CN106119337A - A kind of Rapid identification peanut varieties method to Aspergillus flavus resistance - Google Patents
A kind of Rapid identification peanut varieties method to Aspergillus flavus resistance Download PDFInfo
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Abstract
The present invention relates to a kind of Rapid identification peanut varieties method to Aspergillus flavus resistance, the present invention is logical carries out surface sterilization by fresh or after drying peanut seed, on peanut seed capsule, manufacture wound after sterilization and connect bacterium, using cotton balls moisturizing after connecing bacterium, be then put into 30 DEG C of incubators and carry out light culture.After light culture one week, by calculating the Aspergillus flavus infection index assessment peanut varieties resistance to Aspergillus flavus.With compared with the method weighing Aspergillus flavus Biomass after connecing bacterium, the present invention can not consider the error of the water content of peanut seed own, evaluation result more accurately and is stablized.
Description
Technical field
The present invention relates to a kind of Rapid identification peanut varieties method to Aspergillus flavus resistance, belong to plant germplasm identification technology
Field.
Background technology
Semen arachidis hypogaeae is industrial crops, oil crop and the export crop that China is important, and China's Semen arachidis hypogaeae Annual planting area connects
Nearly 5,000,000 hectares, annual production about 17,000,000 tons.The cultivated area of China's Semen arachidis hypogaeae accounts for the 20% of the total cultivated area in the world, occupies generation
The 2nd, boundary；Yield accounts for about the 40% of Gross World Product, occupies the 1st, the world.But Semen arachidis hypogaeae field management, results, dry in the sun,
Storage, transport and the course of processing are subject to the pollution of Aspergillus flavus and produce aflatoxin, the health of the mankind is caused
Harm greatly, and have a strong impact on the quality of China's Semen arachidis hypogaeae and goods thereof, become the barrier of China's Semen arachidis hypogaeae green trade.Along with
Aflatoxin contamination in the raising of people's Consciousness of food security, Semen arachidis hypogaeae and goods thereof has become as a serious grain peace
Full problem.
Aflatoxin is mainly produced by the secondary metabolism that Aspergillus flavus is similar with one group of chemical constitution that Herba balanophorae involucratae aspergillosis produces
Thing is constituted, and the most separated identifying includes B1, B2, G1, G2, M1, M2Deng 12 kinds of hypotypes, with aflatoxin in natural food
B1The most common, its toxicity and carcinogenecity are the strongest, the significantly larger than toxicity of cyanide, arsenide and organic agricultural chemicals.All
In aflatoxin, the toxicity of B is the strongest, and its toxicity is equivalent to 10 times of potassium cyanide, 68 times of arsenicum.The harm of aflatoxin
Property is to have people and animal livers tissue destruction, may result in hepatocarcinoma the most dead time serious.The detection of aflatoxin
Method includes: thin layer chromatography (TLC), high performance liquid chromatography (HPLC), microtrabeculae screening method, enzyme linked immunosorbent assay
(ELISA), multifunctional column cleanup-post-column derivatization-high performance liquid chromatography, immune affinity column purification-fluorophotometer method, exempt from
Epidemic disease affinity column purification-post-column derivatization-high performance liquid chromatography etc..The pre-treating method of various detection methods is not quite similar, inspection
The index the most differences such as survey mode is different, detection limit.
Aspergillus flavus is very wide in distributed in nature, is normally present in soil and other materials, infects the kind of grain, oil crop
Food, various and feedstuff etc..Peanut pod and Semen arachidis hypogaeae are the main contaminated objects of Aspergillus flavus, and toxigenic bacterium strain infecting peanut is also
Producing aflatoxin in seed core, the edible Semen arachidis hypogaeae infected causes potential threat can to people and animals' life.At high temperature, high humidity
Under the conditions of, Semen arachidis hypogaeae is highly susceptible to the pollution of Aspergillus flavus.In recent years, people often rely mainly on prevention, and strengthening is being gathered in the crops, basks seeds, transported
Fungus-proof measure defeated, that store in processing and the link such as sale, and take mould to select, physical absorption, heating detoxification, ammonia are stifling,
The methods such as alkali refining detoxification, photoelectricity rejecting remove aflatoxin, but these methods exist, and cost is high, medicament remains and detoxification is the most thorough
The end, change the shortcoming such as local flavor of (Groundnut products), from economy and ecological consideration, promote and plant the peanut varieties anti-to Aspergillus flavus height
It is to prevent aflatoxin pollution of peanuts essential measure.Therefore, the accurately and quickly qualification to peanut varieties Aspergillus flavus resistance is right
Most important in the evaluation of peanut varieties.
Past identifies that the Semen arachidis hypogaeae method to the resistance of Aspergillus flavus infection is mainly by following several:
(1) by after artificial vaccination, it is utilized as thin layer chromatography and the content of high performance liquid chromatography detection Aspergillus flavus, should
Method operation is complicated, sample detection costly, high to equipment requirements, it is impossible to the resistance mirror being efficiently applied in peanut breeding research
Fixed；
(2) quantity and degree estimating Aspergillus flavus infection kernel by detecting by an unaided eye after artificial vaccination, this method is grasped
Make simple, but error more greatly can not the effectively evaluating Semen arachidis hypogaeae resistance to Aspergillus flavus infection；(3) by artificial vaccination post analysis
Aflatoxin content, protease inhibitory activity, lipoxidase activity, chitinase activity etc. evaluate Semen arachidis hypogaeae to Aspergillus flavus
Resistance, the method is more accurate, but process is complex, actual application and Difficulty；(4) connect by yellow inulinase bacterial strain
Cultivate peanut and carry out light culture after seed, then use metrical instrument analytical balance, the seed after light culture is weighed, finally calculates
Infection by Aspergillus flavus index also carries out evaluation of resistance.But in the method, the increment of Aspergillus flavus is by different Semen arachidis hypogaeae kinds
Son is weighed before infecting and after infecting and to be determined, due to peanut seed this may be to commenting in the difference infecting before and after's water content
Valency result interferes, thus Stability and veracity is worthy of consideration；(5) reality of some field test Aspergillus flavus resistances of forefathers
Test, not only operating difficulties, waste time and energy, and peanut varieties to Aspergillus flavus resistance expression with different year, plantation place, sowing
Season, the soil texture and plant byturns the change of type and fluctuate, stability is the strongest.
Such as, Chinese patent literature CN1556391A (application number 200310111682.5) discloses one and cultivates peanut to yellow bent
Mycete produces the method for poison Resistance Identification." peanut aspergillus flavus anti aflatoxin production preperty identification method is characterized in that being realized by following steps: 1)
Peanut seed surface sterilization and rehydration；2) peanut seed inoculation: with the healthy Semen arachidis hypogaeae of aspergillus flavus strain inoculation；3) cultivate: after inoculation
Incubator is cultivated 7 days；4) aflatoxin is extracted: toasted, grinding, then with methanol extraction aflatoxin liquid；5) poison
Element measures: add ammonia mercury solution, measures fluorescent value, with between AFB1 and fluorescent value on spectrofluorophotometer
Standard curve calculates AFB1 content and evaluates resistance class." the method need baking, grind, again with methanol extract
Aflatoxin liquid and it needs to add ammonia mercury solution, calculates the content of AFB1 by standard curve.Operation complexity,
Sample detection costly, detection during use multiple toxic reagent, misoperation easily lead to detect environment pollution, because of
This promotion and application relative difficulty；
Chinese patent literature CN102818740A (application number 201210332642.2) discloses one and cultivates peanut to Aspergillus flavus
The authentication method of the resistance infected and application thereof.This invention is to carry out light culture, then after yellow inulinase inoculation peanut seed
Use metrical instrument analytical balance, the seed after light culture is weighed, finally calculate infection by Aspergillus flavus index and carry out resistance and comment
Valency.Due to peanut seed, evaluation result may be interfered by this in the difference infecting before and after's water content, thus the method
Stability and veracity be worthy of consideration.
Summary of the invention
The present invention is directed to the existing qualification Semen arachidis hypogaeae defect to Aspergillus flavus infection resistance method, it is provided that one is quickly, accurately reflect
The method determining the anti-sense kind of aspergillus flavus.The method compensate for existing aspergillus flavus Resistance Identification method cycle length,
Complex operation, the deficiency of Aspergillus flavus infection situation cannot be observed by qualitative, quantitative, can under relatively simple experiment condition effectively
Identify the peanut varieties resistance to Aspergillus flavus, simple to operate, Resistance Identification low cost, experimental result degree of accuracy is high.
Technical scheme is as follows:
A kind of Rapid identification peanut varieties method to Aspergillus flavus resistance, step is as follows:
(1) Aspergillus flavus actication of culture, prepares aspergillus spore suspension；
(2) seed of after planting 80～100 days is peelled off peel, take the intact seed of seed coat as sample, enter with mercuric chloride
Row surface sterilization；Then on seed, prepare the wound removing seed coat of (5.5～6.5mm) × (4.5～5.5mm)；
(3) on the wound of seed, cover sterile cotton bar, cotton sliver drips aspergillus spore suspension prepared by step (1)
Liquid 10～30 microlitre, is placed in sterile petri dish；
(4) sterile petri dish is placed in incubator carries out light culture, take cotton sliver off after cultivating 20～28 hours, be further cultured for
After 132～156 hours, statistics different peanut varieties infects Aspergillus flavus degree, calculates its infestation index, whether judges this kind
Aspergillus flavus resisting, infestation index's computing formula is as follows:
Infestation index={ (0.1n+0.2n+0.5n+0.8n+1.0n)/N} × 100
Wherein: n is the kernel grain number of each rank microbiological contamination；N is for for trying total grain number；
(5) varietal resistance criterion is: infestation index ＜'s 5.0 is high anti-kind；Infestation index >=5.0, and ＜
10.0 be in anti-kind；Infestation index >=10.0, and ＜ 30.0 is middle sense kind；Infestation index >=30.0, and ＜ 50.0
It it is susceptible variety；Infestation index >=50.0 are high sense kinds.
Aspergillus flavus actication of culture in described step (1) is streak culture activation Aspergillus flavus on PDA solid medium
Kind, activation condition is to cultivate 6～7 days in the dark culturing case of 30 DEG C；
Every liter of component of PDA solid medium is as follows:
Rhizoma Solani tuber osi 300g, glucose 20g, agar 20g, pH value is natural.
In described step (1), the preparation process of aspergillus spore suspension is as follows:
Being rinsed by the Aspergillus flavus mycelia covering with PDA solid culture primary surface with sterile purified water, sterilized water is resuspended, adjusts
Whole bacterial concentration is 106Individual/ml.
Seed in described step (2) is the fresh seeds of healthy peanut plant.
Sterilization in described step (2), step is as follows:
Sample is placed in the mercuric chloride solution that concentration is 0.1%, 50～150r/min sterilization 10min, then rushes with distilled water
Wash three times, when rinsing the 3rd time, under conditions of 50～150r/min, rinse 5min.
Wound described in described step (2) is 6mm × 5mm；It is further preferred that described wound is positioned at Semen arachidis hypogaeae two panels
The centre position of cotyledon.This position is removed seed coat and can be increased and infect efficiency.
In described step (2), the quantity of every kind of sample is at least 15.
In described step (3), aseptic cotton sliver size is identical with wound size.
Each culture dish at least 5 samples in described step (3).
Sterile petri dish in described step (3) is toilet paper to fold two-layer be cut into circle and be laid in culture dish, sterilizing,
Each culture dish adds sterile purified water, makes the water content in culture dish reach 80%.Water content reaches 80% and is conducive to yellow song
Mould growth；The fold that moistening toilet paper is formed, can stop rocking of peanut seed, has the most both played peanut seed solid
The tailor-made growth ensureing Aspergillus flavus with moistening environment in holding culture dish again.
The temperature of the light culture described in described step (4) is 30 DEG C, and the time is 7 days.This temperature is the suitableeest of Aspergillus flavus
Growth temperature, beneficially Aspergillus flavus quickly infect peanut seed.
Take cotton sliver off after described step (4) is cultivated 24 hours, be further cultured for 144 hours.
Cultivation 20 described in step (4)～take cotton sliver reason after 28 hours off and be to cover cotton sliver 20～make for 28 hours
Aspergillus spore suspension and peanut seed are fully contacted to infect completely, and taking off after 20～28 hours is to allow the Huang grown
Aspergillosis is fully contacted with peanut seed, beneficially the growth of Aspergillus flavus, and facilitates observation to infect efficiency；
In described step (4), the grade scale of kernel microbiological contamination rank is: 0 grade [0.0]: kernel is normal；I level [0.1]: kernel
Grow mould thing and account for kind of a clothing area less than 1/10；II level [0.2]: kernel grows mustiness thing and accounts for kind of clothing area more than 1/10 and be less than
1/5；III level [0.5]: kernel grows mustiness thing and accounts for kind of clothing area more than 1/5 and less than 1/2；IV level [0.8]: kernel grows
Mustiness thing accounts for kind of clothing area more than 1/2 and less than 4/5；V level [1.0]: kernel surface is covered with mustiness thing.
Beneficial effect
1 compares with certain methods in the past, such as thin layer chromatography, high performance liquid chromatography, artificial vaccination post analysis Huang
The detection methods such as aspertoxin content, protease inhibitory activity, lipoxidase activity, chitinase activity, the present invention couple
Equipment requirements is high, and testing cost is low；
2, the method for the artificial inoculation of comparing over, the present invention devises the link manufacturing wound, improves Aspergillus flavus and connect
The efficiency of bacterium；
3 estimate to identify that Aspergillus flavus is invaded the method for resistance and compares with perusal, and the present invention utilizes Aspergillus flavus infection index to reflect
Determine the resistance of kind, more science, accurately；
4 with connect bacterium after weigh Aspergillus flavus Biomass method compare, the present invention can not consider that peanut seed itself contains
The error of the water yield, evaluation result more accurately and is stablized.
Accompanying drawing explanation
Fig. 1 peanut varieties Guangdong oil 20 connects the photo after bacterium is cultivated；
Fig. 2 peanut varieties Tifrunner connects the photo after bacterium is cultivated；
Fig. 3 peanut varieties Shandong spends No. 14 to connect the photo after bacterium is cultivated；
Fig. 4 peanut varieties Ji spends No. 11 to connect the photo after bacterium is cultivated；
Fig. 5 peanut varieties is spent No. 5 connect the photo after bacterium is cultivated；
Fig. 6 peanut varieties Henan spends No. 6 to connect the photo after bacterium is cultivated；
Fig. 7 peanut varieties is spent No. 6 connect the photo after bacterium is cultivated；
Specific embodiments
Below in conjunction with Figure of description and embodiment, technical scheme is described further, but the present invention is protected
The scope of protecting is not limited to this.
Experimental technique in following embodiment, if no special instructions, is this area conventional method.
Biomaterial
Peanut varieties Guangdong oil 20, Tifrunner, Shandong spend No. 14, Ji spend No. 11, middle spend No. 5, Henan spends No. 6, middle spends No. 6.
Aspergillus flavus strain described in embodiment is purchased from Inst. of Oil Crops, Chinese Academy of Agriculture, numbering SDAFZ1.
Embodiment 1: the cultivation of Aspergillus flavus and the collection of spore
(1) Aspergillus flavus strain is rule on PDA solid medium activation culture, cultivate in the dark culturing case of 30 DEG C
6 days；
Every liter of component of PDA solid medium is as follows:
Rhizoma Solani tuber osi 300g, glucose 20g, agar 20g, pH value is natural.
(2) after the full media surface of Aspergillus flavus filament length, being rinsed with sterile purified water, sterilized water is resuspended,
Calculate its concentration with blood counting chamber, spore concentration is adjusted to 106Individual/ml, standby as aspergillus spore suspension.
Embodiment 2: the process of peanut seed
(1) take peanut varieties Guangdong oil 20, Tifrunner, Shandong spend No. 14, Ji spend No. 11, middle spend No. 5, Henan spends No. 6, middle spends 6
Each 15 of the seed of number healthy peanut plant, peels off peel, takes the intact seed of seed coat as sample, is sub-packed in conical flask
(if the seed dried, need to be soaked in sterilized water by seed and fully water suction)；
(2) adding the mercuric chloride sterilization 10min of mass concentration 0.1%, period, 100r/min rocked, then with distilled water flushing three
Time, when rinsing the 3rd time, 100r/min rinses 5min, then all manufactures the wound of 6mm × 5mm on seed, removes wound part
Seed coat.
Embodiment 3: inoculation Aspergillus flavus
(1) the wound overlying of seed covers an equal amount of aseptic cotton sliver in example 2, drips embodiment on cotton sliver
Aspergillus spore suspension 20 microlitre of preparation in 1, the culture dish being sub-packed in numbered sterilizing is cultivated；
(2) absorbent paper folding two-layer be cut into circle and be laid in culture dish, each culture dish adds the aseptic distillation of 5ml
Water, moist toilet paper, make the water content in culture dish reach 80%, each culture dish puts five seeds, and each sample sets 3 weights
Multiple；
(3) culture dish is placed in incubator, temperature be 30 DEG C carry out light culture 24 hours after take cotton sliver off, more secretly train
Support and cultivate 144 hours.
Embodiment 4: the statistics of infestation index and calculating
(1) add up the infection Aspergillus flavus degree of Semen arachidis hypogaeae, calculate its infestation index, judge this kind whether aspergillus flavus resisting.
Infestation index={ (0.1n+0.2n+0.5n+0.8n+1.0n)/N} × 100
Wherein: n is the kernel grain number of each rank microbiological contamination；N is for for trying total grain number.
Varietal resistance criterion is: infestation index ＜'s 5.0 is high anti-kind；Infestation index >=5.0, and ＜ 10.0
Anti-kind in being；Infestation index >=10.0, and ＜ 30.0 is middle sense kind；Infestation index >=30.0, and ＜'s 50.0 is susceptible
Kind；Infestation index >=50.0 are high sense kinds.
Kernel microbiological contamination grade scale with the relation of kernel symptom is:
Embodiment 5: the qualification of peanut varieties resistance
After inoculation Aspergillus flavus, cultivate 6 days in 30 DEG C of constant incubators, observe Guangdong oil 20, Tifrunner, Shandong spend No. 14,
Ji spend No. 11, middle spend No. 5, Henan spend No. 6, the middle infection conditions spending No. 6 each kinds, result is as shown in Fig. 1～7.In major part Guangdong
Oil 20 and the middle the surface of the seed spending No. 5 do not observe Aspergillus flavus mycelia, and the surface of major part Tifrunner seed is the longest
Expire the mycelia of Aspergillus flavus, shown Guangdong oil 20 and middle spend No. 5 the resistances of Aspergillus flavus are eager to excel than Tifrunner.
The Disease Resistance Identification result of table 1 peanut varieties
Interpretation of result
According to kernel microbiological contamination grade scale, add up above-mentioned peanut varieties kernel microbiological contamination rank after connecing bacterium 6 days, calculate and infect
Index, identifies the above-mentioned peanut varieties resistance to Aspergillus flavus, and qualification result is as shown in table 1.In spend No. 5 and Guangdong oil 20 belongs to high anti-
Aspergillus flavus kind；Tifrunner belongs to high sense kind；In spend No. 6 to belong in anti-kind；Ji spends No. 11, Shandong spends No. 14 and Henan spends 6
Number belong to middle sense kind.
The Resistance Identification result of peanut varieties and previous investigation (document 1:Guo B, Chen X, Dang P, et
al.Peanut gene expression profiling in developing seeds at different
reproduction stages during Aspergillus parasiticus,infection[J].Bmc
Developmental Biology,2008,8(1):1-16.；Document 2:Guo, B., Fedorova, N.D., Chen, X., Wan,
C.H.,Wang,W.,Nierman,W.C.,Bhatnagar,D.and Yu,J.(2011)Gene expression
profiling and identification of resistance genes to Aspergillus flavus
infection in peanut through EST and microarray strategies.Toxins(Basel),3,
737-753. document 3: Guo Jingpei. Resistance Identification and the anti-infection mechanism of Aspergillus flavus are studied [D] by peanut varieties. and Henan is pedagogical
University's master thesis, 2012.) consistent, show that method of the present invention has higher accuracy.
Compared with conventional method, the present invention, under relatively simple experiment condition, can effectively identify peanut varieties
Resistance to Aspergillus flavus infection, is especially suitable for making peanut breeding material batch screening, Resistance Identification low cost.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, the change made under other any spirit without departing from the present invention and principle, modify, substitute, combine, simplify,
All should be the substitute mode of equivalence, within being included in protection scope of the present invention.
Claims (10)
1. the Rapid identification peanut varieties method to Aspergillus flavus resistance, it is characterised in that step is as follows:
(1) Aspergillus flavus actication of culture, prepares aspergillus spore suspension；
(2) seed of after planting 80～100 days is peelled off peel, take the intact seed of seed coat as sample, sterilization；Then planting
The wound removing seed coat of (5.5～6.5mm) × (4.5～5.5mm) is prepared on son；
(3) on the wound of seed, cover sterile cotton bar, cotton sliver drips aspergillus spore suspension 10 prepared by step (1)
～30 μ l, it is placed in sterile petri dish；
(4) sterile petri dish is placed in incubator carries out light culture, take cotton sliver off after cultivating 20～28 hours, be further cultured for 132
～after 156 hours, statistics different peanut varieties infects Aspergillus flavus degree, calculates its infestation index, judges whether this kind resists
Aspergillus flavus, infestation index's computing formula is as follows:
Infestation index={ (0.1n+0.2n+0.5n+0.8n+1.0n)/N} × 100
Wherein: n is the kernel grain number of each rank microbiological contamination；N is for for trying total grain number；
(5) varietal resistance criterion is: infestation index ＜'s 5.0 is high anti-kind；Infestation index >=5.0, and ＜ 10.0
Anti-kind in being；Infestation index >=10.0, and ＜ 30.0 is middle sense kind；Infestation index >=30.0, and ＜'s 50.0 is susceptible
Kind；Infestation index >=50.0 are high sense kinds.
2. the method for claim 1, it is characterised in that the Aspergillus flavus actication of culture in described step (1) is solid at PDA
Streak culture activation Aspergillus flavus strain in body culture medium, activation condition is to cultivate 6～7 days in the dark culturing case of 30 DEG C；
Every liter of component of PDA solid medium is as follows:
Rhizoma Solani tuber osi 300g, glucose 20g, agar 20g, pH value is natural.
3. the method for claim 1, it is characterised in that the preparation step of aspergillus spore suspension in described step (1)
Rapid as follows:
Being rinsed by the Aspergillus flavus mycelia covering with PDA solid culture primary surface with sterile purified water, sterilized water is resuspended, adjusts bacterium
Liquid concentration is 106Individual/ml.
4. the method for claim 1, it is characterised in that the seed in described step (2) is the new of healthy peanut plant
Fresh seed；
Preferably, the sterilization in described step (2), step is as follows:
Sample is placed in the mercuric chloride solution that concentration is 0.1%, 50～150r/min sterilization 10min, then with distilled water flushing three
Time, when rinsing the 3rd time, under conditions of 50～150r/min, rinse 5min.
5. the method for claim 1, it is characterised in that the wound described in described step (2) is 6mm × 5mm；Enter one
Preferably, described wound is positioned at the centre position of Semen arachidis hypogaeae two panels cotyledon to step；
Preferably, in described step (2), the quantity of every kind of sample is at least 15.
6. the method for claim 1, it is characterised in that aseptic cotton sliver size and wound size phase in described step (3)
With；
Each sterile petri dish at least 5 samples in described step (3).
7. the method for claim 1, it is characterised in that the sterile petri dish in described step (3) is to be rolled over by toilet paper
Folded two-layer is cut into circle and is laid in culture dish, sterilizing, and each culture dish adds sterile purified water, makes the water content in culture dish reach
To 80%.
8. the method for claim 1, it is characterised in that the temperature of the light culture described in described step (4) is 30 DEG C,
Time is 7 days.
9. the method for claim 1, it is characterised in that take cotton sliver off after cultivating 24 hours in described step (4), then train
Support 144h.
10. the method for claim 1, it is characterised in that in described step (5), kernel microbiological contamination grade scales at different levels are: 0
Level [0.0]: kernel is normal；I level [0.1]: kernel grows mould thing and accounts for kind of a clothing area less than 1/10；II level [0.2]: kernel grows mould
Shape thing accounts for kind of clothing area more than 1/10 and less than 1/5；III level [0.5]: kernel grows mustiness thing and accounts for kind of clothing area more than 1/5
And less than 1/2；IV level [0.8]: kernel grows mustiness thing and accounts for kind of clothing area more than 1/2 and less than 4/5；V level [1.0]: plant
Core surface is covered with mustiness thing.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107258309A (en) * | 2017-07-21 | 2017-10-20 | 邯郸市农业科学院 | A kind of cotton variety annidation, the breeding method of yielding stability |
| CN107843595A (en) * | 2017-10-24 | 2018-03-27 | 南京苏试广博环境可靠性实验室有限公司 | Mould test method |
| CN109001385A (en) * | 2018-06-15 | 2018-12-14 | 中国农业科学院油料作物研究所 | The indoor appraising method and its application of a kind of peanut pod to Aspergillus flavus infection resistance |
| CN111979293A (en) * | 2020-08-28 | 2020-11-24 | 河南省农业科学院 | Indoor accurate identification method for resistance of peanut kernels to aspergillus flavus infection |
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| CN1556391A (en) * | 2003-12-30 | 2004-12-22 | 中国农业科学院油料作物研究所 | Peanut aspergillus flavus anti aflatoxin production preperty identification method |
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| CN107843595B (en) * | 2017-10-24 | 2020-06-26 | 南京苏试广博环境可靠性实验室有限公司 | Mould test method |
| CN109001385A (en) * | 2018-06-15 | 2018-12-14 | 中国农业科学院油料作物研究所 | The indoor appraising method and its application of a kind of peanut pod to Aspergillus flavus infection resistance |
| CN111979293A (en) * | 2020-08-28 | 2020-11-24 | 河南省农业科学院 | Indoor accurate identification method for resistance of peanut kernels to aspergillus flavus infection |
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