CN103718698B - Method for distinguishing salt tolerance of wheat in germination period - Google Patents
Method for distinguishing salt tolerance of wheat in germination period Download PDFInfo
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Abstract
The invention discloses a method for distinguishing the salt tolerance of wheat in the germination period, which belongs to the field of agriculture. The method comprises the following steps: treating wheat seeds by using artificially prepared seawater, respectively measuring the relative germination rate in 40% artificial seawater, the relative germination rate in 65% artificial seawater, the relative germination potential in 40% artificial seawater, the relative germination potential in 65% artificial seawater and the relative wheat bud length in 40% artificial seawater by taking the ratios of a treated group to a reference group as the salt tolerance indexes, calculating the salt tolerance index of a wheat variety by using a main component analysis method, and grading and evaluating the salt tolerance in the germination period according to the values of the salt tolerance indexes. The method is approximate to the practical production in coastal mud flat agriculture, and the influence of genetic background on the researched properties is eliminated. The method is simple and convenient, the salt tolerance of the wheat in the germination period is comprehensively reflected, and the method can be rapidly and effectively applied to exploration of the salt-tolerance wheat seed quality and breeding of novel varieties.
Description
one, technical field
The present invention designs a kind of authentication method of a kind of wheat malt phase salt resistance newly, belongs to agriculture field.
two, background technology
The soil salinization is one of main abiotic stress factor affecting crop yield in the world, and the saline land of China accounts for 20% of arable area, constitutes serious threat to agricultural production.In numerous modes that saline land utilizes, Screening to use salt-tolerant plant new varieties are one of most economical effective methods in improvement saline land.
Wheat is the important cereal crops of China, and the research evaluated its Tolerant salt is more deep, but majority is (as NaCl and Na with single salt (as NaCl) or two salt
2cO
3) analog salt coerces, the correlative study closer to the seawater stress of Coastal beach practical application does not then almost have.The wheat malt phase refers to that intraseminal embryo starts restoration ecosystem and forms the process of seedling under the condition that environment is applicable.Bud phase salt tolerant is the prerequisite characteristic of salty area wheat breed, is the basis ensureing that wheat is emerged.Existing result of study shows, salt stress is different from the growth effect mechanism after sprouting to seed germination, and therefore, wheat malt phase Tolerant salt should carry out overall merit in conjunction with the elongation ability of bud after the sprouting ability of seed and sprouting.But this single index of statistics percentage of seedgermination after mostly only adopting single salt (NaCl) to process in current wheat malt phase Tolerant salt, comparatively comprehensively cannot evaluate wheat malt phase salt resistance.
three, summary of the invention
Technical problem
The object of the invention is to the deficiency for existing in existing wheat malt phase Tolerant salt, a kind of wheat malt phase Salt-Tolerance Identification method of new wide adaptability is provided, not only make qualification result more reliable, and easy being easy to of method is grasped.
Technical scheme
A kind of wheat malt phase Salt-Tolerance Identification evaluation method, comprises the following steps:
(1) mensuration of wheat germination rate and germination vigor
According to every premium on currency containing 26.726 g NaCl, 2.26 g MgCl
2, 3.248 g MgSO
4, 1.153 g CaCl
2, 0.198 g NaHCO
3artificial seawater is configured with 0.721 g KCl, according to volume by artificial seawater solution dilution to 40% and 65% of original content, with deionized water compare, 40% and 65% artificial seawater process wheat seed, be placed in illumination box 25 DEG C of constant temperature, illumination 16h germinates, the germination rate of one week " Invest, Then Investigate " control group and processed group, and calculate wheat samples 40% artificial seawater relative germination rate to be measured, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate:
Potentiality of seed (%)=(M
1/ M) × 100%
Percentage of seedgermination (%)=(M
2/ M) × 100%
In formula: M
1---the normal germination grain number in germination vigor number of days;
M
2---all normal germination grain numbers;
M---for planting experimentally seed number
Relative germination rate (%)=process germination rate/contrast germination rate × 100%
Relative germination rate (%)=process germination vigor/contrast germination vigor × 100%
(2) mensuration that bud phase salt stress process bud is long
Said method is adopted to configure 40% artificial seawater, compare and 40% artificial seawater process wheat seed by deionized water, be placed in illumination box 25 DEG C of constant temperature, illumination 16h germinates, within 5 days, measure control group afterwards and processed group bud long, and it is long to calculate the relative bud of wheat samples 40% artificial seawater to be measured:
Relative bud long (%)=process bud length/contrast bud long × 100%
(3) wheat malt phase salt resistance comprehensive identification
Measure 40% artificial seawater relative germination rate respectively according to above method, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate are long with the relative bud of 40% artificial seawater, be numbered in order: A, B, C, D, E; Calculate wheat samples salt tolerance index to be measured:
Salt tolerance index X=A*0.251189+B*0.261645+C*0.248657+D*0.256304+E*0.198353;
(4) wheat malt phase salt resistance grade scale:
| Rank | Salt resistance | Salt tolerance index (%) |
| 1 | High resistance to | ≥80.00 |
| 2 | Salt tolerant | 60.01-80.00 |
| 3 | In resistance to | 40.01-60.00 |
| 4 | Responsive | 20.01-40.00 |
| 5 | High sense | 0-20.00 |
Beneficial effect
The present invention utilizes the principal component analysis (PCA) in Chemical Measurement to analyze, can discrete data normalization process, significantly to evaluate the similitude of sample.Correlation matrix is listed according to correlation coefficient, obtain characteristic root and corresponding eigen vector thereof, for excretion is less and disturb larger factor, improve and analyze accurate rate, 1 larger characteristic root is chosen from 5 characteristic roots, the contribution rate of principal component is 77.02%, is enough to the variation tendency that these data are described, meets the requirement of principal component analysis.
According to the scoring systems matrix of principal component, the score formula of each factor can be obtained accordingly: salt tolerance index X=A*0.251189+B*0.261645+C*0.248657+D*0.256304+E*0.198353.
The method overcome index in traditional authentication method many but do not concentrate, comprehensive difference, identification result is inaccurate, the shortcomings such as salt tolerant prediction can not be carried out, and adopt the Relative ratio method of index can reflect the extent of injury of plant more objectively, eliminate the ecology existed between kind and grow difference, therefore the method well solves the more difficult problem of wheat malt phase salt resistance ability overall merit.This method is systemic, comprehensive by force, method is easy and simple to handle, wide adaptability, be easy to grasp, the selection of salt tolerance of whole system is all under controlled condition, make the selection result more reliable, indoor limited space can be utilized to carry out the Preliminary screening of lot of materials, accelerate wheat plant salt tolerant breeding process.
Innovation of the present invention have following some:
The first, with the Seawater Treatment plant of manually preparing, actual closer to Coastal beach agricultural production;
The second, using the relative ratio of the trait phenotypes value after salt stress process and the phenotypic value under collating condition as salt-tolerance index, eliminate phenotypic difference caused by genetic background between wheat germplasm;
3rd, adopt principal component analysis (PCA) to calculate the salt tolerant efficiency of each kind, improve the accuracy that salt tolerant is evaluated;
4th, according to bud phase salt tolerance index, overall merit is carried out to salt tolerance of wheat, and divided rank.
four, accompanying drawing explanation
Fig. 1 is wheat malt phase screening schematic diagram;
five, embodiment
Embodiment:
To participate in the experiment material: the domestic landrace of preserving from Jiangsu Province Agriculture Science Institute wheat variety resources project team, improved variety, innovation strain and external introduced variety, have chosen all kinds of Wheat Germplasm Resources 302 parts.
The qualification of 1 wheat germination rate and germination vigor
According to every premium on currency containing 26.726 g NaCl, 2.26 g MgCl
2, 3.248 g MgSO
4, 1.153 g CaCl
2, 0.198 g NaHCO
3artificial seawater is configured with 0.721 g KCl.Wheat seed soaks 0.5 h through the liquor natrii hypochloritis of 10%, deionized water is cleaned, blot surface moisture after, being positioned over the diameter being covered with double-layer filter paper is in the culture dish of 9 cm.According to volume by artificial seawater solution dilution to 40% and 65% of original content, with deionized water (contrast), 40% and 65% artificial seawater process wheat seed, each process 50 seeds, 3 repetitions.Be placed in illumination box to germinate (25 DEG C of constant temperature, illumination 16h), the germination rate of one week " Invest, Then Investigate " control group and processed group, and calculate wheat samples relative germination rate to be measured and germination vigor.
Potentiality of seed (%)=(M
1/ M) × 100%
Percentage of seedgermination (%)=(M
2/ M) × 100%
In formula: M
1---the normal germination grain number in germination vigor number of days;
M
2---all normal germination grain numbers;
M---for planting experimentally seed number
Relative germination rate (%)=process germination rate/contrast germination rate × 100%
Relative germination rate (%)=process germination vigor/contrast germination vigor × 100%
The impact that the salt stress process of 2 bud phases is long on bud
Said method is adopted to configure 40% artificial seawater, with deionized water (contrast) and 40% artificial seawater process wheat seed, each process 50 seeds, 3 repetitions.Be placed in illumination box and germinate (25 DEG C of constant temperature, illumination 16h).Within 5 days, measure control group afterwards and processed group bud long, and it is long to calculate the relative bud of wheat samples to be measured.
Relative bud long (%)=process bud length/contrast bud long × 100%
3 wheat malt phase salt resistance comprehensive identification methods
Random selecting 302 wheat breeds measure 40% artificial seawater relative germination rate respectively according to above method, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate are long with the relative bud of 40% artificial seawater, are numbered in order: A, B, C, D, E;
The principal component analysis (PCA) in Chemical Measurement is utilized to analyze, can discrete data normalization process, significantly to evaluate the similitude of sample.Correlation matrix is listed according to correlation coefficient, obtain characteristic root and corresponding eigen vector thereof, for excretion is less and disturb larger factor, improve and analyze accurate rate, 1 larger characteristic root is chosen from 5 characteristic roots, the contribution rate of principal component is 77.02%, is enough to the variation tendency that these data are described, meets the requirement of principal component analysis.
According to the scoring systems matrix of principal component, the score formula of each factor can be obtained accordingly: salt tolerance index X=A*0.251189+B*0.261645+C*0.248657+D*0.256304+E*0.198353.Calculate wheat samples salt tolerance index to be measured.
Wheat malt phase salt resistance grade scale:
| Rank | Salt resistance | Salt tolerance index (%) |
| 1 | High resistance to | ≥80.00 |
| 2 | Salt tolerant | 60.01-80.00 |
| 3 | In resistance to | 40.01-60.00 |
| 4 | Responsive | 20.01-40.00 |
| 5 | High sense | 0-20.00 |
Cultivate according to aforesaid way, determine 5 salt-tolerance index of wheat malt phase, basic statistics component analysis (table 1) has been carried out to its phenotypic data, and carried out the classification and valuation (table 2) of salt resistance according to the bud phase salt resistance of this technology to these materials.40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate are long with the relative bud of 40% artificial seawater
The basic statistics component analysis of table 1 wheat malt phase Salt Tolerance Index
| 40% artificial seawater relative germination rate | 65% artificial seawater relative germination rate | 40% artificial seawater relative germination rate | 65% artificial seawater relative germination rate | The relative bud of 40% artificial seawater is long | |
| Mean value | 82.22% | 54.00% | 78.08% | 42.01% | 27.05% |
| Luffing | 18.22%~99.98% | 0.00% ~ 98.06% | 0.00%~99.91% | 0.00%~98.94% | 2.67% ~ 47.94% |
| Standard deviation | 14.03% | 26.95% | 19.71% | 29.07% | 9.53% |
| The coefficient of variation | 0.17 | 0.50 | 0.25 | 0.69 | 0.35 |
The bud phase Salt-Tolerance Identification result of table 2 302 parts of Wheat Germplasm Resources
| Salt tolerant grade | Kind number | Percentage (%) |
| 1 | 114 | 37.75% |
| 2 | 104 | 34.44% |
| 3 | 52 | 17.22% |
| 4 | 27 | 8.94% |
| 5 | 5 | 1.66% |
| Add up to | 302 | 100% |
As shown in Table 1 and Table 2, the present embodiment has carried out bud phase Salt-Tolerance Identification to 302 parts of Wheat Germplasm Resources, finds that all material five salt-tolerance index differences of participating in the experiment are obvious, is suitable for the overall merit of wheat malt phase salt resistance.In 302 parts of Wheat Germplasm Resources of participating in the experiment, to identify in 1 grade high Salt-tolerant Wheat kind matter, 114 parts, 2 grades salt tolerants 104 parts, 3 grades resistance to 52 parts, 4 grades sensitivities 27 parts and 5 grades high responsive 5 parts altogether, account for 37.75% of wheat germplasm of participating in the experiment, 34.44%, 17.22%, 8.94%, 1.66% respectively.This and material of participating in the experiment are mainly derived from the landrace in jiangsu coast county (city) and external introduced variety mostly to be drought resistance and salt tolerance kind consistent, further illustrate the bud phase salt resistance that this technology can be used for identifying Wheat Germplasm Resources.
Innovation of the present invention have following some:
1. with the Seawater Treatment plant of manually preparing, actual closer to Coastal beach agricultural production;
2., using the relative ratio of the trait phenotypes value after salt stress process and the phenotypic value under collating condition as salt-tolerance index, eliminate phenotypic difference caused by genetic background between wheat germplasm;
3. adopt principal component analysis (PCA) to calculate the salt tolerant efficiency of each kind, improve the accuracy that salt tolerant is evaluated;
4. according to bud phase salt tolerance index, overall merit has been carried out to salt tolerance of wheat, and divided rank.
Claims (1)
1. an authentication method for wheat malt phase salt resistance, comprises the following steps:
(1) mensuration of wheat germination rate and germination vigor
According to every premium on currency containing 26.726 g NaCl, 2.26 g MgCl
2, 3.248 g MgSO
4, 1.153 g CaCl
2, 0.198 g NaHCO
3artificial seawater is configured with 0.721 g KCl, according to volume by artificial seawater solution dilution to 40% and 65% of original content, with deionized water compare, 40% and 65% artificial seawater process wheat seed, be placed in illumination box 25 DEG C of constant temperature, illumination 16h germinates, the germination rate of one week " Invest, Then Investigate " control group and processed group, and calculate wheat samples 40% artificial seawater relative germination rate to be measured, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate:
Potentiality of seed (%)=(M
1/ M) × 100%
Percentage of seedgermination (%)=(M
2/ M) × 100%
In formula: M
1---the normal germination grain number in germination vigor number of days;
M
2---all normal germination grain numbers;
M---for planting experimentally seed number
Relative germination rate (%)=process germination rate/contrast germination rate × 100%
Relative germination rate (%)=process germination vigor/contrast germination vigor × 100%;
(2) mensuration that bud phase salt stress process bud is long
Said method is adopted to configure 40% artificial seawater, compare and 40% artificial seawater process wheat seed by deionized water, be placed in illumination box 25 DEG C of constant temperature, illumination 16h germinates, within 5 days, measure control group afterwards and processed group bud long, and it is long to calculate the relative bud of wheat samples 40% artificial seawater to be measured:
Relative bud long (%)=process bud length/contrast bud long × 100%
(3) wheat malt phase salt resistance comprehensive identification
Measure 40% artificial seawater relative germination rate respectively according to above method, 65% artificial seawater relative germination rate, 40% artificial seawater relative germination rate, 65% artificial seawater relative germination rate are long with the relative bud of 40% artificial seawater, be numbered in order: A, B, C, D, E; Calculate wheat samples salt tolerance index to be measured:
Salt tolerance index X=A*0.251189+B*0.261645+C*0.248657+D*0.256304+E*0.198353;
(4) wheat malt phase salt resistance grade scale:
。
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| CN104255119B (en) * | 2014-09-29 | 2016-02-24 | 甘肃农业大学 | Different Nutrient Elements of feeding is improving the application method in salt tolerance of corn |
| CN105612851B (en) * | 2014-11-04 | 2018-03-13 | 中国农业大学 | One kind carries out Evaluation of Salt Tolerance or screening Salt-tolerant Wheat method based on potassium current to wheat |
| CN104472059B (en) * | 2014-12-26 | 2016-06-01 | 江西省农业科学院 | Method for identifying salt tolerance of asparagus |
| CN105359856A (en) * | 2015-11-11 | 2016-03-02 | 中国科学院遗传与发育生物学研究所 | Method and device for batch identification of wheat salt tolerance |
| CN106359096A (en) * | 2016-09-06 | 2017-02-01 | 中国农业科学院郑州果树研究所 | Authenticating method for salt-tolerant kiwi fruit germplasm resource |
| CN106385872B (en) * | 2016-09-07 | 2020-01-17 | 河北大学 | Method for evaluating salt tolerance of seeds of herbaceous local cover plant |
| CN106416864B (en) * | 2016-09-30 | 2020-02-21 | 江苏丘陵地区镇江农业科学研究所 | Rapid identification method for salt tolerance of rice in seedling stage |
| CN106576966A (en) * | 2016-12-13 | 2017-04-26 | 山西省农业科学院高粱研究所 | Method of identifying salt tolerance of grain sorghum |
| CN107172907A (en) * | 2017-03-09 | 2017-09-19 | 贵州省烟草科学研究院 | A kind of seed plant sprouting stage Salt-Tolerance Identification method |
| CN108207170A (en) * | 2018-01-31 | 2018-06-29 | 江苏沿海地区农业科学研究所 | A kind of integrated evaluating method of garlic sprouting stage salt tolerance |
| CN110121978B (en) * | 2019-06-04 | 2021-04-13 | 青岛农业大学 | Method for identifying salt tolerance of different wheat varieties in germination period |
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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 |
| CN103238396B (en) * | 2013-05-28 | 2015-04-29 | 甘肃农业大学 | Screening method of salt resisting germplasms of corn |
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