CN105018566A - Verification method of resistance to mycosphaerella leaf spot of sweet potato - Google Patents
Verification method of resistance to mycosphaerella leaf spot of sweet potato Download PDFInfo
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- CN105018566A CN105018566A CN201510450340.9A CN201510450340A CN105018566A CN 105018566 A CN105018566 A CN 105018566A CN 201510450340 A CN201510450340 A CN 201510450340A CN 105018566 A CN105018566 A CN 105018566A
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Abstract
The invention discloses a verification method of resistance to mycosphaerella leaf spot of sweet potato, belonging to a crop disease and pest resistance verification method. The method is scientific, and has the advantages of simple operational process and low requirements for test conditions. The method comprises the following steps: selecting 3 medium-size smooth-surface no-rot potato blocks to be verified, carefully cleaning, airing, numbering, disinfecting with 75% alcohol, inoculating into a 1*10<6>/mL brown spot spore suspension which is previously prepared from sterilized water, moisturizing with sterilized wet gauze, culturing for 10 days while keeping at the room temperature of 26 DEG C, detecting the rotten diameter of each test potato block, calculating the disease-resistance performance percentage, and judging the resistance to mycosphaerella leaf spot of each tested potato block according to the disease-resistance performance percentage.
Description
Technical field
The present invention relates to a kind of corps diseases Resistance Identification method, be exactly a kind of reaping hook microbial sweet potato brown spot Resistance Identification method specifically, belong to agriculture plant protection field.
Background technology
Sweet potato brown spot (sweet potato brown spot), infected by sickle-like bacteria (Fusarium spp) and cause, for one of the Major Diseases of growth of sweet potato phase and storage period, it is a kind of new expression occurred recent years, the generaI investigation of 2012-2015 sweet potato disease finds, this disease is extensively distributed in national each sweet potato production area.This disease main manifestations on sweet potato potato block is that circle, Vandyke brown are to black scab, slightly cave in, there is slight bitter taste, the similar sweet potato black rot of symptom, field and shelf time all occur, and potato block inedibility after morbidity, and normal and other diseases of shelf time occur jointly, make entirely to store potato block to rot, cause serious in mistake of damaging due to crushing.
Sweet potato brown spot Fusarium Pathogen ubiquity in soil, therefore chemopreventive effects is not obvious, and the selection and popularization of disease-resistant variety is the Main Means controlling this Occurrence and development of disease, and the disease resistance of therefore accurate and effective evaluation kind seems and is even more important.At present, more to the Resistence research of the diseases such as sweet potato black rot, oxyuriasis and soft rot, but because sweet potato brown spot is the de novo disease of sweet potato recent years, the Resistance Identification research of different sweet potato variety to brown spot is quite deficient, at present, there is not yet the sweet potato brown spot Resistance Identification method of standard.
Summary of the invention
The object of this invention is to provide a kind of sweet potato brown spot Resistance Identification method.The advantages such as it is easy that the method has operating process, easy to learn, time saving and energy saving, can determine the anti-perceptual degree of potato block to sweet potato brown spot fast and accurately, for sweet potato breeding for disease resistance screening bulk materials provides technical support.
The present invention realizes with following technical scheme: a kind of sweet potato brown spot Resistance Identification method, 75% alcohol disinfecting potato block, adopts 0.5cm long inoculation syringe needle, dips sweet potato brown patch germ spore, inoculation potato block, cultivate in 26 DEG C of incubators, the moisturizing of sterilizing wet gauze, cultivate 10 days, measure lesion diameter, calculate disease-resistant performance percentage, evaluate the resistance of sweet potato potato block Resistance to Cercospora, specifically carry out as follows:
1. pathogenicbacteria separation and preparation: gather morbidity potato block from field, utilize separate tissue and single spore separation method bacterial isolate bacterium, at 26 DEG C, cultivation 7 days on potato dextrose agar (PDA) flat board, for subsequent use;
2. potato block of participating in the experiment prepares: when gathering in the crops, and each sweet potato variety field is chosen 3 pieces of median sizes, smooth surface, without hindering rotten potato block, carefully being cleaned, drying, and numbering is stand-by after the alcohol disinfecting of 75%;
3. the preparation of inoculation liquid: by under the conidium aseptic water washing that grows on PDA flat board, being mixed with concentration with sterilized water is 1 × 10
6the spore suspension of individual/ml is for subsequent use;
4. inoculated identification: the capacity that is divided in by the spore suspension prepared is in the sterile beaker of 100ml, then No. 8 medical needles are got, 0.5cm is truncated to pliers, dip the spore suspension prepared, needle punching is longitudinally inoculated on sweet potato potato block, and every potato block inoculation three row, often arranges 5 points, once, syringe needle need dip bacterium liquid in every acupuncture.Postvaccinal potato block sterile gauze is covered moisturizing, and every day changes clothes gauze, cultivates in 26 DEG C of incubators.
(5) research and appraisal: inoculate after 10 days, take out potato block, and scab is circular, measure lesion diameter, with in No. hundred, anti-kind triumph for check variety, calculate disease-resistant performance percentage as follows, then carry out the anti-sense evaluating characteristics of kind according to disease-resistant performance percentage;
The anti-sense evaluating characteristics of described sweet potato brown spot is divided into Pyatyi, and judgement criteria at different levels is:
High resistance: disease-resistant performance percentage≤40;
Disease-resistant: 40 disease-resistant performance percentage≤80 of <;
In anti-: 80 disease-resistant performance percentage≤120 of <;
Susceptible: 120 disease-resistant performance percentage≤160 of <;
High sense: the disease-resistant performance percentage of 160 <.
Advantage of the present invention is, the method is with potato block for material, and indoor inoculation sweet potato brown patch germ identifies anti-perception, infects the practical situation of sweet potato brown spot close to field sweet potato potato block.This authentication method science, time saving and energy saving, simple to operate, easy to learn, less demanding to experimental installation, each breeding units all can carry out qualification work.This authentication method can provide technical support for the qualification in early stage of sweet potato breeding for disease resistance, rejects part susceptible variety, for later stage screening varieties alleviates human and material resources.Meanwhile, indoor sterilizing is inoculated, and is easy to test conditions and controls, also enhance the accuracy of result.Promoting the use of of the method, for screening disease-resistant variety with instruct the significant and realistic function of sweet potato breeding for disease resistance.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment
A kind of sweet potato brown spot Resistance Identification method, we carry out the test of brown spot Resistance Identification to the potato block of 16 sweet potato varieties that harvesting time gathers, and carry out as follows:
1. pathogenicbacteria separation and preparation: gather morbidity potato block from experimental plot, sweet potato research centre, the Chinese Academy of Agricultural Sciences Xuzhou, utilize separate tissue and single spore separation method bacterial isolate bacterium, and determine through Koch's Postulates checking.At 26 DEG C, cultivation 7 days on potato dextrose agar (PDA) flat board, for subsequent use;
2. potato block of participating in the experiment prepares: when gathering in the crops, select 16 sweet potato varieties, and each sweet potato variety is chosen 3 pieces of median sizes, smooth surface, without hindering rotten potato block, carefully being cleaned, drying, and numbering is stand-by after the alcohol disinfecting of 75%;
3. the preparation of inoculation liquid: by under the conidium aseptic water washing that grows on PDA flat board, being mixed with concentration with sterilized water is 1 × 10
6the spore suspension of individual/ml is for subsequent use;
4. inoculated identification: the capacity that is divided in by the spore suspension prepared is in the sterile beaker of 100ml, then adopt and get No. 8 medical needles, 0.5cm is truncated to pliers, dip the spore suspension prepared, needle punching is longitudinally inoculated on sweet potato potato block, and every potato block inoculation three row, often arranges 5 points, once, syringe needle need dip bacterium liquid in every acupuncture.By the moisturizing of postvaccinal potato block sterile gauze, every day changes clothes gauze, cultivates in 26 DEG C of incubators.
5. (5) research and appraisal: inoculate after 10 days, take out potato block, and scab is circular, measure lesion diameter, with in No. hundred, anti-kind triumph for check variety, calculate disease-resistant performance percentage as follows, then carry out the anti-sense evaluating characteristics of kind according to disease-resistant performance percentage;
The anti-sense evaluating characteristics of described sweet potato brown spot is divided into Pyatyi, and judgement criteria at different levels is:
High resistance: disease-resistant performance percentage≤40;
Disease-resistant: 40 disease-resistant performance percentage≤80 of <;
In anti-: 80 disease-resistant performance percentage≤120 of <;
Susceptible: 120 disease-resistant performance percentage≤160 of <;
High sense: the disease-resistant performance percentage of 160 <.
Table 1: sweet potato potato block is to sweet potato brown spot Resistance Identification result
* HR: high resistance, high resistance; R: anti-, resistance; MR: in anti-, middle resistance; HS: high sense, high susceptibility; S: sense, susceptibility; MS: middle sense, middle susceptibility
Conclusion:
As can be seen from Table 1: the potato block of inoculation brown patch germ is placed in 26 DEG C of biochemical cultivation cases and cultivates 10 days " Invest, Then Investigate " scab morbidity diameters, and calculate disease-resistant performance percentage and evaluate, wherein: the resistance rank of lucky Xu No. 1 and Xu's potato 25 pairs of sweet potato brown spots is " disease-resistant "; Xu's potato 29, business's potato 19, the triumph resistance ranks of kind to sweet potato brown spot such as No. hundred be " in resist "; The resistance rank of cigarette potato 25 and wide potato 87 pairs of sweet potato brown spots is " susceptible "; The varietal resistance ranks such as Xu's potato 26, No. 5, rock potato, Xu Yu potato 34, Xu Yu potato 35, Xu's potato 33, Xu's potato 28, No. 9, imperial potato, Ji potato 25 and chestnut are fragrant are " high sense ", belong to high sense kind.
Claims (1)
1. a sweet potato brown spot Resistance Identification method, it is characterized in that: with 75% alcohol disinfecting potato block, adopt 0.5cm long inoculation syringe needle, dip sweet potato brown patch germ spore, inoculation potato block, cultivate in 26 DEG C of incubators, the moisturizing of sterilizing wet gauze, cultivate 10 days, measure lesion diameter, calculate disease-resistant performance percentage, evaluate the resistance of sweet potato potato block Resistance to Cercospora, specifically carry out as follows:
(1) pathogenicbacteria separation and preparation: field gathers morbidity potato block, utilizes separate tissue and single spore separation method bacterial isolate bacterium, cultivates 7 days at 26 DEG C on potato dextrose agar PDA flat board, for subsequent use;
(2) potato block of participating in the experiment prepares: when gathering in the crops, and each sweet potato variety field is chosen 3 pieces of median sizes, smooth surface, without hindering rotten potato block, carefully being cleaned, drying, and numbering is stand-by after the alcohol disinfecting of 75%;
(3) preparation of inoculation liquid: by under the conidium aseptic water washing that grows on PDA flat board, being mixed with concentration with sterilized water is 1 × 10
6the spore suspension of individual/ml is for subsequent use;
(4) inoculated identification: the capacity that is divided in by the spore suspension prepared is in the sterile beaker of 100ml, then No. 8 medical needles are adopted, 0.5cm is truncated to pliers, dip the spore suspension prepared, needle punching is longitudinally inoculated on sweet potato potato block, and every potato block inoculation three row, often arranges 5 points, once, syringe needle need dip bacterium liquid in every acupuncture; By the moisturizing of postvaccinal potato block sterile gauze, every day changes clothes gauze, cultivates in 26 DEG C of incubators;
(5) research and appraisal: inoculate after 10 days, take out potato block, and scab is circular, measure lesion diameter, with in No. hundred, anti-kind triumph for check variety, calculate disease-resistant performance percentage as follows, then carry out the anti-sense evaluating characteristics of kind according to disease-resistant performance percentage;
The anti-sense evaluating characteristics of described sweet potato brown spot is divided into Pyatyi, and judgement criteria at different levels is:
High resistance: disease-resistant performance percentage≤40;
Disease-resistant: 40 disease-resistant performance percentage≤80 of <;
In anti-: 80 disease-resistant performance percentage≤120 of <;
Susceptible: 120 disease-resistant performance percentage≤160 of <;
High sense: the disease-resistant performance percentage of 160 <.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105875231A (en) * | 2016-04-12 | 2016-08-24 | 江苏徐州甘薯研究中心 | Indoor invasiveness-resisting determination method for sweet potato stem nematode |
| CN106635830A (en) * | 2016-11-22 | 2017-05-10 | 江苏省农业科学院 | Culture method of alternaria brassicicola used for identifying resistance to black spot of sweet potatoes |
| CN109507368A (en) * | 2018-11-16 | 2019-03-22 | 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 accurate Resistance Identification method of seedling stage sweet potato black rot |
| CN112522359A (en) * | 2020-12-09 | 2021-03-19 | 广东省农业科学院作物研究所 | Resistance identification method for sweet potato scab for stem tip vegetables |
| CN112725367A (en) * | 2021-03-01 | 2021-04-30 | 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) | Sweet potato sucrose invertase gene IbINV and application thereof |
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2015
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Non-Patent Citations (5)
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| 周佳明: "甘薯黑斑病抗性鉴定不同分级方法效果研究", 《四川师范学院学报》 * |
| 周佳明: "甘薯黑斑病抗性鉴定方法研究", 《西南农业学报》 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105875231A (en) * | 2016-04-12 | 2016-08-24 | 江苏徐州甘薯研究中心 | Indoor invasiveness-resisting determination method for sweet potato stem nematode |
| CN106635830A (en) * | 2016-11-22 | 2017-05-10 | 江苏省农业科学院 | Culture method of alternaria brassicicola used for identifying resistance to black spot of sweet potatoes |
| CN109507368A (en) * | 2018-11-16 | 2019-03-22 | 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 accurate Resistance Identification method of seedling stage sweet potato black rot |
| CN112522359A (en) * | 2020-12-09 | 2021-03-19 | 广东省农业科学院作物研究所 | Resistance identification method for sweet potato scab for stem tip vegetables |
| CN112725367A (en) * | 2021-03-01 | 2021-04-30 | 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) | Sweet potato sucrose invertase gene IbINV and application thereof |
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