CN102612962B - Method for identifying resistance of sweet potatoes in seedling stage to black spot - Google Patents
Method for identifying resistance of sweet potatoes in seedling stage to black spot Download PDFInfo
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- CN102612962B CN102612962B CN 201210095052 CN201210095052A CN102612962B CN 102612962 B CN102612962 B CN 102612962B CN 201210095052 CN201210095052 CN 201210095052 CN 201210095052 A CN201210095052 A CN 201210095052A CN 102612962 B CN102612962 B CN 102612962B
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Abstract
The invention relates to a method for identifying resistance of crops to plant diseases and insect pests, in particular to a method for identifying resistance of sweet potatoes in a seedling stage to black spot. The method is scientific, and easy to implement, and has low requirements on experimental conditions. The method comprises the following steps of: taking shoot-tip seedlings of sweet potatoes which grow consistently in a seedling bed or a field, cutting off the lowest 2-3 leaves, cleaning by using a disinfectant fluid, putting into 1*10<4>/ml black spot spore suspension which is prepared by using sterile water in advance, culturing at room temperature of between 25 and 27DEG C for 6 days, investigating harm degrees in which experimental sweet potato seedlings are infected by the black spot, calculating each repeated average harm level, and judging the infection resistance of the experimental sweet potato seedlings to the black spot according to the harm level.
Description
Technical field
The present invention relates to a kind of corps diseases Resistance Identification method, is exactly a kind of sweet potato black rot seedling resistance authentication method specifically, belongs to the agriculture plant protection field.
Background technology
Sweet potato black rot is the important fungal disease of a kind of main harm sweet potato root system and basal part of stem, and all there is generation in each sweet potato producing region, the world.Nineteen thirty-seven is imported China into, and this disease occurs for existing 26 provinces, municipality directly under the Central Government, autonomous regions, in the sweet potato seedbed, land for growing field crops and shelf time all can cause harm, is one of the three large serious plant disease in northern China potato district.After the potato seedling is infected, produce circle or the little blackspot of fusiformis of depression in stem, after enlarge gradually, around potato seedling base portion pin shape darkly, acrial part yellow leaf or the blackening of seedling base portion is rotted.Potato transplantation of seedlings land for growing field crops 1-2 is after week in spite of illness, and the radical leaves jaundice comes off, when seriously ill, and butt rot, remaining fibrous vascular bundle only, the potato seedling is withered.After shelf time potato piece was injured, scab was just the black point, was expanded into gradually circle or fusiformis scab, and the middle black thorn that produces is concurrent with other fungus and bacteriums, easily causes rotten.The production loss that China every year causes because of this disease is in 10% left and right.
Studies show that, the selection and popularization of disease-resistant variety is the Main Means of controlling this disease.Therefore, estimating effectively, fast and accurately varietal resistance seems and is even more important.Resistance Identification research to sweet potato black rot is material mainly with the potato piece, and the potato seedling is less to the Resistance Identification research of black spot.(1990) such as Qiu Rui sickles adopt the climing entry needle inoculated identification of land for growing field crops potato, but due to the difference of the climing growing way of potato between different cultivars and wood fibre cellulose content, easily cause the error of result.
Summary of the invention
The objective of the invention is provides a kind of sweet potato black rot seedling resistance authentication method for overcoming above-mentioned the deficiencies in the prior art part.After the method was removed the bottom petiole by the top seedling of consistent sweet potato that will grow, it was 1 * 10 that immersion prepares concentration
4In the black spot spore suspension of individual/ml, can tell fast different cultivars potato seedling after six days to the steps such as anti-sense degree of black spot, can determine fast and accurately that the potato seedling is to the resistance level of black spot, have easy, easy to learn, the advantage such as time saving and energy saving is 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 black rot seedling resistance authentication method is characterized in that: carry out as follows:
(1) germ separates and cultivates: gather morbidity potato piece from the field, isolate alternaria with the fungi separating method, cultivated 7 days on potato dextrose agar, and standby;
(2) the potato seedling of participating in the experiment is prepared: from the seedbed or 6 consistent top seedling of sweet potato of growth of field clip, be advisable with 5-6 internode depending on potato seedling internode length, cut bottom two blades from petiole base after taking back laboratory, with aqua sterilisa, potato seedling basal part of stem is cleaned up, standby;
(3) preparation of inoculation bacterium liquid: will be in the conidium that grows on potato dextrose agar with under aseptic water washing, being mixed with concentration with sterile water is 1 * 10
4The spore suspension of individual/ml is standby;
(4) inoculated identification: the spore suspension for preparing is divided in the aseptic triangular flask that capacity is 250ml, then will put down gently in triangular flask for examination potato seedling, 3 of every kinds, be 3 repetitions, cultivate under 25-27 ℃, establish simultaneously blank sterile water for contrast, add with sterile water after the consumption of triangular flask culture fluid;
(5) research and appraisal: the 6 days following part black spot of " Invest, Then Investigate " potato seedling water surface incidences, and the potato seedling is divided into 0-4 level totally 5 grades according to occurring degree, according to be injured grade with kind to the resistance of black spot be divided into disease-resistant, in anti-and susceptible three ranks;
(6) the potato seedling grade scale of being injured: 0 grade: can't see illness; 1 grade: have fragmentary minor illness spot to occur, scab number≤20; 2 grades: scab is larger, and scab counts 20 of ﹥, but not in flakes; 3 grades: scab is more, and spot is linked to be sheet, length≤2cm; 4 grades: scab in flakes, and length ﹥ 2cm in flakes;
(7) average onset grade=(repeating I rank+repetition II rank+repetition III rank)/3; Disease-resistant rank is determined: disease-resistant: mined grade≤1.0; In anti-: 1.0＜mined grade＜3.0; Susceptible: mined grade 〉=3.0.
Advantage of the present invention is: the method take the seedbed or land for growing field crops potato seedling stem apex be material, the indoor inoculation alternaria is identified disease resistance can show different cultivars potato seedling accurately, fast, intuitively to the resistance level of sweet potato black rot.The method infects the actual conditions of black spot closer to the field sweet potato's seedlings, simple to operate, easy to learn, less demanding to experimental facilities, each breeding units all can be carried out evaluation work.This authentication method can be identified in earlier stage for the sweet potato breeding for disease resistance provides technical support, rejects the part susceptible variety, for the later stage screening varieties alleviates human and material resources.Simultaneously, take the seedbed or land for growing field crops potato seedling stem apex be expert evidence, can reduce to greatest extent the otherness of drawing materials, reduce experimental error.The indoor sterilizing inoculation is easy to experimental condition and controls, and has also strengthened the accuracy of result.Promoting the use of of the method is for the screening disease-resistant variety with instruct the significant and realistic function of sweet potato breeding for disease resistance.
Embodiment
The invention will be further described below in conjunction with embodiment:
Embodiment,
Sweet potato black rot seedling resistance authentication method, we carry out the test of black spot Resistance Identification to the potato seedling of 13 sweet potato varieties that the seedbed gathers, and carry out as follows:
(1) germ separates and cultivates: gather morbidity potato piece from the field, isolate alternaria with the fungi separating method, cultivated 7 days on potato dextrose agar, and standby;
(2) the potato seedling of participating in the experiment is prepared: gather No. hundred, North China 52-45, Nanjing 92, Xu's potato 18, Zhejiang Xu No. 1, Xu 273, Ji agriculture 22, Yantai 252, Xu 02-9-11, Xu 02-16-17, peaceful 55-1, Xu 22-1, Xu 25-2, triumph totally 13 sweet potato varieties (being) potato seedling from the seedbed, every kind is 6 consistent top seedling of sweet potato of growth of clip respectively, be advisable with 5-6 internode depending on potato seedling internode length, cut bottom 2-3 sheet blade from petiole base after taking back laboratory, with aqua sterilisa, potato seedling basal part of stem is cleaned up, standby;
(3) preparation of inoculation bacterium liquid: will be in the black spot conidium that grows on potato dextrose agar with under aseptic water washing, being mixed with concentration with sterile water is 1 * 10
4The spore suspension of individual/ml is standby;
(4) inoculated identification: the spore suspension for preparing is divided in the aseptic triangular flask that capacity is 250ml, then will put down gently in triangular flask for examination potato seedling, 3 of every kinds, be 3 repetitions, cultivate under 25-27 ℃, establish simultaneously blank sterile water for contrast, fill it up with sterile water after culture fluid consumption;
(5) research and appraisal as a result: the 6 days following part black spot of " Invest, Then Investigate " potato seedling water surface incidences, and according to occurring degree, the potato seedling is divided into 0-4 level totally 5 grades, according to be injured grade with kind to the resistance of black spot be divided into disease-resistant, in anti-and susceptible three ranks;
The potato seedling grade scale of being injured:
0 grade: can't see illness;
1 grade: have fragmentary minor illness spot to occur, scab number≤20;
2 grades: scab is larger, and scab counts 20 of ﹥, but not in flakes;
3 grades: scab is more, and spot is linked to be sheet, length≤2cm;
4 and: scab in flakes, and length ﹥ 2cm in flakes;
Average onset grade=(repeating I rank+repetition II rank+repetition III rank)/3;
Disease-resistant rank is determined: disease-resistant: mined grade≤1.0; In anti-: 1.0＜mined grade＜3.0; Susceptible: mined grade 〉=3.0.
Test for identification the results are shown in Table 1:
Table 1 sweet potato black spot in seedling stage Resistance Identification result
Conclusion: as can be seen from Table 1, in 14 sweet potato varieties identifying, Xu 02-9-11, Nanjing 92, North China 52-45 and Xu's potato 18 totally 4 kinds (being) potato seedling are " disease-resistant " to the resistance rank of black spot; Xu potato 25-2, Xu's potato 22-1, Zhejiang Xu No. 1, Xu 273 and Xu 02-16-17 totally 5 kinds (being) potato seedling are " in anti-" to the resistance rank of black spot; Ji agriculture 22, No. hundred, triumph, peaceful 55-1 and Yantai 252 the resistance rank of totally 4 product potato seed seedling black spots are " susceptible ".
Claims (1)
1. sweet potato black rot seedling resistance authentication method is characterized in that: carry out as follows:
(1) germ separates and cultivates: gather morbidity potato piece from the field, isolate alternaria with the fungi separating method, cultivated 7 days on potato dextrose agar, and standby;
(2) the potato seedling of participating in the experiment is prepared: from the seedbed or 6 consistent top seedling of sweet potato of growth of field clip, be advisable with 5-6 internode depending on potato seedling internode length, cut bottom two blades from petiole base after taking back laboratory, with aqua sterilisa, potato seedling basal part of stem is cleaned up, standby;
(3) preparation of inoculation bacterium liquid: will be in the conidium that grows on potato dextrose agar with under aseptic water washing, being mixed with concentration with sterile water is 1 * 10
4The spore suspension of individual/ml is standby;
(4) inoculated identification: the spore suspension for preparing is divided in the aseptic triangular flask that capacity is 250ml, then will put down gently in triangular flask for examination potato seedling, 3 of every kinds, be 3 repetitions, cultivate under 25-27 ℃, establish simultaneously blank sterile water for contrast, add with sterile water after the consumption of triangular flask culture fluid;
(5) research and appraisal: the 6 days following part black spot of " Invest, Then Investigate " potato seedling water surface incidences, and the potato seedling is divided into 0-4 level totally 5 grades according to occurring degree, according to be injured grade with kind to the resistance of black spot be divided into disease-resistant, in anti-and susceptible three ranks;
(6) the potato seedling grade scale of being injured: 0 grade: can't see illness; 1 grade: have fragmentary minor illness spot to occur, scab number≤20; 2 grades: scab is larger, and scab counts 20 of ﹥, but not in flakes; 3 grades: scab is more, and spot is linked to be sheet, length≤2cm; 4 grades: scab in flakes, and length ﹥ 2cm in flakes;
(7) average onset grade=(repeating I rank+repetition II rank+repetition III rank)/3; Disease-resistant rank is determined: disease-resistant: mined grade≤1.0; In anti-: 1.0＜mined grade＜3.0; Susceptible: mined grade 〉=3.0.
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| CN106031352B (en) * | 2015-03-11 | 2018-12-04 | 李保华 | Method for evaluating control effect of bactericide on apple floral organ infected diseases |
| CN105010126B (en) * | 2015-07-06 | 2017-09-26 | 山东省农业科学院作物研究所 | A kind of quick breeding method of anti-black spot sweet potato variety |
| CN107258417A (en) * | 2016-04-06 | 2017-10-20 | 万素梅 | The occurrence regularity of jujube alternaria |
| 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 |
| CN109392646A (en) * | 2018-11-23 | 2019-03-01 | 云南省烟草农业科学研究院 | A kind of investigation method of tobacco black shank resistance |
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