CN102487742A - Method for identifying rice blast resistance of rice - Google Patents
Method for identifying rice blast resistance of rice Download PDFInfo
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- CN102487742A CN102487742A CN2011103966744A CN201110396674A CN102487742A CN 102487742 A CN102487742 A CN 102487742A CN 2011103966744 A CN2011103966744 A CN 2011103966744A CN 201110396674 A CN201110396674 A CN 201110396674A CN 102487742 A CN102487742 A CN 102487742A
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Abstract
A method for identifying rice blast resistance of rice comprises the steps of scissoring three rice stems in a large bract period from a rice growing field, bringing the rice stems back to a laboratory, scissoring 2cm from the upper portion of each stem section and 5-6cm from the lower portion of each stem section to obtain one rice stem with the whole length of approximate 7-8cm, putting the three rice stems into culture dishes with two layers of filter paper, adding sterile water of 1mL into each culture dish, washing cultivated rice blast fungus spores with the sterile water, obtaining concentration of 106 pcs/ml, dropping spore suspending liquid of 8 muL to stem section sites of each rice stem through a liquid moving gun, putting the culture dishes into a illuminating incubator at the temperature of 26 DEG C, wrapping the culture dishes with transparent films to keep humidity in the culture dishes, cultivating for ten days in light and dark alternating modes to survey disease conditions, dividing survey disease levels into three levels, regarding the first level as disease-resistant cultivars, regarding the second level as susceptible cultivars, and regarding the third level as highly susceptible cultivars. The method optimizes the concentration and inoculation amount of inoculated rice blast fungus spores and establishes simple, convenient and accurate grading standards.
Description
Technical field
Patent of the present invention relates to a kind of method of rice blast resistance being identified through the rice stem stalk that exsomatizes, and is mainly used in the disease-resistant screening in rice varieties and the breeding of new variety process.
Background technology
Rice blast (Magnaporthe oryzae) is the severe diseases of potential threat on the Rice Production, and all there is generation the countries and regions of 85 the paddy rice plantations in the existing whole world.Annual paddy rice is because the loss that rice blast causes accounts for 10%～15% of gross yield, with a toll of multi-million dollar.All there is generation in China north and south rice district, the popular area of disease, heavy reached at 40%-50%, heavy especially field piece even No kernels or seeds are gathered, as in a year of scarcity.
Aspect disease control, proposed to use and chemical control is the comprehensive preventive health measures of key technology with resistant variety reasonable, these measures are to prevent and treat rice blast, guarantee that China's grain-production safety brought into play enormous function effectively.
The resistance authentication method of existing rice blast; Particularly the resistance of fringe pest is identified; Compare with the resistance authentication method of leaf pest; The difficulty that the fringe pest are identified is because the notable difference at different cultivars heading stage and long etc. from the breeding time that is seeded into heading stage so far, does not have unified artificial infection authentication method both at home and abroad as yet.Because some defective is not separately generally adopted, the more method of usefulness is to carry out the live body injection inoculation in booting stage at present; Bacterial concentration is about 1 * 105 spore/mL; Injection volume is 1.0 mL, and 20～30 fringe buds of each kind inoculation are investigated incidence stage of yellow ripeness to paddy rice.
Summary of the invention
The objective of the invention is to solve the weak point that exists in the above-mentioned existing method, provide a kind of easy, effectively, rice blast resistance authentication method accurately.
In order to reach above purpose, the present invention adopts following technical scheme:
Clip is in the paddy rice cane of big bud phase from paddy rice plantation field, takes back the laboratory, clip cane joint top 2cm; Bottom 5～6cm; Always be about the long cane of 7～8cm, 3 rhizome bars are put into the culture dish of double medium filtration paper, every ware adds the sterile water of 1mL; With cultured magnaporthe grisea spore with aseptic washing after, transferring to concentration is 10
6Individual/ml, save the spore suspension that people 8 μ L are dripped in the site with liquid-transfering gun at the cane of every rhizome bar.Culture dish is put into 26 ℃ illumination box, and encapsulate with clear films, to keep the humidity in the culture dish, alternation of light and darkness is cultivated 10 days investigation state of an illness.Grade scale is following:
1 grade: scab length is less than 0.5cm
2 grades: scab length is 0.6cm～1.5cm
3 grades: scab length is greater than 1.5cm
1 grade of the sick level of investigation be disease-resistant variety, 2 grades be susceptible variety, 3 grades be that height is felt kind.
The inventive method than existing method innovation part is:
Clip is in the paddy rice cane of big bud phase from paddy rice plantation field; Exposed the Zao 3-4 of 4～8cm days than spike length in the existing method; The breeding time of clip paddy rice cane is different, and the big bud phase cane than the Zao 3-4 of existing method days is susceptible disease more, and experimental result is more genuine and believable.The rice blast spore concentration and the inoculum concentration of inoculation have been optimized.Founded easier, grade scale accurately.
Embodiment
Clip is in the paddy rice cane of big bud phase from paddy rice plantation field, and clip is about the long cane of 7cm, cane joint top 2cm, and bottom 5cm puts into 3 rhizome bars in the culture dish (every ware adds the sterile water of 1mL) of double medium filtration paper; Cultured magnaporthe grisea spore is washed with sterile water, and transferring to concentration is 106/ml, drips the spore suspension of people 8 μ L in the cane joint site of every rhizome bar with liquid-transfering gun.Culture dish is put into 26 ℃ illumination box, and encapsulate with clear films, to keep the humidity in the culture dish, alternation of light and darkness is cultivated 10 days investigation state of an illness.Grade scale is following: 1 grade: scab length is less than 0.5cm; 2 grades: scab length is 0.6cm～1.5cm; 3 grades: scab length is greater than 1.5cm; 1 grade of the sick level of investigation be disease-resistant variety, 2 grades be susceptible variety, 3 grades be that height is felt kind.
The contrast experiment is following:
With Jiangxi Province's cultivated area in recent years big and resistance difference tangible 9 early rice varieties and Lijiang xintuanheigu (contrast) be material, adopted the two kinds of methods in following ground that the fringe pest resistance of rice varieties is identified.First kind: ecological multiple spot is seeding method in batches, is promptly selecting 5 different test countiesies (point) of ecological condition across the entire province, and each kind is sowed (live) in three batches, and be 5 days interval, investigates disease tassel yield stage of yellow ripeness in paddy rice under field conditions (factors); Second kind: live body fringe bud injection inoculation method, promptly in the paddy rice big bud phase, injection concentration be the magnaporthe grisea spore liquid 1mL of 1 * 105/mL in the fringe bud, stage of yellow ripeness by the investigation of 0～9 grade standard;
1, ecological multiple spot seeding method in batches
Select the different Xinfeng, 5 counties of ecotope, Yifeng, calm and peaceful, Wuyuan and Yongxiu County to make an experiment in the whole province.Each county all selects rice blast Chang Faqu to make an experiment.Same kind is divided 3 batches of sowings, and be 5 days interval, and various places are sowed to the first tenday period of a month Mo at the beginning of 4 months according to weather.Sowing is adopted live.Every kind sowing area is 5m
2, seeding quantity is 15g, interval 33.3 cm between the sub-district, and each kind is arranged by sequence number.The experimental field is not except that preventing and treating the rice blast, and all the other are by normal farming operation, and suitable many nitrogen fertilizer applications, bring out rice blast.In paddy rice stage of yellow ripeness, investigate the fringe pest incidence of disease of each kind, method and resistance are divided according to methods such as Sun Shuyuan.
2, live body fringe bud injection inoculation method
In each sub-district, selected the rice bud of broken chest (spike of rice does not expose yet) just June 22, with each bacterial strain spore liquid of syringe injection 1mL, same bacterial strain is injected 3 rice buds on each kind.Investigate the state of an illness stage of yellow ripeness, method and standard are pressed according to methods such as Sun Shuyuan.
Result of the test:
List the qualification result of above-mentioned three kinds of methods in table 1 at present.Do 2 explanations, first, in three batches of sowing tests of 9 main breeds in Yifeng and Yongxiu County all not the fringe pest take place, therefore, in the table 1 ecological multiple spot in batches the disease tassel yield of seeding method be the mean value of Xinfeng, calm and peaceful and three locations, Wuyuan test.The second, exsomatize sick level in fringe section drop inoculation method and the live body fringe bud injection inoculation method is the mean value of 8 inoculation morbidities.Can know from table 2, in 10 kinds that supply examination, adopt ecological multiple spot seeding method in batches; The disease-resistant variety of identifying (anti-in comprising) has 6, adopts the stem stalk drop inoculation of exsomatizing legal, and the disease-resistant variety of evaluation has 5; Adopt live body fringe bud injection inoculation method, the disease-resistant variety of evaluation has 1, and this shows; The morbidity of live body fringe bud injection inoculation method is the heaviest in these three kinds of methods, and ecological multiple spot the result that identifies of seeding method and stripped fringe section drop inoculation method is close in batches.From concrete qualification result; In ecological multiple spot 6 disease-resistant varieties that seeding method is identified in batches; The result that 5 kinds wherein adopt the fringe section drop inoculation method that exsomatizes to identify also shows as disease-resistant (anti-in comprising), only except the land two excellent 996, is accredited as middle sense; This shows that the qualification result of these two kinds of methods is quite to coincide.Because ecological multiple spot seeding method in batches is the resistance qualification result of land for growing field crops nature, be objectively, in view of the above, we think that the result of the stripped fringe section drop inoculation method of the present invention has higher confidence level.Compare, live body fringe bud injection inoculation method can not distinguish effectively kind in anti-, middle sense and susceptibility.
The result of three kinds of authentication methods of table 1 relatively
Claims (1)
1. rice blast resistance authentication method is characterized in that: plant the paddy rice cane that the field clip is in the big bud phase from paddy rice, take back the laboratory; Clip cane joint top 2cm, bottom 5～6cm always is about the long cane of 7～8cm; 3 rhizome bars are put into the culture dish of double medium filtration paper; Every ware adds the sterile water of 1mL, with cultured magnaporthe grisea spore with aseptic washing after, transferring to concentration is 10
6Individual/ml, save the spore suspension that people 8 μ L are dripped in the site with liquid-transfering gun at the cane of every rhizome bar, culture dish is put into 26 ℃ illumination box; And encapsulate with clear films; To keep the humidity in the culture dish, alternation of light and darkness is cultivated 10 days investigation state of an illness, and the sick level of investigation grade scale is following:
1 grade: scab length is less than 0.5cm
2 grades: scab length is 0.6cm～1.5cm
3 grades: scab length is greater than 1.5cm
1 grade be disease-resistant variety, 2 grades be susceptible variety, 3 grades be high sense kind.
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Cited By (13)
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| CN103205501A (en) * | 2013-04-19 | 2013-07-17 | 云南省农业科学院农业环境资源研究所 | Method for identifying rice blast-resistant gene of wild rice |
| CN103229674A (en) * | 2013-05-21 | 2013-08-07 | 江苏胜田农业科技发展有限公司 | Method for simply and rapidly identifying resistance of pyricularia grisea |
| CN104521592A (en) * | 2015-01-04 | 2015-04-22 | 连云港市农业科学院 | Large-scale quantitative and strengthened inoculation authenticating method for paddy black-streaked dwarf disease resistance |
| CN105145303A (en) * | 2015-09-10 | 2015-12-16 | 广西大学 | Method for rapidly inducing rice blast of pure sand seedlings |
| CN105191680A (en) * | 2015-11-02 | 2015-12-30 | 湖北省农业科学院植保土肥研究所 | Indoor rice variety Magnaporthe oryzae resistance identification method |
| CN105557347A (en) * | 2014-10-13 | 2016-05-11 | 江西省农业科学院植物保护研究所 | Gray mold resistance identified seedling stage inoculation method for capsicum |
| CN105613072A (en) * | 2016-01-20 | 2016-06-01 | 安徽理想种业有限公司 | Identification method of resistance to rice blast |
| CN106212142A (en) * | 2016-09-27 | 2016-12-14 | 福建省农业科学院水稻研究所 | The in-vitro verification method of bacterial blight of rice |
| CN108060088A (en) * | 2018-01-11 | 2018-05-22 | 江西省农业科学院植物保护研究所 | A kind of culture medium for being used for Pyricularia oryzae culture, producing spore and preparation method thereof |
| CN108070530A (en) * | 2018-02-05 | 2018-05-25 | 中国水稻研究所 | A kind of method that illumination cultivation frame using improvement carries out rice blast inoculated identification |
| CN110628871A (en) * | 2019-10-30 | 2019-12-31 | 江西省农业科学院植物保护研究所 | Method for identifying rice blast resistance of wild rice |
| CN112553085A (en) * | 2020-11-27 | 2021-03-26 | 云南农业大学 | Non-toxic rice blast strain for preventing and controlling rice blast and application thereof |
| CN113151531A (en) * | 2021-01-11 | 2021-07-23 | 江西省农业科学院植物保护研究所 | Screening method and identification method of blast rice neck blast resistance identification bacterial line |
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| CN103205501A (en) * | 2013-04-19 | 2013-07-17 | 云南省农业科学院农业环境资源研究所 | Method for identifying rice blast-resistant gene of wild rice |
| CN103229674A (en) * | 2013-05-21 | 2013-08-07 | 江苏胜田农业科技发展有限公司 | Method for simply and rapidly identifying resistance of pyricularia grisea |
| CN105557347B (en) * | 2014-10-13 | 2018-11-20 | 江西省农业科学院植物保护研究所 | The Seedling Inoculation method of Botrytis cinerea Disease Resistance Identification |
| CN105557347A (en) * | 2014-10-13 | 2016-05-11 | 江西省农业科学院植物保护研究所 | Gray mold resistance identified seedling stage inoculation method for capsicum |
| CN104521592A (en) * | 2015-01-04 | 2015-04-22 | 连云港市农业科学院 | Large-scale quantitative and strengthened inoculation authenticating method for paddy black-streaked dwarf disease resistance |
| CN104521592B (en) * | 2015-01-04 | 2016-09-07 | 连云港市农业科学院 | Inoculation identification method is quantitatively strengthened in the scale of a kind of black streaked dwarf virus of rice resistance |
| CN105145303A (en) * | 2015-09-10 | 2015-12-16 | 广西大学 | Method for rapidly inducing rice blast of pure sand seedlings |
| CN105145303B (en) * | 2015-09-10 | 2017-11-03 | 广西大学 | A kind of method that pure sandy soil nursery fast induces rice blast |
| CN105191680A (en) * | 2015-11-02 | 2015-12-30 | 湖北省农业科学院植保土肥研究所 | Indoor rice variety Magnaporthe oryzae resistance identification method |
| CN105191680B (en) * | 2015-11-02 | 2019-04-05 | 湖北省农业科学院植保土肥研究所 | A kind of interior rice varieties blast resistance identification method |
| CN105613072A (en) * | 2016-01-20 | 2016-06-01 | 安徽理想种业有限公司 | Identification method of resistance to rice blast |
| CN106212142A (en) * | 2016-09-27 | 2016-12-14 | 福建省农业科学院水稻研究所 | The in-vitro verification method of bacterial blight of rice |
| CN108060088A (en) * | 2018-01-11 | 2018-05-22 | 江西省农业科学院植物保护研究所 | A kind of culture medium for being used for Pyricularia oryzae culture, producing spore and preparation method thereof |
| CN108060088B (en) * | 2018-01-11 | 2021-03-16 | 江西省农业科学院植物保护研究所 | Culture medium for magnaporthe grisea culture and spore production and preparation method thereof |
| CN108070530A (en) * | 2018-02-05 | 2018-05-25 | 中国水稻研究所 | A kind of method that illumination cultivation frame using improvement carries out rice blast inoculated identification |
| CN110628871A (en) * | 2019-10-30 | 2019-12-31 | 江西省农业科学院植物保护研究所 | Method for identifying rice blast resistance of wild rice |
| CN110628871B (en) * | 2019-10-30 | 2023-09-15 | 江西省农业科学院植物保护研究所 | Method for identifying rice blast resistance of wild rice |
| CN112553085A (en) * | 2020-11-27 | 2021-03-26 | 云南农业大学 | Non-toxic rice blast strain for preventing and controlling rice blast and application thereof |
| CN112553085B (en) * | 2020-11-27 | 2022-11-01 | 云南农业大学 | Nontoxic rice blast strain for preventing and controlling rice blast and application thereof |
| CN113151531A (en) * | 2021-01-11 | 2021-07-23 | 江西省农业科学院植物保护研究所 | Screening method and identification method of blast rice neck blast resistance identification bacterial line |
| CN113151531B (en) * | 2021-01-11 | 2023-09-29 | 江西省农业科学院植物保护研究所 | Screening method and identification method for rice blast spike neck blast resistance identification bacterial strain |
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Application publication date: 20120613 |