CN115280932A - Seed priming method for improving salt tolerance of alfalfa - Google Patents
Seed priming method for improving salt tolerance of alfalfa Download PDFInfo
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- KGDJAQAMSDMZCD-UHFFFAOYSA-M hydrogen carbonate lanthanum(3+) oxygen(2-) Chemical compound C([O-])(O)=O.[O-2].[La+3] KGDJAQAMSDMZCD-UHFFFAOYSA-M 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 46
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- 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
- 239000002105 nanoparticle Substances 0.000 claims description 17
- 238000001035 drying Methods 0.000 claims description 11
- 239000002245 particle Substances 0.000 claims description 11
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- 238000012258 culturing Methods 0.000 claims description 10
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- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 230000000249 desinfective effect Effects 0.000 claims description 8
- 238000005286 illumination Methods 0.000 claims description 7
- 229910017569 La2(CO3)3 Inorganic materials 0.000 claims description 6
- 229960001633 lanthanum carbonate Drugs 0.000 claims description 6
- 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 description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 abstract description 15
- 239000002689 soil Substances 0.000 abstract description 12
- 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 abstract description 9
- 241000196324 Embryophyta Species 0.000 abstract description 7
- 238000009395 breeding Methods 0.000 abstract description 4
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- 239000002250 absorbent Substances 0.000 description 14
- 230000002745 absorbent Effects 0.000 description 14
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- 239000010452 phosphate Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
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- 229910052746 lanthanum Inorganic materials 0.000 description 3
- FZLIPJUXYLNCLC-UHFFFAOYSA-N lanthanum atom Chemical compound [La] FZLIPJUXYLNCLC-UHFFFAOYSA-N 0.000 description 3
- 244000025254 Cannabis sativa Species 0.000 description 2
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- IXVMHGVQKLDRKH-KNBKMWSGSA-N brassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@@H](O)[C@H](O)[C@@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-KNBKMWSGSA-N 0.000 description 1
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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
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- 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 carbonic acid lanthanum oxide [ La 2 O 2 (CO 3 )]And (3) carrying out priming culture on the alfalfa seeds by using the nano priming solution, collecting and treating the seeds, priming the alfalfa seeds and carrying out seed germination culture. According to the method, the carbonic acid lanthanum oxide nano solution is used for initiating the alfalfa seeds, so that the germination rate and the germination potential of the alfalfa seeds under salt stress are obviously 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 problem of hurry of the pasture salt-tolerant cultivation land; has important significance for the wide-range promotion and cultivation of the alfalfa in the soil salinization areaProvides a basis for the application of the nanometer lanthanum oxide carbonate in pasture plants, especially alfalfa and improving the salt tolerance of the pasture plants.
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
The global saline-alkali soil is about 1 × 10 per year 6 ～1.5×10 6 hm 2 The speed of the method is increased, and the salinized and secondary salinized lands in China have 4.0 multiplied by 10 7 hm 2 Above, with deterioration of ecological environment and unreasonable development and utilization, soil salinization will further expand. Alfalfa (C)Medicago sativa) The leguminous alfalfa is high in yield, quality, nutritional value and ecological value, palatability and digestibility, belongs to perennial herbage, and is known as the king of herbage. The alfalfa is widely distributed, high in yield, rich and comprehensive in nutritional ingredients, especially high in vitamin content and crude protein content, is a good protein feed source for high-yield dairy cows, has a high forage grass feed development value, is developed in roots and stems, can increase soil porosity, improve soil water storage capacity, reduce soil volume weight, and has a remarkable effect in keeping water and soil. Compared with other pasture, the pasture has higher salt tolerance and the function of improving soil salinization, and is an ideal high-quality pasture for repairing and utilizing the salinized soil. The alfalfa has great application and development values, and scientific research, application and popularization of the alfalfa are urgently needed to be increased.
Seed priming, also known as seed osmoregulation, is a pre-sowing seed pretreatment technique that allows seeds to slowly absorb water under controlled conditions (e.g., slow, quantitative water uptake and gradual return to dryness) to prepare the seeds physiologically for germination without causing damage. At present, research reports show that the seed germination capacity of alfalfa under salt stress can be improved by utilizing a seed priming technology. Analysis of test results of the jujuanjuan shows that the brassinolide initiation under the condition of salt stress improves the germination quantity, speed and quality of alfalfa seeds, the sand initiation also obviously improves the germination vigor and germination rate of alfalfa varieties, meanwhile, the average germination time of the three varieties is shortened, and the sand initiation effect is also highlighted on the germination index and vitality index. However, no nano-initiation technology has been tested to improve the salt tolerance of alfalfa.
Rare earth lanthanum is widely used in exhaust gas catalysts, agricultural films, optical fibers, electrode materials and other applications. Lanthanum has a toxic excitatory effect, which means that the physiological processes of the organism, especially the growth and development of plants, are stimulated by toxic substances. The nanometer lanthanum carbonate oxide is a new research field because the characteristics of the rare earth lanthanum are combined with the characteristics of the nanometer material. More and more research is beginning to focus on La 2 O 2 (CO 3 ) NPs potential environmental risks, but at present, the application of the nano lanthanum oxide carbonate in the germination of grass seeds is not reported, the influence of the nano lanthanum oxide carbonate on plants is greatly related to the concentration, the treatment time and plant materials of the nano lanthanum oxide carbonate, and the difference of the concentration and the treatment time of the medicament can cause the difference of the promotion or inhibition effect on the plants.
The existing alfalfa development technology is slow in advance, long in breeding working period and low in germination rate, the research of improving the salt tolerance of alfalfa seeds by utilizing a nano-initiation technology is fresh, particularly the research of improving the salt tolerance of alfalfa seeds by utilizing the nano-lanthanum carbonate initiation technology is not reported, the existing nano-initiation technology does not reach an extremely obvious degree on the improvement of the salt tolerance of alfalfa, and the alleviation degree of salt damage is very limited. Based on this, it is highly desirable to provide a priming method for improving the salt tolerance of alfalfa seeds by those skilled in the art.
Disclosure of Invention
Aiming at the problems, the invention 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 the alfalfa in the prior art.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention relates to a seed priming method for improving salt tolerance of alfalfa, which specifically comprises the following steps:
s1: preparing nanometer priming solution by mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs,99.99% high purity, 50 nm) was dispersed in deionized water by ultrasonic vibration (100w, 40khz);
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting sunken full seeds, and sterilizing, washing and airing the seeds for later use;
s3: the initiation process comprises the steps of putting seeds into a centrifuge tube, adding a nano initiation solution to immerse the seeds, shaking the initiation solution to enable the initiation solution to be fully contacted with the seeds, putting the seeds into a dark incubator for culture, pouring out a suspension after the culture is finished, sucking water by using absorbent paper, uniformly and flatly spreading the suspension on clean absorbent paper, and drying the seeds at room temperature for subsequent germination;
s4: and (3) germinating the seeds, namely germinating and culturing the seeds initiated in the step (S3) by adopting a paper germinating method, adding a proper amount of culture solution during germination and culturing, placing the seeds in an artificial climate incubator for continuous culture, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
Further, in the step S1, the purity of the lanthanum carbonate oxide nanoparticles 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 nanometer priming solution carbonic acid lanthanum oxide in the step S1 is 0-500mg/L.
Furthermore, the concentration of the nanometer priming solution lanthanum oxide carbonate in the step S1 is 10mg/L.
Further, in the step S2, 75% ethanol is adopted for disinfection, the disinfection time is 30S, and distilled water is adopted for washing for 4-5 times.
Further, the temperature of the dark incubator in the step S3 is constant at 5-15 ℃, the incubation time is 12-36h, and the drying time is 48h.
Furthermore, the temperature of the dark incubator in the step S3 is constant at 10 ℃, and the incubation time is 24h.
Further, the germination on paper method in the step S4 is to place the seeds in a culture dish spread with filter paper.
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 is 16h each day, the dark time is 8h, and the temperature is 25 ℃.
Compared with the prior art, the invention has the beneficial effects that:
according to the method, the carbonic acid lanthanum oxide nanoparticle solution is used for treating the alfalfa seeds, and then the seeds are dried, so that the germination rate and the germination potential of the alfalfa seeds under the salt stress are obviously improved, the salt tolerance of the alfalfa is obviously enhanced, the high-efficiency and quick salt damage relieving capability is shown, and the breeding period is shortened. By adopting a seed initiation technology, the process of cultivating the alfalfa in a soil salinization environment is greatly shortened, and the urgent need of pasture salt-tolerant cultivation fields is better solved; the method has important significance for the wide-range popularization and cultivation of the alfalfa in the soil salinization area, fills up the data blank of the application of the nano material in the alfalfa seed initiation technology, and provides a basis for the application of the nano lanthanum carbonate oxide in pasture plants, particularly the alfalfa and improving the salt tolerance of the nano lanthanum carbonate oxide.
Drawings
FIG. 1 shows La concentrations of different concentrations 2 O 2 (CO 3 ) Influence of NPs initiation on germination potential of alfalfa seeds under salt stress;
FIG. 2 shows La concentrations of different concentrations 2 O 2 (CO 3 ) Influence of NPs initiation on germination rate of alfalfa seeds under salt stress;
FIG. 3 shows La concentrations 2 O 2 (CO 3 ) Influence of NPs initiation on germination indexes of alfalfa seeds under salt stress;
FIG. 4 shows La concentrations of different concentrations 2 O 2 (CO 3 ) Influence of NPs initiation on germination peaks of alfalfa seeds under salt stress;
FIG. 5 shows La concentrations of different concentrations 2 O 2 (CO 3 ) Influence of NPs priming on the root length and seedling length of alfalfa germinating seeds under salt stress.
Detailed Description
In order that the objects and advantages of the invention will be more clearly understood, the invention is further described in detail below with reference to examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The instruments, reagents, materials and the like used in the following examples are conventional instruments, reagents, materials and the like in the prior art and are commercially available in a normal manner unless otherwise specified. Unless otherwise specified, the experimental methods, detection methods, and the like in the following examples are conventional experimental methods, detection methods, and the like in the prior art.
Example 1
The seed priming method for improving the salt tolerance of alfalfa provided by the embodiment specifically comprises the following steps:
s1: preparing nanometer priming solution by mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (99.99 percent high purity, 50 nm) is dispersed in deionized water through ultrasonic vibration (100W, 40kHz), and ultrasonic vibration is carried out for 30min to prepare 10mg/L carbonic acid lanthanum oxide nano priming solution; wherein the lanthanum oxide carbonate nanoparticles are purchased from Ron (rhawn) corporation;
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting the sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifugal tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the seeds into a 10 ℃ constant-temperature dark incubator to be cultured for 24 hours, pouring out suspension after the culture is finished, sucking water by using absorbent paper, evenly and flatly spreading the seeds on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating the seeds, performing 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 climate incubator for continuous culture for 10 days, wherein the illumination time of each day in the artificial incubator is 16h, the dark time is 8h, the temperature is 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white exposed parts as a germination standard.
Example 2
The seed priming method for improving the salt tolerance of alfalfa specifically comprises the following steps:
s1: preparing nanometer priming solution, and mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (nitric phosphate), 99.99 percent high purity and 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40kHz), and ultrasonic vibration is carried out for 30min to prepare 50mg/L carbonic acid lanthanum oxide nano priming solution; wherein the lanthanum oxide carbonate nanoparticles are purchased from Rohn (rhawn);
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting the sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifuge tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the mixture into a constant-temperature dark incubator at 5 ℃ for culturing for 36 hours, pouring out a suspension after the culture is finished, sucking water by using absorbent paper, uniformly and evenly spreading the suspension on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating the seeds, performing 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 climate incubator for continuous culture for 10 days, wherein the illumination time of each day in the artificial incubator is 16h, the dark time is 8h, the temperature is 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white exposed parts as a germination standard.
Example 3
The seed priming method for improving the salt tolerance of alfalfa specifically comprises the following steps:
s1: preparing nanometer priming solution by mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (nitric phosphate), 99.99 percent high purity and 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40kHz), and ultrasonic vibration is carried out for 30min to prepare 100mg/L carbonic acid lanthanum oxide nano priming solution; wherein the lanthanum oxide carbonate nanoparticles are purchased from Ron (rhawn) corporation;
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting the sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifuge tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the mixture into a 15 ℃ constant-temperature dark incubator to be cultured for 12 hours, pouring out a suspension after the culture is finished, sucking water by using absorbent paper, evenly and flatly spreading the mixture on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating 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 climate incubator for continuous culture for 10 days, wherein the illumination time of each day in the artificial incubator is 16h, the dark time is 8h, the temperature is 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
Example 4
The seed priming method for improving the salt tolerance of alfalfa specifically comprises the following steps:
s1: preparing nanometer priming solution, and mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (nitric phosphate), 99.99 percent high purity and 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40kHz), and ultrasonic vibration is carried out for 30min to prepare 200mg/L carbonic acid lanthanum oxide nano priming solution; wherein the lanthanum oxide carbonate nanoparticles are purchased from Ron (rhawn) corporation;
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifuge tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the mixture into a constant-temperature dark incubator at 13 ℃ for culturing for 20 hours, pouring out a suspension after the culture is finished, sucking water by using absorbent paper, uniformly and evenly spreading the suspension on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating 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 climate incubator for continuous culture for 10 days, wherein the illumination time of each day in the artificial incubator is 16h, the dark time is 8h, the temperature is 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
Example 5
The seed priming method for improving the salt tolerance of alfalfa provided by the embodiment specifically comprises the following steps:
s1: preparing nanometer priming solution by mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (99.99 percent high purity, 50 nm) is dispersed in deionized water through ultrasonic vibration (100W, 40kHz), ultrasonic vibration is carried out for 30min, 500mg/L carbonic acid lanthanum oxide nanometer priming solution is prepared,wherein the lanthanum oxide carbonate nanoparticles are purchased from Rohn (rhawn);
s2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting the sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifugal tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the seeds into a constant-temperature dark incubator at 8 ℃ for culturing for 18 hours, pouring out suspension after the culture is finished, sucking water by using absorbent paper, evenly and flatly spreading the seeds on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating the seeds initiated in the step (S3) by adopting a paper germination method, placing the seeds initiated by different nano initiation solutions 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 climate incubator for continuous culture for 10 days, wherein the illumination time of the artificial incubator every day is 16h, the dark time is 8h, the temperature is 25 ℃, adding distilled water regularly every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
Example 6
The seed priming method for improving the salt tolerance of alfalfa provided by the embodiment comprises the following specific steps:
s1: preparing nanometer priming solution, and mixing lanthanum oxide carbonate nanoparticles (La) 2 O 2 (CO 3 ) NPs (nitric phosphate), 99.99 percent high purity and 50 nm) are dispersed in deionized water through ultrasonic vibration (100W, 40kHz), and ultrasonic vibration is carried out for 30min to prepare 0mg/L lanthanum oxide carbonate nano priming solution.
S2: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting the sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s3: the initiation process comprises the steps of putting seeds into a 10ml centrifuge tube, adding 2ml of nano initiation liquid to immerse the seeds, shaking the initiation liquid to enable the initiation liquid to be fully contacted with the seeds, putting the mixture into a 9 ℃ constant-temperature dark incubator for culture for 30 hours, pouring out a suspension after the culture is finished, sucking water by using absorbent paper, evenly and flatly spreading the mixture on clean absorbent paper, and drying the seeds for 48 hours at room temperature for a subsequent germination test;
s4: and (2) germinating the seeds, namely germinating and culturing the seeds initiated in the step (S3) by adopting a paper germinating method, placing the seeds subjected to initiation treatment by different nano initiating solutions in a culture dish paved with 2 layers of filter paper, adding 5ml of 200mM NaCl culture solution during germination and culturing, placing the seeds in an artificial climate incubator for continuous culture for 10 days, wherein the artificial incubator has the light time of 16h every day, the dark time of 8h and the temperature of 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
Comparative example 1
The comparative example is used as a CK group, and the germination of the alfalfa salt tolerance seeds of the comparative example specifically comprises the following steps:
s1: collecting and treating seeds, namely removing impurities and shrunken alfalfa seeds for later use, selecting 200 alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting sinking full seeds, disinfecting with 75% ethanol for 30s, washing with distilled water for 4-5 times, and airing for later use;
s2: germinating the seeds, performing germination culture on the selected alfalfa seeds 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 climate incubator for continuous culture for 10 days, wherein the illumination time of each day in the artificial incubator is 16h, the dark time is 8h, the temperature is 25 ℃, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
The germination rates, germination vigors, germination indexes and germination peak values of examples 1 to 6 and comparative example 1 were calculated, respectively, the root lengths and seedling lengths were measured, and the average values were obtained by parallel measurement 5 times, respectively, and the measurement results are shown in FIGS. 1 to 5.
Referring to the attached drawings 1-5, compared with comparative example 1 (CK), the germination potential of the alfalfa seeds after being initiated by the nano lanthanum oxide carbonate initiation solution with the concentration of 10mg/L at 10 ℃ is improved to 58 percent from the control, the germination rate is improved to 72 percent from the control 10 percent, the germination index is improved to 20.31 from the control 2.08, the germination peak value is improved to 12.5 from the control 2, the root length is improved to 23.41mm from the control 11.79 mm, the seedling length is improved to 15.51mm from the control 9.75mm, the germination rate and the germination speed of the alfalfa seeds under salt stress are obviously improved, and the influence of salt damage on the growth of the seedlings is relieved.
In conclusion, the seed initiation method for improving the salt tolerance of the alfalfa is characterized in that 0-500mg/L lanthanum oxide carbonate nanoparticles (La) are used 2 O 2 (CO 3 ) NPs,50 nm) solution is used for treating the alfalfa seeds for 24 hours in the dark at constant temperature, and then the seeds are dried, 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. The method has important significance for the wide-range popularization and cultivation of the alfalfa in the soil salinization area, and provides a basis for the application of the nano lanthanum oxide carbonate in pasture, particularly the alfalfa, and the improvement of the salt tolerance of the nano lanthanum oxide carbonate.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A seed priming method for improving salt tolerance of alfalfa is characterized by comprising the following steps: the method comprises the following steps:
s1: preparing a nano priming solution, namely dispersing lanthanum oxide carbonate nano particles in deionized water through ultrasonic vibration;
s2: collecting and processing seeds, selecting alfalfa seeds with uniform size, full particles and uniform color, washing the seeds with clear water, selecting sunken full seeds, disinfecting, washing and drying;
s3: the initiation process comprises the steps of putting seeds into a centrifuge tube, adding a nano initiation solution to immerse the seeds, fully contacting the seeds, putting the seeds into a dark incubator for culture, pouring out a suspension after the culture is finished, and drying the seeds at room temperature;
s4: and (4) germinating the seeds, namely germinating and culturing the seeds initiated in the step (S3) by adopting a paper germination method, adding a proper amount of culture solution during germination and culturing, placing the seeds into an incubator for continuous culturing, periodically adding distilled water every day, and recording the germination number of the seeds every day by taking the white dew as a germination standard.
2. The method of claim 1, wherein the method comprises the steps of: the purity of the lanthanum carbonate oxide nanoparticles 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 method of claim 1, wherein the method comprises the steps of: the concentration of the nanometer initiation liquid lanthanum oxide carbonate in the step S1 is 0-500mg/L.
4. The method of claim 3, wherein the method comprises: the concentration of the nanometer initiation liquid lanthanum oxide carbonate in the step S1 is 10mg/L.
5. The method of claim 1, wherein the method comprises the steps of: and in the step S2, 75% ethanol is adopted for disinfection, the disinfection time is 30S, and distilled water is adopted for washing for 4-5 times.
6. The method of claim 1, wherein the method comprises the steps of: and in the step S3, the temperature of the dark incubator is kept at 5-15 ℃, the culture time is 12-36h, and the drying time is 48h.
7. The method of claim 7, wherein the method comprises: and in the step S3, the temperature of the dark incubator is 10 ℃, the temperature is constant, and the culture time is 24 hours.
8. The method of claim 1, wherein the method comprises the steps of: the germination on paper method in step S4 is to place seeds in a culture dish with filter paper.
9. The method of claim 8, wherein the method comprises: the filter paper is 2 layers.
10. The method of claim 1, wherein the method comprises the steps of: the culture solution in the step S4 is 200mM NaCl solution, the culture time in the incubator is 10 days, the illumination time per day is 16h, and the temperature is 25 ℃.
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