Disclosure of Invention
The invention aims to provide a culture medium for improving germination rate and a preparation method thereof. Solves the problems of insufficient nutrition, easy pest and disease damage and less regeneration stubbles of the pea seedlings for a long time.
Specifically, the invention is realized by the following technical scheme:
a preparation method of a culture medium for improving germination rate comprises the following steps: adding nitrogen-containing fertilizer, inorganic phosphate and magnesium sulfate into water, heating at 50-60 deg.C for 1-2 hr, adding aluminosilicate, heating at 60-80 deg.C for 2-3 hr, filtering while hot, and drying the solid to obtain the final product.
Further preferably, the preparation method of the culture medium for improving the germination rate comprises the following steps: adding a nitrogen-containing fertilizer, inorganic phosphate accounting for 20-50% of the mass of the nitrogen-containing fertilizer and magnesium sulfate accounting for 20-50% of the mass of the nitrogen-containing fertilizer into water 1-1.2 times of the mass of the nitrogen-containing fertilizer, heating at 50-60 ℃ for 1-2 hours, adding aluminosilicate accounting for 70-90% of the mass of the nitrogen-containing fertilizer, heating at 60-80 ℃ for 2-3 hours, filtering while hot, taking out solids, and drying at 150 ℃ and 180 ℃ for 0.5-2 hours to obtain the fertilizer.
The nitrogen-containing fertilizer is a guanylurea derivative.
The structural formula of the guanylurea derivative is as follows:
the inorganic phosphate is ammonium phosphate or diammonium hydrogen phosphate.
The aluminosilicate is montmorillonite or zeolite or a mixture of the montmorillonite and the zeolite in any proportion.
The invention also discloses a culture medium for improving the germination rate, which is prepared by adopting the preparation method of the culture medium for improving the germination rate.
The invention also discloses application of the culture medium for improving the germination rate in high-yield pea seedling planting.
The concrete application is as follows:
the high-yield pea seedling planting method comprises the following steps:
(1) screening seeds: screening peas with grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and no deformity;
(2) sterilizing, namely soaking peas in a sterilizing solution for 24-36 hours, and washing the peas once with distilled water to obtain sterile peas;
the using amount of the sterilizing liquid is 5-10 times of the weight of peas;
(3) mixing seeds: adding seeds of Papilionaceae plants except semen Pisi Sativi into sterile semen Pisi Sativi, and adding 5-10 seeds of Papilionaceae plants except semen Pisi Sativi into each 100 sterile semen Pisi Sativi to obtain mixed seeds;
the seeds of the Papilionaceae plant except peas are sterilized before incorporation by the same method as that used to obtain sterile peas;
(4) accelerating germination: preparing 10-30 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding mixed seeds, spreading all the seeds without overlapping, germinating for 2-10 days at 20-25 ℃ and relative humidity of 75-85% in a dark condition, and removing buds of Papilionaceae plants except peas to obtain pea buds;
(5) culturing: transferring the pea sprouts to a seedling raising tray containing a culture medium, and culturing for 5-12 days to obtain the nutritional pea seedlings.
Preferably, the seeds of the Papilionaceae plants except peas are any one of kidney beans, chickpeas and soybeans.
Preferably, the sterilizing liquid is prepared by mixing sodium hypochlorite, 4-phenylbenzimidazole derivatives and water according to a mass ratio of (0.05-0.3) to (0.05-0.4) to 100.
Preferably, the structural formula of the 4-phenylbenzimidazole derivative is:
wherein R ═ methyl, hydroxyl, or sulfonic acid groups.
When R is hydroxyl, the specific structural formula is as follows:
the invention has the advantages that: a novel culture substrate is developed, the culture substrate adsorbs guanylurea derivatives, the guanylurea derivatives have double effects of sterilization and nutrition supply to pea seedlings, nitrate is slowly released by the pea seedlings during cultivation, the nitrate is slowly adsorbed by aluminosilicate in the culture substrate and is slowly replaced by the originally adsorbed guanylurea derivatives, a slow release effect is achieved, and the number of harvested stubbles of the pea seedlings is increased. The culture medium can be recycled to adsorb nitrogenous fertilizer again, so that the cost is saved.
Detailed Description
The present invention will be further described in detail with reference to specific examples, which are provided for illustration only and are not intended to limit the invention.
Specifically, the instrument or compound sources or CAS numbers referred to in the following examples are as follows:
pea, Latin literature name Pisum sativum Linn, variety is green pea.
Calcium-based montmorillonite, purchased from 50 mesh from manufacturer of huge rock mineral products in Lingshui county.
Zeolite, available from Okinawa Gnetsu environmental protection materials Co., Ltd, 50 mesh.
Fluorescent lamp, power 18W, luminous flux 1800 lm.
Sodium hypochlorite, CAS number: 7681-52-9.
2-methyl-4-phenylbenzimidazole, CAS No.: 110179-50-5.
2-sulfonic-4-phenylbenzimidazole, CAS No.: 1027179-57-2.
Soybean, latin literature name Glycine max (Linn.) merr, soybean variety soybean.
Phaseolus vulgaris, Latin literature name Phaseolus vulgaris, and the variety is white flower-frame bean.
Chickpeas, the Latin literature name Cicer arietinum, a variety of Wush chickpeas.
N-acetyl-5 methoxytryptamine, CAS No.: 8041-44-9.
Diamine hydrogen phosphate, CAS No.: 7783-28-0.
Magnesium sulfate, CAS No.: 7487-88-9.
CAS number: 1075753-20-6.
2-methyl-4-phenylbenzimidazole, CAS No.: 110179-50-5.
2-sulfonic-4-phenylbenzimidazole, CAS No.: 1027179-57-2.
Example 1
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is 2-methyl-4-phenylbenzimidazole;
(3) mixing seeds: adding soybean into sterile pea, adding 7 soybean into each 100 sterile pea to obtain mixed seed;
the soybeans were sterilized prior to incorporation by the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing soybean sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
the culture medium for improving the germination rate is prepared by adding nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water 1.1 times of the weight of the nitrogen-containing fertilizer, heating at 60 deg.C for 1 hr, adding
Adding zeolite 80 wt% of the nitrogenous fertilizer, heating at 70 deg.C for 2 hr, hot sieving with 300 mesh sieve, collecting solid, and drying at 170 deg.C for 1 hr to obtain the final product;
the fertilizer containing nitrogen is
Example 2
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is 2-methyl-4-phenylbenzimidazole;
(3) mixing seeds: adding beans into sterile peas, wherein 7 beans are added into each 100 sterile peas to obtain mixed seeds;
the beans are sterilized prior to incorporation by the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing bean sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
the culture medium for improving the germination rate is prepared by adding nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water 1.1 times of the weight of the nitrogen-containing fertilizer, heating at 60 deg.C for 1 hr, adding
Adding zeolite 80 wt% of the nitrogenous fertilizer, heating at 70 deg.C for 2 hr, hot sieving with 300 mesh sieve, collecting solid, and drying at 170 deg.C for 1 hr to obtain the final product;
the fertilizer containing nitrogen is
Example 3
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is 2-methyl-4-phenylbenzimidazole;
(3) mixing seeds: adding chickpeas into sterile peas, and adding 7 chickpeas into each 100 sterile peas to obtain mixed seeds;
the chickpeas were sterilized prior to incorporation using the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing chickpea sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
the culture medium for improving the germination rate is prepared by adding nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water 1.1 times of the weight of the nitrogen-containing fertilizer, heating at 60 deg.C for 1 hr, adding
Adding zeolite 80 wt% of the nitrogenous fertilizer, heating at 70 deg.C for 2 hr, hot sieving with 300 mesh sieve, collecting solid, and drying at 170 deg.C for 1 hr to obtain the final product;
the fertilizer containing nitrogen is
Example 4
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is 2-methyl-4-phenylbenzimidazole;
(3) mixing seeds: adding chickpeas into sterile peas, and adding 7 chickpeas into each 100 sterile peas to obtain mixed seeds;
the chickpeas were sterilized prior to incorporation using the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing chickpea sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
adding a nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water which is 1.1 times of the nitrogen-containing fertilizer, heating at 60 ℃ for 1 hour, adding calcium-based montmorillonite, wherein the adding amount of the calcium-based montmorillonite is 80% of the nitrogen-containing fertilizer, heating at 70 ℃ for 2 hours, sieving with a 300-mesh sieve while the calcium-based montmorillonite is hot, taking the solid, and drying at 170 ℃ for 1 hour to obtain the culture medium for improving the budding rate;
the fertilizer containing nitrogen is
Example 5
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is 2-methyl-4-phenylbenzimidazole;
(3) mixing seeds: adding chickpeas into sterile peas, and adding 7 chickpeas into each 100 sterile peas to obtain mixed seeds;
the chickpeas were sterilized prior to incorporation using the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing chickpea sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
adding a nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water which is 1.1 times of the nitrogen-containing fertilizer by mass, heating at 60 ℃ for 1 hour, adding aluminosilicate with the addition amount being 80% of the nitrogen-containing fertilizer by mass, heating at 70 ℃ for 2 hours, sieving with a 300-mesh sieve while hot, taking the solid, and drying at 170 ℃ for 1 hour to obtain the culture medium for improving the budding rate;
the fertilizer containing nitrogen is
Said aluminosilicate being
The mass ratio of zeolite to calcium-based montmorillonite is 2.5: 1.
Example 6
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the structural formula of the 4-phenylbenzimidazole derivative is as follows:
(3) mixing seeds: adding chickpeas into sterile peas, and adding 7 chickpeas into each 100 sterile peas to obtain mixed seeds;
the chickpeas were sterilized prior to incorporation using the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing chickpea sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
adding a nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water which is 1.1 times of the nitrogen-containing fertilizer by mass, heating at 60 ℃ for 1 hour, adding aluminosilicate with the addition amount being 80% of the nitrogen-containing fertilizer by mass, heating at 70 ℃ for 2 hours, sieving with a 300-mesh sieve while hot, taking the solid, and drying at 170 ℃ for 1 hour to obtain the culture medium for improving the budding rate;
the fertilizer containing nitrogen is
Said aluminosilicate being
The mass ratio of zeolite to calcium-based montmorillonite is 2.5: 1.
Example 7
A high-yield pea seedling planting method comprises the following planting steps:
(1) screening seeds: screening peas with the grain weight of 0.12-0.20g, no worm damage, no broken residue, no mildew and rot and no deformity, wherein the average grain weight is 0.15 g;
(2) sterilizing, namely soaking the screened peas in sterilizing liquid for 24 hours, and washing the peas once with distilled water with the same mass as the sterilizing liquid to obtain sterile peas;
the using amount of the sterilizing liquid is 5 times of the weight of peas;
the sterilizing liquid is prepared by mixing sodium hypochlorite, a 4-phenylbenzimidazole derivative and water according to a mass ratio of 0.1:0.2: 100;
the 4-phenylbenzimidazole derivative is as follows: 2-sulfo-4-phenylbenzimidazole;
(3) mixing seeds: adding chickpeas into sterile peas, and adding 7 chickpeas into each 100 sterile peas to obtain mixed seeds;
the chickpeas were sterilized prior to incorporation using the same method as the aseptic peas obtained;
(4) accelerating germination: preparing 15 mu mol/L N-acetyl-5 methoxyl tryptamine aqueous solution, adding the mixed seeds, spreading all the seeds without overlapping, germinating for 7 days at 20 ℃ under the condition of relative humidity of 80% and keeping out of the sun, and removing chickpea sprouts to obtain pea sprouts;
(5) culturing: transferring pea sprouts into a seedling raising tray containing a culture medium for improving the sprouting rate, wherein the thickness of the culture medium for improving the sprouting rate is 2cm, continuously illuminating for 12 hours every day by adopting common fluorescent lamp illumination, and culturing for 7 days at the temperature of 25 ℃ and the relative humidity of 60% to obtain the nutritional pea seedlings;
adding a nitrogen-containing fertilizer, 30% of diammonium hydrogen phosphate and 25% of magnesium sulfate into water which is 1.1 times of the nitrogen-containing fertilizer by mass, heating at 60 ℃ for 1 hour, adding aluminosilicate with the addition amount being 80% of the nitrogen-containing fertilizer by mass, heating at 70 ℃ for 2 hours, sieving with a 300-mesh sieve while hot, taking the solid, and drying at 170 ℃ for 1 hour to obtain the culture medium for improving the budding rate;
the fertilizer containing nitrogen is
Said aluminosilicate being
The mass ratio of zeolite to calcium-based montmorillonite is 2.5: 1.
Test example 1
Statistics of germination rate
For each 1000 pea seedlings planted according to the method of the culture medium of the invention as described in examples 1-7, the germination rates of three, five and seven crops were counted. The pea seedlings are still cultured for 5 days according to the original conditions after being harvested, and the pea seedlings are considered to be not germinated if the height of the germinated pea seedlings is less than 5cm or the pea seedlings are not germinated within 5 days, and the pea seedlings are removed. The counting base number is 1000 when the germination rate is calculated.
The statistical results are shown in table 1.
TABLE 1 sprouting Rate
|
Germination percentage in three crops (%)
|
Bud ratio in the fifth crop (%)
|
Bud ratio in seven crops (%)
|
Example 1
|
95.4
|
65.4
|
42.6
|
Example 2
|
96.3
|
66.2
|
45.9
|
Example 3
|
97.0
|
73.9
|
55.0
|
Example 4
|
97.7
|
73.5
|
54.4
|
Example 5
|
98.8
|
80.7
|
68.4
|
Example 6
|
99.3
|
82.5
|
71.5
|
Example 7
|
99.4
|
84.3
|
79.5 |
As can be seen from the above table, the pea seedling planting method provided by the invention has a high sprouting rate, the conventional planting method generally can only produce one crop, and the more advanced method in the prior art generally produces three crops, but the method provided by the invention generally can produce five crops, and at most 77.5% of the crops can produce seven crops, which is far beyond the prior art.
Test example 2
Determination of pea seedling total phenol content
The total phenol content of the pea seedlings cultivated by the culture medium of the invention in the examples 1-7 is measured by Folin-Ciocalteu colorimetry.
The test results are shown in table 2.
TABLE 2 Total phenol content
|
Total phenol content (mg/g)
|
Example 1
|
686.4
|
Example 2
|
692.3
|
Example 3
|
725.3
|
Example 4
|
739.0
|
Example 5
|
794.8
|
Example 6
|
812.9
|
Example 7
|
846.2 |
As can be seen from the table, the pea seedlings planted by the pea seedling planting method provided with the culture medium have good nutritional value, and the total phenol content is more than 650 mg/g. Wherein the planting method provided by the embodiment 7 is particularly optimal, and the total phenol content of the obtained first crop pea seedlings can be as high as 846.2 mg/g.