisture stand-by.
Sterilizing methods is: 121 DEG C of moist heat sterilizations 30 minutes.
The acid water pH value of described concentrated hydrochloric acid configuration is 6.0.
The present invention provides a kind of nutrient matrix of application in plasmodiophora brassicae expanding propagation, should the nutrition of application in plasmodiophora brassicae expanding propagation
Matrix components is 100% quartz sand.
Quartz sand particle size is 1mm～2mm.
The present invention provides the application in plasmodiophora brassicae inoculated identification is bred of a kind of plasmodiophora brassicae inoculation nutrient matrix.
Plasmodiophora brassicae inoculation nutrient matrix that the present invention provides and preparation method thereof, by the comparison to 6 kinds of culture matrixes,
The formula of No. 6 culture matrixes found i.e. includes according to volume ratio: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～
1.2, the sickness rate susceptible host reaches 100%, disease index reaches 69.6 (more than the threshold values of disease index 50), is that one can
Good culture matrix for plasmodiophora brassicae inoculated identification；In plasmodiophora brassicae inoculation test, susceptible host falls ill to show and stablizes, energy
Enough Screening and Identification carrying out plasmodiophora brassicae Race Identification and the anti-clubroot of kind；Plasmodiophora brassicae inoculation nutrient matrix according to
Volume ratio includes: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2, uses the nutrient matrix of this proportioning formula to connect
Planting plasmodiophora brassicae, the sickness rate of susceptible variety reaches 100%, and morbidity is stable；
The sickness rate of host's Seedling of No. 2 culture matrix formula that is 100% quartz sands is relatively low, the rear batallion but its host's Seedling is fallen ill
The well-grown supporting body easily forms big single tumor, and No. 2 culture medium can be used to the expanding propagation as plasmodiophora brassicae, easy
Use to the biggest lump.
Below in conjunction with test, the application principle of the present invention is further described.
The present invention is by 6 kinds of nutrient matrix formula of test and 7 plasmodiophora brassicae inoculum densities (resting spore suspension concentration)
Impact on plasmodiophora brassicae inoculated identification effect.Result of the test shows that No. 2 (quartz sands) are sought with No. 6 (peat soil-Vermiculitum-perlite)
The process supporting matrix formulations all has clubroot disease symptom, and other processes (1,3,4, No. 5 process) and does not falls ill.Wherein No. 6 battalion
The process clubroot sickness rate supporting matrix formulations reaches 100%, and disease index reaches 69.6, is a kind of preferably plasmodiophora brassicae inoculation battalion
Supporting matrix formulations, its sickness rate is high, morbidity stably can be utilized for the discriminating of plasmodiophora brassicae biological strain and anti-clubroot breeding material
The evaluation and screening of material；The knee that No. 2 vigorous easy formation of Vegetative growth processed is bigger, can be used as the expanding propagation of plasmodiophora brassicae
Nutrient matrix.The sickness rate of clubroot all shows rising, Qi Zhongjie along with the increase sickness rate of inoculum density with disease index
Planting concentration is 1 × 106、1×107With 1 × 108The process of spores/ml 50dai sickness rate after inoculation all reaches 100%, and
And clubroot morbidity is stably preferable inoculum density.
Specifically comprising the following steps that of test
1 materials and methods
1.1 test materials and appointed condition
Plasmodiophora brassicae derives from Ke Cun Wang Gui China of Yixian County of Anhui Province field cabbage type rape clubroot diseased plant, through Williams physiology
Microspecies differential host is accredited as No. 4 microspecies, and inoculation host material is that Chinese cabbage susceptible variety " fast-growing 2 " is (by the Chinese Academy of Agricultural Sciences
Flowers provide with vegetable institute of the present invention doctor Zhang Hui).Artificial vaccination test is carried out in weather greenhouse, and illumination controlled as daytime
16h, 8h at night；Temperature controls to be daytime 24 DEG C, night 18 DEG C, and humid control is 80%.
1.2 nutrient matrix formula inoculation tests
The main nutritious soil of nutrient matrix composition, peat soil, quartz sand, Vermiculitum, perlite etc. 5 kinds, EXPERIMENTAL DESIGN 6 kinds battalion
Supporting matrix formulations to process, the proportioning of its nutrient matrix is following (table 1).High-pressure sterilizing pot is first used before being used by the nutrient matrix of configuration
Sterilization treatment (121 DEG C, 30min).The nutrient matrix of sterilizing is distributed in the seedlings nursing plate of 50 holes (every hole 100ml), and is placed on
Irrigate with sour water (pH6.0) on supporting pallet；Susceptible variety " fast-growing 2 " with after 70% (v/v) ethanol disinfection 10s with aseptic
Deionized water (ddH2O) concussion rinsing 2 times, then every hole sows 3, and each test processes 3 times and repeats, and often repeats 30 Seedlings.
7d of emerging inoculates, and drawing inoculum density with plastic dropper is 1 × 107The resting spore suspension 1ml of spores/ml is inoculated in Seedling
Daughter root portion.Pouring in pallet with sour water (pH6.0 configures with concentrated hydrochloric acid) after inoculation, inoculation Seedling puts into artificial climate greenhouse；Inoculation
Keep moisture in Nutrition Soil sufficient in latter 2 weeks." Invest, Then Investigate " disease morbidity in 6 weeks.
Table 1, nutrient matrix formula
| Numbering | Process |
| 1 | 100% Vermiculitum |
| 2 | 100% quartz sand |
| 3 | Vermiculitum/quartz sand 1:1 |
| 4 | Vermiculitum/quartz sand/Nutrition Soil 1:1:1 |
| 5 | Vermiculitum/Nutrition Soil 1:1 |
| 6 | Peat soil/Vermiculitum/perlite 1:1:1 |
1.3 plasmodiophora brassicae inoculum density tests
Plasmodiophora brassicae resting spore concentration sets 1 × 10 respectively2、1×103、1×104、1×105、1×106、1×107、1×
1087 inoculum densities such as spores/ml process (table 2).The nutrient matrix used is the battalion in nutrient matrix formula inoculation test
Support No. 6 formula of substrate, be mainly composed of peat soil, Vermiculitum, perlite according to 1:1:1 than arrange uniform mixing, nutrient matrix is through going out
Bacterium is distributed in round plastic nutritive cube (diameter 15cm, high 12cm) after processing.The sowing of inoculation Seedling (fast-growing 2) processes and connects
The method of kind processes 3 times and repeats with nutrient matrix formula inoculation test, each nutritive cube 10 Seedlings of sowing, each test, often repeat
30 Seedlings.After inoculation, 15dai or 20dai sampling uses the plasmodiophora brassicae of observation by light microscope host Gen Mao to infect situation, inoculates 6
Week " Invest, Then Investigate " evaluation clubroot incidence.
Table 2, the inoculum density of resting spore suspension
| Process number | The concentration (spores/ml) of resting spore suspension |
| 1 | 1×102 |
| 2 | 1×103 |
| 3 | 1×104 |
| 4 | 1×105 |
| 5 | 1×106 |
| 6 | 1×107 |
| 7 | 1×108 |
Prepared by 1.4 plasmodiophora brassicae inoculation liquid
Collect resting spore and use (2013) methods such as Feng, from refrigerator-freezer, take out the Brassica campestris L knee 1 at being stored at-20 DEG C
Individual (about 30 grams), put 1h at room temperature, the knee after softening of thawing are put into (nine sun cooking machine JYL-C16D) in blender,
Add the sterile deionized water (ddH of 100ml2O) stirring.After knee fully blends, mixed liquor is with after 8 layers of filtered through gauze, obtains
Plasmodiophora brassicae resting spore suspension is centrifuged 5 minutes at 50g, collects supernatant (containing resting spore), therefrom samples 1ml in supernatant
Dilute 50 times, check plasmodiophora brassicae resting spore quantity with medical blood counting chamber and calculate concentration (1 milliliter of dormancy of resting spore
Resting spore quantity/80 × 400 × 10 in the little lattice in number=80 of resting spore in spore suspension4× 50), then use
Sterile deionized water (ddH2O) concentration of spore suspension it is transferred to desired concn insert in 4 DEG C of refrigerators and save backup.
1.5 optical microscope
20d (dai) after inoculation, observes plasmodiophora brassicae with optical microscope (Olympus BX51) and infects host's situation, each
Repeat to take 3 Seedlings, root knife blade is cut into the root segment of 5mm length, respectively with optics microscopical 20 times and 40 times of object lens
Observe the situation of secondary protoplasm group quantity in Root hair infection rate and cell, it is thus achieved that the meansigma methods conduct of data of 3 young plants
The data value repeated.
1.6 knee investigation process with data
42d (dai) after inoculation, the incidence of investigation record Brassica campestris L clubroot, investigation standard uses 0 to 3 grade and carrys out classification,
Seedling stage Brassica campestris L clubroot grade scale: 0 grade is that root is anosis, and 1 grade has tiny tumor for root, and 2 grades have median tumor for root,
3 grades have abnormal big tumor for root.Calculate diseased plant rate and disease index respectively.Diseased plant rate %=morbidity strain number/investigate total strain number
×100；Disease index=∑ (diseased plant number × appropriate level)/(investigating total strain number × highest level) × 100.Use Excell soft
Part repairing experiment initial data also carries out correlation regression statistical analysis and maps；Utilize SAS9.1 software to the test data side of carrying out
Difference analysis and significance analysis.
2 results and analysis
Plasmodiophora brassicae is infected the impact of susceptible host by 2.1 nutrient matrixes
The result of the present invention using 6 kinds of different nutrient matrixes to carry out plasmodiophora brassicae effect of inoculation shows Different Nutrition substrate
Process clubroot incidence and differ greatly (table 3).Test inoculation investigated criteria for classification according to the clubroot of 0 to 3 grade after 6 weeks,
The incidence of investigation inoculation host's Seedling Brassica campestris L clubroot, only No. 2 process (100% quartz sand culture matrix) and No. 6 process
(peat soil/Vermiculitum/perlite 1:1:1) inoculation host's Seedling has clubroot to fall ill, and other No. 1, No. 3, No. 4, No. 5 processes inoculation and posts
Main Seedling is not fallen ill.Wherein No. 6 host's Seedling sickness rate processed reach 100%, and disease index reaches 69.6, is that preferable clubroot connects
The nutrient matrix planted.No. 2 process inoculation host's Seedling clubroot sickness rate is 52.4%.
From the point of view of the growth potential of young plant, No. 1 to No. 5 cultivation from the point of view of host's Seedling growth potential that 6 kinds of medium matrixs are cultivated
The host Miao Shengchang of substrate is vigorous, and No. 2 host's Seedlings processed do not have difference with the host's Seedling processed of not falling ill on growth potential, but
Being that the host's Seedling growth potential on No. 6 nutrient matrixes is more weak, host's Seedling is significantly less than host's Seedling of other process do not fallen ill, this
Phenomenon is the absorbability result by the limitations affect of old complaint lump of nutrient and moisture due to morbidity strain root.At No. 6
Reason compares with No. 2 process of same morbidity, and the young plants of No. 2 process are significantly greater than No. 6 process, from the knee development characteristics of diseased plant
Seeing, there are many lumps in No. 6 roots processed, also have lump to occur on fibrous root, present morning of falling ill, the feature of disease weight；And No. 2
The diseased plant root processed presents single bigger lump.
Table 3, the clubroot incidence of 6 kind of nutrient matrix
| Numbering | Process | Diseased plant rate % | Disease index |
| 1 | 100% Vermiculitum | 0.0 | 0.0 |
| 2 | 100% quartz sand | 52.4 | 36.6 |
| 3 | Vermiculitum/quartz sand 1:1 | 0.0 | 0.0 |
| 4 | Vermiculitum/quartz sand/Nutrition Soil 1:1:1 | 0.0 | 0.0 |
| 5 | Vermiculitum/Nutrition Soil 1:1 | 0.0 | 0.0 |
| 6 | Peat soil/Vermiculitum/perlite 1:1:1 | 100.0 | 69.6 |
The impact on plasmodiophora brassicae inoculated identification of 2.2 inoculum densities
To after the resting spore inoculation of suspension liquid susceptible host 20dai of 7 kinds of inoculum densities optical microscope plasmodiophora brassicae
The result infected shows: the root hair that 20dai all processes after inoculation all has plasmodiophora brassicae to infect, between different inoculum densities processes
Root hair infection rate significant difference, and Root hair infection rate raises along with the increase of inoculum density, and inoculum density is 1 × 107With 1
×108The process 20dai Root hair infection rate of spores/ml reaches or close to 100%, and inoculum density is 1 × 105、1×
106、1×107With 1 × 108The process of spores/ml 20dai after inoculation has occurred in that knee, but low inoculum density
Process and knee does not the most occur.Secondary protoplasm group number difference between 20dai different vaccination concentration processes after inoculation is the most notable.
After inoculation, the survey showed that for the classification of 50dai clubroot: the process of 7 kinds of inoculum densities all has knee, different
Inoculum density processes intercurrent disease rate and all reaches significant difference with disease index.Sickness rate is 12% to 100%, along with inoculation is dense
Sickness rate and the disease index of the increase susceptible variety of degree all show rising, and wherein inoculum density is 1 × 106、1×107With 1 ×
108The process of spores/ml 50dai sickness rate after inoculation all reaches 100%；The process of low inoculum density then has not to be sent out in a large number
Diseased plant, wherein resting spore concentration 1 × 102The process sickness rate of spores/ml is less than 20%.
This test is by the comparison to 6 kinds of culture matrixes, and the formula of No. 6 culture matrixes of discovery is in the morbidity of susceptible host
Rate reaches 100%, disease index reaches 69.6 (more than the threshold values of disease index 50), is a kind of to can be used for clubroot inoculated identification
Good culture matrix.The morbidity stability showed in plasmodiophora brassicae inoculation test due to it, this nutrient matrix formula can
It is used for carrying out the Screening and Identification of Race Identification and varietal resistance.The sickness rate of host's Seedling of No. 2 culture matrix formula is relatively low,
But after the morbidity of its host's Seedling, the well-grown of trophosome easily forms big single tumor, and therefore No. 2 culture medium can be used
The expanding propagation of the plasmodiophora brassicae being used as in the present invention, is readily obtained single big knee with these No. 2 culture medium expanding propagation plasmodiophora brassicaes.Knee
The concentration of bacterium inoculated identification is with 1 × 106To 1 × 107Spores/ml is advisable, and low concentration sickness rate is relatively low, spore concentration too Gao Ze
There will be morbidity and be early prematurely formed dead Seedling.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Any amendment, equivalent and the improvement etc. made within god and principle, should be included within the scope of the present invention.
Claims (7)
1. a plasmodiophora brassicae inoculation nutrient matrix, it is characterised in that this plasmodiophora brassicae inoculation by nutrient matrix component according to volume
Ratio is: peat soil: Vermiculitum: perlite=1:0.8～1.2:0.8～1.2.
2. the preparation method of a plasmodiophora brassicae inoculation nutrient matrix as claimed in claim 1, it is characterised in that described plasmodiophora brassicae
The preparation method of inoculation nutrient matrix comprises the following steps:
Peat soil, Vermiculitum, perlite are mixed according to volume ratio 1:0.8～1.2:0.8～1.2 ratio uniform, and by preparation
Nutrient matrix is distributed in sterilizing bag；
The sterilizing bag that will be equipped with nutrient matrix is put in autoclave, sterilizing；
Nutrient matrix after sterilized process is distributed in round plastic nutritive cube and is compacted；
Configure acid water with concentrated hydrochloric acid, the pallet bottom nutritive cube pours into the acid water with concentrated hydrochloric acid configuration, makes Nutrient medium
Matter soil is impregnated with acid moisture stand-by.
3. the preparation method of plasmodiophora brassicae inoculation nutrient matrix as claimed in claim 2, it is characterised in that sterilizing methods is:
121 DEG C of moist heat sterilizations 30 minutes.
4. the preparation method of plasmodiophora brassicae inoculation nutrient matrix as claimed in claim 2, it is characterised in that described concentrated hydrochloric acid
The acid water pH value of configuration is 6.0.
5. the preparation method of a plasmodiophora brassicae inoculation nutrient matrix as claimed in claim 2 prepare in plasmodiophora brassicae expanding propagation
The nutrient matrix of application, it is characterised in that should the nutrient matrix component of application be 100% quartz sand in plasmodiophora brassicae expanding propagation.
6. the nutrient matrix of application in plasmodiophora brassicae plasmodiophora brassicae expanding propagation as claimed in claim 5, it is characterised in that quartz
Sand grains footpath is 1mm～2mm.
7. a plasmodiophora brassicae inoculation nutrient matrix as claimed in claim 1 application in plasmodiophora brassicae inoculated identification is bred.
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| CN107058460A (en) * | 2017-04-25 | 2017-08-18 | 江西省农业科学院植物保护研究所 | The inoculation identification method of crop in cruciferae clubroot |
| CN107466720A (en) * | 2017-08-25 | 2017-12-15 | 李国梁 | A kind of breeding method of the nontoxic apple seedling of high dwarfing stock |
| CN110178579A (en) * | 2019-07-02 | 2019-08-30 | 沈阳农业大学 | A kind of Cruciferae clubroot identification method |
| CN110487615A (en) * | 2019-08-29 | 2019-11-22 | 沈阳农业大学 | A kind of composite fluorescence colouring method for identifying ten Zi Hua section plant clubroots |
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| CN107058460A (en) * | 2017-04-25 | 2017-08-18 | 江西省农业科学院植物保护研究所 | The inoculation identification method of crop in cruciferae clubroot |
| CN107466720A (en) * | 2017-08-25 | 2017-12-15 | 李国梁 | A kind of breeding method of the nontoxic apple seedling of high dwarfing stock |
| CN110178579A (en) * | 2019-07-02 | 2019-08-30 | 沈阳农业大学 | A kind of Cruciferae clubroot identification method |
| CN110178579B (en) * | 2019-07-02 | 2021-09-24 | 沈阳农业大学 | Cruciferous clubroot identification method |
| CN110487615A (en) * | 2019-08-29 | 2019-11-22 | 沈阳农业大学 | A kind of composite fluorescence colouring method for identifying ten Zi Hua section plant clubroots |
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