CN107172907A - A kind of seed plant sprouting stage Salt-Tolerance Identification method - Google Patents
A kind of seed plant sprouting stage Salt-Tolerance Identification method Download PDFInfo
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- CN107172907A CN107172907A CN201710138050.XA CN201710138050A CN107172907A CN 107172907 A CN107172907 A CN 107172907A CN 201710138050 A CN201710138050 A CN 201710138050A CN 107172907 A CN107172907 A CN 107172907A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
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- Soil Sciences (AREA)
- Environmental Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides a kind of seed plant sprouting stage Salt-Tolerance Identification method, the screening of configuration, seed including artificial seawater solution, the sterilization processing of seed, the osmotic adjustment of seed, the low temperature drying method of seed, the processing of seed salt stress, count germination percentage and calculate salt tolerance index, to being capable of salt resistant character under comprehensive assessment plant natural conditions.The artificial seawater solution collocation method that the present invention is used is relative to the existing artificial seawater in the identification of plant resistance to pressure at present, salt component is closer to nature seawater, and qualification result can the salt resistance ability of different cultivars compares between progress species in the case of unified metric unit and in species.
Description
Technical field
The invention belongs to plant physiology field, and in particular to a kind of seed plant sprouting stage Salt-Tolerance Identification method.
Background technology
The soil salinization is global environmental problem, is just existed always before occurring from the mankind and cultivation culture, but people
The farming operation of class significantly exacerbates this process.The arable land in the whole world about 20%, 50% irrigated land are by salinization of soil at present
Influence, Chinese saline and alkaline soil is up to 99,130,000 hm2, the land area that Agriculture, forestry And Animal Husbandry land used is endangered by salinization of soil has 36,300,000 hm2, wherein
Plough 9,210,000 hm2.Huang-Huai-Hai plain, loess plateau and inland area, Northwest, China are mainly distributed on, Northeast plain and coastal area also have
The salt-soda soil of large area.The salinity of soil middle and high concentration causes osmotic stress so that ionic equilibrium is broken in plant, causes oxygen
Change damage, the crops planted on arable land are often resulted in the significantly underproduction, or even total crop failure.In order to adapt to salination environment, plant
Internal portion has evolved a set of complicated genetic system so that plant can survive under a certain degree of salt stress environment
And produce offspring, to ensure that species continue.
Plant is to the reaction of Soil salinity stress and improves the research of its salt resistance ability and has had since human civilization just constantly quilt by oneself
Explore, but it should be founded from modern biology by system research and count.The index identified plant salt endurance is in international and national
Research is also more, and these authentication methods are divided into Direct Identification and the class of INDIRECTLY IDENTIFIED two.The bud of current authentication method more options plant
Phase, seedling stage are measured, and measurement index is percentage of seedgermination, germinating energy, survival rate of seedling or other physiological and biochemical indexs.At this
The salt species selected in a little researchs is different, has the single salt stress of selective chlorination sodium, also there is selection artificial seawater (artificial
Sea water, ASW) many salt stress environments of simulation.Even if the processing of mono-salt NaCl Stress is selected, but the concentration of salt solution of selection
Also there are a variety of results, processing environment has water planting and earth culture, and selection artificial seawater simulates the research of many salt stresses, salt species and concentration
Difference research difference is also larger.This can not be compared between resulting in qualification result, and it is more difficult to reuse.
The content of the invention
In view of this, the invention provides a kind of seed plant sprouting stage Salt-Tolerance Identification method, including artificial seawater solution
Configuration, the screening of seed, the sterilization processing of seed, the osmotic adjustment of seed, the low temperature drying method of seed, seed salt stress
Handle, count germination percentage and calculate salt tolerance index, to being capable of salt resistant character under comprehensive assessment plant natural conditions.The present invention
The artificial seawater solution collocation method of use more connects relative to the existing artificial seawater in the identification of plant resistance to pressure at present, salt component
It is bordering on nature seawater, the salt tolerant that qualification result can be between progress species in the case of unified metric unit with different cultivars in species
Ability compares.
The invention provides a kind of seed plant sprouting stage Salt-Tolerance Identification method, comprise the following steps：Configure artificial seawater
Solution, the screening of seed, the sterilization processing of seed, the osmotic adjustment of seed, the processing of seed salt stress, count germination percentage and calculate
Salt tolerance index.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the artificial seawater solution includes following
Component：
Sodium chloride (NaCl), sodium sulphate (Na2SO4), potassium chloride (KCl), sodium acid carbonate (NaHCO3), KBr (KBr),
Boric acid (H3BO3), sodium fluoride (NaF), magnesium chloride hexahydrate (MgCl2.6H2O), calcium chloride dihydrate (CaCl2.2H2O), six water chlorination
Strontium (SrCl2.6H2O), pure water (H2O)。
It is highly preferred that in the seed plant sprouting stage Salt-Tolerance Identification method of the present invention, the standard recipe of the artificial seawater
For：
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the screening technique of the seed is：Choose
Full grains, consistent, the ripe vegetable seeds that can be sprouted uniform in size, except mould impurity elimination.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the bactericidal treatments of the seed are
Choose sodium hypochlorite (NaClO) solution of 10% mass concentration, immersion seed 10 minutes, then pure water rinsing 3~5 times.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, seed osmotic adjustment method is selection 100
Potassium nitrate (the KNO of mM/l (mmol/L)3) solution, immersion seed 2 hours, then pure water rinsing 3~5 times.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the low temperature drying method of seed is will be wet
Profit seed is positioned over temperature setting for 25 degrees Celsius (DEG C), opens in the air dry oven of ventilation function 12 hours, takes off seed
Water is dried.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the seed salt stress processing method is
The culture dish of different capabilities is selected, seed is laid in culture dish bottom, the artificial seawater salting liquid of various concentrations is added, will train
Foster ware is placed in phjytotron or illumination box, and temperature is maintained at 25 degrees Celsius (DEG C), and light dark period is 12h/12h,
Air humidity is 75% or so, is placed 7~10 days.It is highly preferred that the concentration of the artificial seawater salting liquid of the various concentrations is
0.0th, 0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0 times of ASW.
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the method for described statistics germination percentage is
The seed handled by salt stress is taken out, number takes the number of chitting piece under each salinity, and the standard of germination is compared to the salt side of body
Compel before processing sub-states, bud exposes or seedling, and the statistics germination percentage is：
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the salt tolerance index IstFor：
Retain 2 decimals, then IstCodomain be [0.00,1.00].
Preferably, in seed plant sprouting stage Salt-Tolerance Identification method of the invention, the seed plant includes but is not limited to
Wheat, rape.
Brief description of the drawings
Fig. 1 is part Variety utilization qualification result figure (part thing (product) the kind salt tolerant in one embodiment of the present of invention
Property qualification result)；
Fig. 2 overall germination figure (rapes under different salinity stress for each material in one embodiment of the present of invention
Kind Qingyou 2 germination under various concentrations artificial seawater solution)；
Fig. 3 bud growing state figure (wheat breed west under different salinity for each material in one embodiment of the present of invention
The germination under various concentrations artificial seawater solution of agriculture 668).
Embodiment
The technical scheme of the Salt-Tolerance Identification of seed plant sprouting stage described in one embodiment of the invention includes：Artificial seawater is molten
The configuration of liquid, the early stage processing of seed, seed salt stress processing counts germination percentage and calculates salt tolerance index.
The configuration of artificial seawater：Described artificial seawater includes sodium chloride (NaCl), sodium sulphate (Na2SO4), potassium chloride
(KCl), sodium acid carbonate (NaHCO3), KBr (KBr), boric acid (H3BO3), sodium fluoride (NaF), magnesium chloride hexahydrate
(MgCl2.6H2O), calcium chloride dihydrate (CaCl2.2H2O), six water strontium chloride (SrCl2.6H2O), pure water (H2O) etc..Wherein concentration
Allocation ratio for each composition in 1.0 artificial seawaters (1.0ASW) sees attached list 1.
The artificial seawater salinity constituent of table 1 and ratio (concentration is 1 artificial seawater salinity, is designated as 1AWS)
Note：Other each concentration artificial seawater solutions, by the proportional dilution of volume
Each salt component in artificial seawater solution is proportionally reduced according to collocation method, be made into concentration for 0.0 (pure water),
0.1st, 0.2,0.3,0.4,0.5,0.6,0.7,0.8, the 0.9, salting liquid of 1.0 times of artificial seawater concentration, totally 11 concentration gradients.
The early stage processing of seed：Choose full grains, consistent plant maturation uniform in size can germination seed, gone except mould
It is miscellaneous, grain number more than 550, it is proposed that 1300.
The sterilization processing of seed, sodium hypochlorite (NaOCl) solution of selection 10%, immersion seed 10 minutes (min), then
Pure water rinsing 3~5 times；
The osmotic adjustment of seed：During because of harvest seed, seed length is on different plants, different parts, and its maturity can not
It is completely the same, in order to ensure that seed initial infiltration gesture is consistent, select the potassium nitrate (KNO with 100 mM/ls (mmol/L)3)
Solution seed soaking 2 hours (h), then clear water flushing 3-5 times, the surface of the seed moisture is blotted with blotting paper, it is desirable to ensure that seed is in
State before showing money or valuables one carries unintentionally.
The low temperature drying of seed：Seed after osmotic adjustment is in moisture state, is unfavorable for storing for a long time, while in this hair
Seed after bright middle osmotic adjustment can not such as be handled with salt stress in time, and seed, which will germinate, to be used, therefore need to be by seed again
Secondary dewater treatment.Processing method is that seed is positioned over into temperature setting is 25 degrees Celsius (DEG C), and the air blast for opening ventilation function is done
12 hours in dry case, dry seed dehydration.
Salt stress processing：The culture dish of appropriate specification size is selected, bottom is covered with 2 metafiltration paper.The specification size of culture dish
Selected according to seed grain number and granular size, it is desirable to which seed can be evenly laid out in culture dish bottom.
(n is not less than 50 to number access mesh n, it is proposed that n takes seed 100) after sterilization, osmotic adjustment processing, is placed in 1 training
Support in ware, need number to take the seed of 11 culture dishes.
Be separately added into 11 culture dishes concentration for 0.0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,
The salting liquid 20ml of 1.0 times of artificial seawater concentration.
Culture dish is placed in phjytotron or illumination box, temperature is maintained at 25 degrees Celsius (DEG C), brightness week
Phase is 12h/12h, and air humidity is 75% or so.
Place 10~15 days, note checking salting liquid in culture dish daily, it is ensured that filter paper is moistened.
Count germination percentage and calculate salt tolerance index
Germination percentage is counted, the seed handled by salt stress is taken out, number takes the number of chitting piece under each salinity, hair
The standard of bud is that bud exposes or seedling compared to salt stress before processing sub-states.Then under each salinity gradient chitting piece number
Mesh is designated as：a0.0、a0.1、a0.2、a0.3、a0.4、a0.5、a0.6、a0.7、 a0.8、a0.9、a1.0；
Selected seed germination percentage calculation formula is
Germination percentage (r) is decimally represented, retains two-decimal.
Calculated with this under each salinity, the germination percentage of seed：r0.0、r0.1、r0.2、r0.3、r0.4、r0.5、r0.6、 r0.7、
r0.8、r0.9、r1.0；
If percentage of seedgermination r is Dependent variable, salinity x is independent variable, in the same species kind in identical environment
Percentage of seedgermination r and salinity x Density Distribution are (parameter θ is the salt resistance ability of the kind)：
R=f (x, θ)
Then germination percentage is distributed as in salinity 0.0ASW-1ASW
Thus it is the numerical value determined by parameter θ, parameter θ ∝ F (x, θ) (x ∈ to know F (x, θ) (x ∈ (0.0,1.0))
(0.0,1.0)).Germination percentage is such as designated as I in salinity 0.0ASW-1ASW distributionsst(be referred to as salt tolerance index), then IstMirror can be used as
Determine an index of salt resistant character, while the property for being distributed function is understood, each salinity in the case where knowing 0.0~1.0 salinity gradient
Under germination percentage, then IstCan approximate evaluation be：
Retain 2 decimals, then IstCodomain be [0.00,1.00].
Test in triplicate, the test error between the repeatability and estimating test of checking test.
Further technical scheme is illustrated below by way of specific embodiment, it should be understood that be only this hair below
Bright exemplary illustration, is not intended to limit the invention scope of the claims.
The rape variety Salt-Tolerance Identification of embodiment 1
S1 configures each concentration artificial seawater solution according to subordinate list 1；
S2 chooses 1 rape material (Qingyou 2), number seeds about 1800.Handled according to following scheme：10% time
Sodium chlorate solution handle 10 minutes->Pure water rinsing 5 times->100mmol/L potassium nitrate solution soak seed 2 hours->Pure water is rushed
Wash 5 times->Air dry oven is dehydrated.
S3 chooses 100*100mm 11, square culture dish, and each culture dish places seed (Qin's oil after S2 step process
No. 2) number 50, compiled according to the artificial seawater concentration of addition as 0.0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,
0.9th, 1.0, each culture dish addition solution 20ml.
S4 repeats step S3 twice.
S5 by culture dish be placed in phjytotron (temperature is maintained at 25 degrees Celsius, and light dark period is 12 hours/12 hours,
Air humidity is 75% or so)
The germination of seed is counted after S615 days, 2 are the results are shown in Table.
S7 calculate rape variety Qingyou 2 three salt tolerance index qualification results be：0.39、0.41、0.40；
The wheat breed of embodiment 2 (is) Salt-Tolerance Identification
Method be the same as Example 1,3 wheat breeds of selection (are) material (9088, continuous 15Z24 are ground in western agriculture 668, Shandong), as a result
It is shown in Table 2：
The part thing (product) of table 2. plants Salt-Tolerance Identification result
Fig. 1 is that part thing (product) plants Salt-Tolerance Identification result, shows that, with the increase of artificial seawater solution concentration, example is planted
The germination percentage of species present under totality by trend, but thing (product) inter-species germination percentage has with artificial seawater solution change in concentration
Notable difference.The salting liquid of the bright single concentration of selection of the chart carries out vegetable seeds Salt-Tolerance Identification simultaneously, can be molten because of selection salt
The difference of liquid concentration, draws opposite result.Such as select the solution of 0.5 artificial seawater concentration, the germination of rape variety Qingyou 2
Relatively each wheat breed (being) of rate is all higher, shows that rape variety Qingyou 2 is better than to the adaptability of salt stress under the concentration
Wheat breed selected by example (is), but under 0.7 artificial seawater concentration, rape variety Qingyou 2 germination percentage is then less than example institute
The germination percentage of wheat breed (being) is selected, so as to draw opposite conclusion.
Fig. 2 is germination of the rape variety Qingyou 2 under various concentrations artificial seawater solution, in artificial seawater concentration
After 0.8, seed is without germination, and in the case where artificial seawater concentration is less than 0.5, and the germination percentage of rape seed exceedes
0.90。
Fig. 3 is germination of the western agriculture 668 of wheat breed under various concentrations artificial seawater solution, compared to the rape variety Qin
Oil 2, wheat breed has seed sprouting under each concentration artificial seawater solution, but sprout upgrowth situation is different, overall to cash
With the increase of artificial seawater concentration, germination percentage reduction, and sprout grows suppressed trend.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of seed plant sprouting stage Salt-Tolerance Identification method, comprises the following steps：Configure artificial seawater (artificial sea
Water, ASW) solution, the screening of seed, the sterilization processing of seed, the osmotic adjustment of seed, the low temperature drying method of seed, plant
Alite Stress treatment, counts germination percentage and calculates salt tolerance index.
2. according to the method described in claim 1, it is characterised in that the artificial seawater solution includes following components：
Sodium chloride (NaCl), sodium sulphate (Na2SO4), potassium chloride (KCl), sodium acid carbonate (NaHCO3), KBr (KBr), boric acid
(H3BO3), sodium fluoride (NaF), magnesium chloride hexahydrate (MgCl2.6H2O), calcium chloride dihydrate (CaCl2.2H2O), six water strontium chloride
(SrCl2.6H2O), pure water (H2O)。
3. method according to claim 2, it is characterised in that the 1 normal concentration artificial seawater (1.0ASW), which is formulated, is：
4. according to the method described in claim 1, it is characterised in that the earlier stage processing method of the seed is：Particle is chosen to satisfy
Full, consistent plant maturation uniform in size can germination seed, except mould impurity elimination.
5. according to the method described in claim 1, it is characterised in that the bactericidal treatments of the seed are 10% mass of selection
Sodium hypochlorite (NaClO) solution of concentration, immersion seed 10 minutes, then pure water rinsing 3~5 times.
6. according to the method described in claim 1, it is characterised in that the osmotic adjustment method of the seed is：Choose 100 mmoles
That/liter (mmol/L) potassium nitrate (KNO3) solution, immersion seed 2 hours, then pure water rinsing 3~5 times.
7. according to the method described in claim 1, it is characterised in that the low temperature drying method of the seed is：Seed will be moistened
Temperature setting is positioned over for 25 degrees Celsius (DEG C), opens in the air dry oven of ventilation function 12 hours, dries seed dehydration.
8. according to the method described in claim 1, it is characterised in that the seed salt stress processing method is selection different capabilities
Culture dish, seed is laid in culture dish bottom, the artificial seawater salting liquid of various concentrations is added, culture dish is placed in people
In work climatic chamber or illumination box, temperature is maintained at 25 degrees Celsius (DEG C), and light dark period is 12 hours/12 hours, and air is wet
Spend for 75% or so, placement 10~15 days.
9. according to the method described in claim 1, it is characterised in that the concentration of the artificial seawater salting liquid of the various concentrations is
0.0th, 0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8,0.9,1.0 times of ASW.
10. according to the method described in claim 1, it is characterised in that the method for described statistics germination percentage is that will be coerced through supersalt
The seed for compeling processing takes out, and number takes the number of chitting piece under each salinity, and the standard of germination is compared to salt stress before processing kind
Sub- state, bud exposes or seedling, and the statistics germination percentage is：
Preferably, the salt tolerance index is：
<mrow>
<msub>
<mi>I</mi>
<mrow>
<mi>s</mi>
<mi>t</mi>
</mrow>
</msub>
<mo>=</mo>
<mn>0.05</mn>
<mo>*</mo>
<mrow>
<mo>(</mo>
<msub>
<mi>r</mi>
<mn>0.0</mn>
</msub>
<mo>+</mo>
<msub>
<mi>r</mi>
<mn>1.0</mn>
</msub>
<mo>)</mo>
</mrow>
<mo>+</mo>
<mn>0.1</mn>
<mo>*</mo>
<munderover>
<mo>&Sigma;</mo>
<mrow>
<mi>i</mi>
<mo>=</mo>
<mn>0.1</mn>
</mrow>
<mn>0.9</mn>
</munderover>
<msub>
<mi>r</mi>
<mi>i</mi>
</msub>
</mrow>
Retain 2 decimals, then IstCodomain be [0.00,1.00]
Preferably, the seed plant includes but is not limited to wheat, rape.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110226382A (en) * | 2019-05-20 | 2019-09-13 | 湖北省农业科学院经济作物研究所 | Nature seawater coerces cotton seed germination phase Tolerant salt method |
| CN110274868A (en) * | 2019-06-22 | 2019-09-24 | 沧州市农林科学院 | A kind of formula of indoor crops salt tolerant Rapid identification |
| CN110583395A (en) * | 2019-09-11 | 2019-12-20 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Method for evaluating salt tolerance of japonica rice seeds under salt stress |
| CN111869368A (en) * | 2020-08-13 | 2020-11-03 | 天津农学院 | Rapid seedling stage evaluation method for salt tolerance of rape seeds |
| CN113030172A (en) * | 2021-03-31 | 2021-06-25 | 中国林业科学研究院 | Method for predicting salt tolerance of plant |
| CN114946319A (en) * | 2022-06-08 | 2022-08-30 | 中国农业科学院棉花研究所 | Method for evaluating drought resistance of upland cotton in germination period by using double-concentration PEG6000 |
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| CN102860159A (en) * | 2012-10-10 | 2013-01-09 | 江苏省农业科学院 | Salt-tolerance determining method for barley at seedling stage |
| CN103718698A (en) * | 2014-01-07 | 2014-04-16 | 江苏省农业科学院 | Method for distinguishing salt tolerance of wheat in germination period |
| CN104472059A (en) * | 2014-12-26 | 2015-04-01 | 江西省农业科学院 | Salt tolerance identification method for asparagus officinalis L. |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102860159A (en) * | 2012-10-10 | 2013-01-09 | 江苏省农业科学院 | Salt-tolerance determining method for barley at seedling stage |
| CN103718698A (en) * | 2014-01-07 | 2014-04-16 | 江苏省农业科学院 | Method for distinguishing salt tolerance of wheat in germination period |
| CN104472059A (en) * | 2014-12-26 | 2015-04-01 | 江西省农业科学院 | Salt tolerance identification method for asparagus officinalis L. |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110226382A (en) * | 2019-05-20 | 2019-09-13 | 湖北省农业科学院经济作物研究所 | Nature seawater coerces cotton seed germination phase Tolerant salt method |
| CN110274868A (en) * | 2019-06-22 | 2019-09-24 | 沧州市农林科学院 | A kind of formula of indoor crops salt tolerant Rapid identification |
| CN110583395A (en) * | 2019-09-11 | 2019-12-20 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Method for evaluating salt tolerance of japonica rice seeds under salt stress |
| CN111869368A (en) * | 2020-08-13 | 2020-11-03 | 天津农学院 | Rapid seedling stage evaluation method for salt tolerance of rape seeds |
| CN113030172A (en) * | 2021-03-31 | 2021-06-25 | 中国林业科学研究院 | Method for predicting salt tolerance of plant |
| CN114946319A (en) * | 2022-06-08 | 2022-08-30 | 中国农业科学院棉花研究所 | Method for evaluating drought resistance of upland cotton in germination period by using double-concentration PEG6000 |
| CN114946319B (en) * | 2022-06-08 | 2024-01-26 | 中国农业科学院棉花研究所 | Method for evaluating drought resistance of upland cotton in germination period by using double-concentration PEG6000 |
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