CN104480191A - Method for rapidly identifying resistance of apple branches and trunks to physalospora piricola nose - Google Patents
Method for rapidly identifying resistance of apple branches and trunks to physalospora piricola nose Download PDFInfo
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- CN104480191A CN104480191A CN201510016515.5A CN201510016515A CN104480191A CN 104480191 A CN104480191 A CN 104480191A CN 201510016515 A CN201510016515 A CN 201510016515A CN 104480191 A CN104480191 A CN 104480191A
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Abstract
The invention relates to a physalospora piricola nose resistance identification method, in particular to a method for rapidly identifying resistance of apple branches and trunks to the physalospora piricola nose. The physalospora piricola nose is inoculated to a PDA culture medium to be cultured so that a physalospora piricola nose culture medium bacterial colony can be obtained; pseudo identified varieties or rootstock young sprout leaves without plant diseases or insect pests at the top end, the front faces of the leaves are stabbed by an inoculating needle, and then the physalospora piricola nose is inoculated; after the bacteria-bearing leaves are cultured for six days, the diameters of disease spots of the leaves are measured according to a crossing method, the disease spot area of the leaves are worked out and compared with the disease spot areas of known resistance grade materials, and the grade of resistance of varieties to be identified or rootstocks to the branch and trunk physalospora piricola nose is determined. According to the method for rapidly identifying the resistance of apple branches and trunks to the physalospora piricola nose, leaves are inoculated, infection conditions suitable for the physalospora piricola nose are created, physalospora piricola nose resistance identification is performed on the infection result, few identification materials are needed, identification time is short, repeatability is high, and the identification method is simple, easy to master and popularize, and low in cost.
Description
Technical field
The present invention relates to a kind of Target spot pathogen Resistance Identification method, be specially Rapid identification apple limb to the method for Target spot pathogen resistance.
Background technology
At present about the method many employing field branch of apple limb to ring spot Resistance Identification and evaluation is inoculated, investigated and fall ill grade and calculate the method for disease index, branch amount needed for the method is many, disease cycle is long, field work amount is large and affect larger by natural environmental condition, cause that Resistance Identification duration, authentication method are loaded down with trivial details, identification result is repeated poor, and easily cause field diseases Spreading and diffusion.
Summary of the invention
For above-mentioned technical problem, the invention provides a kind of method of ring spot Resistance Identification, make apple variety or the qualification of stock to ring spot resistance is simple, the used time is short, qualification result is reproducible, concrete technical scheme is:
Rapid identification apple limb, to the method for Target spot pathogen resistance, comprises the following steps:
(1) PDA substratum makes
Potato is cleaned peeling, be cut into small pieces and put into the pot little fire that adds water and boil 0.5 hour, filter with double gauze, add in filtrate, agar and glucose, add water constant volume, potato: glucose: agar: the ratio that distilled water adds is 200g:20g:12g:1000mL; Be divided in test tube or triangular flask while hot, 121 DEG C of sterilizings 30 minutes, make PDA substratum;
(2) Target spot pathogen is cultivated
Target spot pathogen is inoculated into by Bechtop on above-mentioned PDA substratum, cultivates in illumination box under 25 DEG C of conditions, cover with after culture dish until mycelia and obtain ring spot substratum bacterium colony, for blade inoculation;
(3) blade is chosen
Choose intend 5-6 sheet under identification of species or stock young sprout top ripe, completely launch and be expert evidence without disease and pest blade, repeat 6 times;
(4) blade inoculation
Dry after the blade aseptic water washing to be identified chosen, wrap up in petiole base with the sterilized non-fat bale of cotton, a upper sterilized water moisturizing is for subsequent use; Stab face of blade with inoculating needle, take off the homogeneous circular bacterium cake of growth with diameter 5mm punch tool from ring spot substratum colony edge is isometrical, center, raw mycelia face is placed in blade puncture wounds;
(5) carry disease germs leaf culture
Filter paper is spread in tray bottom, the water of pallet height about 1/3rd is added in pallet, bar shaped foam is placed in pallet, be positioned in pallet by postvaccinal blade, make the petiole being wrapped in absorbent cotton be dipped in sterilized water, vane frame is on bar shaped foam, pallet mouth is covered with preservative film, be placed in incubator and carry out dark culturing, culture condition temperature is 25 DEG C, and humidity is 100%;
(6) Resistance Identification
Cultivation the 6th day, right-angled intersection method is adopted to measure leaf spot lesion diameter, with scab transverse diameter and vertical footpath product representation lesion area, the lesion area of blade and the lesion area of known resistance class material contrast, and determine that kind to be identified or stock are to the resistance class of Trunk canker.
Wherein known resistance class material is the lesion area of cockscomb, and standard lesion area is 8.5mm
2, determine that kind to be identified belonging to blade or stock are to the resistance class of Trunk canker, concrete standard is:
X=leaf spot lesion area/8.5mm
2.
X<1.1, represents kind to be identified or stock high resistance Trunk canker;
1.1≤X<1.3, represents anti-Trunk canker in kind to be identified or stock;
1.3≤X<1.7, represents in kind to be identified or stock and feels Trunk canker;
X >=1.7, represent kind to be identified or stock height sense Trunk canker.
Rapid identification apple limb provided by the invention, to the method for Target spot pathogen resistance, by blade inoculation, is created suitable Target spot pathogen infectious condition, is carried out ring spot Resistance Identification to result of infection; Qualification material requested is few, the used time is short, repeatable high, and authentication method is simple and easy to grasp, and cost is low, easily promote.
Embodiment
The specific embodiment of the present invention is described in conjunction with the embodiments.
June-July twice is to different apple varieties and stock material sampling, Resistance Identification has been carried out by the inventive method, selected apple variety and stock are the near different resistant materials by the qualification of field limb inoculation ring spot, the method that its standard of perfection founds antagonism characteristic of disease grade classification with reference to Yan Zhen divides disease index, highly disease-resistant grade (0.00%-10.00%); The disease-resistant grade of moderate (10.01%-25.00%); Moderate Infection Grades (25.01%-40.00%); Height Infection Grades (> 40.01%).
Rapid identification apple limb, to the method for Target spot pathogen resistance, comprises the following steps:
(1) PDA substratum makes
Potato is cleaned peeling, be cut into small pieces and put into the pot little fire that adds water and boil 0.5 hour, filter with double gauze, add in filtrate, agar and glucose, add water constant volume, potato: glucose: agar: the ratio that distilled water adds is 200g:20g:12g:1000mL; Be divided in test tube or triangular flask while hot, 121 DEG C of sterilizings 30 minutes, make PDA substratum;
(2) Target spot pathogen is cultivated
Target spot pathogen is inoculated into by Bechtop on above-mentioned PDA substratum, cultivates in illumination box under 25 DEG C of conditions, cover with after culture dish until mycelia and obtain ring spot substratum bacterium colony, for blade inoculation;
(3) blade is chosen
Choose 13-50, MM106,13-9, Qiao Najin, 13-40, Dou Nan, 77-34, Liquan brachyplast, Qiao Na gold 8 intend 5-6 sheet under identification of species or stock young sprout top ripe, completely launch and be expert evidence without disease and pest blade, repeat 6 times;
(4) blade inoculation
Dry after the blade aseptic water washing to be identified chosen, wrap up in petiole base with the sterilized non-fat bale of cotton, a upper sterilized water moisturizing is for subsequent use; Stab face of blade with inoculating needle, take off the homogeneous circular bacterium cake of growth with diameter 5mm punch tool from ring spot substratum colony edge is isometrical, center, raw mycelia face is placed in blade puncture wounds;
(5) carry disease germs leaf culture
Filter paper is spread in tray bottom, the water of pallet height about 1/3rd is added in pallet, bar shaped foam is placed in pallet, be positioned in pallet by postvaccinal blade, make the petiole being wrapped in absorbent cotton be dipped in sterilized water, vane frame is on bar shaped foam, pallet mouth is covered with preservative film, be placed in incubator and carry out dark culturing, culture condition temperature is 25 DEG C, and humidity is 100%;
(6) Resistance Identification
Cultivation the 6th day, right-angled intersection method is adopted to measure leaf spot lesion diameter, with scab transverse diameter and vertical footpath product representation lesion area, the lesion area of blade and the lesion area of known resistance class material contrast, and determine that kind to be identified or stock are to the resistance class of Trunk canker.
Known resistance class material is the lesion area of cockscomb, and standard is 8.5mm
2, determine that kind to be identified belonging to blade or stock are to the resistance class of Trunk canker, concrete standard is:
X=leaf spot lesion area/8.5mm
2.
X<1.1, represents kind to be identified or stock high resistance Trunk canker;
1.1≤X<1.3, represents anti-Trunk canker in kind to be identified or stock;
1.3≤X<1.7, represents in kind to be identified or stock and feels Trunk canker;
X >=1.7, represent kind to be identified or stock height sense Trunk canker.
Twice test result is in table 1, table 2, the inventive method qualification limb is tried material all shows consistent to the qualification result of ring rot of apple resistance and known field limb inoculated identification result 13-50, MM106,13-9,13-40, Dou Nan, 77-34, Liquan brachyplast 7, only in Qiao Na golden apple kind, this qualification result is anti-in being, and limb inoculated identification result is middle sense, the two is difference to some extent.Therefore think that this authentication method is practical.
Table 1 branch inoculation Resistance Identification and present method blade inoculation authentication method contrast (the 1st sub-sampling)
Table 2 branch inoculation Resistance Identification and present method blade inoculation authentication method contrast (the 2nd sub-sampling)
Claims (2)
1. Rapid identification apple limb is to the method for Target spot pathogen resistance, it is characterized in that, comprises the following steps:
(1) PDA substratum makes
Potato is cleaned peeling, be cut into small pieces and put into the pot little fire that adds water and boil 0.5 hour, filter with double gauze, add in filtrate, agar and glucose, add water constant volume, potato: glucose: agar: the ratio that distilled water adds is 200g:20g:12g:1000mL; Be divided in test tube or triangular flask while hot, 121 DEG C of sterilizings 30 minutes, make PDA substratum;
(2) Target spot pathogen is cultivated
Target spot pathogen is inoculated into by Bechtop on above-mentioned PDA substratum, cultivates in illumination box under 25 DEG C of conditions, cover with after culture dish until mycelia and obtain ring spot substratum bacterium colony, for blade inoculation;
(3) blade is chosen
Choose intend 5-6 sheet under identification of species or stock young sprout top ripe, completely launch and be expert evidence without disease and pest blade, repeat 6 times;
(4) blade inoculation
Dry after the blade aseptic water washing to be identified chosen, wrap up in petiole base with the sterilized non-fat bale of cotton, a upper sterilized water moisturizing is for subsequent use; Stab face of blade with inoculating needle, take off the homogeneous circular bacterium cake of growth with diameter 5mm punch tool from ring spot substratum colony edge is isometrical, the center in raw mycelia face is placed in blade puncture wounds;
(5) carry disease germs leaf culture
Filter paper is spread in tray bottom, the water of pallet height about 1/3rd is added in pallet, bar shaped foam is placed in pallet, be positioned in pallet by postvaccinal blade, make the petiole being wrapped in absorbent cotton be dipped in sterilized water, vane frame is on bar shaped foam, pallet mouth is covered with preservative film, be placed in incubator and carry out dark culturing, culture condition temperature is 25 DEG C, and humidity is 100%;
(6) Resistance Identification
Cultivation the 6th day, adopt right-angled intersection method to measure leaf spot lesion diameter, with scab transverse diameter and vertical footpath product representation lesion area, the lesion area of blade and the lesion area of known resistance class material contrasted, and determine identification of species or stock resistance class.
2. Rapid identification apple limb according to claim 1 is to the method for Target spot pathogen resistance, it is characterized in that: known resistance class material is the lesion area of cockscomb, and standard lesion area is 8.5mm
2.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104962605A (en) * | 2015-07-01 | 2015-10-07 | 中国农业科学院蔬菜花卉研究所 | Screening method of medicinal agents for preventing bacterial soft rot of vegetables |
| CN105165604A (en) * | 2015-10-15 | 2015-12-23 | 中国农业科学院作物科学研究所 | Method for identifying resistance of soybeans to soybean phytophthora blight and application of method in breeding |
| CN106554986A (en) * | 2015-09-28 | 2017-04-05 | 李保华 | It is a kind of to detect the thecasporous method of Botryosphaeria berengeriana f. sp |
| CN111345209A (en) * | 2020-03-27 | 2020-06-30 | 云南省农业科学院热带亚热带经济作物研究所 | Method for identifying field disease resistance of cassava to root rot |
| CN111471741A (en) * | 2020-06-10 | 2020-07-31 | 南京农业大学 | Method for rapidly identifying ring spot resistance of pear trees |
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| CN101864481A (en) * | 2009-09-29 | 2010-10-20 | 青岛农业大学 | SCAR molecule marking method for detecting apple tree trunk ring spot resistance gene |
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2015
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Patent Citations (2)
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| CN101864481A (en) * | 2009-09-29 | 2010-10-20 | 青岛农业大学 | SCAR molecule marking method for detecting apple tree trunk ring spot resistance gene |
| CN104152534A (en) * | 2014-08-08 | 2014-11-19 | 四川省农业科学院经济作物育种栽培研究所 | Method for quickly measuring resistance of eggplant verticillium wilt at seedling stage |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104962605A (en) * | 2015-07-01 | 2015-10-07 | 中国农业科学院蔬菜花卉研究所 | Screening method of medicinal agents for preventing bacterial soft rot of vegetables |
| CN106554986A (en) * | 2015-09-28 | 2017-04-05 | 李保华 | It is a kind of to detect the thecasporous method of Botryosphaeria berengeriana f. sp |
| CN105165604A (en) * | 2015-10-15 | 2015-12-23 | 中国农业科学院作物科学研究所 | Method for identifying resistance of soybeans to soybean phytophthora blight and application of method in breeding |
| CN105165604B (en) * | 2015-10-15 | 2017-08-25 | 中国农业科学院作物科学研究所 | A kind of application for identifying soybean to the method for soybean blight resistance and its in breeding |
| CN111345209A (en) * | 2020-03-27 | 2020-06-30 | 云南省农业科学院热带亚热带经济作物研究所 | Method for identifying field disease resistance of cassava to root rot |
| CN111471741A (en) * | 2020-06-10 | 2020-07-31 | 南京农业大学 | Method for rapidly identifying ring spot resistance of pear trees |
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