CN101603073B - Field identification method for peanut aspergillus flavus infection resistance - Google Patents
Field identification method for peanut aspergillus flavus infection resistance Download PDFInfo
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- CN101603073B CN101603073B CN 200910159347 CN200910159347A CN101603073B CN 101603073 B CN101603073 B CN 101603073B CN 200910159347 CN200910159347 CN 200910159347 CN 200910159347 A CN200910159347 A CN 200910159347A CN 101603073 B CN101603073 B CN 101603073B
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- peanut
- aspergillus flavus
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- 235000020232 peanut Nutrition 0.000 title claims abstract description 60
- 235000010777 Arachis hypogaea Nutrition 0.000 title claims abstract description 59
- 235000017060 Arachis glabrata Nutrition 0.000 title claims abstract description 56
- 235000018262 Arachis monticola Nutrition 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 25
- 208000036641 Aspergillus infections Diseases 0.000 title claims abstract description 10
- 241001553178 Arachis glabrata Species 0.000 title 1
- 244000105624 Arachis hypogaea Species 0.000 claims abstract description 62
- 241000228197 Aspergillus flavus Species 0.000 claims abstract description 34
- 240000008042 Zea mays Species 0.000 claims abstract description 19
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 19
- 235000005822 corn Nutrition 0.000 claims abstract description 19
- 238000011081 inoculation Methods 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000003306 harvesting Methods 0.000 claims abstract description 10
- 239000002689 soil Substances 0.000 claims abstract description 3
- 241000228212 Aspergillus Species 0.000 claims description 7
- 239000007921 spray Substances 0.000 claims description 7
- 240000007594 Oryza sativa Species 0.000 claims description 5
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 239000003337 fertilizer Substances 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 238000009395 breeding Methods 0.000 claims description 4
- 230000001488 breeding effect Effects 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 230000008485 antagonism Effects 0.000 claims description 2
- 210000000582 semen Anatomy 0.000 claims description 2
- 206010067482 No adverse event Diseases 0.000 abstract description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 description 7
- 229930195730 Aflatoxin Natural products 0.000 description 6
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 6
- 239000005409 aflatoxin Substances 0.000 description 6
- 238000012797 qualification Methods 0.000 description 6
- 238000011109 contamination Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 3
- 239000004568 cement Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 235000003276 Apios tuberosa Nutrition 0.000 description 1
- 235000010744 Arachis villosulicarpa Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004033 diameter control Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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Abstract
The invention provides a field identification method for peanut aspergillus flavus infection resistance, which comprises the following steps: 1) crushing corn until the diameter thereof is between 3 and 5 millimeters, and producing aspergillus flavus spores according to 50 to 75 grams of corn/m<2> of peanut inoculation aspergillus flavus; 2) stopping watering 5 to 6 weeks before harvesting peanuts, making ditches of which the depth is about 2 to 3 centimeters among peanut rows before the inoculation, broadcasting the aspergillus flavus spores in the ditches among the rows for three times before harvesting the peanuts, and covering the ditches with soil for 2 to 3 millimeters; and 3) using a watering can to water the near-earth surface evenly after the inoculation according to 100 to 150 grams/m<2> to create an environment of which the temperature and the moisture are suitable for the propagation of the aspergillus flavus. Compared with the prior art, the method is the most similar field peanut growing environment, and has accurate identification result and high stability; and the used devices can be used for a plurality of years with only once investment, and generate no adverse effects on environmental pollution and human body health.
Description
Invention field
The present invention relates to agricultural technology field, relate in particular to a kind of field identification method of peanut aspergillus flavus infection resistance.
Background technology
Peanut is the main oil crops of China and cash crop, China's peanut per unit area yield, gross output and export volume all rank first, the peanut cultivation area accounts for 19% of world's total cultivated area, occupy the second place of the world, the export trade accounts for the world 50%, occupy first of the world main peanut export State, average annual gross output accounts for 41% of world's gross output, and China is world production, the maximum country of outlet peanut.
Aflatoxin contamination has seriously restricted the optimum Sustainable development of China's peanut industry in recent years, has become the worldwide problem that has a strong impact on peanut production, processing, trade and food safety.The Yellow River and Huai He River Haiti district is China peanut main producing region, and ultimate production accounts for more than 50% of the whole nation, and export volume only Shandong just accounts for about 70%.But in recent years, because the influence of factors such as shortage aspergillus flavus resisting kind and results, processing causes the living main producing region of the Yellow River and Huai He River seaflower aflatoxin contamination to be and increases the weight of trend year by year, had a strong impact on the development of China's peanut export trade and local economy.Just because of the aflatoxin contamination problem, the trading partner European Union of China's peanut maximum closed the gate of import China peanut in 2009 substantially, and this is to cause one of basic reason that domestic peanut raw material and goods price significantly dive in the recent period.Therefore, solve the flavus pollution problem, guarantee peanut food safety, strengthen China's peanut competitiveness in the international market, make increasing peasant income, enterprise's synergy, the peanut production of promotion China more attains a new height and has seemed particularly urgent.
Aflatoxin pollution of peanuts is the problem of more complicated, wherein before the results arid or continuously long and shelling damage overcast and rainy, results back time of drying etc. be to cause the major cause of the Yellow River and Huai He River marine products district aflatoxin pollution of peanuts in recent years, but the reason of most critical is to lack high anti-and even immune flavus to infect and produces malicious peanut varieties.In recent years from groundnut germplasm and the seed selection of conventional hybridization technological approaches part aspergillus flavus resisting kind, but the aspergillus flavus resisting ability that detects these kinds is many based on indoor detection, major cause is that indoor detection cost is low, the time is short relatively, but indoor detection significant disadvantages is to represent field environment, resistance instability fully.The peanut varieties field aspergillus flavus resisting Performance Testing technology that this patent is announced then can effectively address the above problem, and improves resistance and identifies efficient, can significantly accelerate breeding process.
Summary of the invention
(goal of the invention) the objective of the invention is to solve peanut varieties field aspergillus flavus infection resistance and identifies that difficulty is big, resistance detects problems such as instability.
(technical scheme) present method provides a kind of authentication method of easy, cheap, efficient detection field peanut varieties aspergillus flavus infection resistance, and it comprises the steps:
Semen Maydis powder is broken to diameter 3-5mm, and is indoor according to 50g-75g corn/m
2Peanut inoculation Aspergillus flavus produces the Aspergillus flavus spore;
5-6 week stop to water before the harvesting peanut.Draw ditch in the ranks in peanut before the inoculation, the about 2-3cm of the degree of depth divides before the preferred 2.5cm, harvesting peanut and three times aspergillus spore is spread fertilizer over the fields in ditch in the ranks, with native covering 2-3mm;
With watering can according to 100g-150g/m
2Evenly spray water is created Aspergillus flavus and is suitable for breeding the humiture environment in postvaccinal near-earth surface.
Wherein, if it is excessive then aspergillus spore amount that carry is on the low side to infect the corn kernel of medium as flavus, infect also insufficient, inconvenient operation when infecting with Aspergillus flavus if broken seed is too small, therefore broken corn powder diameter control is advisable at 3-5mm, preferred 4-5mm.
According to an embodiment of the invention, the inoculum size of described Aspergillus flavus is preferably 55-70g corn/m
2Peanut, more preferably 60-65g corn/m
2Peanut.
According to an embodiment of the invention, before harvesting peanut the 35th day, 25 days, 15 days aspergillus spore spread fertilizer over the fields in ditch in the ranks, cover 2.5mm with soil.
According to an embodiment of the invention, preferably according to 120g-140g/m
2Evenly spray water is in postvaccinal near-earth surface, more preferably according to 125g-135g/m
2Evenly spray water is in postvaccinal near-earth surface.
According to an embodiment of the invention, described aspergillus flavus strain is after 30 ℃ of czapek's solutions are cultivated 7 days, and being made into concentration with sterilized water is 1 * 10
8The spore suspension of/mL, inoculation advance behind the corn dregs of rice standby.
The temperature that aspergillus flavus growth of the present invention is suited is 28-38 ℃, and relative humidity is 85%.
(invention effect) domestic existing peanut varieties aspergillus flavus infection resistance method is accredited as the master with indoor substantially, and significant disadvantage is that resistance is identified and can not be represented field environment, poor for applicability.The then main easily generation aflatoxin contamination peanut home environment of selecting is identified in domestic and international existing peanut aspergillus flavus resisting field, words to be tried are said that kind plants in aforementioned region, rely on physical environment and carry out the evaluation of flavus resistance, but this authentication method existence is subjected to natural environment influence heavy, defective such as qualification result poor stability, cost height, efficient are low, the present invention has then remedied above-mentioned shortcoming, has that cost is low, high-level efficiency, stability is strong and be suitable for advantage such as evaluation in batches.
Compared with prior art, the present invention is close to field peanut growing environment, and qualification result is accurate, stability is high; Equipment used is once invested and can be used for many years, and environmental pollution, HUMAN HEALTH have no adverse effects.
Embodiment
Peanut varieties to be identified is planted in the cement pit that has the canopy lid, all the water mudding is tight in around the cement pit and bottom, fill medium loam, the plantation aspergillus flavus infection resistance draws from the peanut cultivating variety J11 of India with to flavus and infects susceptible peanut cultivating variety golden flower 1012 (Arachis hypogaea L.);
Artificial or the machine pulverizing of corn, diameter 3-5mm, indoor according to 50g-75g corn/m
2Peanut inoculation Aspergillus flavus (Aspergillus Flavus) produces the Aspergillus flavus spore;
Preceding 6 weeks of harvesting peanut stop to water the Artificial Control rainwater.Draw ditch in the ranks in peanut before the inoculation, the about 2-3cm of the degree of depth, harvesting peanut preceding 35 days, 25 days, 15 days spreads fertilizer over the fields aspergillus spore in ditch in the ranks, with native covering 2-3mm;
With watering can according to 100g-150g/m
2Evenly spray water is created Aspergillus flavus and is suitable for breeding the humiture environment in postvaccinal near-earth surface.
Infecting Aspergillus flavus situation antagonism according to peanut varieties during results estimates.
Experimental period: 2005-2007, every kind is established contrast, arid+Aspergillus flavus is infected 2 processing, and three repetitions are averaged.
Peanut varieties: used peanut (Arachis hypogaea L.) kind is golden flower 1012, J11.
Strains tested and cultivation:
Aspergillus flavus strain (Aspergillus Flavus) (preserve in the laboratory, can buy from biotech firm).Bacterial strain is after 30 ℃ of czapek's solutions are cultivated 7 days, and being made into concentration with sterilized water is 1 * 10
8The spore suspension of/mL, inoculation advance behind the corn dregs of rice standby.
Treatment process:
Golden flower 1012, J11 inoculation before results has the corn of aspergillus spore, respectively at results sampling in preceding 0 day, 7 days, 9 days, 11 days, 13 days, 15 days.
The cultivation of Aspergillus flavus, field inoculation and sampling:
Aspergillus flavus strain is after 30 ℃ of czapek's solutions are cultivated 7 days, and being made into concentration with sterilized water is 1 * 10
6The spore suspension of/mL, inoculation advance behind the corn dregs of rice standby.
Peanut varieties golden flower 1012, J11 are inoculated in the peanut root in the preceding corn dregs of rice that will inoculate flavus in 30 days of results in the sample plot, and inoculation in per 10 days is once inoculated 3 times altogether.
Table 1 Virginia type peanut kind in 2005 J11 resource qualification result relatively
| Infection rate (%) | Accuracy rate (%) | Stability (%) | |
| Routine techniques | 70 | 60 | 60 |
| The invention technology | 90 | 90 | 95 |
Find out that from last table the invention technology is more accurate than routine techniques qualification result, stability is stronger.
Table 2 peanut varieties golden flower 1012 resource qualification results in 2006 relatively
| Infection rate (%) | Accuracy rate (%) | Stability (%) | |
| Routine techniques | 75 | 70 | 70 |
| The invention technology | 95 | 95 | 100 |
Table 3 peanut varieties J11, golden flower 1012 cultivar identification results in 2007 compare
Find out that from last table the invention technology is more accurate at pearl beans type peanut varieties resource aspergillus flavus infection resistance qualification result than routine techniques, stability is stronger.
Claims (10)
1. field identification method for peanut aspergillus flavus infection resistance, it comprises the steps:
1) Semen Maydis powder is broken to diameter 3-5mm, and is indoor according to 50g-75g corn/m
2Peanut inoculation Aspergillus flavus produces the Aspergillus flavus spore;
2) 5-6 week stops to water before the harvesting peanut, draw ditch in the ranks in peanut before the inoculation, divide before the degree of depth 2-3cm, harvesting peanut and three times aspergillus spore spread fertilizer over the fields in ditch in the ranks, with soil covering 2-3mm;
3) use watering can according to 100g-150g/m
2Evenly spray water is created the humiture environment that Aspergillus flavus is suitable for breeding in postvaccinal near-earth surface, infects Aspergillus flavus situation antagonism according to peanut varieties during results and estimates.
2. authentication method according to claim 1 is characterized in that, described corn is through machinery or manually be crushed to 4-5mm.
3. authentication method according to claim 1 is characterized in that, the inoculum size of described Aspergillus flavus is 55-70g corn/m
2Peanut.
4. authentication method according to claim 3 is characterized in that, the inoculum size of described Aspergillus flavus is 60-65g corn/m
2Peanut.
5. authentication method according to claim 1 is characterized in that, before harvesting peanut the 35th day, 25 days, 15 days aspergillus spore is spread fertilizer over the fields in ditch in the ranks.
6. authentication method according to claim 1 is characterized in that, according to 120g-140g/m
2Evenly spray water is in postvaccinal near-earth surface.
7. authentication method according to claim 6 is characterized in that, according to 125g-135g/m
2Evenly spray water is in postvaccinal near-earth surface.
8. authentication method according to claim 1 is characterized in that, described aspergillus flavus strain is after 30 ℃ of czapek's solutions are cultivated 7 days, and being made into concentration with sterilized water is 1 * 10
8The spore suspension of/ml, inoculation advance behind the corn dregs of rice standby.
9. authentication method according to claim 1 is characterized in that, the temperature that described aspergillus flavus growth is suited is 28-38 ℃, and relative humidity is 85%.
10. authentication method according to claim 1 is characterized in that, described peanut varieties is golden flower 1012, J11.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200910159347 CN101603073B (en) | 2009-07-10 | 2009-07-10 | Field identification method for peanut aspergillus flavus infection resistance |
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|---|---|---|---|
| CN 200910159347 CN101603073B (en) | 2009-07-10 | 2009-07-10 | Field identification method for peanut aspergillus flavus infection resistance |
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| CN101603073A CN101603073A (en) | 2009-12-16 |
| CN101603073B true CN101603073B (en) | 2013-09-11 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104711316A (en) * | 2014-12-12 | 2015-06-17 | 山东省花生研究所 | Identification method for peanut rod rot resistance of peanut germplasm resources |
| CN105340603B (en) * | 2015-11-09 | 2018-06-22 | 山东省花生研究所 | A kind of method of the aspergillus flavus Field infection of peanut seed |
| CN105230376A (en) * | 2015-11-12 | 2016-01-13 | 青岛友诚高新技术有限公司 | Method of field infected peanuts |
| CN107870224A (en) * | 2017-11-03 | 2018-04-03 | 安徽省农业科学院烟草研究所 | A kind of field identification method for evaluating corn aspergillus flavus infection resistance ability |
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2009
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Non-Patent Citations (3)
| Title |
|---|
| 卢代华等.籼稻杂交水稻对稻曲病的田间抗性差异.《植物保护学报》.2008,第35卷(第4期),289-294. * |
| 唐兆秀等.国外抗黄曲霉花生品种资源的引进鉴定研究.《福建农业学报》.1999,第14卷(第3期),7-9. * |
| 袁虹霞等.花生品种(系)对叶斑病的抗性鉴定.《河南农业科学》.2004,(第12期),35-38. * |
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| CN101603073A (en) | 2009-12-16 |
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