CN101990819A - Fusarium graminearum caused rootstock rot seedling-stage resistance identification method - Google Patents
Fusarium graminearum caused rootstock rot seedling-stage resistance identification method Download PDFInfo
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Abstract
The invention relates to a fusarium graminearum caused rootstock rot seedling-stage resistance identification method comprising the following steps of sprouting measured wheat varieties or series seeds after sterilizing, and then inoculating activated fusarium graminearum; spreading the wheat seeds inoculated with the fusarium graminearum in a sterilized wet tissue, wrapping up, and keeping wet and culturing for 1 to 3 days under the shielding conditions, and then naturally growing for 11 to 13 days under natural lighting to form seedlings; immersing edges of the wet tissue in distilled water after the package layer of the surface of the wet tissue is dried during the natural growing period; and finally identifying the rootstock rot of the seedlings, and using the result as fusarium graminearum caused disease indexes of the measured wheat varieties or series seeds.
Description
Technical field
The present invention relates to the disease resistance authentication method of a plant species, especially a kind of wheat root neck rot resistance in seedling stage authentication method that is caused by Fusarium graminearum belongs to the plant protection technology field.
Background technology
A plurality of microspecies of sickle-like bacteria can cause the generation of wheat scab (Fusarium Head Blight is called for short FHB) and root neck rot (Crown Rot is called for short CR), are one of main pathogenic bacteria that influence Wheat Production.Infect wheatear portion and the wheat scab that causes by Fusarium graminearum, fusarium culmorum etc., it is the important disease of warm rainy and weather humid region wheat, mainly be popular in Canada, the U.S., South America, Europe, African southern areas and China, become a kind of worldwide wheat diseases.China is the country of global wheat scab injured area maximum, and middle and lower reach of Yangtze River winter wheat district and northeast spring wheat district are the main generation areas of head blight, and annual injured area surpasses 7,000,000 hm
2, Fusarium graminearum is the advantage reaping hook bacterial classification that causes present wheat scab popular, is subjected to the influence of global environmental change in recent years, head blight oriented the Yellow River and Huai He River basin and other areas spread trend.
Studies show that, many sickle-like bacteria such as fusarium culmorums (F.culmorum) that cause wheat scab, fusarium avenaceum (F.avenaceum), Fusarium graminearum (F.graminearum), fusarium acuminatum (F.acuminatum) also can cause the wheat root neck rot, wherein especially pathogenic the strongest with Fusarium graminearum, the wheat root neck rot is in Australia, Europe, North America, South America, the West Asia, the morbidity of wheat plantings such as area such as north African and South Africa area is serious, can cause wheat plant short and small, the minimizing of tillering, seed wrinkle is flat even not solid and cause production loss, local wheat genetic breeding and pathologists' extensive concern (Mitter etc. have been subjected to, Plant Pathology, 2006,55:433-441).U.S.'s wheat root neck rot mainly betides each states of northwest Pacific such as the warmer Idaho State of weather, Oregon, the State of Washington, the state of Colorado, the Wyoming State and Texas.In Australia, Fusarium graminearum and false Fusarium graminearum (F.pseudograminearum) are the main pathogenic bacteria that cause wheat scab and root neck rot, the root neck rot is the second largest disease that is only second to leaf blight, in the Queensland state, New South Wales, Nan'ao, western Australia be particularly serious, common wheat output is generally lost 30%-70%, all not anti-this disease of durum wheat (durum wheat) kind, the annual loss that is caused by the wheat root neck rot surpasses 6,000 ten thousand Australian Dollars; Same suitable Tanaka of environmental condition, head blight and root neck rot can take place simultaneously (Mitter etc., Plant Pathology, 2006,55:433-441).In the popular some areas of China's wheat scab, as middle and lower reach of Yangtze River winter wheat district, possess the pathogenetic condition of wheat root neckrot, found the wheat root neck rot plant that causes by Fusarium graminearum.Simultaneously, the trend that spreads of the oriented the Yellow River and Huai He River winter wheat of the microbial wheat scab of China's reaping hook district in recent years, the frequency and the severity that take place strengthen day by day, the weather conditions of Yellow River-Huai River region wheat late growth stage help the generation of root neck rot, therefore, should pay attention to and strengthen correlative study by the microbial wheat root neck rot of reaping hook.
The anti-source of screening root neck rot be the precondition of carrying out research of root neck rot genetics of resistance and breeding of new variety, and the foundation of simple and effective wheat root neck rot resistance authenticate technology system is the key of carrying out disease-resistant germ plasm resource screening and molecular genetic research.At present, the wheat Fusarium graminearum root neck rot resistance authentication method of having reported mainly is divided into artificial infection and sick garden is inoculated naturally.Most of artificial infection methods are inoculated at seed or seedling stage, are becoming the strain phase that incidence is estimated.It is to carry out the disease evaluation in the maturing stage that sick garden is inoculated naturally.
(Inoculation methods for assessing resistance in wheat to crown rot caused by Fusarium gramineum group 1 such as Dodman, Australian Journal of Agricultural Research, 1987,38:473-486) three kinds of methods that the wheat root neck rot is identified have been reported, i.e. seed inoculation, soil inoculation and basal part of stem inoculation.The incidence of disease of seed inoculation method and severity are all lower, and this method causes that emergence rate reduces simultaneously.The soil inoculation soil that mainly carries disease germs is directly inoculated, or wheat is sprinkled upon in soil surface or the seed furrow with the seed that carries disease germs after emerging, and the incidence of disease of this method and severity are all higher.The basal part of stem inoculation mainly contains two types, and the one, spore liquid is sprayed at the plants stems base portion, the anti-invasive of disease-resistant variety is significantly better than susceptible variety; The 2nd, spore liquid is injected at basal part of stem, the anti-invasive of this method disease-resistant variety and susceptible variety does not have notable difference.(Resistance to crown rot in wheat identified through an improved method for screening adult plants.Australasian Plant Pathology such as Wallwork, 2004,33:1-7) reported that seed inoculates, becomes the strain phase to carry out resistance authentication method (soil inoculation method of carrying disease germs) in the soil that carries disease germs, this method is longer qualification cycle, required space is bigger, in the maturing stage disease is estimated.(A high-throughput glasshouse bioassay to detect crown resistance in wheat germplasm.Plant Pathology such as Mitter, 2006,55:433-441) reported in seedling stage spore suspension has been dropped in basal part of stem, inoculate back 35 days and carry out the method that resistance is identified, it is referred to as spore suspension drop inoculation method, this method is not easy to operation, is seeded in the easy landing of spore suspension of plant base portion.(A simple method for the assessment for crown rot disease severity in wheat seedlings inoculated with Fusarium pseudograminearum.Journal of Phytopathology such as Li, 2008,156 (11-12): 751-754) reported the soil inoculation method (chitting piece inoculation method) of a kind of neck rot, the chitting piece of inoculating is planted in the polypots that sterile soil is housed, in greenhouse or phytotron, grow, when the wilting symptom appears in plant, water, inoculate back 35 days and carry out the evaluation of disease severity.
The said method of report such as Wallwork, Mitter and Li is all inoculated in seedling stage or seed, becomes the strain phase to carry out evaluation of resistance, and the inoculation method of Li etc. is than the easier operation of method of Mitter and Wallwork etc.Simultaneously, the severity of the method for Li etc. morbidity is than higher, and disease severity and field maturing stage disease show good relationship.This several method is all grown (more than 35 days) from being inoculated into the required time of evaluation of resistance, and the resistance that shows is strain-forming period resistance, needs soil to inoculate or grow simultaneously, and workload is bigger.Therefore in root neck rot resistance is identified, press for exploitation, set up easy, the wheat root neck rot resistance seedling authenticate technology system effectively and rapidly of a cover, under the ambient controlled condition, adopt the resistance difference between suitable inoculation method, the rational state of an illness evaluation criterion research kind, in the short period of time bulk materials is identified, for rapidly and efficiently root neck rot breeding for disease resistance and genetic research lays the foundation.
Carry out resistance evaluation seedling stage at the root neck rot that causes by Fusarium graminearum at wheat, be not reported at present.
Summary of the invention
The objective of the invention is to: at present by the wheat Fusarium graminearum cause wheat root neck rot resistance identify exist be subjected to environmental influence easily, basically be by seed, soil, inoculation in seedling stage, there are a series of problems such as morbidity is insufficient, qualification time is long, workload is big, provide a kind of at wheat root neck rot resistance in the seedling stage authentication method that causes by Fusarium graminearum.
The object of the present invention is achieved like this: a kind of wheat root neck rot resistance in seedling stage authentication method that is caused by Fusarium graminearum, it is characterized in that: at first in culture dish, germinate after the seed disinfection with tested wheat breed or strain, then the Fusarium graminearum after the inoculation activation; The wheat seed of inoculation Fusarium graminearum is layered in the hygenic towelette of the bacterium of going out, preserve moisture under the lucifuge condition and cultivated 1～3 day in the parcel back, self-sow 11～13 days under the natural lighting condition then, form seedling, during self-sow, when the surperficial integument of paper handkerchief occurs when dry, place distilled water to soak the ora terminalis of paper handkerchief; At last the root neck rot of seedling is identified the disease index of the root neck rot that the result is caused by Fusarium graminearum as tested wheat breed or strain.
In the present invention: the Fusarium graminearum after the activation is meant: the Fusarium graminearum bacterial classification is placed on the PDA solid culture medium activate, cultivate the back and regulate bacterial concentration to 1 * 10 with hemocytometer
6Individual spore/mL; Seed disinfection is meant: after the seed of tested wheat breed or strain is used the 2min that sterilizes in the 1%NaClO solution, use aseptic water washing again 3 times; Germinate in the culture dish and be meant: in being lined with the culture dish of two-layer wet filter paper, under 22 ℃ of temperature condition, germinate; Fusarium graminearum after the inoculation activation is meant: when bud reaches 4～6mm, be that 0.1% Tween 20, concentration are 1 * 10 containing volume ratio
6Inoculate 2min in the Fusarium graminearum suspension of individual spore/mL.
In the present invention: place on the PDA solid culture medium activation to be meant the Fusarium graminearum bacterial classification: the Fusarium graminearum bacterial classification to be placed on the 1/4 intensity PDA solid culture medium activate; 1/4 intensity PDA solid culture medium is by 10g potato dextrose agar powder, and the 15g agar powder adds 1L distilled water, forms behind 121 ℃ of autoclaving 20min.
In the present invention: the described wheat seed that will inoculate Fusarium graminearum is layered in the hygenic towelette of the bacterium of going out, preserve moisture to cultivate under the lucifuge condition and be meant in the parcel back: the paper handkerchief that will sterilize is tiled in through disinfecting on the vinyl disc, make it moistening with distilled water, with the wheat seed of inoculating 10 same breed of Fusarium graminearum or strain linearly shape be layered on the paper handkerchief, consistent and the sensing paper handkerchief edge of the direction of growth of bud, the distance at bud and paper handkerchief edge is 1.8～2.5cm, paper handkerchief is from one end to the other side rolled along the orientation of seed, put into the foam box of lucifuge, add distilled water and make paper roll fully moistening, the cultivation of under 22 ℃ of environment, preserving moisture; Describedly be meant: under 22 ℃, natural lighting every day 12h condition, grow at self-sow under the natural lighting condition, during self-sow, when the surperficial integument of paper handkerchief occurs when dry, place distilled water to soak 10s the ora terminalis of the paper handkerchief of seed rhizosphere one end; Described root neck rot to seedling identifies, is that 10 tested seeds with every paper handkerchief are a repetition, and the evaluation of the tested wheat seed of each kind or strain is at least based on the testing result of three repetitions of parallel test.
In the present invention: during the root neck rot of seedling identified, the disease level is divided into the 0-5 level by the disease severity, wherein:
0 grade is not had disease symptom for leaf sheath;
1 grade is that the first leaf sheath scab length is less than 1.0cm;
2 grades is the first leaf sheath scab length 1.0-2.0cm;
3 grades be the first leaf sheath scab length greater than 2.0cm, the illness of wilting does not appear in seedling;
4 grades the wilting illness occurs for seedling;
5 grades is seedling death;
Disease index=[∑ (state of an illness rank * this grade strain number)/(investigating total strain number * the highest sick level)] * 100;
With with the mean value of each disease index that repeats of parallel test in the tested wheat seed of kind or strain disease index as the final evaluation of this wheat breed or strain;
The standard of disease index is 7 ranks: the index of immunity is 0; The index of high anti-HR is＜20; The index of anti-R is 20.01～40.0; In the index of anti-MR be 40.01～60.0; The index of middle sense MS is 60.01～70.0; The index of sense S is 70.01～80.0; The index of high sense HS is 80.01～100.0.
The invention has the advantages that: (1) qualification cycle is short, from being inoculated into the disease evaluation, be about 15 days, with respect to seed directly inoculate, methods (generally needing 35-60 days) such as inoculation, drop inoculation in seedling stage are carried disease germs in chitting piece inoculation, soil, shortened the test period greatly, thereby can in the regular hour, carry out resistance evaluation in seedling stage to large quantities of wheat lines.(2) this method all is in the stage in seedling stage from being seeded to the disease evaluation, is a kind of method of identifying wheat root neck rot resistance in seedling stage.(3) the required space of experiment is little, and after the inoculation, continued growth in uncovered plastic casing or foam box is until carrying out the disease evaluation.(4) do not need soil, reduce the workload of bringing because of soil sterilization and inoculation.(5) under environmental conditions such as laboratory, greenhouse, phytotron, all can operate.(6) goodness of fit height of the sick garden of the resistance qualification result of this method acquisition and field qualification result.
Description of drawings
Fig. 1 is a photo of postvaccinal tested wheat seed being arranged parcel in paper handkerchief;
Fig. 2 is that paper handkerchief launches, observe the seedling incidence photo.
Embodiment
Embodiment 1
By identification of species: select 17 parts of wheat breeds or the strain introduced from Australia for use, they are known because of Fusarium graminearum causes that the field of root neck rot becomes strain phase disease resistance to be, and (network address is the data source of disease resistance in Queensland, Australia 2007,2008 and wheat breed introduction in 2009
Http:// www.dpi.qld.gov.au/265121.htm), and document Mitter etc., Plant Pathology, 2006,55:433-441; Li etc., Journal of Phytopathology, 2008,156 (11-12): 751-754.
Qualification process:
The preparation method of Fusarium graminearum suspension: Fusarium graminearum is F15; provided by the plant protection of academy of agricultural sciences, Jiangsu Province that [Fusarium graminearum bacterial classification F15 delivers (Ren Lijuan etc. on document; anti-banded sclerotial blight of wheat and head blight QTL Position Research; the wheat crops journal; 2007,27 (3): 416-420)].With Fusarium graminearum bacterial classification F15 on the superclean bench on 1/4 intensity PDA solid culture medium activation culture (1/4 intensity PDA solid culture medium collocation method is: 10g potato dextrose agar powder, the 15g agar powder, add 1L distilled water, 121 ℃ of autoclaving 20min), at 22 ℃, under the light application time 12h condition, cultivate after 7 days mycelia is applied on the same intensity PDA medium, continuation is at 22 ℃, cultivated 10 days under the light application time 12h condition, scrape off the media surface mycelia, continue again to cultivate after 7 days, on super-clean bench, use distilled water wash, three layers of sterile gauze filter, and regulate bacterial concentration to 1 * 10 with hemocytometer
6Individual spore/mL.
Wheat seed germinates down in 22 ℃ in being lined with the culture dish of two-layer wet filter paper after the 1%NaClO solution disinfection cleans, when bud reaches the 5mm left and right sides, contain 0.1% (volume ratio) Tween 20, concentration is 1 * 10
6Inoculate 2min in the Fusarium graminearum suspension of individual spore/mL.
With the paper handkerchief (24cm * 24cm) be tiled in that sterilized through on the square plastic dish of disinfecting, make it moistening with an amount of distilled water, with the wheat seed of 10 inoculations linearly shape be layered on the paper handkerchief, consistent and the sensing paper handkerchief edge of the direction of growth of bud, the distance at bud and paper handkerchief edge is 2cm, paper handkerchief is from one end to the other side rolled (see figure 1) along the orientation of seed, put into the foam box of lucifuge, each kind triplicate.Each repeats to put at random the foam box of lucifuge, adding distilled water makes paper roll fully moistening, preserve moisture at 22 ℃, lucifuge condition and to cultivate 48h, under 22 ℃, light application time 12h condition, grow then, when the outer appearance of paper roll is dry, the paper handkerchief other end is soaked 10s in distilled water, inoculate back 14 days with paper handkerchief expansion (Fig. 2) and carry out the disease level identification by individual plant.
In the evaluation, by the disease severity disease level is divided into the 0-5 level: 0 grade is not had disease symptom for leaf sheath; 1 grade is that the first leaf sheath scab length is less than 1.0cm; 2 grades is the first leaf sheath scab length 1.0-2.0cm; 3 grades be the first leaf sheath scab length greater than 2.0cm, the illness of wilting does not appear in seedling; 4 grades the wilting illness occurs for seedling; 5 grades is seedling death.
Calculate disease index according to the disease opinion rating: disease index=[∑ (state of an illness rank * this grade strain number)/(investigating total strain number * the highest sick level)] * 100%, calculate the average disease index that obtains 3 repetitions.Standard according to the disease resistance disease index: the index of immunity is 0, the index of high anti-(HR) is＜20, the index of anti-(R) is 20.01～40.0, in the index of anti-(MR) be 40.01～60.0, the index of middle sense (MS) is 60.01～70.0, the index of sense (S) is 70.01～80.0, and the index of high sense (HS) is 80.01～100.0.
As can be seen from Table 1,17 parts of wheat lines coverage rate aspect root neck rot resistance is bigger, wherein, 2 parts of varietal resistances reach anti-(R) level, and 3 parts of varietal resistances resist (MR) in being, 4 parts of kinds are middle sense (MS), 6 parts of kinds are sense (S), and 2 parts of kinds are high sense (HS).
By the comparison of table 1, the resistance qualification result and the public of visible this method are known, adopt conventional method to have the higher goodness of fit at field strain-forming period resistance qualification result.
17 parts of wheat breeds of table 1 resistance in seedling stage inoculation qualification result and strain-forming period resistance
Embodiment 2
Adopt the authentication step of embodiment 1, select 41 unknown wheat breeds or strain,, number respectively in the evaluation and be defined as JS-1～JS-41 in regular turn for the ease of 41 wheat breeds or strain are distinguished.
In order to check the repeatability of this method, 41 wheat breeds or strain have been designed 2 independently evaluations at different time respectively, comprise 3 repetitions in each the evaluation, each qualification result is based on the mean value of 3 repetitions.The average disease index and the evaluation of resistance of 2 tests are listed in table 2, and 2 result of the tests are carried out correlation analysis, and the correlation coefficient of twice disease index is 0.91, and the repeatable higher of resistance authentication method in this seedling stage is described.
2 tests of 41 wheat breeds of table 2 or strain resistance in seedling stage inoculation qualification result
More than each embodiment be not to concrete restriction of the present invention.
Claims (5)
1. wheat root neck rot resistance in a seedling stage authentication method that is caused by Fusarium graminearum is characterized in that: at first germinate in culture dish after the seed disinfection with tested wheat breed or strain, then the Fusarium graminearum after the inoculation activation; The wheat seed of inoculation Fusarium graminearum is layered in the hygenic towelette of the bacterium of going out, preserve moisture under the lucifuge condition and cultivated 1～3 day in the parcel back, self-sow 11～13 days under the natural lighting condition then, form seedling, during self-sow, when the surperficial integument of paper handkerchief occurs when dry, place distilled water to soak the ora terminalis of paper handkerchief; At last the root neck rot of seedling is identified the disease index of the root neck rot that the result is caused by Fusarium graminearum as tested wheat breed or strain.
2. wheat root neck rot resistance in the seedling stage authentication method that causes by Fusarium graminearum according to claim 1, it is characterized in that: the Fusarium graminearum after the activation is meant: the Fusarium graminearum bacterial classification is placed on the PDA solid culture medium activate, cultivate the back and regulate bacterial concentration to 1 * 10 with hemocytometer
6Individual spore/mL; Seed disinfection is meant: after the seed of tested wheat breed or strain is used the 2min that sterilizes in the 1%NaClO solution, use aseptic water washing again 3 times; Germinate in the culture dish and be meant: in being lined with the culture dish of two-layer wet filter paper, under 22 ℃ of temperature condition, germinate; Fusarium graminearum after the inoculation activation is meant: when bud reaches 4～6mm, be that 0.1% Tween 20, concentration are 1 * 10 containing volume ratio
6Inoculate 2min in the Fusarium graminearum suspension of individual spore/mL.
3. wheat root neck rot resistance in the seedling stage authentication method that is caused by Fusarium graminearum according to claim 2 is characterized in that: the Fusarium graminearum bacterial classification is placed to activate on the PDA solid culture medium be meant: the Fusarium graminearum bacterial classification is placed on the 1/4 intensity PDA solid culture medium activate; 1/4 intensity PDA solid culture medium is by 10g potato dextrose agar powder, and the 15g agar powder adds 1L distilled water, forms behind 121 ℃ of autoclaving 20min.
4. according to claim 1 or 2 or 3 described wheat root neck rot resistance in the seedling stage authentication methods that cause by Fusarium graminearum, it is characterized in that: the described wheat seed that will inoculate Fusarium graminearum is layered in the hygenic towelette of the bacterium of going out, preserve moisture to cultivate under the lucifuge condition and be meant in the parcel back: the paper handkerchief that will sterilize is tiled in through disinfecting on the vinyl disc, make it moistening with distilled water, with the wheat seed of inoculating 10 same breed of Fusarium graminearum or strain linearly shape be layered on the paper handkerchief, consistent and the sensing paper handkerchief edge of the direction of growth of bud, the distance at bud and paper handkerchief edge is 1.8～2.5cm, paper handkerchief is from one end to the other side rolled along the orientation of seed, put into the foam box of lucifuge, add distilled water and make paper roll fully moistening, the cultivation of under 22 ℃ of environment, preserving moisture; Describedly be meant: under 22 ℃, natural lighting every day 12h condition, grow at self-sow under the natural lighting condition, during self-sow, when the surperficial integument of paper handkerchief occurs when dry, place distilled water to soak 10s the ora terminalis of the paper handkerchief of seed rhizosphere one end; Described root neck rot to seedling identifies, is that 10 tested seeds with every paper handkerchief are a repetition, and the evaluation of the tested wheat seed of each kind or strain is at least based on the testing result of three repetitions of parallel test.
5. wheat root neck rot resistance in the seedling stage authentication method that is caused by Fusarium graminearum according to claim 4 is characterized in that: during the root neck rot of seedling is identified, the disease level is divided into the 0-5 level by the disease severity, wherein:
0 grade is not had disease symptom for leaf sheath;
1 grade is that the first leaf sheath scab length is less than 1.0cm;
2 grades is the first leaf sheath scab length 1.0-2.0cm;
3 grades be the first leaf sheath scab length greater than 2.0cm, the illness of wilting does not appear in seedling;
4 grades the wilting illness occurs for seedling;
5 grades is seedling death;
Disease index=[∑ (state of an illness rank * this grade strain number)/(investigating total strain number * the highest sick level)] * 100;
With with the mean value of each disease index that repeats of parallel test in the tested wheat seed of kind or strain disease index as the final evaluation of this wheat breed or strain;
The standard of disease index is 7 ranks: the index of immunity is 0; The index of high anti-HR is＜20; The index of anti-R is 20.01～40.0; In the index of anti-MR be 40.01～60.0; The index of middle sense MS is 60.01～70.0; The index of sense S is 70.01～80.0; The index of high sense HS is 80.01～100.0.
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| CN102220407A (en) * | 2011-05-12 | 2011-10-19 | 云南农业大学 | Method for identifying gray mold resistance of gerbera |
| CN103621333A (en) * | 2013-12-02 | 2014-03-12 | 江苏省农业科学院 | Method for identifying resistance of wheat take-all disease |
| CN104737669A (en) * | 2015-04-22 | 2015-07-01 | 成都大学 | Bud stress resistance detection device and bud stress resistance detection method |
| CN104770175A (en) * | 2015-04-15 | 2015-07-15 | 广东省农业科学院作物研究所 | Method for inoculating fresh corn indoors by using pathogenic fusarium verticillioides |
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| CN101323875A (en) * | 2008-07-24 | 2008-12-17 | 江苏省农业科学院 | Wheat fusarium stalk rot seedlings inoculation identification method |
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| CN109197379B (en) * | 2018-11-12 | 2020-11-13 | 贵州大学 | Efficient single-spore propagation method for arbuscular mycorrhizal fungi |
| CN117210601A (en) * | 2023-09-25 | 2023-12-12 | 扬州大学 | Method for rapidly, simply and conveniently identifying wheat scab resistance with high flux |
| CN117210601B (en) * | 2023-09-25 | 2024-05-03 | 扬州大学 | Method for rapidly, simply and conveniently identifying wheat scab resistance with high flux |
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