CN113940205A - Method for identifying salt tolerance of holly at seedling stage - Google Patents
Method for identifying salt tolerance of holly at seedling stage Download PDFInfo
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- CN113940205A CN113940205A CN202111220475.8A CN202111220475A CN113940205A CN 113940205 A CN113940205 A CN 113940205A CN 202111220475 A CN202111220475 A CN 202111220475A CN 113940205 A CN113940205 A CN 113940205A
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- leaves
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- 230000015784 hyperosmotic salinity response Effects 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 23
- 235000003332 Ilex aquifolium Nutrition 0.000 title claims abstract description 15
- 235000002296 Ilex sandwicensis Nutrition 0.000 title claims abstract description 15
- 235000002294 Ilex volkensiana Nutrition 0.000 title claims abstract description 15
- 241000209035 Ilex Species 0.000 claims abstract description 29
- 150000003839 salts Chemical class 0.000 claims abstract description 15
- 241001299553 Ilex chinensis Species 0.000 claims abstract description 12
- 235000003366 Ilex purpurea Nutrition 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 235000003325 Ilex Nutrition 0.000 claims description 11
- 239000008367 deionised water Substances 0.000 claims description 10
- 229910021641 deionized water Inorganic materials 0.000 claims description 10
- 241000196324 Embryophyta Species 0.000 claims description 7
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000012266 salt solution Substances 0.000 claims description 5
- 238000004080 punching Methods 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 241000238631 Hexapoda Species 0.000 claims description 2
- 241000607479 Yersinia pestis Species 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 210000003462 vein Anatomy 0.000 claims description 2
- 235000012431 wafers Nutrition 0.000 claims 2
- 238000011109 contamination Methods 0.000 claims 1
- 238000012216 screening Methods 0.000 abstract description 2
- 239000000463 material Substances 0.000 description 11
- 239000002689 soil Substances 0.000 description 5
- 235000002710 Ilex cornuta Nutrition 0.000 description 4
- 241001310146 Ilex cornuta Species 0.000 description 4
- 235000010326 Osmanthus heterophyllus Nutrition 0.000 description 4
- 239000003513 alkali Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 235000007297 Gaultheria procumbens Nutrition 0.000 description 2
- 241000736246 Pyrola Species 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- OSWPMRLSEDHDFF-UHFFFAOYSA-N methyl salicylate Chemical compound COC(=O)C1=CC=CC=C1O OSWPMRLSEDHDFF-UHFFFAOYSA-N 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 241000270708 Testudinidae Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N17/00—Investigating resistance of materials to the weather, to corrosion, or to light
- G01N17/02—Electrochemical measuring systems for weathering, corrosion or corrosion-protection measurement
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Botany (AREA)
- Forests & Forestry (AREA)
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to a method for identifying salt tolerance of ilex chinensis in a seedling stage, which can judge the salt tolerance of a variety by measuring the relative conductivity of ilex chinensis leaves under salt stress. The method for quickly and efficiently identifying the salt tolerance of the holly in the seedling stage can effectively reflect the salt tolerance of the holly; the method is simple, convenient and quick, has strong operability, is not influenced by the growth period of the holly and the external environment, can effectively reduce the workload of field salt tolerance identification, can also be used for screening the ecological adaptability of the holly, and has wide application prospect.
Description
Technical Field
The invention relates to the field of biotechnology and agricultural engineering, in particular to a method for early identifying salt tolerance of holly.
Background
The area distribution of saline-alkali soil in China is wide, the total area of coastal beaches of Jiangsu currently occupies the first position of China, and suitable salt-tolerant tree species with high economic value are screened out in order to improve the condition of saline-alkali soil in China. The holly plant resource is rich, and the holly plant resource has higher economic value and ornamental value, and has great significance in researching the applicability of the holly plant resource in saline-alkali soil. At present, the research on the evaluation of the salt tolerance of the holly is less. The reports of plum snow, et al (2014) indicate that glossy wintergreen has strong stress resistance to cold, drought, and saline-alkaline environments. Studies by Cheng et al (2017) show that the salt tolerance of the tortoise shell wintergreen is very good. Therefore, the holly plant has certain potential in breeding the salt-tolerant tree species. However, the identification of the salt tolerance of the ilex has the problems of long time, large workload, large environmental influence and the like.
When the plant is damaged by salt, the cell membrane is damaged, the membrane permeability is increased, so that the electrolyte in the cell is leaked out, and the conductivity of the cell leaching solution is increased, namely the degree of the increase of the membrane permeability is related to the strength of the salt tolerance of the plant. Therefore, by comparing the degree of increase of the leaf cell membrane permeability of different varieties under the same salt concentration, the strong and weak stress resistance between varieties can be reflected.
Disclosure of Invention
The invention aims to provide a method for quickly and efficiently identifying the salt tolerance of ilex chinensis in the seedling stage. According to the method, the damage degree of the cell membrane of the leaves is judged according to the relative conductivity of the leaves under salt stress, so that the difference of salt tolerance of the materials to be detected is judged, and the salt tolerance of the holly can be truly reflected.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a method for identifying the salt tolerance of ilex chinensis in a seedling stage comprises the following culture steps:
(1) putting the ilex chinensis seedlings to be detected into 50-100 mM sodium chloride solution, and carrying out salt solution treatment;
(2) putting a certain amount of deionized water into a clean glass container, and measuring the conductivity EC of the deionized water by using a conductivity meter0；
(3) Selecting 8-12 functional leaves, punching 8-12 round holes with the diameter of 0.8cm on the leaves by using a handheld puncher, and putting the obtained leaf wafer into the deionized water prepared in the step (2);
(4) putting the solution obtained in the step (3) in water bath at 37 ℃ for 50-70 min, and measuring the conductivity EC by using a conductivity meter1；
(5) Placing the solution obtained in the step (4) in water bath at 100 ℃ for 10-20 min, taking out, cooling to room temperature, shaking uniformly, and measuring the conductivity EC by using a conductivity meter2；
(6) Calculating relative blade conductivity(％)＝[(EC1-EC0)/(EC2-EC0)]×100％；
(7) According to the relative conductivity of the leaves, carrying out salt tolerance classification on the ilex to be detected, wherein the relative conductivity is less than or equal to 20 percent and is salt-tolerant; the relative conductivity of the leaves is more than 20 percent and less than or equal to 35 percent, and the salt tolerance is moderate; the relative conductivity of the leaves is more than 35 percent and less than or equal to 50 percent, and the leaves are salt-resistant; the relative conductivity of the leaves is more than 50 percent, and the leaves are extremely salt-resistant.
Preferably, in the step (1), the number of the plants of the holly is more than 3, the height of the plants is 10cm +/-2 cm, and the length of the root system is 5 cm.
Preferably, in step (1), the culture conditions are 25 +/-2 ℃, the relative humidity is 80% +/-5%, and the illumination time is 14 h.
Further, in the step (1), the salt treatment time is 25 d.
Furthermore, in the step (2), glassware needs to be washed and dried by deionized water in advance.
Further, in the step (3), the leaves are selected to be functional leaves which are similar in leaf age and are healthy and free of diseases and insect pests at the same position, and main veins of the leaves are avoided during punching.
Furthermore, in the step (3), the leaves are not required to be touched by hands so as to avoid pollution.
Further, in step (4), the solution needs to be shaken frequently during the water bath to soak the cell fluid at the wound site.
Further, in the step (4), after the conductivity is measured, the electrode is cleaned.
Further, in the step (5), the solution is taken out and cooled to 20-25 ℃.
Compared with the prior art, the invention has the beneficial effects that:
the method is simple, convenient and quick, has strong operability, is not influenced by the growth period of the holly and the external environment, can effectively reflect the salt tolerance of the holly, can effectively reduce the workload of field salt tolerance identification, can also be used for screening the ecological adaptability of the holly, and has wide application prospect.
Detailed Description
The above-mentioned contents of the present invention are further described in detail by way of examples below, but it should not be understood that the scope of the above-mentioned subject matter of the present invention is limited to the following examples, and any technique realized based on the above-mentioned contents of the present invention falls within the scope of the present invention.
Test materials: the total amount of the test materials is 6 parts, including 2 parts of thornless Chinese holly and golden Chinese holly of good salt resistance, and 4 parts of seed line materials with the serial number of DQ1-DQ 4.
Preparing a salt solution: preparing 18L of 100mM sodium chloride solution, weighing 105.20g of sodium chloride, adding water to a constant volume of 18L, and subpackaging the prepared high-salt solution into 1L triangular bottles, wherein each bottle contains 1L of sodium chloride for later use;
the method comprises the following operation steps: taking out the seedlings of the holly from the soil, cleaning soil at the root, taking 9 seedlings from each material, putting the roots of the taken seedlings into a subpackaged 1L of sodium chloride solution, setting 3 plants in each bottle to be the same as 1 material, repeating the setting of 1 material for 3 times, and treating for 25 days under the culture conditions of the temperature of 25 +/-2 ℃, the relative humidity of 80 +/-5 percent and the illumination time of 14 hours. Selecting functional leaves of the ilex to be tested, punching 10 round pieces of slices with the diameter of 0.8cm on the leaves by using a handheld puncher, putting the obtained slice round pieces into a glass vessel filled with 20ml of deionized water, and measuring the conductivity EC of the deionized water by using a conductivity meter in advance0Then placing the solution containing the leaf in water bath at 37 deg.C for 60min, and measuring the conductivity EC1(ii) a Then placing the solution in water bath at 100 deg.C for 15min, taking out, cooling, shaking, and measuring conductivity EC2。
And (4) counting results:
(1) relative conductivity of the leaves after 25d salt solution treatment.
Table 16 electrical conductivity of identified materials
(2) And (4) according to the relative conductivity survey result of the leaves, carrying out salt tolerance classification on the ilex to be detected.
Utilizing the conductivity of the leaves of the materials in the table 1 to classify the salt tolerance of the ilex to be tested, wherein the relative conductivity is less than or equal to 20 percent, and the ilex is salt tolerant; the relative conductivity of the leaves is more than 20 percent and less than or equal to 35 percent, and the salt tolerance is moderate; the relative conductivity of the leaves is more than 35 percent and less than or equal to 50 percent, and the leaves are salt-resistant; the relative conductivity of the leaves is more than 50 percent, and the leaves are extremely salt-resistant.
TABLE 26 salt tolerance Performance of the materials
According to the results of the test, the Chinese holly and golden Chinese holly have moderate salt tolerance, which is consistent with the published results of the former. The method can be used for accurately identifying the salt resistance of different materials, and is simple and convenient and easy to operate.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention in any way, and any person skilled in the art can make any simple modification, equivalent replacement, and improvement on the above embodiment without departing from the technical spirit of the present invention, and still fall within the protection scope of the technical solution of the present invention.
Claims (10)
1. A method for identifying the salt tolerance of ilex in seedling stage is characterized in that: the culture steps are as follows:
(1) putting the ilex chinensis seedlings to be detected into 50-100 mM sodium chloride solution, and carrying out salt solution treatment;
(2) putting a certain amount of deionized water into a clean glass container, and measuring the conductivity EC of the deionized water by using a conductivity meter0；
(3) Selecting 8-12 functional leaves, punching 10 wafers with the diameter of 0.8cm on the leaves by using a handheld puncher, and putting the obtained wafer of the leaves into the deionized water prepared in the step (2);
(4) putting the solution obtained in the step (3) in water bath at 37 ℃ for 50-70 min, and measuring the conductivity EC by using a conductivity meter1；
(5) Putting the solution obtained in the step (4) in a water bath at 100 ℃ for 10-20 min, taking out, cooling to room temperature, shakingAfter being homogenized, conductivity EC is measured by a conductivity meter2；
(6) Calculating relative leaf conductivity (%) - (EC)1-EC0)/(EC2-EC0)]×100％；
(7) According to the relative conductivity of the leaves, carrying out salt tolerance classification on the ilex to be detected, wherein the relative conductivity is less than or equal to 20 percent and is salt-tolerant; the relative conductivity of the leaves is more than 20 percent and less than or equal to 35 percent, and the salt tolerance is moderate; the relative conductivity of the leaves is more than 35 percent and less than or equal to 50 percent, and the leaves are salt-resistant; the relative conductivity of the leaves is more than 50 percent, and the leaves are extremely salt-resistant.
2. The method for identifying the salt tolerance of the ilex chinensis in the seedling stage as claimed in claim 1, wherein in the step (1), the number of the ilex chinensis plants is more than 3, the plant height is 10cm +/-2 cm, and the root length is 5> cm.
3. The method for identifying the salt tolerance of ilex chinensis in the seedling stage of claim 1, wherein in the step (1), the culture conditions are that the temperature is 25 +/-2 ℃, the relative humidity is 80% +/-5% and the illumination time is 14 h.
4. The method for identifying the salt tolerance of ilex chinensis in the seedling stage of claim 1, wherein in the step (1), the salt treatment time is 25 days.
5. The method for identifying the salt tolerance of ilex chinensis in the seedling stage as claimed in claim 1, wherein in the step (2), glassware is washed with deionized water and dried in advance.
6. The method for identifying the salt tolerance of the holly leaves in the seedling stage as claimed in claim 1, wherein in the step (3), the leaves are selected to be healthy and functional leaves with similar leaf age and no plant diseases and insect pests at the same part, and main veins of the leaves are avoided when the holes are punched.
7. The method for identifying salt tolerance of ilex in seedling stage as claimed in claim 1, wherein in step (3), the leaves are not touched by hand to avoid contamination.
8. The method for identifying salt tolerance of ilex in seedling stage as claimed in claim 1, wherein in step (4), the solution is shaken frequently during the water bath to soak the cell fluid at the wound.
9. The method for identifying the salt tolerance of ilex in the seedling stage of claim 1, wherein in the step (4), after the conductivity is measured, the electrodes are cleaned.
10. The method for identifying the salt tolerance of ilex chinensis in the seedling stage as claimed in claim 1, wherein in the step (5), the solution is taken out and cooled to 20-25 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111220475.8A CN113940205A (en) | 2021-10-20 | 2021-10-20 | Method for identifying salt tolerance of holly at seedling stage |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111220475.8A CN113940205A (en) | 2021-10-20 | 2021-10-20 | Method for identifying salt tolerance of holly at seedling stage |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785383A (en) * | 2010-03-17 | 2010-07-28 | 吉林省农业科学院 | Method for screening corn salt/alkali-tolerance germplasm |
| CN103392405A (en) * | 2013-07-04 | 2013-11-20 | 甘肃农业大学 | Simple screening method for salt-tolerance maize inbred lines |
| CN106688685A (en) * | 2016-12-30 | 2017-05-24 | 四川农业大学 | Method of relieving high temperature stress of kiwi fruit trees |
| CN107889742A (en) * | 2017-11-24 | 2018-04-10 | 四川农业大学 | The method that haloduric corn kind is cultivated using corn allopolyploid |
-
2021
- 2021-10-20 CN CN202111220475.8A patent/CN113940205A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785383A (en) * | 2010-03-17 | 2010-07-28 | 吉林省农业科学院 | Method for screening corn salt/alkali-tolerance germplasm |
| CN103392405A (en) * | 2013-07-04 | 2013-11-20 | 甘肃农业大学 | Simple screening method for salt-tolerance maize inbred lines |
| CN106688685A (en) * | 2016-12-30 | 2017-05-24 | 四川农业大学 | Method of relieving high temperature stress of kiwi fruit trees |
| CN107889742A (en) * | 2017-11-24 | 2018-04-10 | 四川农业大学 | The method that haloduric corn kind is cultivated using corn allopolyploid |
Non-Patent Citations (1)
| Title |
|---|
| 刘婷等: "13种植物耐盐性的离体快速分析", 《天津农学院学报》 * |
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