CN104255449A - Indoor auxiliary sweet potato drought tolerance identification method - Google Patents
Indoor auxiliary sweet potato drought tolerance identification method Download PDFInfo
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- CN104255449A CN104255449A CN201410439300.XA CN201410439300A CN104255449A CN 104255449 A CN104255449 A CN 104255449A CN 201410439300 A CN201410439300 A CN 201410439300A CN 104255449 A CN104255449 A CN 104255449A
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Abstract
The invention relates to a sweet potato drought tolerance identification method, and in particular relates to an indoor auxiliary sweet potato drought tolerance identification method, belonging to the technical field of sweet potato drought tolerance identification. The method comprises the following steps: by selecting a test tube seedling stem section which is subjected to subculture for more than or just 50 days as an explant, cutting a single section, inoculating the explant into a high-permeation identification culture medium, performing inoculation treatment for 30 days, subsequently surveying the relative root growth number coefficients of different varieties, and judging the drought tolerance grade of the varieties according to the relative root growth number coefficients of different varieties, wherein the drought tolerance of the variety of which the relative root growth number coefficient is greater than or equal to 1.0 is high, the drought tolerance of the variety of which the relative root growth number coefficient is greater than 0.5 and less than 1.0 is medium, and the drought tolerance of the variety of which the relative root growth number coefficient is smaller than or equal to 0.5 is relatively poor. The method is simple to operate, is flexible and convenient and applicable to screening on a great number of materials, can relieve the workload of field drought tolerance identification and can improve the working efficiency.
Description
Technical field
The present invention relates to a kind of sweet potato drought resistance authentication method, be specifically related to a kind of method of indoor assistant identification sweet potato drought resistance, belong to sweet potato drought resistance authenticate technology field.
Background technology
The object of sweet potato drought resistance qualification is mainly screened the kind of suitable drought-hit area cultivation and promotes seed selection that is drought-enduring, the special new varieties of high-quality, high yield from the sweet potato variety be bred as.The diversification crop that sweet potato is integrated as novel energy raw material and rapidoprint, nervous for current arable land, for accomplishing " not striving grain with people; do not strive ground with grain ", and sweet potato is the cereal crops that a kind of adaptability and resistance are stronger, the marginal lands such as inland is saline and alkaline, arid and hilly and mountainous land are the new world that following Sweet Potato Industry is expanded.For promoting screening, the qualification of the materials such as the drought-enduring resource of sweet potato, forefathers by the process of rain cover simulating drought, carry out the qualification of sweet potato drought resistance, but this all needs to spend a large amount of manpower and materials, workload is very large, and due to processing region limited, be difficult to the primary dcreening operation, the qualification that meet lot of materials.And to be coerced by research and under control treatment condition, the relation of the change such as oxidize-resistant physiology index and the drought-enduring correlation of sweet potato, owing to being difficult to directly to judge sweet potato drought resistance by minority physical signs, and the detection of physical signs, workload is also larger, the many factors such as weather all likely have influence on its accuracy detected and repeatability, thus impact is to the judgement to drought resistance result.In a word, low the extreme influence screening of drought-enduring sweet potato material and the process of drought-enduring breeding of new variety of drought-enduring determination rates.
Summary of the invention
The object of the invention is to overcome above-mentioned existing methods not enough, a kind of method of indoor assistant identification sweet potato drought resistance is provided, the method has fast, efficiently, be not subject to other environmental influences extraneous, ensure that environmental condition residing for expert evidence is consistent, can the drought resistance of actual response material.
The present invention realizes with following technical scheme: a kind of method of indoor assistant identification sweet potato drought resistance, be explant by getting the sweet potato test-tube plantlet of squamous subculture >=50d, cut single segmental, being inoculated in height, to ooze qualification medium (be 100g/L according to W/V, aseptically, PEG 8000 is dissolved in 1/2MS liquid nutrient medium (through high-temperature sterilization but do not add agar powder), then will be 1:1.5 according to V/V containing the 1/2MS liquid nutrient medium of PEG 8000, be distributed into 1/2MS solid culture medium culture dish, balance 24h, then liquid nutrient medium remaining above solid culture medium is removed, the 1/2MS solid culture medium of such configuration is qualification medium), inoculation 30d " Invest, Then Investigate " is taken root number relatively, calculate number system number of relatively taking root, according to the drought resistance classification of several coefficient decision kind of relatively taking root.Specifically carry out as follows:
(1) configuration of 1/2MS solid culture medium: MS minimal medium is a large amount of, trace element reduces by half, and adds sucrose 30g/L, Phytagel 5g/L, adjusts PH 5.7 before sterilizing; Being distributed into diameter after sterilizing is 90mm culture dish, cools stand-by.
(2) configure and highly ooze qualification medium: first, be 100g/L according to W/V, aseptically, PEG 8000 be dissolved in through high-temperature sterilization but do not add the 1/2MS liquid nutrient medium of agar powder, then the 1/2MS liquid nutrient medium containing PEG 8000 is incorporated with the culture dish of 1/2MS solid culture medium, balance 24h, then removes liquid nutrient medium remaining above solid culture medium, for subsequent use; Described is 1:1.5 containing the 1/2MS liquid nutrient medium of PEG 8000 and the V/V ratio of 1/2MS solid culture medium;
(3) stem section inoculation: choose squamous subculture >=50d different cultivars test-tube plantlet material, cut single segmental, be inoculated in height and ooze qualification medium culture;
(4) number system number of relatively taking root is investigated: after inoculation 30d, investigate the number of taking root of each process material and corresponding contrast, calculate number system number of relatively taking root; Number system of relatively taking root number=high osmotic treatment kind number/check variety of taking root is taken root number;
(5) relatively take root the kind of number system number >=1.0, drought resistance is strong; 0.5 < is taken root the kind of number system number < 1.0 relatively, and drought resistance is medium; The kind of number system of relatively taking root number≤0.5, drought resistance is poor.
Advantage of the present invention is: the method is workable, simple, and efficient quick is not subject to seasonal restrictions, and can get rid of the impact of external environmental condition difference on drought-enduring qualification result, is the effective ways that primary dcreening operation drought resistance sweet potato variety is assisted in a kind of indoor.
Embodiment
Further illustrate method of the present invention and effect below in conjunction with embodiment, but do not limit the scope of the invention.
Embodiment 1,
1. test material and method
1.1 test materials: test selected materials be drought-enduring qualification base through Xinjiang, within continuous 2 years, be accredited as drought resistance strong, in, 6 parts of materials differing from.Wherein:
Drought resistance is strong: ten thousand purples 56, wide potato 87;
Drought resistance is medium: Xushen21 well, Xu 20-1;
Drought resistance is poor: peaceful purple potato No. 1, day purple 0602.
1.2 test methods:
The configuration of 1/2MS solid culture medium: MS minimal medium macroelement and trace element reduce by half, adds sucrose 30g/L, Phytagel 5g/L, adjusts PH 5.7 before sterilizing; Being distributed into diameter after sterilizing is 90mm culture dish, cools stand-by.
Height oozes the configuration of qualification medium: first, be 50g/L, 100g/L, 200g/L, 300g/L according to W/V, aseptically, PEG 8000 is dissolved in 1/2MS liquid nutrient medium (through high-temperature sterilization but do not add agar powder), then by being 1:1.5 containing the 1/2MS liquid nutrient medium of PEG 8000 according to V/V, 1/2MS solid culture medium culture dish is distributed into, balance 24h, then liquid nutrient medium remaining above solid culture medium is removed, for subsequent use.
Stem section is inoculated: choose squamous subculture 30d, 40d, 50d, 60d different cultivars test-tube plantlet material, cut single segmental, be inoculated in height and ooze qualification medium culture.
1.3 investigation and data statistics: after inoculation 30d, investigate the number of taking root of each process material and corresponding contrast, calculate and relatively to take root number system number, number system of relatively taking root number=high osmotic treatment kind number/check variety of taking root is taken root number.
2. results and analysis
2.1 different squamous subculture time test-tube plantlets ooze the reaction of coercing to height
Choose the test-tube plantlet stem section of the day purple 0602 different squamous subculture time of drought resistance difference, be inoculated in that variable concentrations is high oozes qualification medium, screen under comparatively Thief zone concentration, day, purple 0602 test-tube plantlet survival rate was the squamous subculture test-tube plantlet material of 100%, result shows: under 50g/L concentration for the treatment of, and the survival rate of different squamous subculture time stem section can reach 100; Along with the raising of concentration for the treatment of, stem section survival rate is on a declining curve, even all dead; Along with stem section Subculture Time extends, and the growth such as stem intersegmental part fiber, there is higher anti-adversity ability, after the stem section of different squamous subculture time is inoculated in that 100g/L is high and oozes medium, the test-tube plantlet stem section survival rate of squamous subculture 30d, 40d is lower than 100%, after squamous subculture is more than or equal to 50d, the survival rate of test-tube plantlet stem section reaches 100%; Improve further if height oozes concentration, test-tube plantlet stem section cannot adapt to this concentration, and survival rate is lower than 50%, and even dehydration is withered dead; Therefore, comparatively speaking, it is the proper explant of high osmotic treatment that squamous subculture is more than or equal to 50d test-tube plantlet stem section.Specifically in table 1.
Table 1: different subculture number of days stem section is to the reaction of difference infiltration process
The screening of 2.2 suitable high osmotic treatment concentration
The different cultivars test-tube plantlet stem section of squamous subculture 50d, after variable concentrations PEG 8000 process 30d, result shows: under 200g/L, 300g/L concentration for the treatment of, the kind that only drought resistance is strong has a small amount of root system to generate, in drought resistance, difference kind all generate without root system, and all there are a large amount of dehydration phenomena of mortality of stem section in individual process, are not suitable for as screening concentration; Under low concentration 50g/L treatment conditions, drought resistance is strong, in kind number of relatively taking root distinguish not too large, be unfavorable for that Sorting Materials is identified; And under 100g/L concentration for the treatment of, the material that drought resistance is different can be distinguished preferably according to its number of relatively taking root, compare the suitable concentration of comparatively high osmotic treatment.In table 2.
Table 2: different disposal concentration to be taken root several impacts relatively on each kind
The performance of 2.3 different drought resistance kinds under lab simulation drought treatment conditions
Kind squamous subculture 50d test-tube plantlet stem sections different for drought resistance being inoculated in contrast (normal 1/2MS medium) and height oozes on qualification medium (100g/L), be embodied in: in control medium, different cultivars is due to genotype difference, Root-ing length is also different, many and the root of No. 1, peaceful purple potato, Xushen21 well root of hair number is long also long, but the stronger kind root of hair number of drought resistance few; And under high osmotic treatment condition, because stress effect is larger, root is long all shorter, but drought resistance is that root of hair number is many compared with strong Performance of cultivar, the less not even root of hair of kind root of hair number that drought resistance is poor, the kind that drought resistance is strong number system number of relatively taking root all is greater than 1.0, and the medium kind of drought resistance takes root number system number relatively between 0.5-1.0, and the kind of drought resistance difference is then less than or equal to 0.5. (specifically seeing the following form 3)
Table 3: relatively the take root investigation such as number of different drought resistance kind under high osmotic treatment condition
The 2.4 3 parts of test materials choosing unknown drought resistance grade carry out the drought-enduring qualification checking of the auxiliary drought-enduring qualification in indoor and field
Choose No. 6, business's potato, river potato 101, Hainan spends three parts of new materials to ooze qualification and field drought resisting qualification checking respectively by 100g/L PEG 8000 is high, result shows (see table 4): the result uniformity through indoor auxiliary Preliminary Identification material and field test is better, the method is suitable for the Preliminary screening of lot of materials, eliminate the poor material of drought resistance early, be convenient to the meticulous qualification for many years of drought resistance good a few materials multiple spot, increase work efficiency, reduce working strength.
The drought-enduring qualification of the drought-enduring assistant identification in indoor and field of table 4 unknown drought resistance material
Claims (2)
1. the method for an indoor assistant identification sweet potato drought resistance, be explant by getting the sweet potato test-tube plantlet of squamous subculture >=50d, cut single segmental, be inoculated in height and ooze qualification medium, inoculation 30d " Invest, Then Investigate " is taken root number relatively, calculate number system number of relatively taking root, according to the drought resistance classification of several coefficient decision kind of relatively taking root; Specifically carry out as follows:
(1) configure 1/2MS solid culture medium: MS minimal medium macroelement and trace element reduce by half, add sucrose 30g/L, Phytagel 5g/L, before sterilizing, adjust PH 5.7; Being distributed into diameter after sterilizing is 90mm culture dish, cools stand-by;
(2) configure and highly ooze qualification medium: first, be 100g/L according to W/V, aseptically, PEG 8000 be dissolved in through high-temperature sterilization but do not add the 1/2MS liquid nutrient medium of agar powder, then the 1/2MS liquid nutrient medium containing PEG 8000 is incorporated with the culture dish of 1/2MS solid culture medium, balance 24h, then removes liquid nutrient medium remaining above solid culture medium, for subsequent use; Described is 1:1.5 containing the 1/2MS liquid nutrient medium of PEG 8000 and the V/V ratio of 1/2MS solid culture medium;
(3) stem section inoculation: the different cultivars test-tube plantlet material choosing squamous subculture >=50d, cuts single segmental, is inoculated in height and oozes qualification medium culture;
(4) number system number of relatively taking root is investigated: after inoculation 30d, investigate the number of taking root of each process material and corresponding contrast, calculate number system number of relatively taking root;
(5) relatively take root the kind of number system number >=1.0, drought resistance is strong; 0.5 < is taken root the kind of number system number < 1.0 relatively, and drought resistance is medium; The kind of number system of relatively taking root number≤0.5, drought resistance is poor.
2. the method for indoor assistant identification sweet potato drought resistance according to claim 1, described number system number=high osmotic treatment kind of relatively taking root number/check variety of taking root is taken root number.
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| CN105010127A (en) * | 2015-07-06 | 2015-11-04 | 山东省农业科学院作物研究所 | Drought-resistant sweet potato variety rapid breeding method |
| CN105145350A (en) * | 2015-07-22 | 2015-12-16 | 青岛农业大学 | Preparation method and application of PEG culture medium with solid-liquid two phases |
| CN105660213A (en) * | 2015-08-27 | 2016-06-15 | 安徽科技学院 | Method for identifying drought tolerance of soybean germination phase by utilizing PEG-6000 |
| CN106538388A (en) * | 2016-11-07 | 2017-03-29 | 中国农业科学院郑州果树研究所 | The authentication method of drought-enduring Actinidia under a kind of conditions of tissue culture |
| CN107046995A (en) * | 2017-04-06 | 2017-08-18 | 江苏徐淮地区徐州农业科学研究所 | A kind of drought-enduring authentication method in sweet potato field |
| CN110495390A (en) * | 2019-07-17 | 2019-11-26 | 广东省农业科学院作物研究所 | A method of identification sweet potato drought tolerance |
| CN110786237A (en) * | 2019-05-09 | 2020-02-14 | 山东农业大学 | Method for improving salt tolerance of chrysanthemum through in vitro culture |
| CN110786236A (en) * | 2019-05-09 | 2020-02-14 | 山东农业大学 | Method for improving drought resistance of chrysanthemum by PEG (polyethylene glycol) in-vitro culture |
| CN112970456A (en) * | 2021-02-05 | 2021-06-18 | 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) | Indoor auxiliary identification method for salt tolerance of sweet potatoes |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105145350A (en) * | 2015-07-22 | 2015-12-16 | 青岛农业大学 | Preparation method and application of PEG culture medium with solid-liquid two phases |
| CN105660213A (en) * | 2015-08-27 | 2016-06-15 | 安徽科技学院 | Method for identifying drought tolerance of soybean germination phase by utilizing PEG-6000 |
| CN106538388A (en) * | 2016-11-07 | 2017-03-29 | 中国农业科学院郑州果树研究所 | The authentication method of drought-enduring Actinidia under a kind of conditions of tissue culture |
| CN106538388B (en) * | 2016-11-07 | 2019-05-07 | 中国农业科学院郑州果树研究所 | The identification method of drought-enduring Actinidia under a kind of conditions of tissue culture |
| CN107046995A (en) * | 2017-04-06 | 2017-08-18 | 江苏徐淮地区徐州农业科学研究所 | A kind of drought-enduring authentication method in sweet potato field |
| CN110786237A (en) * | 2019-05-09 | 2020-02-14 | 山东农业大学 | Method for improving salt tolerance of chrysanthemum through in vitro culture |
| CN110786236A (en) * | 2019-05-09 | 2020-02-14 | 山东农业大学 | Method for improving drought resistance of chrysanthemum by PEG (polyethylene glycol) in-vitro culture |
| CN110495390A (en) * | 2019-07-17 | 2019-11-26 | 广东省农业科学院作物研究所 | A method of identification sweet potato drought tolerance |
| CN112970456A (en) * | 2021-02-05 | 2021-06-18 | 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) | Indoor auxiliary identification method for salt tolerance of sweet potatoes |
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