CN103621333A - Method for identifying resistance of wheat take-all disease - Google Patents
Method for identifying resistance of wheat take-all disease Download PDFInfo
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- CN103621333A CN103621333A CN201310638545.0A CN201310638545A CN103621333A CN 103621333 A CN103621333 A CN 103621333A CN 201310638545 A CN201310638545 A CN 201310638545A CN 103621333 A CN103621333 A CN 103621333A
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Abstract
The invention relates to a method for identifying the resistance of the wheat take-all disease. The method comprises the following steps of adopting a disease-susceptiblevariety of yangmai 158 as a control variety, carrying out identification of the resistance of the wheat take-all disease on the wheat to be measured, i.e., disinfecting seeds of the wheat to be measured and the yangmai 158, and putting the seeds in a culture dish for pregermination; selecting and moving a seedling with three germinated seed roots and the root length being 1.5-2.5cm into the culture dish padded with water-absorbing filtering paper; grafting the middle part of each seed root into a bacterial disk with strongly-pathogenic bacterial strain with gaeumannomyces graminis var. tritici, culturing for five days, measuring the length of lesionson the wheat to be measured and the yangmai 158, and calculating the severity of the wheat take-all disease of the wheat to be measured by comparison with the yangmai 158. The method has the advantages of short identification period, simple and convenient operation, small occupied space and high accuracy.
Description
Technical field
The present invention relates to the disease Resistance Identification method of a Plants, especially a kind of take-all Resistance Identification method, belongs to plant protection technology field.
Background technology
Take-all (Take-all), is a kind of typical root soil-borne disease, mainly by gaeumannomyce wheat variety (Gaeumannomycesgraminisvar.Tritici is called for short Ggt), is caused.Amount reproduction after germ mycelia intrusion wheat strain root, destroys root tissue cell, stops up root canal, make plant endotrophic and the moisture can not normal transport, cause the wheat strain minimizing of tillering, plant yellow be downgraded even withered, when wheat comes to the ripening period, there is dead ears symptom.Germ throughout the growing season can be infected wheat, the most obvious with strain phase symptom.In damp soil, the external hyphae amount reproduction of Pathogen of Take-all, is wrapped in stem foot surface, form one deck black mycelia sheath, and it is darker more to approach base portion color, shape has sticked a black plaster like the basal part of stem at wheat, therefore, full rot is also referred to as " black leg ".
Take-all found when a plurality of country, have report afterwards, and be on the rise in South Australia early than 1852.Within 1931, take-all is found in China Zhejiang Province for the first time, national Ge Mai district also finds this disease in succession subsequently, full rot all has in various degree and occurs in 22 provinces such as China Jiangsu, Anhui, Henan at present, wherein serious with ground generations such as Henan, Shandong, within 2012, Henan Province's onset area reaches 4,500,000 mu.Full rot can cause the wheatland underproduction 20%～50% of being injured, and severe patient even has no harvest.After morbidity, the seed of seed farm can not utilize, at present take-all in Henan, a plurality of provinces such as Jiangsu, Anhui are listed in supplementary quarantine disease.
In production, the main method of agricultural measures and chemical control that adopts is carried out prevention and control to take-all, but these method costs are high, and effect is undesirable, and can cause negative effect to environment.Breeding resistant variety is a kind of method of economic environmental protection and reliable control plant disease.The screening anti-source of take-all, is the precondition of full rot disease-resistant variety seed selection and genetics of resistance research, and foundation easy, economic, effectively Resistance Identification method is the key of disease-resistant Screening of Germplasm and genetic research.
The domestic and international method in indoor evaluation take-all has multiple at present, as test tube method, pot-culture method, polyethylene is cover method etc. tiltedly, but these methods are substantially similar, its essence is exactly at container (test tube, Pen Portland, polyethylene bagging etc.) in, pack matrix (vermiculite into, river sand etc.), in matrix, add pathogen (bacterium dish, bacterium cake, sick wheat etc.), then sow wheat thereon and on wheat seed, cover again one deck matrix, be placed on temperature control wet, under the conditions such as illumination, cultivate, after wheat is fully fallen ill, root is washed out, again according to old complaint rate, root system lesion area accounts for the indexs such as ratio of the root system gross area and assesses the disease resistance of wheat breed to full rot.These indoor appraising methods condition control with operability aspect compare and there is obvious superiority with field test method, but they still need huge workload (separation of the sterilizing, filling, root system that comprises matrix from matrix and rinsing etc.), in addition from be seeded into resistance evaluation for up to more than 4 weeks, result acceptor's viewing of simultaneously assessing by These parameters rings larger.Therefore, for take-all, need to set up a set of easy, Resistance Identification system fast and accurately, under controlled environmental condition, utilize suitable inoculation method and objective state of an illness evaluation criterion, at short notice bulk materials is identified, for breeding for disease resistance and the disease-resistant Mechanism Study of take-all lays the foundation.
Summary of the invention
For current take-all, identify that workload is large, the time long, qualification result acceptor viewing rings the series of problems such as larger, a kind of easy, take-all Resistance Identification method is fast and accurately provided.
The object of the present invention is achieved like this:
A take-all Resistance Identification method, is characterized in that: by cultivars to disease susceptible Yangmai No.158, be check variety, wheat breed to be measured is carried out to take-all Resistance Identification, concrete grammar is:
A) the strong Pathogenicity Strains 25 ℃ of growths on PDA flat board of gaeumannomyce wheat variety (Gaeumannomyces graminis var.tritici) are punched at plate edge with the card punch of diameter 3mm for 7 days afterwards, obtain bacterium dish;
B) vernalization in culture dish will be placed on after the seed disinfection of wheat to be measured and Yangmai No.158; After 2 days, choose and send 3 seminal roots and root length moves on in the culture dish that is lined with absorbent filter the seedling of 1.5-2.5cm; Described vernalization refers to: in culture dish, pad 2 layers of wet filter paper, seed broadcast equably on filter paper and ventral groove down, at 25 ℃, cultivate;
C) in the middle part access steps A of each seminal root) the bacterium dish that obtains;
D) cultivate the scab length of measuring afterwards for 5 days on wheat to be measured and Yangmai No.158 root at 25 ℃;
Calculate the severity of take-all to be measured, severity=wheat scab length ÷ contrast to be measured wheat scab length * 100;
Take-all resistance standard is 5 ranks: immunity (I), severity=0; Anti-(R), 0 < severity < 30; In anti-(MR), 30≤severity < 60; Middle sense (MS), 60≤severity < 90; Sense (S), severity >=90.
In the present invention, the strong Pathogenicity Strains of described gaeumannomyce wheat variety is G1037 or G10 or G80.
In the present invention, described sterilization refers to soaks wheat seed after 2 minutes and uses aseptic water washing 3 times again in 1%NaClO solution; Described vernalization refers to: in culture dish, pad 2 layers of wet filter paper, with the seed of kind broadcast equably on filter paper and ventral groove down, 30/ware, at 25 ℃, cultivate; Seedling moves on in the culture dish that is lined with absorbent filter and refers to, with the seedling of 4 seeds of kind, is evenly placed in the middle of culture dish, and their root is spoke axle shape outwardly and arranges.
In the present invention, using the mean value of scab length on all wheat roots in each culture dish as a repetition, get the mean value of four repetitions as the scab length of wheat to be measured or Yangmai No.158.
In the present invention, the diameter of described culture dish is 9cm.
The invention has the advantages that: (1) is short qualification cycle, overall process only needs 7 days, and traditional method needs about 4 weeks; (2) easy and simple to handle, do not need with culture matrixes such as vermiculites, river sand, also without by root system separation and rinsing from matrix, thereby saved matrix sterilizing, loaded and washed a large amount of work such as root; (3) take up room littlely, in laboratory, just can complete, required condition is easy to control; (4) accuracy is high, and qualification result is determined by the scab length directly measuring, rather than determines by the estimation state of an illness, thereby acceptor's viewing rings less.
Embodiment
The full rot resistant determination that embodiment 1 Jing Zhou rye, general wheat are No. 5
Resistance Identification method is as follows:
A) preparation of gaeumannomyces graminis bacterium dish
Strong this bacterial strain of Pathogenicity Strains G1037(of gaeumannomyces graminis Yi You our unit is published) 25 ℃ of cultivation 7-15 days on PDA flat board, now mycelia has been covered with flat board, with the card punch of diameter 3mm, at plate edge, punches, and forms bacterium dish.
B) treat the preparation of inoculation material
The seed of Jing Zhou rye, general wheat No. 5 and Yangmai No.158 is soaked after 2 minutes and uses aseptic water washing 3 times again in 1%NaClO solution; In 9cm culture dish, pad 2 layers of wet filter paper, filter paper water suction is fully but without ponding, the seed after above-mentioned sterilization is broadcast equably on filter paper and ventral groove down, 30, every ware, cultivates 2 days for 25 ℃; Select the seed that sends young shoot and 3 seminal roots and the long 1.5-2.5cm of root, move on in the 9cm culture dish that is lined with absorbent filter 4, every ware; 4 seeds are placed in the middle of culture dish, make its ventral groove down and root is arranged towards surrounding, 12 seminal roots of 4 seeds are spoke axle shape and arrange;
C) inoculation and cultivation
The bacterium dish obtaining by transfer needle picking steps A, faces down its mycelia to be attached to the middle part of seminal root, and 12 roots in culture dish respectively connect a bacterium dish, culture dish lid on inoculation bonnet;
D) incidence investigation
The cultivation of 25 ℃ is after 5 days, with ruler, measure the length of black on the root of wheat to be measured and Yangmai No.158 on each root that connects bacterium dish or brown scab, in each culture dish, the mean value of the scab length of 12 roots is a repetition, each kind is established four repetitions, get the mean value of four repetitions as the scab length of wheat to be measured or Yangmai No.158, according to the severity of the scab length computation state of an illness.
Severity=wheat scab length ÷ contrast to be measured wheat scab length * 100;
Take-all resistance standard is 5 ranks: immunity (I), severity=0; Anti-(R), 0 < severity < 30; In anti-(MR), 30≤severity < 60; Middle sense (MS), 60≤severity < 90; Sense (S), severity >=90.
Investigation result: Jing Zhou rye root scab average length is 0.08cm, and severity is 4.1; No. 5 root scab average lengths of general wheat are 1.14cm, and severity is 59.1; Yangmai No.158 root scab average length is 1.93cm, and severity is 100.According to above-mentioned resistance grade scale, the resistance grade of Jing Zhou rye is anti-(R), and general wheat resists (MR) in being for No. 5, and Yangmai No.158 is sense (S).
In existing bibliographical information, as the research > > (Wang Meinan etc. of < < wheat crops to gaeumannomyces graminis disease resistance, Journal of Northwest Sci Tech University of Agriculture and Forestry, 2001, 29(3): 98-100), < < gaeumannomyces graminis Pathogenic Tests and Variety Disease-resistance Journal of Sex Research > > (Wang Ning etc., plant genetic resources journal, 2012, 13(3): 478-483), think that rye has significant resistance to full rot, general wheat has higher resistance to full rot No. 5, Yangmai No.158 is susceptible variety, this is consistent with the full rot resistance result that method of the present invention is identified.
The full rot Resistance Identification of 2 51 wheat breeds of embodiment
Adopt the method for embodiment 1, take Yangmai No.158 as check variety, identify the resistance of 51 wheat breeds to full rot.For the reliability of check the method, in the different time, these kinds have been carried out to 2 times and identified.
Take-all resistance standard is 5 ranks, immunity (I), severity=0; Anti-(R), 0 < severity < 30; In anti-(MR), 30≤severity < 60; Middle sense (MS), 60≤severity < 90; Sense (S), severity >=90.Result is as shown in table 1.
The result of 2 Resistance Identifications of table 151 wheat breed
The result of identifying for 2 times is more consistent, correlation coefficient is 0.8192, and this authentication method is described
Repeatability better.
Each embodiment is not limitation of the present invention above.
Claims (5)
1. a take-all Resistance Identification method, is characterized in that: by cultivars to disease susceptible Yangmai No.158, be check variety, wheat breed to be measured is carried out to take-all Resistance Identification, concrete grammar is:
A) by gaeumannomyce wheat variety (
gaeumannomyces graminis var. tritici) strong Pathogenicity Strains 25 ℃ of growths on PDA flat board punch at plate edge with the card punch of diameter 3mm for 7 days afterwards, acquisition bacterium dish;
B) vernalization in culture dish will be placed on after the seed disinfection of wheat to be measured and Yangmai No.158; After 2 days, choose and send 3 seminal roots and root length moves on in the culture dish that is lined with absorbent filter the seedling of 1.5-2.5cm; Described vernalization refers to: in culture dish, pad 2 layers of wet filter paper, seed broadcast equably on filter paper and ventral groove down, at 25 ℃, cultivate;
C) in the middle part access steps A of each seminal root) the bacterium dish that obtains;
D) cultivate the scab length of measuring afterwards for 5 days on wheat to be measured and Yangmai No.158 root at 25 ℃;
Calculate the severity of take-all to be measured, severity=wheat scab length ÷ contrast to be measured wheat scab length * 100;
Take-all resistance standard is 5 ranks: immunity (I), severity=0; Anti-(R), 0 < severity < 30; In anti-(MR), 30≤severity < 60; Middle sense (MS), 60≤severity < 90; Sense (S), severity >=90.
2. wheat full rot Resistance Identification method according to claim 1, is characterized in that, the strong Pathogenicity Strains of described gaeumannomyce wheat variety is G1037 or G10 or G80.
3. according to take-all Resistance Identification method described in claim 1 or 2, it is characterized in that, described sterilization is used aseptic water washing 3 times after referring to wheat seed being soaked to 2 minutes in 1% NaClO solution again; Described vernalization refers to: in culture dish, pad 2 layers of wet filter paper, with the seed of kind broadcast equably on filter paper and ventral groove down, 30/ware, at 25 ℃, cultivate; Seedling moves on in the culture dish that is lined with absorbent filter and refers to, with the seedling of 4 seeds of kind, is evenly placed in the middle of culture dish, and their root is spoke axle shape outwardly and arranges.
4. take-all Resistance Identification method according to claim 3, is characterized in that, using the mean value of scab length on all wheat roots in each culture dish as a repetition, gets the mean value of four repetitions as the scab length of wheat to be measured or Yangmai No.158.
5. take-all Resistance Identification method according to claim 4, is characterized in that, the diameter of described culture dish is 9cm.
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| CN107810682A (en) * | 2017-09-28 | 2018-03-20 | 江苏省农业科学院 | For the indoor biometricses method of wheat sharp eyespot pesticide control |
| CN110117637A (en) * | 2019-05-06 | 2019-08-13 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of garlic leaf blight Resistance Identification method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107810682A (en) * | 2017-09-28 | 2018-03-20 | 江苏省农业科学院 | For the indoor biometricses method of wheat sharp eyespot pesticide control |
| CN107810682B (en) * | 2017-09-28 | 2021-08-06 | 江苏省农业科学院 | Indoor bioassay method for wheat sharp eyespot control agent |
| CN110117637A (en) * | 2019-05-06 | 2019-08-13 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of garlic leaf blight Resistance Identification method |
| CN110117637B (en) * | 2019-05-06 | 2023-03-14 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Identification method for resistance to garlic leaf blight |
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