CN114793530A - Method for promoting germination and growth of semen cassiae seeds by utilizing seaweed extract - Google Patents
Method for promoting germination and growth of semen cassiae seeds by utilizing seaweed extract Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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Abstract
The invention discloses a method for promoting germination and growth of cassia seed seeds by using seaweed extract. Belongs to the technical field of biology, and comprises the following specific operation steps: step (1), preparing raw materials; step (2), preparing an enteromorpha extracting solution; step (3), performing seed germination experiments; step (4), carrying out a cassia seed seedling growth experiment; and (5) carrying out data statistics and analysis. The seaweed extract is a safe and potential multifunctional agricultural biofertilizer; large algal blooms such as green tide, golden tide and the like burst to provide sufficient raw materials for preparing the seaweed extract; the invention provides a new way for further development and utilization of green tide outbreak enteromorpha; the method for preparing the seaweed extract has low cost, and can prepare a large amount of seaweed extract in a short time; compared with the use of chemical fertilizers and pesticides, the seaweed extract is green and has no residue.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to a method for promoting germination of semen cassiae seeds; in particular to a method for promoting the germination and growth of cassia seeds by utilizing seaweed extract.
Background
The cassia seed is used as a common traditional Chinese medicine, is sweet, salty, bitter and slightly cold in taste; has effects of clearing heat, improving eyesight, loosening bowel to relieve constipation, etc., and can be used for treating conjunctival congestion, pain, photophobia, lacrimation, headache, giddiness, dim eyesight, and constipation; modern medical research shows that cassia seed has the effects of reducing blood pressure, regulating lipid, protecting liver, improving renal function and the like, and is a representative medicine for treating cardiovascular and cerebrovascular diseases, hyperlipidemia and constipation in the traditional Chinese medicine; in recent years, with intensive studies on chemical components, pharmacological effects, clinical effects and the like of cassia seeds, cassia seeds are widely used in the fields of medicines and foods. The method for promoting seed germination comprises using plant growth regulator such as gibberellin (GA3), indoleacetic acid (IAA), naphthylacetic acid (NAA), 6-benzyladenine (6-BA), Kinetin (KT), etc.; environmental factors such as temperature and salinity promote seed germination.
Kelp is rich in various compounds such as proteins, lipids, phytohormones, carbohydrates, amino acids, osmoprotectants, antibacterial compounds and minerals; the application of the seaweed extract to agriculture is greatly stimulated due to the demand of organic agriculture and organic food; such as promoting seed germination, enhancing the health and growth of plants, improving water and nutrient absorption, resisting freezing and salt, biologically preventing and resisting plant pathogenic organisms, repairing pollutants in polluted soil, applying fertilizers and the like.
The maximum residual limit of 548 pesticides in food is set in the latest release relevant policy of China, wherein 28 pesticides are plant growth regulators, and the total amount of the plant growth regulators accounts for more than 50% of the registered plant growth regulators in China. The plant growth regulator is frequently used in the crop planting process, and is unreasonably used for a long time and has the characteristics of slow degradation and easy residue of part of the plant growth regulator, so that the residue of the plant growth regulator is inevitably caused. At present, the residual status of the plant growth regulator in crops has been reported more, and the detection rate of paclobutrazol is generally higher. The seaweed extract is used as a novel green multifunctional biological fertilizer, so that the use and the residue of chemical fertilizer and pesticide can be reduced, and a new way is provided for further development and utilization of the ulva and green tide outbreak enteromorpha.
The seaweed extract can be used as a safe and potential multifunctional agricultural biofertilizer, and the seaweed extract is used as the biofertilizer, so that the adverse effect of main nutrient deficiency can be reduced, and the use of chemical fertilizers is reduced; meanwhile, a new way is provided for further development and utilization of green tide outbreak enteromorpha.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a method for promoting germination and growth of cassia seeds by using seaweed extract; the seaweed extract can be used as a safe and potential multifunctional agricultural biofertilizer, and the seaweed extract is used as the biofertilizer, so that the adverse effect of main nutrient deficiency can be reduced, and the use of chemical fertilizers is reduced; meanwhile, a new way is provided for further development and utilization of green tide outbreak enteromorpha.
The technical scheme is as follows: the invention relates to a method for promoting germination and growth of semen cassiae seeds by utilizing seaweed extract, which utilizes the seaweed extract to promote the germination of the semen cassiae seeds and comprises the following specific operation steps:
step (1), preparing raw materials; taking seaweed (enteromorpha and ulva) as donor plant materials and semen Cassiae seeds as test materials for standby;
step (2), preparing a seaweed extract, namely crushing collected seaweed by adopting a physical method to extract the seaweed extract, and specifically, the method comprises the following steps: washing collected seaweed with clear water, washing with pure water for 3 times, drying in a 70 deg.C oven to constant weight, and breaking the dried seaweed into powder with a wall breaking machine to obtain seaweed powder; adding 1000mL of 50% ethanol into 100g of seaweed powder, performing ultrasonic extraction for 2h by using an ultrasonic cleaning machine, and standing at 4 ℃ for 24 h; centrifuging at 4250rpm for 15 min, collecting supernatant, and filtering with Whatman No. 1 filter paper; extracting the lower layer powder residue once by the method, and combining the two extraction filtrates; evaporating the filtrate to 100mL in a rotary evaporator to obtain 1g/mL seaweed extract, sequentially diluting in gradient to obtain 0.4g/mL, 0.2g/mL, 0.1g/mL and 0.05g/mL seaweed extract, and refrigerating at 4 deg.C for use;
step (3), a seed germination experiment, the operation method is as follows:
selecting semen Cassiae seeds, breaking dormancy with 4% sodium hypochlorite, and soaking in the extracted seaweed extractive solution;
specifically, the method comprises the following steps: selecting full and healthy semen Cassiae seeds, soaking and sterilizing with 4% sodium hypochlorite for 10min, and cleaning with pure water for 3-4 times; respectively adding the cleaned seeds into the seaweed extract with four concentrations and pure water as a control group, soaking for 24h, and cleaning with pure water after 5cm of seeds are submerged; placing the processed semen Cassiae seeds in a culture dish padded with two layers of filter paper, cleaning the culture dish in advance, sterilizing in an autoclave, and drying in an oven; adding 0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract and pure water into 100 seeds per culture dish, and repeating each treatment for 3 times; then respectively supplementing proper volumes of seaweed extract and pure water according to the evaporation amount every day, and replacing filter paper every three days; the culture conditions include day and night temperature of 25 deg.C/20 deg.C, photoperiod of 14h/10h, and light intensity of 100 μmol photons m -2 s -1 .；
Recording the germination condition of the seeds at a fixed time every day, and determining that the germination quantity of the seeds does not increase any more after continuous 48 hours as the end of the seed germination experiment; measuring germination vigor, germination index, vitality index and germination rate; the correlation calculation formula is as follows:
and (3) germination potential: the percentage of the number of seeds germinated in the initial recording period to the number of test seeds;
germination index ═ Σ (G) t /D t ) Wherein: g t The number of sprouts in t time, D t Corresponding germination days;
viability index ═ germination index × individual plant weight (g);
germination percentage (%) — germination number ÷ inoculation number × 100;
step (4), performing a cassia seed seedling growth experiment, namely planting cassia seeds soaked by the seaweed extracting solution in a seedling culture disc, irrigating for 2 times every day, and after the experiment begins for two weeks, selecting cassia seeds with consistent growth vigor for each treatment to perform chlorophyll, plant height and root length measurement;
soaking semen Cassiae in seaweed extractive solution for 24h, irrigating for 10ml each plant 2 times per day, setting 3 repetitions for each treatment, and randomly arranging for 3 plug trays; after two weeks from the beginning of the test, 10 cassia seeds seedlings with consistent growth vigor are selected for each treatment to carry out the measurement of each index;
Chla(mg/g FW)＝13.95*OD 665 -6.88*OD 649 ；
Chlb(mg/g FW)＝24.96*OD 649 -7.32*OD 665 ；
plant height (cm): the ruler measures the distance from the top of the seedling to the root when the seedling stands open to the ground;
major root length (cm): measuring the distance from the rhizome to the root tip of the main root by a vernier caliper;
(5) data statistics and analysis; all data are sorted and preliminarily analyzed by excel; one-way ANOVA analysis of variance and LSD multiple comparisons and the Waller-Duncan test were performed using SPSS and histograms were plotted using Origin.
Has the advantages that: compared with the prior art, the invention has the characteristics that: 1. the seaweed extract is a safe and potential multifunctional agricultural biofertilizer; 2. large algal blooms such as green tide, golden tide and the like burst to provide sufficient raw materials for preparing the seaweed extract; 3. a new way is provided for further development and utilization of green tide outbreak enteromorpha; 4. the cost for preparing the seaweed extract is low, and a large amount of seaweed extract can be prepared in a short time; 5. compared with the use of chemical fertilizers and pesticides, the seaweed extract is green and has no residue.
Drawings
FIG. 1 is a flow chart of the preparation of the present invention;
FIG. 2 is a graph comparing the effect of different seaweed extracts on Chla of the leaves of a cassia plant according to the present invention;
FIG. 3 is a graph comparing the effect of different concentrations of seaweed extract on the leaf Chlb of a cassia seed plant according to the present invention;
FIG. 4 is a graph comparing the effect of different concentrations of seaweed extract on the height of a cassia seed plant in the examples of the present invention;
FIG. 5 is a graph showing the effect of different concentrations of seaweed extract on the root length of a cassia seed plant in the examples of the present invention.
Detailed Description
The invention is further described with reference to the following figures and examples.
The invention relates to a method for promoting germination and growth of cassia seeds by using seaweed extract, which utilizes the seaweed extract to promote germination of the cassia seeds and comprises the following specific operation steps:
step (1), preparing raw materials; taking seaweed (Enteromorpha prolifera and Ulva lactuca) as donor plant material, and semen Cassiae seed as test material;
step (2), preparing a seaweed extract, namely crushing collected seaweed by adopting a physical method to extract the seaweed extract, and specifically, the method comprises the following steps: washing collected seaweed with clear water, washing with pure water for 3 times, drying in a 70 deg.C oven to constant weight, and breaking the dried seaweed into powder with a wall breaking machine to obtain seaweed powder; adding 1000mL of 50% ethanol into 100g of seaweed meal, performing ultrasonic extraction for 2h by using an ultrasonic cleaning machine, and standing at 4 ℃ for 24 h; centrifuging at 4250rpm for 15 min, collecting supernatant, and filtering with Whatman No. 1 filter paper; extracting the lower layer powder residue once by the method, and combining the two extraction filtrates; evaporating the filtrate to 100mL in a rotary evaporator to obtain 1g/mL seaweed extract, sequentially diluting in gradient to obtain 0.4g/mL, 0.2g/mL, 0.1g/mL and 0.05g/mL seaweed extract, and refrigerating at 4 deg.C for use;
step (3), a seed germination experiment, the operation method is as follows:
selecting semen Cassiae seed, breaking dormancy with 4% sodium hypochlorite, and soaking in the extracted seaweed extractive solution.
Specifically, the method comprises the following steps: selecting full and healthy semen Cassiae seeds, soaking and sterilizing with 4% sodium hypochlorite for 10min, and cleaning with pure water for 3-4 times; respectively adding the cleaned seeds into the seaweed extract with four concentrations and pure water as a control group, soaking for 24h, and cleaning with pure water after 5cm of seeds are submerged; placing the processed semen Cassiae seeds in a culture dish padded with two layers of filter paper, cleaning the culture dish in advance, sterilizing in an autoclave, and drying in an oven; 100 seeds per culture dish are added0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract, pure water, each treatment 3 times; then respectively supplementing proper volumes of seaweed extract and pure water according to the evaporation amount every day, and replacing filter paper every three days; the culture conditions include day and night temperature of 25 deg.C/20 deg.C, photoperiod of 14h/10h, and light intensity of 100 μmol photons m -2 s -1 .；
Recording the germination condition of the seeds at a fixed time every day, and determining that the germination quantity of the seeds does not increase any more after continuous 48 hours as the end of the seed germination experiment; measuring the germination vigor, the germination index, the vitality index and the germination rate; the correlation calculation formula is as follows:
and (3) germination potential: the percentage of the number of seeds germinated in the initial recording period to the number of test seeds;
germination index ═ Σ (G) t /D t ) Wherein: g t The number of sprouts in t time, D t Corresponding germination days;
viability index ═ germination index × individual plant weight (g);
germination percentage (%) — germination number ÷ inoculation number × 100;
step (4), performing a cassia seed seedling growth experiment, namely planting cassia seeds soaked by the seaweed extracting solution in a seedling culture disc, irrigating for 2 times every day, and after the experiment begins for two weeks, selecting cassia seeds with consistent growth vigor for each treatment to perform chlorophyll, plant height and root length measurement;
soaking semen Cassiae in seaweed extractive solution for 24h, irrigating for 10ml each plant 2 times per day, setting 3 repetitions for each treatment, and randomly arranging for 3 plug trays; after two weeks from the beginning of the test, 10 cassia seeds seedlings with consistent growth vigor are selected for each treatment to carry out the measurement of each index;
Chla(mg/g FW)＝13.95*OD 665 -6.88*OD 649 ；
Chlb(mg/g FW)＝24.96*OD 649 -7.32*OD 665 ；
plant height (cm): the ruler measures the distance from the top of the seedling to the root when the seedling stands open to the ground;
major root length (cm): measuring the distance from the rhizome to the root tip of the main root by a vernier caliper;
(5) data statistics and analysis; all data are sorted and preliminarily analyzed by excel; performing one-way ANOVA analysis of variance and LSD multiple comparison and Waller-Duncan test by using SPSS, and drawing a histogram by using Origin; as shown in table 1 and table 2,
TABLE 1 measurement results of pH and conductivity of seaweed extracts with different concentrations
The pH of the seaweed extract is mostly lower than that of the control group, and the conductivity of the seaweed extract shows an obvious multiple relation along with the increase of the concentration multiple of the seaweed extract, wherein the conductivity of the seaweed extract with any concentration is greater than that of the control group.
TABLE 2 Effect of different concentrations of seaweed extracts on the germination of cassia seeds
Compared with a control group, the germination potential of the cassia seed seeds can be improved by other seaweed extracting solutions with different concentrations except that the 0.4 percent ulva extracting solution has no obvious difference; compared with a control group, the enteromorpha extracting solutions with different concentrations can improve the germination index of the cassia seeds, the 0.05 percent ulva extracting solution can obviously improve the germination index of the cassia seeds, and the 0.1 percent, 0.2 percent and 0.4 percent have no obvious difference compared with the control group; in conclusion, the effect of the enteromorpha extract on promoting the germination of the semen cassiae is better than that of the ulva extract, wherein the effect of 0.05 percent of the enteromorpha extract on promoting the germination of the semen cassiae is the best.
As shown in fig. 2-3, as can be seen from fig. 2 and 3, when compared with the clear water control group, the application of the seaweed extract to irrigate the cassia seed seedlings can effectively promote the generation of the chlorophyll content, and the seaweed extract tends to increase first and then decrease with the increase of the concentration of the seaweed extract, and the chla content increases 128.10%, 98.53%, 248.12%, 183.49%, 79.72%, 200.83%, 127.87% and 91.67% respectively, and is significantly related to the control group, compared with the control group, the chla content increases 128.10%, 98.53%, 248.12%, 183.49%, and the chlb content increases 79.72%, 200.83%, 127.87% and 91.67%, respectively.
As can be seen from FIG. 4, compared with the control group, the plant height of the applied seaweed extract is larger than that of the control group, but most of the plant heights are not significantly different, wherein the plant heights of 0.05% of the enteromorpha extract, 0.1% of the enteromorpha extract, 0.2% of the enteromorpha extract and 19.14% of the ulva extract are respectively increased by 25.8%, 14.49% and 19.14% compared with the control group, and the plant heights are significantly different.
As can be seen from FIG. 5, compared with the control group, the root growth of the seaweed extract solution is significantly increased by 0.05% of the Enteromorpha prolifera extract solution from the root growth, and the seaweed extract solutions with other concentrations have no significant difference.
According to the combination of a seed germination test and a seedling growth test of the cassia seeds, the enteromorpha extract and the ulva extract have certain promotion effects on seed germination and seedling growth of the cassia seeds, wherein the concentration range of the seaweed extract is properly applied to the seaweed extract is between 0.05 and 0.2 percent.
The innovation points of the invention are as follows: 1. the preparation of the seaweed extract only needs 50% ethanol extraction except physical crushing, and the process is simple; 2. the seaweed extract has influence on the cassia seed, and the optimal application concentration is 0.05-0.2% by combining germination index and growth index; 3. a new way is provided for further development and utilization of green tide outbreak enteromorpha; 4. the seaweed extract can be used as a method for promoting seed germination, so that the use of chemical fertilizers and pesticides can be reduced.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may be made by those skilled in the art without departing from the principle of the invention.
Claims (5)
1. A method for promoting germination and growth of cassia seeds by utilizing seaweed extract is characterized by comprising the following specific operation steps of:
step (1), preparing raw materials;
step (2), preparing seaweed extract;
step (3), performing seed germination experiments;
and (4) carrying out a cassia seed seedling growth experiment.
2. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,
in step (1), the preparation of the material is specifically:
the seaweed is taken as donor plant material, and the cassia seed is taken as test material for standby.
3. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,
in the step (2), the collected seaweed is physically crushed to extract seaweed extract.
4. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,
in the step (3), the specific operation method of the seed germination experiment is as follows:
selecting semen Cassiae seed, breaking dormancy with 4% sodium hypochlorite, and soaking in the extracted seaweed extractive solution.
5. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,
in the step (4), the cassia seed seedling growth experiment specifically comprises:
and (3) planting the cassia seeds soaked in the seaweed extracting solution in a seedling culture tray, irrigating for 2 times every day, and after two weeks of the test, selecting cassia seeds with consistent growth vigor for each treatment to measure chlorophyll, plant height and root length.
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| US5201930A (en) * | 1991-09-30 | 1993-04-13 | Aqua-10 Corporation | Plant growth product |
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| CN111165116A (en) * | 2020-01-09 | 2020-05-19 | 浙江农林大学 | Alginic acid treatment method for improving seed vitality |
| US20200337315A1 (en) * | 2017-11-17 | 2020-10-29 | Bernard PORT-LOUIS | Seaweed composition |
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| US5201930A (en) * | 1991-09-30 | 1993-04-13 | Aqua-10 Corporation | Plant growth product |
| US20150011392A1 (en) * | 2012-02-13 | 2015-01-08 | Syngenta Participations Ag | Plant growth regulation |
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Application publication date: 20220729 |