CN104255449B - Indoor auxiliary sweet potato drought tolerance identification method - Google Patents
Indoor auxiliary sweet potato drought tolerance identification method Download PDFInfo
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- CN104255449B CN104255449B CN201410439300.XA CN201410439300A CN104255449B CN 104255449 B CN104255449 B CN 104255449B CN 201410439300 A CN201410439300 A CN 201410439300A CN 104255449 B CN104255449 B CN 104255449B
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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 Rhizoma Dioscoreae esculentae drought tolerance authentication method is and in particular to a kind of interior auxiliary identifies Rhizoma Dioscoreae esculentae drought tolerance
Method, belongs to Rhizoma Dioscoreae esculentae drought tolerance identification technology field.
Background technology
The purpose of Rhizoma Dioscoreae esculentae drought tolerance identification mainly screens the kind of suitable drought-hit area cultivation from the sweet potato variety of incubation
With drought-enduring, the special new varieties of high-quality, high yield the selection-breeding of promotion.Rhizoma Dioscoreae esculentae is integrated as novel energy raw material and rapidoprint
Diversification crop, for current anxiety of ploughing, for accomplishing " not striving grain with people, do not strive ground with grain ", and Rhizoma Dioscoreae esculentae is a kind of adaptation
Property and the stronger cereal crops of resistance, the marginal land such as inland is saline and alkaline, arid and hilly and mountainous land are that following Sweet Potato Industry is expanded
New world.For promoting screening, the identification of the materials such as the drought-enduring resource of Rhizoma Dioscoreae esculentae, forefathers are processed by rain cover simulating drought, carry out
Rhizoma Dioscoreae esculentae drought tolerance is identified, but this is required for spending a large amount of manpower and materials, and workload is very big, and because processing region has
Limit is it is difficult to meet the primary dcreening operation of lot of materials, identification.And coerced by research and control treatment under the conditions of, oxidize-resistant physiology index
Deng the relation of change and the drought-enduring dependency of Rhizoma Dioscoreae esculentae, due to being difficult to directly judge Rhizoma Dioscoreae esculentae drought tolerance with minority physical signs and raw
The detection of reason index, also ratio is larger for workload, and the many factors such as weather are likely to have influence on accuracy and the repetition of its detection
Property, thus impact is to the judgement to drought tolerance result.In a word, drought-enduring determination rates lowly significantly impact drought-enduring Rhizoma Dioscoreae esculentae material
Screening and drought-enduring breeding of new variety process.
Content of the invention
The invention aims to overcoming above-mentioned existing methods not enough, provide a kind of interior auxiliary identification Rhizoma Dioscoreae esculentae drought-enduring
Property method, the method has quick, efficient, is not subject to other environmental influences extraneous it is ensured that expert evidence local environment bar
Part is consistent, can actual response material drought tolerance.
The present invention is to be realized with following technical scheme: the method that a kind of interior auxiliary identifies Rhizoma Dioscoreae esculentae drought tolerance, by taking
The Rhizoma Dioscoreae esculentae test tube seedling of successive transfer culture >=50d is explant, cuts single segmental, is inoculated in hypertonic identification culture medium and (according to w/v is
100g/l, aseptically, by Macrogol 8000 be dissolved in 1/2ms fluid medium (through high temperature sterilize but be not added with fine jade
Cosmetics), then the 1/2ms fluid medium containing Macrogol 8000 is 1:1.5 according to v/v, is distributed into 1/2ms solid culture
Base culture dish, balances 24h, then removes fluid medium remaining above solid medium, the 1/2ms solid being configured so that
Culture medium is identification culture medium), the relative number of taking root of investigation after inoculation 30d, calculate number system number of relatively taking root, according to number system of relatively taking root
Number judges the drought tolerance classification of kind.Specifically carry out as follows:
(1) configuration of 1/2ms solid medium: ms minimal medium is a large amount of, trace element halves, adds sucrose 30g/l,
Phytagel 5g/l, adjusts ph 5.7 before sterilizing；It is distributed into a diameter of 90mm culture dish, cooling is stand-by after sterilizing.
(2) configure hypertonic identification culture medium: first, be 100g/l according to w/v, aseptically, by Polyethylene Glycol
8000 are dissolved in through high temperature sterilize but are not added with the 1/2ms fluid medium of agar powder, then by 1/ containing Macrogol 8000
2ms fluid medium is incorporated with the culture dish of 1/2ms solid medium, balances 24h, then will be remaining above solid medium
Fluid medium is removed, standby；The described 1/2ms fluid medium containing Macrogol 8000 and 1/2ms solid medium
V/v is than for 1:1.5；
(3) stem section inoculation: choose successive transfer culture >=50d different cultivars test tube seedling material, cut single segmental, be inoculated in hypertonic
Identification culture medium culturing；
(4) number system number of relatively taking root is investigated: after inoculation 30d, investigation is each to process material and the corresponding number of taking root compareing, and calculates
Relative number system number of taking root；Number system number=high osmotic treatment kind of relatively taking root number/check variety of taking root is taken root number；
(5) kind of relative number system number >=1.0 of taking root, drought tolerance is strong；The kind of the relative number system number ＜ 1.0 of taking root of 0.5 ＜,
Drought tolerance is medium；The kind of relative number system number≤0.5 of taking root, drought tolerance is poor.
The invention has the advantage that the method is workable, simple, efficient quick, it is not subject to seasonal restrictions, and can
The exclusion impact to drought-enduring qualification result for the external environmental condition difference, is a kind of having of interior auxiliary primary dcreening operation drought tolerance sweet potato variety
Efficacious prescriptions method.
Specific embodiment
Further illustrate the method for the present invention and effect with reference to embodiment, but do not limit the scope of the invention.
Embodiment 1,
1. test material and method
1.1 test materials: test selected materials are through Xinjiang drought-enduring identification base, are accredited as drought tolerance in continuous 2 years
By force, 6 parts of materials in, poor.Wherein:
Drought tolerance is strong: ten thousand purples 56, wide potato 87；
Drought tolerance is medium: Xushen21 well, Xu 20-1；
Drought tolerance is poor: peaceful Rhizoma Steudnerae Henryanae No. 1, day purple 0602.
1.2 test methods:
The configuration of 1/2ms solid medium: ms minimal medium a great number of elements and trace element halve, adds sucrose 30g/
L, phytagel 5g/l, adjusts ph 5.7 before sterilizing；It is distributed into a diameter of 90mm culture dish, cooling is stand-by after sterilizing.
The configuration of hypertonic identification culture medium: first, be 50g/l, 100g/l, 200g/l, 300g/l according to w/v, aseptic
Under the conditions of, Macrogol 8000 is dissolved in 1/2ms fluid medium (through high temperature sterilize but be not added with agar powder), then will contain
The 1/2ms fluid medium of Macrogol 8000 is 1:1.5 according to v/v, is distributed into 1/2ms solid medium culture dish, balance
Then fluid medium remaining above solid medium is removed by 24h, standby.
Stem section is inoculated: chooses successive transfer culture 30d, 40d, 50d, 60d different cultivars test tube seedling material, cuts single segmental, connect
Plant in hypertonic identification culture medium culturing.
1.3 investigation and data statisticss: after inoculation 30d, each number of taking root processing material and corresponding comparison of investigation, calculate relatively
Number system of taking root number, number system number=high osmotic treatment kind of relatively taking root number/check variety of taking root is taken root number.
2. result and analysis
The 2.1 different reactions to hyperosmotic stress for the successive transfer culture time test tube seedling
Choose the test tube seedling stem section of the day purple 0602 different successive transfer culture times of drought tolerance difference, be inoculated in variable concentrations hypertonic
Identification culture medium, under compared with Thief zone concentration, day, purple 0602 test tube shoot survival percent was 100% successive transfer culture test tube seedling material for screening
Material, result shows: under 50g/l concentration for the treatment of, the survival rate of different successive transfer culture time stem sections is up to 100；Dense with processing
The raising of degree, stem section survival rate is on a declining curve, or even all dead；Extend with stem section Subculture Time, and inside stem section
Fiber etc. is developed, and has higher anti-adversity ability, after the stem section of different successive transfer culture times is inoculated in 100g/l hypertonic culture medium,
The test tube seedling stem section survival rate of successive transfer culture 30d, 40d is less than 100%, after successive transfer culture is more than or equal to 50d, test tube seedling stem section
Survival rate to 100%；If hypertonic concentration improves further, test tube seedling stem section will be unable to adapt to this concentration, survives
Rate is less than 50%, or even the withered death of dehydration；Therefore, comparatively speaking, successive transfer culture is more than or equal to 50d test tube seedling stem section is height
Ooze the suitable explant of process.Specifically it is shown in Table 1.
Table 1: the reaction that different subculture natural law stem sections are processed to different infiltrations
The screening of 2.2 suitable high osmotic treatment concentration
The different cultivars test tube seedling stem section of successive transfer culture 50d, after variable concentrations Macrogol 8000 processes 30d, result
Show: under 200g/l, 300g/l concentration for the treatment of, only a strong kind of drought tolerance has a small amount of root system to generate, in drought tolerance, poor
Kind all no root system generate, and individual process in all have stem section a large amount of dehydration phenomena of mortality, be not suitable for as screening concentration；?
Under low concentration 50g/l treatment conditions, drought tolerance is strong, in kind number of relatively taking root distinguish less big, be unfavorable for Sorting Materials
Identification；And under 100g/l concentration for the treatment of, the different material of drought tolerance preferably can be distinguished according to its number of relatively taking root, compare
The suitable concentration of more high osmotic treatment.It is shown in Table 2.
Table 2: different disposal concentration relative several impacts of taking root to each kind
2.3 different drought tolerance kinds simulate the performance under non-irrigated treatment conditions indoors
By kind successive transfer culture 50d test tube seedling stem sections different for drought tolerance be inoculated in comparison (normal 1/2ms culture medium) and
In hypertonic identification culture medium (100g/l), it is embodied in: in control medium, different cultivars is different due to genotype, sends out
Root length is also different, and peaceful Rhizoma Steudnerae Henryanae 1, Xushen21 well root of hair number are many and root length is also long, but the stronger kind root of hair of drought tolerance
Number is simultaneously few；And under the conditions of high osmotic treatment, because stress effect is larger, root length is all shorter, but the stronger kind table of drought tolerance
It is now that root of hair number is many, the poor less not even root of hair of kind root of hair number of drought tolerance, the relative number system of taking root of the strong kind of drought tolerance
Number is all higher than 1.0, and the medium kind of drought tolerance takes root number system number between 0.5-1.0 relatively, and the kind of drought tolerance difference is then
Less than or equal to 0.5. (being specifically shown in table 3 below)
Table 3: the investigation such as number of relatively taking root under the conditions of high osmotic treatment for the different drought tolerance kinds
3 parts of test materials of the 2.4 unknown drought tolerance grades of selection carry out the drought-enduring identification of indoor auxiliary and the drought-enduring identification in field
Checking
Selection business's potato 6, river potato 101, Hainan spend three parts of new materials to pass through the hypertonic mirror of 100g/l Macrogol 8000 respectively
Fixed and field drought resisting identification verifies, result shows (being shown in Table 4): through the result of indoor auxiliary Preliminary Identification material and field test
Concordance is preferable, the preliminary screening of the suitable lot of materials of the method, eliminates the poor material of drought tolerance early, is easy to drought tolerance relatively
The fine identification for many years of good a few materials multiple spot, improves work efficiency, reduces working strength.
The interior drought-enduring auxiliary identification of the unknown drought tolerance material of table 4 and the drought-enduring identification in field
Claims (1)
1. the method that a kind of interior auxiliary identifies Rhizoma Dioscoreae esculentae drought tolerance, is explant by the Rhizoma Dioscoreae esculentae test tube seedling taking successive transfer culture >=50d
Body, cuts single segmental, is inoculated in hypertonic identification culture medium, and the relative number of taking root of investigation after inoculation 30d calculates number system of relatively taking root
Number, according to the drought tolerance classification of several coefficient decision kinds of relatively taking root；Specifically carry out as follows:
(1) configure 1/2ms solid medium: ms minimal medium a great number of elements and trace element halve, add sucrose 30g/l,
Phytagel 5g/l, adjusts ph 5.7 before sterilizing；It is distributed into a diameter of 90mm culture dish, cooling is stand-by after sterilizing；
(2) configure hypertonic identification culture medium: first, it is 100g/l according to w/v, aseptically, Macrogol 8000 is molten
Xie Yu through high temperature sterilize but is not added with the 1/2ms fluid medium of agar powder, then by the 1/2ms liquid containing Macrogol 8000
Culture medium is incorporated with the culture dish of 1/2ms solid medium, balances 24h, then by liquid training remaining above solid medium
Foster base is removed, standby；The described 1/2ms fluid medium containing Macrogol 8000 with the v/v ratio of 1/2ms solid medium is
1:1.5；
(3) stem section inoculation: choose the different cultivars test tube seedling material of successive transfer culture >=50d, cut single segmental, be inoculated in hypertonic mirror
Determine culture medium culturing；
(4) number system number of relatively taking root is investigated: after inoculation 30d, investigation is each to process material and the corresponding number of taking root compareing, and calculates relatively
Number system of taking root number；
(5) kind of relative number system number >=1.0 of taking root, drought tolerance is strong；The kind of the relative number system number ＜ 1.0 of taking root of 0.5 ＜, drought-enduring
Property is medium；The kind of relative number system number≤0.5 of taking root, drought tolerance is poor；
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 |
| CN105145350B (en) * | 2015-07-22 | 2017-09-15 | 青岛农业大学 | A kind of preparation method and application of solid-liquid two-phase PEG culture mediums |
| 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 |
| 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 |
| CN107046995B (en) * | 2017-04-06 | 2020-09-11 | 江苏徐淮地区徐州农业科学研究所 | Sweet potato field drought tolerance identification method |
| 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 |
| 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 |
| 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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| CA2656258A1 (en) * | 2006-06-20 | 2007-12-27 | Wageningen Universiteit | Use of plant chromatin remodeling genes for modulating plant architecture and growth |
| CN102498791A (en) * | 2011-10-13 | 2012-06-20 | 安徽省农业科学院水稻研究所 | Fast screening method of drought-tolerant rice |
| CN103004324A (en) * | 2012-12-24 | 2013-04-03 | 山东省花生研究所 | Method for quickly identifying and choosing drought-resistant peanuts indoors |
| CN103283457B (en) * | 2013-05-31 | 2015-05-27 | 云南农业大学 | Drought-resisting batata seedling culture method |
| CN103343139B (en) * | 2013-06-27 | 2014-06-11 | 杭州昂众生物科技有限公司 | Novel method for enhancing drought resistance of plants |
| CN103636494A (en) * | 2013-11-22 | 2014-03-19 | 简玉君 | Drought resistance identification method of virus-free potato test-tube seedling |
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