CN115280932B - Seed initiation method for improving salt tolerance of alfalfa - Google Patents
Seed initiation method for improving salt tolerance of alfalfa Download PDFInfo
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- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 title claims abstract description 81
- 241000219823 Medicago Species 0.000 title claims abstract description 80
- 230000000977 initiatory effect Effects 0.000 title claims abstract description 65
- 238000000034 method Methods 0.000 title claims abstract description 48
- 230000015784 hyperosmotic salinity response Effects 0.000 title claims abstract description 35
- 230000035784 germination Effects 0.000 claims abstract description 72
- 229910017569 La2(CO3)3 Inorganic materials 0.000 claims abstract description 35
- NZPIUJUFIFZSPW-UHFFFAOYSA-H lanthanum carbonate Chemical compound [La+3].[La+3].[O-]C([O-])=O.[O-]C([O-])=O.[O-]C([O-])=O NZPIUJUFIFZSPW-UHFFFAOYSA-H 0.000 claims abstract description 35
- 229960001633 lanthanum carbonate Drugs 0.000 claims abstract description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 49
- 239000007788 liquid Substances 0.000 claims description 46
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- 239000012153 distilled water Substances 0.000 claims description 18
- 238000011010 flushing procedure Methods 0.000 claims description 18
- 239000002105 nanoparticle Substances 0.000 claims description 17
- 230000037452 priming Effects 0.000 claims description 17
- 230000001954 sterilising effect Effects 0.000 claims description 13
- 239000002245 particle Substances 0.000 claims description 10
- 238000012545 processing Methods 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 239000000725 suspension Substances 0.000 claims description 9
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 238000005286 illumination Methods 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 4
- 230000003203 everyday effect Effects 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 2
- 230000007226 seed germination Effects 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 230000001502 supplementing effect Effects 0.000 claims 1
- 150000003839 salts Chemical class 0.000 abstract description 15
- 239000002689 soil Substances 0.000 abstract description 12
- 241000196324 Embryophyta Species 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 abstract description 5
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 239000002250 absorbent Substances 0.000 description 12
- 230000002745 absorbent Effects 0.000 description 12
- 238000012258 culturing Methods 0.000 description 12
- 239000012535 impurity Substances 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000011160 research Methods 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 3
- 229910052746 lanthanum Inorganic materials 0.000 description 3
- FZLIPJUXYLNCLC-UHFFFAOYSA-N lanthanum atom Chemical compound [La] FZLIPJUXYLNCLC-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- YAGMEFLENMBNJT-UHFFFAOYSA-N [O-2].[La+3].C(O)(O)=O.[O-2].[O-2].[La+3] Chemical compound [O-2].[La+3].C(O)(O)=O.[O-2].[O-2].[La+3] YAGMEFLENMBNJT-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 229910052761 rare earth metal Inorganic materials 0.000 description 2
- 150000002910 rare earth metals Chemical class 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 241000380130 Ehrharta erecta Species 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 150000001647 brassinosteroids Chemical class 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000003 effect on germination Effects 0.000 description 1
- 239000007772 electrode material Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- KGDJAQAMSDMZCD-UHFFFAOYSA-M hydrogen carbonate lanthanum(3+) oxygen(2-) Chemical compound C([O-])(O)=O.[O-2].[La+3] KGDJAQAMSDMZCD-UHFFFAOYSA-M 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- 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
-
- 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
- A01C1/02—Germinating apparatus; Determining germination capacity of seeds or the like
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Environmental Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physiology (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention discloses a seed initiation method for improving salt tolerance of alfalfa, and belongs to the technical field of pasture cultivation. The method of the invention prepares lanthanum carbonate [ La ] 2 O 2 (CO 3 )]The nanometer initiating liquid-seed collection-treatment-initiation of alfalfa seeds-seed germination culture is used for initiating culture of alfalfa seeds. According to the method, the lanthanum carbonate oxide nano solution is used for initiating the alfalfa seeds, so that the germination rate and the germination vigor of the alfalfa seeds under salt stress are remarkably improved, the salt tolerance of the alfalfa is enhanced, the efficient and rapid salt damage relieving capability is shown, and the breeding period is shortened. The method of the invention better solves the urgent need of salt-tolerant pasture cultivation land; has important significance for large-scale popularization, cultivation and planting of alfalfa in soil salinization areas, and provides basis for application of nano lanthanum carbonate oxide to pasture plants, especially alfalfa and improvement of salt tolerance of the nano lanthanum carbonate oxide.
Description
Technical Field
The invention relates to the technical field of pasture cultivation, in particular to a seed initiation method for improving salt tolerance of alfalfa.
Background
Global saline-alkali soil is being 1×10 per year 6 ～1.5×10 6 hm 2 The speed of (2) is increased, and the Chinese salinized and secondary salinized lands have 4.0X10 percent 7 hm 2 Above, with deterioration and irrational ecological environmentThe development and utilization of the soil are further expanded. Alfalfa @Medicago sativa) Is leguminous alfalfa with high yield, high quality, high nutritive value and ecological value, good palatability and high digestibility, belongs to perennial grass and is known as 'grass king'. The alfalfa is widely distributed, high in yield, rich and comprehensive in nutrition components, particularly high in vitamin content and crude protein content, is a good protein feed source for high-yield cows, has a large forage feed development value, is developed in rhizome, can increase the soil porosity, improve the soil water storage capacity, reduce the soil volume weight and has remarkable effect in water and soil conservation. Compared with other pastures, the pastures have higher salt tolerance and the effect of improving soil salinization, thus being an ideal high-quality pastures for repairing and utilizing salinized soil. The alfalfa has great application and development values, and scientific research and application, popularization and planting of the alfalfa are needed to be increased.
Seed priming, also known as seed osmotic regulation, refers to a pre-sowing seed pretreatment technique that slowly absorbs water under controlled conditions (e.g., slow quantitative water absorption, gradual drying back, etc.), prepares the seeds for germination in advance, and does not cause injury. At present, research reports that the seed germination capacity of alfalfa under salt stress can be improved by utilizing a seed initiation technology. The analysis of the test result of Jie Xiujuan shows that brassinosteroids trigger to improve the quantity, speed and quality of germination of alfalfa seeds under the condition of salt stress, sand trigger also obviously improves the germination vigor and germination rate of alfalfa varieties, shortens the average germination time of the three varieties, and also obviously shows the sand trigger effect on germination indexes and vitality indexes. However, no experiment has been performed to explore the improvement of alfalfa salt tolerance by nano-initiation technology.
Rare earth lanthanum is widely used in exhaust gas catalysts, agricultural films, optical fibers, electrode materials and other applications. Lanthanum has a toxicant excitation effect, which means that physiological processes of organisms, especially growth and development of plants, are stimulated by toxic substances. It is formed by combining the characteristics of rare earth lanthanum and the characteristics of nano materials by nano lanthanum carbonate oxideIs a new research field. More and more research is beginning to focus on La 2 O 2 (CO 3 ) The potential environmental risk of NPs, but the application of nano lanthanum carbonate oxide in the germination of pasture seeds is not reported at present, the influence of the nano lanthanum carbonate oxide on plants is greatly related to the concentration, the treatment time and the plant materials, and the difference of the promotion or inhibition effect on the plants can be caused by the different medicament concentration and the treatment time.
The development technology of the existing alfalfa is slow in promotion, long in breeding working period and low in germination rate, researches for improving the salt tolerance of alfalfa seeds by utilizing a nano initiation technology are few, particularly, researches for improving the salt tolerance of alfalfa seeds by initiating with nano lanthanum carbonate oxide are not reported, the improvement of the salt tolerance of the alfalfa by the existing nano initiation technology is not very remarkable, and the alleviation degree of salt damage is very limited. Based on this, it is highly desirable for those skilled in the art to provide an priming method for improving salt tolerance of alfalfa seeds.
Disclosure of Invention
The invention aims at the problems and provides a seed priming method for improving the salt tolerance of alfalfa. Solves the problems of long breeding period, low germination rate and low salt tolerance of alfalfa in the prior art.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the invention relates to a seed initiation method for improving salt tolerance of alfalfa, which specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) were dispersed in deionized water by ultrasonic vibration (100 w,40 khz);
s2: collecting and processing seeds, namely removing impurities and shrunken seeds of alfalfa for later use, selecting alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing, flushing and airing for later use;
s3: the method comprises the steps of initiating, placing seeds into a centrifuge tube, adding nano initiating liquid to submerge the seeds, shaking the initiating liquid to enable the initiating liquid to be fully contacted with the seeds, placing the seeds into a dark incubator for culturing, pouring out suspension after culturing, using water absorbing paper to absorb water, uniformly spreading the suspension on clean water absorbing paper, and drying the seeds back at room temperature for subsequent germination;
s4: and (3) germinating the seeds, wherein the seeds initiated in the step (S3) are germinated by adopting a paper germination method, a proper amount of culture solution is added during germination culture, the seeds are placed in an artificial climate incubator for continuous culture, distilled water is periodically added every day, and the number of the germinated seeds per day is recorded by taking dew as a germination standard.
Further, in the step S1, the purity of the lanthanum carbonate oxide nano particles is 99.99%, the particle size is 50nm, the ultrasonic vibration frequency is 40kHz, and the ultrasonic vibration time is 30min.
Further, the concentration of the nano initiating liquid carbonic acid lanthanum oxide in the step S1 is 0-500mg/L.
Further, the concentration of the nano initiating liquid carbonic acid lanthanum oxide in the step S1 is 10mg/L.
Further, in the step S2, 75% ethanol is adopted for sterilization, the sterilization time is 30S, and distilled water is adopted for washing 4-5 times.
Further, the temperature of the dark incubator in the step S3 is constant at 5-15 ℃, the culture time is 12-36h, and the drying time is 48h.
Further, the temperature of the dark incubator in the step S3 is constant at 10 ℃, and the incubation time is 24 hours.
Further, the germination method on paper in the step S4 is to place seeds in a culture dish with filter paper laid thereon.
Still further, the filter paper is 2 layers.
Further, the culture solution in the step S4 is 200mM NaCl solution, the culture time in the incubator is 10 d, the light time per day is 16 hours, the dark time is 8 hours, and the temperature is 25 ℃.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the alfalfa seeds are treated by using the lanthanum carbonate oxide nanoparticle solution, and then the seeds are dried back, so that the germination rate and the germination vigor of the alfalfa seeds under salt stress are remarkably improved, the salt tolerance of the alfalfa is remarkably enhanced, the efficient and rapid salt damage relieving capability is shown, and the breeding period is shortened. By adopting the seed initiation technology, the cultivation process of the alfalfa in the soil salinization environment is greatly shortened, and the urgent need of pasture salt-tolerant cultivation land is well solved; the method has important significance for large-scale popularization, cultivation and planting of the alfalfa in the soil salinization area, fills the data blank of the application of the nano material in the aspect of the alfalfa seed initiation technology, and provides a basis for the application of the nano lanthanum carbonate oxide in pasture plants, especially the alfalfa, and the improvement of the salt tolerance of the nano lanthanum carbonate oxide.
Drawings
FIG. 1 shows La at various concentrations 2 O 2 (CO 3 ) NPs induce influence on germination vigor of alfalfa seeds under salt stress;
FIG. 2 is a graph of La at various concentrations 2 O 2 (CO 3 ) NPs induce influence on germination rate of alfalfa seeds under salt stress;
FIG. 3 shows La at various concentrations 2 O 2 (CO 3 ) NPs induce influence on germination index of alfalfa seeds under salt stress;
FIG. 4 shows La at various concentrations 2 O 2 (CO 3 ) NPs induce influence on germination peak value of alfalfa seeds under salt stress;
FIG. 5 shows La at various concentrations 2 O 2 (CO 3 ) NPs trigger effects on alfalfa germination seed root length and seedling length under salt stress.
Detailed Description
The present invention will be described in further detail with reference to examples in order to make the objects and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
The instruments, reagents, materials, etc. used in the following examples are conventional instruments, reagents, materials, etc. known in the art, and are commercially available. The experimental methods, detection methods, and the like in the following examples are conventional experimental methods, detection methods, and the like existing in the prior art unless otherwise specified.
Example 1
The seed priming method for improving the salt tolerance of alfalfa in the embodiment specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40 kHz), and ultrasonic vibration is carried out for 30min, so as to prepare 10mg/L lanthanum carbonate oxide nano initiating liquid; wherein the lanthanum carbonate oxide nanoparticles are purchased from rohn (rhawn);
s2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to fully contact with the seeds, the seeds are placed in a 10 ℃ constant temperature dark incubator to be cultured for 24 hours, suspension liquid is poured out after the culture is finished, water is absorbed by using absorbent paper and evenly spread on clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination test;
s4: and (3) germinating the seeds, carrying out germination culture on the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination culture, placing the seeds in an artificial climatic incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the darkness time is 8 hours, the temperature is 25 ℃, and distilled water is periodically added per day, so that the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Example 2
The seed priming method for improving the salt tolerance of alfalfa in the embodiment specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40 kHz), and ultrasonic vibration is carried out for 30min, so as to prepare 50mg/L lanthanum carbonate oxide nano initiating liquid; wherein the lanthanum carbonate oxide nanoparticles are purchased from rohn (rhawn);
s2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to be fully contacted with the seeds, the seeds are placed in a constant temperature and dark incubator at 5 ℃ for culturing for 36 hours, suspension liquid is poured out after the culturing is finished, water is absorbed by using absorbent paper and evenly spread on clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination test;
s4: and (3) germinating the seeds, carrying out germination culture on the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination culture, placing the seeds in an artificial climatic incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the darkness time is 8 hours, the temperature is 25 ℃, and distilled water is periodically added per day, so that the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Example 3
The seed priming method for improving the salt tolerance of alfalfa in the embodiment specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) are dispersed in deionized water by ultrasonic vibration (100W, 40 kHz)In water, carrying out ultrasonic vibration for 30min to prepare 100mg/L lanthanum carbonate oxide nano initiating liquid; wherein the lanthanum carbonate oxide nanoparticles are purchased from rohn (rhawn);
s2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to be fully contacted with the seeds, the seeds are placed in a constant temperature and dark incubator at 15 ℃ for culturing for 12 hours, suspension liquid is poured out after the culturing is finished, water is absorbed by using absorbent paper and evenly spread on the clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination test;
s4: and (3) germinating the seeds, carrying out germination culture on the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination culture, placing the seeds in an artificial climatic incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the darkness time is 8 hours, the temperature is 25 ℃, and distilled water is periodically added per day, so that the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Example 4
The seed priming method for improving the salt tolerance of alfalfa in the embodiment specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40 kHz), and ultrasonic vibration is carried out for 30min, so as to prepare 200mg/L lanthanum carbonate oxide nano initiating liquid; wherein the lanthanum carbonate oxide nanoparticles are purchased from rohn (rhawn);
s2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to fully contact with the seeds, the seeds are placed in a 13 ℃ constant temperature dark incubator to be cultured for 20 hours, suspension liquid is poured out after the culture is finished, water is absorbed by using absorbent paper and evenly spread on clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination tests;
s4: and (3) germinating the seeds, carrying out germination culture on the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination culture, placing the seeds in an artificial climatic incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the darkness time is 8 hours, the temperature is 25 ℃, and distilled water is periodically added per day, so that the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Example 5
The seed priming method for improving the salt tolerance of alfalfa in the embodiment specifically comprises the following steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) were dispersed in deionized water by ultrasonic vibration (100 w,40 khz), for 30min, to prepare 500mg/L lanthanum carbonate oxide nano-initiator, wherein lanthanum carbonate oxide nanoparticles were purchased from rohn (rhawn);
s2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to fully contact with the seeds, the seeds are placed in a constant temperature and dark incubator at 8 ℃ for culturing for 18 hours, suspension liquid is poured out after the culturing is finished, water is absorbed by using absorbent paper and evenly spread on clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination test;
s4: and (3) germinating the seeds, namely germinating and culturing the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds subjected to initiation treatment of different nano initiation liquids in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination and culturing, placing the culture solution in an artificial climate incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the dark time is 8 hours, the temperature is 25 ℃, distilled water is periodically added per day, and the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Example 6
The seed priming method for improving the salt tolerance of alfalfa comprises the following specific steps:
s1: preparing nano initiating liquid, adding lanthanum carbonate oxide nano particles (La 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) are dispersed in deionized water by ultrasonic vibration (100W, 40 kHz), and ultrasonic vibration is carried out for 30min to prepare 0mg/L lanthanum carbonate oxide nano initiating liquid.
S2: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process, the seeds are placed in a 10ml centrifuge tube, then 2ml of nano initiating liquid is added to submerge the seeds, the initiating liquid is shaken to enable the initiating liquid to be fully contacted with the seeds, the seeds are placed in a 9 ℃ constant temperature dark incubator to be cultured for 30 hours, suspension liquid is poured out after the culture is finished, water is absorbed by using absorbent paper and evenly spread on clean absorbent paper, and the seeds are dried back for 48 hours at room temperature for subsequent germination tests;
s4: and (3) germinating the seeds, namely germinating and culturing the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds subjected to initiation treatment of different nano initiation liquids in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination and culturing, placing the culture solution in an artificial climate incubator for continuous culture for 10 d, wherein the illumination time per day in the artificial incubator is 16 hours, the dark time is 8 hours, the temperature is 25 ℃, distilled water is periodically added per day, and the number of the germinated seeds per day is recorded by taking the exposure as a germination standard.
Comparative example 1
The comparative example is used as a CK group, and the alfalfa salt tolerance seeds of the comparative example germinate, and specifically comprises the following steps:
s1: collecting and processing seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds which are uniform in size, full in particles and uniform in color, flushing the seeds with clear water, selecting submerged full seeds, sterilizing with 75% ethanol for 30 seconds, flushing with distilled water for 4-5 times, and airing for later use;
s2: the seeds germinate, the picked alfalfa seeds are germinated and cultivated by a paper germination method, the seeds are placed in a culture dish paved with 2 layers of filter paper, 5ml of 200mM NaCl culture solution is added during germination and cultivation, the seeds are placed in an artificial climate incubator for continuous cultivation for 10 days, the illumination time of each day in the artificial incubator is 16 hours, the darkness time is 8 hours, the temperature is 25 ℃, distilled water is periodically added every day, and the germination number of the seeds per day is recorded by taking the exposure as a germination standard.
The germination percentage, germination potential, germination index, germination peak value, root length and seedling length of examples 1 to 6 and comparative example 1 were calculated, and the root length and seedling length were measured in parallel for 5 times, and the average value was obtained, and the measurement results are shown in fig. 1 to 5.
Referring to fig. 1-5, it can be seen that, compared with the comparative example 1 (CK), the germination vigor of alfalfa seeds is increased from 4% to 58% in comparison with the control, the germination rate is increased from 10% to 72% in comparison with the control, the germination index is increased from 2.08 to 20.31 in comparison with the control, the germination peak value is increased from 2 to 12.5 in comparison with the control, the root length is increased from 11.79 mm to 23.41mm, the seedling length is increased from 9.75mm to 15.51mm in comparison with the control, the germination rate and germination speed of alfalfa seeds under salt stress are remarkably increased, and the influence of salt damage on the growth of seedlings is relieved.
In summary, the invention relates to a seed initiation method for improving the salt tolerance of alfalfa by using 0-500mg/L of carbonate lanthanum oxide nano particles (La 2 O 2 (CO 3 ) NPs,50 nm) the solution is used for treating alfalfa seeds 24h under constant temperature and darkness, and then the seeds are dried back, so that the germination rate and the germination vigor of the alfalfa seeds under salt stress are remarkably improved, and the salt tolerance of the alfalfa is remarkably enhanced. Has important significance for large-scale popularization, cultivation and planting of alfalfa in soil salinization areas, and provides basis for application of nano lanthanum carbonate oxide to pasture, especially alfalfa, and improvement of salt tolerance of the nano lanthanum carbonate oxide.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (8)
1. A seed priming method for improving salt tolerance of alfalfa is characterized in that: the method comprises the following steps:
s1: preparing nano initiating liquid, namely dispersing lanthanum carbonate oxide nano particles into deionized water through ultrasonic vibration to obtain 10mg/L lanthanum carbonate oxide nano initiating liquid;
s2: collecting and processing seeds, selecting alfalfa seeds with uniform size, plump grains and uniform color, flushing the seeds with clear water, selecting submerged plump seeds, sterilizing, flushing and airing;
s3: the initiation process, the seed is put into centrifuge tube, add nano initiation liquid to submerge the seed, and fully contact with the seed, put into dark incubator to culture, pour out suspension after culture, and the seed is dried back under room temperature;
s4: and (3) seed germination, namely performing germination culture on the seeds initiated in the step (S3) by adopting a paper germination method, adding a proper amount of culture solution during germination culture, placing the seeds in an incubator for continuous culture, and periodically supplementing distilled water every day, wherein the number of the seeds germinated every day is recorded by taking the dew as a germination standard.
2. The seed priming method for improving salt tolerance of alfalfa according to claim 1, wherein: the purity of the lanthanum carbonate oxide nano particles in the step S1 is 99.99%, the particle size is 50nm, the ultrasonic vibration frequency is 40kHz, and the ultrasonic vibration time is 30min.
3. The seed priming method for improving salt tolerance of alfalfa according to claim 1, wherein: and in the step S2, 75% ethanol is adopted for sterilization, the sterilization time is 30S, and distilled water is adopted for washing 4-5 times.
4. The seed priming method for improving salt tolerance of alfalfa according to claim 1, wherein: the temperature of the dark incubator in the step S3 is kept constant at 5-15 ℃, the culture time is 12-36h, and the drying time is 48h.
5. The seed priming method for improving salt tolerance of alfalfa of claim 4, wherein: the temperature of the dark incubator in the step S3 is constant at 10 ℃, and the incubation time is 24 hours.
6. The seed priming method for improving salt tolerance of alfalfa according to claim 1, wherein: the germination method on paper in the step S4 is to place seeds in a culture dish paved with filter paper.
7. The seed priming method for improving salt tolerance of alfalfa of claim 6, wherein: the filter paper is 2 layers.
8. The seed priming method for improving salt tolerance of alfalfa according to claim 1, wherein: the culture solution in the step S4 is 200mM NaCl solution, the culture time in the incubator is 10 d, the illumination time per day is 16 hours, and the temperature is 25 ℃.
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