CN113277889B - Algae-bacterium composite nutrient solution and preparation method and application thereof - Google Patents

Algae-bacterium composite nutrient solution and preparation method and application thereof Download PDF

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CN113277889B
CN113277889B CN202110699970.5A CN202110699970A CN113277889B CN 113277889 B CN113277889 B CN 113277889B CN 202110699970 A CN202110699970 A CN 202110699970A CN 113277889 B CN113277889 B CN 113277889B
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崔岩
罗光宏
陈叶
刘海燕
边建文
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Hexi University
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Abstract

The invention relates to the technical field of plant nutrient solutions, in particular to an algae-bacterium composite nutrient solution and a preparation method and application thereof. An algae-bacterium composite nutrient solution is prepared from the following raw materials in parts by mass: 800-1200 parts of debarya liquid, 10-50 parts of composite bacillus liquid, 10-50 parts of pseudomonas liquid and 10-50 parts of photosynthetic bacteria liquid. The invention can obviously improve the yield, quality and resistance of crops, especially can obviously improve the growth vigor of pepper and tomato seeds and the growth promoting effect of seedlings, and can reduce the use of chemical fertilizers and pesticides, thereby achieving two purposes.

Description

Algae-bacterium composite nutrient solution and preparation method and application thereof
Technical Field
The invention relates to the technical field of plant nutrient solutions, in particular to an algae-bacterium composite nutrient solution and a preparation method and application thereof.
Background
China is a big agricultural country, and in order to meet the increasing needs of population, chemical fertilizers are largely used in agricultural production for the last decades to promote the yield and income increase of agricultural products, so that soil, water and agricultural products are polluted, and microorganisms which are dependent on the growth and survival of plants in the soil are killed, so that the ecological balance of farmlands in high-yield areas of many crops is disordered, the more diseases and insect pests are treated, the more problems of farmland pollution, water pollution, quality reduction of agricultural products and the like are serious, and irreversible influence is caused on the environment.
Along with the continuous improvement of living standard, people pay more and more attention to healthy diet and balanced nutrition, and from the first time, only paying attention to big fish meat, drinking milk and supplementing calcium, to the current time, paying attention to healthy diet which is non-transgenic and has no pesticide residue, and paying attention to balanced nutrition of vitamins and trace elements. Therefore, in the current vegetable and fruit planting, a large amount of chemical fertilizers and pesticides are required to be avoided for killing insects, otherwise, the potential harm to human bodies and the environment is caused.
The active phycomycete cell nutrient solution is one kind of green organic biological fertilizer, and is prepared with soil micro algae as main material and through adding composite active bacteria, mixing and culturing to promote the accumulation of bioactive matter. The nutrient solution fixes N in the air through the metabolism activity of microorganisms 2 /CO 2 The increase of the effective nitrogen/organic matter content of the soil is realized, a healthy soil microbial community structure is constructed, the soil fertility is improved, and the sustainable utilization of the soil is realized. The fertilizer plays a good role in increasing crop yield, improving fruit quality, enhancing crop resistance, reducing the use of pesticides and fertilizers and protecting ecological balance.
Therefore, the problem to be solved by the skilled in the art is how to provide an algae activated cell nutrient solution, which can significantly improve the yield quality and resistance of crops and reduce the use of chemical fertilizers and pesticides.
Disclosure of Invention
The invention aims to provide an algae-bacteria composite nutrient solution, and a preparation method and application thereof, the invention not only can improve the germination rate, the germination vigor, the germination index and the vitality index of tomato and pepper seeds, but also can obviously promote the growth of tomato and pepper seedlings, and the use of the invention can reduce the use of chemical fertilizers and pesticides, thereby playing a good role in protecting ecological balance.
In order to achieve the above object, the present invention provides the following technical solutions:
an algae bacterium composite nutrient solution is prepared from the following raw materials in parts by mass: 800-1200 parts of debarya liquid, 10-50 parts of composite bacillus liquid, 10-50 parts of pseudomonas liquid and 10-50 parts of photosynthetic bacteria liquid.
Preferably, the debarked algae liquid OD 680 The value is 0.8 to 1.2; OD of composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid 600 The value is 0.8 to 1.2.
Preferably, the bacillus subtilis, the bacillus licheniformis and the bacillus megaterium are adopted as the bacillus subtilis; the number ratio of the bacillus subtilis to the bacillus licheniformis to the bacillus megaterium is 1-3: 1-3.
The invention also provides a preparation method of the algae-bacterium composite nutrient solution, which comprises the following steps:
(1) Respectively culturing the debaryomyces, the composite bacillus, the pseudomonas and the photosynthetic bacteria by using special culture media until the logarithmic growth phase to obtain a debaryomyces liquid, a composite bacillus liquid, a pseudomonas liquid and a photosynthetic bacteria liquid;
(2) Respectively centrifuging the chlamydomonas debarkii, the composite bacillus, the pseudomonas and the photosynthetic bacteria liquid obtained in the step (1) to discard supernatant, respectively adding the same volume of the specific culture medium for the chlamydomonas debarkii for cleaning, resuspending, adjusting OD value, mixing, and culturing for 15-25 days at 25-27 ℃.
Preferably, the special culture medium for culturing the debarked algae is Tris 2.42g 4 Cl 0.4g，MgSO 4 ·7H 2 O 0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to constant volume to 1L; the culture temperature of the chlamydomonas debarkii is 19-21 ℃, and the pH value is 6.4-6.6.
Preferably, the special culture medium for culturing the bacillus compositus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose and KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 28-32 ℃, and the pH value is 6.8-7.2.
Preferably, the culture medium special for culturing the pseudomonas is 10g/L of tryptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 1L of distilled water; the culture temperature of the pseudomonas is 26-30 ℃, and the pH value is 6.8-7.0.
Preferably, the special culture medium for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 O0.25g, acetic acid 0.5g, yeast extract 0.2g, distilled water 1L; the culture temperature of the photosynthetic bacteria is 27-29 ℃, and the pH value is 7.1-7.3.
Preferably, the centrifugal conditions of the chlamydomonas debarkii liquid in the step (2) are as follows: 4500-5500 r/min, 4-6 min; the centrifugation conditions of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid are as follows: 7500-8500 r/min, 4-6 min.
Furthermore, the invention also provides application of the algae-bacterium composite nutrient solution in cultivation of hot pepper or tomatoes.
Furthermore, the phycomycete composite nutrient solution needs to be diluted when in use. In the cultivation of pepper, the effect is best when the pepper is diluted to 100-200 times of solution for use. In the cultivation of tomatoes, the effect is best when the tomato cultivation liquid is diluted to 100-300 times.
Compared with the prior art, the invention has the following beneficial effects:
the invention takes soil microalgae as a main material, adds a composite active strain, and performs mixed culture on the soil microalgae and the composite active strain to promote the accumulation of bioactive substances. The nutrient solution fixes N in the air through the metabolism activity of microorganisms 2 /CO 2 The increase of the effective nitrogen/organic matter content of the soil is realized, a healthy soil microbial community structure is constructed, the soil fertility is improved, and the sustainable utilization of the soil is realized. The fertilizer plays a good role in increasing crop yield, improving fruit quality, enhancing crop resistance, reducing pesticide and fertilizer application and protecting ecological balance.
2. The invention can obviously improve the germination rate, the germination vigor, the germination index and the vitality index of the pepper and tomato seeds and has obvious promotion effect on the growth of pepper and tomato seedlings.
3. Culturing microalgae: the adopted microalgae species are Chlamydomonas debarkii which are automatically separated from a soil habitat in a laboratory. The culture medium is simplified TAP culture medium, and is sterilized at 121 deg.C under high pressure for 20min, cooled for use, and subjected to microalgae amplification culture. The solid culture medium is added with 1.5 to 2.0 percent of agar powder. Under the aseptic condition, a single algae colony on the solid culture medium is picked out by an inoculating loop and inoculated into a 12-hole plate, and the 12-hole plate is placed in a light incubator for culture for 3 to 4 days; transferring to a 250ml conical flask containing 100ml of culture solution for culture for 12-15 days; transferring into 1000ml conical flask containing 500ml culture solution, culturing for 10-15 days, and performing step-by-step enlarged culture to obtain desired algae biomass. Shaking the algae for 2 times every day to prevent the algae from precipitating and agglomerating.
4. And (3) culturing bacteria: the bacterial species used include Bacillus compositus, photosynthetic bacteria and Pseudomonas. The strains can rapidly propagate and colonize in plant rhizosphere, body surface or in vivo and in soil in large quantity, effectively repel, prevent and interfere colonization and infection of plant pathogenic microorganisms on plants, thereby achieving the effects of bacteriostasis and disease prevention, can secrete active substances, activate a plant defense system, enhance the immunity and disease resistance of crops, have good effects on regulating plant growth, preventing and treating plant diseases and insect pests and regulating soil micro-ecological balance, and have certain effects of fixing nitrogen and fixing CO 2 Of the cell.
Detailed Description
The invention provides an algae-bacteria composite nutrient solution which is prepared from the following raw materials in parts by weight: 800-1200 parts of debarya liquid, 10-50 parts of composite bacillus liquid, 10-50 parts of pseudomonas liquid and 10-50 parts of photosynthetic bacteria liquid;
preferably 850-1150 parts of debaryomyces liquid, 20-40 parts of composite bacillus liquid, 20-40 parts of pseudomonas liquid and 20-40 parts of photosynthetic bacteria liquid;
further preferably 900 to 1100 parts of debarked algae solution, 25 to 35 parts of composite bacillus solution, 25 to 35 parts of pseudomonas solution and 25 to 35 parts of photosynthetic bacteria solution;
more preferably 1000 parts of chlamydomonas debarkii solution, 30 parts of composite bacillus solution, 30 parts of pseudomonas solution and 30 parts of photosynthetic bacteria solution.
In the present inventionThe Duabanga solution OD 680 The value is 0.8 to 1.2; preferably 0.9 to 1.1, and more preferably 1.0.
In the present invention, the OD of the composite bacillus liquid, the Pseudomonas liquid and the photosynthetic bacteria liquid 600 The value is 0.8 to 1.2; preferably 0.9 to 1.1, and more preferably 1.0.
In the invention, the composite bacillus is bacillus subtilis, bacillus licheniformis and bacillus megaterium; the number ratio of the bacillus subtilis to the bacillus licheniformis to the bacillus megaterium is 1-3: 1-3; preferably 1-2: 1-2; further preferably 1: 1.
The invention also provides a preparation method of the algae-bacteria composite nutrient solution, which comprises the following steps:
(1) Respectively culturing the debaryomyces, the composite bacillus, the pseudomonas and the photosynthetic bacteria by using special culture media until the logarithmic growth phase to obtain a debaryomyces liquid, a composite bacillus liquid, a pseudomonas liquid and a photosynthetic bacteria liquid;
(2) Respectively centrifuging the chlamydomonas debarkii, the composite bacillus, the pseudomonas and the photosynthetic bacteria liquid obtained in the step (1) to discard supernatant, respectively adding the same volume of a special culture medium for the chlamydomonas debarkii for cleaning, resuspending, adjusting OD value, mixing, and culturing for 15-25 days at 25-27 ℃; preferably 25.5-26.5 ℃, and culturing for 17-23 days; further preferably at 26 ℃, culturing for 19-21 days; more preferably at 26 ℃ for 20 days.
Preferably, the special culture medium for culturing the debarkinson is Tris 2.42g 4 Cl 0.4g，MgSO 4 ·7H 2 O 0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to a constant volume of 1L; the culture temperature of the chlamydomonas debarkii is 19-21 ℃, and the pH value is 6.4-6.6.
In the invention, the special culture medium for culturing the compound bacillus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose and KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 28-32 ℃, and the pH value is 6.8-7.2; preferably 29-31 ℃, pH6.9-7.1; further preferably 30 ℃ and pH7.0.
In the invention, the culture medium special for culturing the pseudomonas is 10g/L of tryptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 1L of distilled water; the culture temperature of the pseudomonas is 26-30 ℃, and the pH value is 6.8-7.0; preferably, the culture temperature is 27-29 ℃, and the pH is 6.9; more preferably, the culture temperature is 28 ℃ and the pH is 6.9.
In the invention, the special culture medium for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 0.25g of O, 0.5g of acetic acid, 0.2g of yeast extract and 1L of distilled water; the culture temperature of the photosynthetic bacteria is 27-29 ℃, and the pH value is 7.1-7.3; preferably, the culture temperature is 28 ℃ and pH7.2.
In the invention, the centrifugal conditions of the Dunaliella bardawil liquid in the step (1) are as follows: 4500-5500 r/min, 4-6 min; preferably 4600 to 5400r/min,5min; further preferably 4800-5200 r/min,5min; more preferably 5000r/min,5min.
In the invention, the centrifugation conditions of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid are as follows: 7500-8500 r/min, 4-6 min; preferably 7700-8300 r/min,5min; further preferably 7900-8100 r/min,5min; more preferably 8000r/min,5min.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
A preparation method of an algae-bacterium composite nutrient solution comprises the following steps:
(1) Respectively culturing the debark algae, the composite bacillus (the number ratio of bacillus subtilis to bacillus licheniformis to bacillus megaterium is 1: 3), the pseudomonas and the photosynthetic bacteria by using special culture media until after logarithmic growth phase to obtain a debark algae liquid, a composite bacillus liquid, a pseudomonas liquid and a photosynthetic bacteria liquid, then respectively centrifuging (the centrifugation condition of the debark algae liquid is 4500r/min and 4min; the centrifugation condition of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid is 7500r/min and 4 min), respectively adding the same volume of the debark algae with the culture media after discarding supernatant, cleaning and re-suspending;
(2) Respectively adjusting OD values (OD of the debaryomyces solution) of the resuspended debaryomyces solution, the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid in the step (1) 680 The value was adjusted to 0.8; OD of composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid 600 Adjusting the value to 0.8), mixing 800 parts of the debark algae solution, 10 parts of the composite bacillus solution, 10 parts of the pseudomonas solution and 10 parts of the photosynthetic bacteria solution according to the proportion, and culturing for 15 days at 25 ℃;
the special culture medium for culturing the debarkinson is Tris 2.42g 4 Cl 0.4g，MgSO 4 ·7H 2 O0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to a constant volume of 1L; the culture temperature of the chlamydomonas debarkii is 19 ℃, and the pH value is 6.4;
the special culture medium for culturing the composite bacillus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose and KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 28 ℃, and the pH value is 6.8;
the culture medium special for culturing the pseudomonas is 10g/L of tryptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 1L of distilled water; the culture temperature of the pseudomonas is 26 ℃, and the pH value is 6.8;
the culture medium special for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 0.25g of O, 0.5g of acetic acid, 0.2g of yeast extract and 1L of distilled water; the culture temperature of the photosynthetic bacteria is 27 ℃, and the pH value is 7.1.
Example 2
A preparation method of an algae-bacteria composite nutrient solution comprises the following steps:
(1) Respectively culturing the debaryomyces delavayi and the composite bacillus (the number ratio of bacillus subtilis to bacillus licheniformis to bacillus megaterium is 3: 1), and culturing the pseudomonas and the photosynthetic bacteria by using special culture media until a logarithmic growth phase is reached to obtain a debaryomyces delavayi liquid, a composite bacillus liquid, a pseudomonas liquid and a photosynthetic bacteria liquid, then respectively centrifuging (the centrifugation condition of the debaryomyces delavayi liquid is 5500r/min and 6min; the centrifugation condition of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid is 8500r/min and 6 min), discarding supernatant, and respectively adding the same volume of the debaryomyces delavayi liquid to clean and resuspend the same volume of the culture media;
(2) Respectively adjusting OD values (OD of the debaryomyces solution) of the resuspended debaryomyces solution, the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid in the step (1) 680 The value was adjusted to 1.2; OD of composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid 600 The value is adjusted to 1.2), 1200 parts of the chlamydomonas debarkii liquid, 50 parts of the composite bacillus liquid, 50 parts of the pseudomonas liquid and 50 parts of the photosynthetic bacteria liquid are mixed according to the proportion, and the mixture is cultured for 25 days at the temperature of 27 ℃;
the special culture medium for culturing the debarkinson is Tris 2.42g 4 Cl 0.4g，MgSO 4 ·7H 2 O0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to constant volume to 1L; the culture temperature of the chlamydomonas debarkii is 21 ℃, and the pH value is 6.6;
the special culture medium for culturing the composite bacillus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose and KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 32 ℃, and the pH value is 7.2;
the culture medium special for culturing the pseudomonas is 10g/L of tryptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 1L of distilled water; the culture temperature of the pseudomonas is 26-30 ℃, and the pH value is 6.8-7.0;
the culture medium special for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 0.25g of O, 0.5g of acetic acid, 0.2g of yeast extract and 1L of distilled water; the culture temperature of the photosynthetic bacteria is 29 ℃, and the pH value is 7.3.
Example 3
A preparation method of an algae-bacteria composite nutrient solution comprises the following steps:
(1) Respectively culturing the debark algae, the composite bacillus (the number ratio of bacillus subtilis to bacillus licheniformis to bacillus megaterium is 1: 1), the pseudomonas and the photosynthetic bacteria by using special culture media until after logarithmic growth phase to obtain debark algae liquid, composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid, then respectively centrifuging (the centrifugation conditions of the debark algae liquid are 5000r/min and 5min; the centrifugation conditions of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid are 8000r/min and 5 min), respectively adding the same volume of the debark algae with the culture media after discarding supernatant, cleaning and resuspending;
(2) Respectively adjusting OD values (OD of the debaryomyces solution) of the resuspended debaryomyces solution, the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid in the step (1) 680 The value is adjusted to 1; OD of composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid 600 The value is adjusted to 1), 800-1200 parts of the debaryomyces solution, 30 parts of the composite bacillus solution, 30 parts of the pseudomonas solution and 30 parts of the photosynthetic bacteria solution are mixed according to the proportion, and the mixture is cultured for 20 days at 26 ℃;
the special culture medium for culturing the debarkinson is Tris 2.42g 4 Cl 0.4g，MgSO 4 ·7H 2 O0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to constant volume to 1L; the culture temperature of the chlamydomonas debarkii is 20 ℃, and the pH value is 6.5;
the special culture medium for culturing the composite bacillus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose and KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 30 ℃, and the pH value is 7.0;
the culture medium special for culturing the pseudomonas is 10g/L of tryptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 1L of distilled water; the culture temperature of the pseudomonas is 28 ℃, and the pH value is 6.9;
the culture medium special for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 0.25g of O, 0.5g of acetic acid, 0.2g of yeast extract and 1L of distilled water; the culture temperature of the photosynthetic bacteria is 28 ℃, and the pH value is 7.2.
Example 4
After culturing the algal fungus composite nutrient solution obtained in example 3 for 20 days, the ratio of the culture medium to the culture medium was adjusted according to 1: 100. 1:300 and 1, 400 were diluted in 4 total proportions and the germinated paper was soaked and treated with distilled water (CK) as a control, each treatment being repeated 3 times.
Rinsing the pepper seeds with distilled water, washing off dirt on the surfaces of the pepper seeds, sucking the pepper seeds with filter paper, wrapping the pepper seeds with clean gauze, then sterilizing the pepper seeds twice with 1 percent potassium permanganate, sterilizing the pepper seeds for 10min for the 1 st time, and washing the pepper seeds for 4 to 6 times with sterile water; sterilizing for 8min at 2 nd time, and washing with sterile water. Selecting healthy receptor seeds with uniform color and luster and size, dibbling 30 seeds in the germination box at equal intervals respectively to ensure that proper intervals are reserved among the seeds, placing the seeds in a constant-temperature incubator at 25 ℃, supplementing water for 1 time by using distilled water every day to supplement lost water, recording the germination number of the seeds every day, and calculating the germination rate, the germination vigor, the germination index and the vitality index of the pepper seeds; after the test is finished 15 days later, the length of all seedlings and the length of roots in each germination box are measured by a graduated scale; the seedling weight and the root weight of the seedlings are measured by an analytical balance.
TABLE 1 Effect of different concentrations of nutrient solutions on pepper seeds
Figure BDA0003129419960000121
As can be seen from table 1, when the pepper seeds are soaked by using the phycomycete composite nutrient solution, the germination rate and the germination vigor of the seeds after 200 times dilution are the highest, respectively 96.7% and 76.3%, which are respectively 12% and 15.9% higher than those of the seeds obtained by CK treatment, and the germination rates of the seeds after 100 times dilution, 300 times dilution and 400 times dilution treatment are respectively 92.2%, 86.7% and 85.6%, which are respectively 7.5%, 2% and 0.9% higher than those of the seeds obtained by CK treatment; the germination potential is 72.4%, 66.8% and 64.7% in sequence, and is respectively increased by 12%, 6.4% and 4.3% compared with CK. For germination rate, the difference between the 100-fold and 200-fold diluted treatment groups and the rest experimental groups reaches a significant level (P < 0.05), but the difference between the 100-fold and 200-fold treated groups is not significant (P > 0.05), and the difference is not significant when the dilution is 300-fold and 400-fold compared with CK (P > 0.05). For the germination potential, the difference of different treatment groups compared with the control group reaches a significant level (P < 0.05).
As can be seen from table 1, when pepper seeds were soaked in the phycomycete mixed nutrient solution, the differences between the treatment groups diluted 100 times, 200 times and 300 times compared with the control group were significant (P < 0.05), and the differences between the treatment groups diluted 100 times, 200 times and 300 times and 400 times compared with the control group were not significant (P > 0.05). Compared with a control, the vitality indexes of the seeds diluted by 100 times, 200 times, 300 times and 400 times are respectively increased by 93.126, 73.589, 76.761 and 17.769, the other treatments except for the 400 times treatment group are all compared with the control, the difference is significant (P < 0.05), the 100 times treatment group is significantly different from the 200 times and 300 times treatment group (P < 0.05), and the 300 times and 400 times treatment group are not significantly different (P > 0.05).
TABLE 2 Effect of different concentrations of nutrient solutions on pepper seedlings
Figure BDA0003129419960000131
As can be seen from Table 2, the seedlings of the pepper treated by 100-fold, 200-fold and 300-fold dilution were increased by 2.9, 2.18 and 0.75mm, respectively, compared with the control group, the difference was significant (P < 0.05), and the seedling of the pepper treated by 400-fold dilution was increased by 0.1mm compared with the control group, without significant difference (P > 0.05). The seedling length of the treated pepper seedlings is obviously better than CK, and the seedling length increase of each treatment is from large to small, namely 100 times, 200 times, 300 times, 400 times and CK in sequence. The higher the concentration of the applied nutrient solution is, the stronger the promotion effect on the growth of the pepper seedlings is.
As can be seen from Table 2, the root lengths of the pepper seedlings diluted 100 times, 200 times and 300 times are respectively increased by 7.11 mm, 5.55 mm and 4.34mm compared with the control, the difference is all significant (P < 0.05), and the 400 times of treatment is increased by 1.67mm compared with the control without significant difference (P > 0.05).
As can be seen from table 2, the roots of the pepper seedlings diluted 100 times, 200 times and 300 times are increased by 0.030, 0.018 and 0.006mm respectively compared with the control, the difference is significant (P < 0.05) compared with the control, the difference between the treatment groups is significant (P < 0.05), and the 400 times of treatment is not significant (P > 0.05) compared with the control.
As can be seen from Table 2, the seedling weight of the pepper seedlings diluted 100 times and 200 times is increased by 0.005g and 0.004g respectively compared with the control, the difference reaches a significant level (P is less than 0.05), and the seedling weight of the pepper seedlings diluted 300 times and 400 times is increased by 0.002g and 0.001g respectively compared with the control, and the difference is not significant (P is more than 0.05). There was no significant difference between the 100-fold and 200-fold treatment groups (P > 0.05).
Example 5
Tomato seed germination and seedling growth experiments:
after culturing for 20 days using the same algal fungus complex nutrient solution as in example 4, the ratio of the complex nutrient solution to the nutrient solution was adjusted to 1: 100. 1:300 and 1, 400 were diluted in 4 total proportions and the germinated paper was soaked and treated with distilled water (CK) as a control, each treatment being repeated 3 times. Rinsing the tomato seeds with distilled water, washing off dirt on the surfaces of the tomato seeds, drying the tomato seeds by using filter paper, wrapping the tomato seeds with clean gauze, then sterilizing the tomato seeds twice with 1 percent potassium permanganate, sterilizing the tomato seeds for 10min for the 1 st time, and washing the tomato seeds for 4 to 6 times with sterile water; sterilizing for 8min at the 2 nd time, and washing with sterile water. Selecting healthy receptor seeds with uniform color and size, dibbling 30 seeds in the germination box at equal intervals respectively to ensure that proper intervals are reserved among the seeds, placing the seeds in a constant-temperature incubator at the temperature of (25 +/-1) DEG C, replenishing distilled water for 1 time every day to replenish lost water, recording the germination number of the seeds every day, and calculating the germination rate, the germination vigor, the germination index and the vitality index of the tomato seeds; after the test is finished 15 days later, the length of all seedlings, the length of roots and the length of hypocotyls in each germination box are measured by a graduated scale; the seedling weight and the root weight of the seedlings are measured by an analytical balance.
TABLE 3 Effect of different concentrations of nutrient solutions on tomato seeds
Figure BDA0003129419960000151
Figure BDA0003129419960000161
As can be seen from Table 3, when the tomato seeds are soaked in the composite nutrient solution of phycomycetes, the germination rate and germination potential of the seeds increase with the increase of the dilution concentration. The germination rate and the germination potential of the seeds subjected to 100-time dilution treatment are the highest and are respectively 86.7% and 66.7%, the germination rate is increased by 12.3% and 21.5% compared with a control group, the germination rates of the seeds subjected to 200-time dilution treatment and 300-time dilution treatment are respectively 84.4% and 82.2%, the germination rates of the seeds subjected to 200-time dilution treatment and 300-time dilution treatment are respectively 10% and 7.8%, and the germination rates of the seeds subjected to 400-time dilution treatment are not different from those of CK. Compared with the control, the germination rate and the germination potential of the seeds of each treatment are different to a significant level (P < 0.05) except for 400 times of treatment.
As can be seen from table 3, when the tomato seeds are soaked in the composite nutrient solution of phycomycetes, the germination indexes of the seeds diluted 100 times, 200 times, 300 times and 400 times are respectively increased by 4.45, 2.600, 1.656 and 1.261 compared with the control, and the difference between different treatments is significant compared with the control, but the difference between the treatment groups diluted 200 times, 300 times and 400 times is not significant (P > 0.05). Compared with a control, the vitality indexes of the seeds diluted by 100 times, 200 times, 300 times and 400 times are respectively increased by 542.523, 472.107, 340.481 and 320.743, the difference of different treatments is significant compared with the control (P < 0.05), the difference of the 100 times treatment group and the 200 times treatment group, the difference of the 200 times treatment group and the 300 times and 400 times treatment group is significant (P < 0.05), and the difference of the 300 times and the 400 times treatment group is not significant (P > 0.05).
TABLE 4 Effect of different concentrations of nutrient solutions on tomato seedlings
Figure BDA0003129419960000171
As can be seen from Table 4, the effect on tomato root length varies for different application concentrations. Compared with a control group, the root lengths of the treatment groups diluted by 100 times, 200 times, 300 times and 400 times are respectively increased by 19.02 mm, 18.04 mm, 17.69 mm and 9.97mm, and the difference of different treatments compared with the control group is significant (P < 0.05), but the difference of the treatment groups diluted by 100 times, 200 times and 300 times is not significant (P > 0.05).
As can be seen from Table 4, the effect on tomato seedling growth varies with the concentration applied. Compared with a control, the seedling lengths of the 100-fold and 200-fold diluted treatment groups are respectively increased by 9.52mm and 6.65mm, and the difference is significant compared with the control (P < 0.05), but the difference is not significant compared with the CK treatment by 300-fold and 400-fold treatments (P > 0.05). The difference was significant (P < 0.05) compared between the 100-fold and 200-fold treatment groups. The promoting effect of the nutrient solution on tomatoes is gradually enhanced along with the increase of the application concentration.
As can be seen from Table 4, the effect on the hypocotyl length of tomato was different for different concentrations applied. Compared with the control group, the seedling lengths of the treatment groups diluted by 100 times, 200 times, 300 times and 400 times are respectively increased by 9mm, 6.18mm, 1.91mm and 1.57mm. The difference was significant (P < 0.05) in both 100-fold and 200-fold diluted treatment groups compared to the control, and significant (P < 0.05) between 100-fold and 200-fold treated groups, but not significant (P > 0.05) between 300-fold and 400-fold diluted treatment groups compared to CK treatment. The low-concentration nutrient solution has limited promoting effect on tomato hypocotyls, but the promoting effect is gradually enhanced along with the increase of the application concentration.
As can be seen from table 4, different concentrations applied have different effects on the root weight of tomato. Compared with the control group, the root weight of the treatment groups diluted by 100 times, 200 times, 300 times and 400 times is respectively increased by 0.006, 0.003 and 0.002g, the difference of the treatment groups diluted by 100 times, 200 times and 300 times is significant (P < 0.05), and the difference between the treatment groups diluted by 100 times, 200 times and 300 times is significant (P < 0.05), but the difference between the treatment groups diluted by 400 times and CK is not significant (P > 0.05). The high-concentration nutrient solution has a remarkable promoting effect on the root weight of the tomato seedlings.
As can be seen from table 4, the seedlings of the tomato seedlings diluted 100 times, 200 times, 300 times and 400 times in the treatment groups increased 0.005g, 0.003g, 0.002g and 0.001g, respectively, compared with the control group, and the treated groups diluted 100 times and 200 times in the control group showed significant differences (P < 0.05), no significant difference between the treated groups diluted 100 times and 200 times (P > 0.05), and no significant difference between the treated groups diluted 300 times, 400 times and the control group (P > 0.05).
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (8)

1. The algae-bacterium composite nutrient solution is characterized by being prepared from the following raw materials in parts by mass: 800-1200 parts of debarya solution, 10-50 parts of composite bacillus solution, 10-50 parts of pseudomonas solution and 10-50 parts of photosynthetic bacteria solution;
the Dunaliella solution OD 680 The value is 0.8 to 1.2; OD of composite bacillus liquid, pseudomonas liquid and photosynthetic bacteria liquid 600 The value is 0.8 to 1.2;
the composite bacillus is bacillus subtilis, bacillus licheniformis and bacillus megaterium; the number ratio of the bacillus subtilis to the bacillus licheniformis to the bacillus megaterium is 1-3: 1-3.
2. The preparation method of the algae-bacteria composite nutrient solution as claimed in claim 1, which is characterized by comprising the following steps:
(1) Respectively culturing the debaryomyces, the composite bacillus, the pseudomonas and the photosynthetic bacteria by using special culture media until the logarithmic growth phase to obtain a debaryomyces liquid, a composite bacillus liquid, a pseudomonas liquid and a photosynthetic bacteria liquid;
(2) Respectively centrifuging the chlamydomonas debarkii, the composite bacillus, the pseudomonas and the photosynthetic bacteria liquid obtained in the step (1) to discard supernatant, respectively adding the same volume of the specific culture medium for the chlamydomonas debarkii for cleaning, resuspending, adjusting OD value, mixing, and culturing for 15-25 days at 25-27 ℃.
3. The method for preparing the algae-bacteria composite nutrient solution as claimed in claim 2, wherein the special culture medium for culturing the debarked algae is Tris 2.42g, NH 4 Cl 0.4g，MgSO 4 ·7H 2 O 0.1g，CaCl 2 ·2H 2 O 0.05g，K 2 HPO 4 0.108g，KH 2 PO 4 0.056g of glacial acetic acid and 1ml of glacial acetic acid, and adding distilled water to a constant volume of 1L; the culture temperature of the chlamydomonas debarkii is 19-21 ℃, and the pH value is 6.4-6.6.
4. The method for preparing the composite nutrient solution for algae bacteria according to claim 3, wherein the special culture medium for culturing the composite bacillus is 0.3g of baking soda, 0.2g of citric acid, 0.3g of yeast extract, 1g of glucose, KH 2 PO 4 0.35g, 1L of distilled water; the culture temperature of the composite bacillus is 28-32 ℃, and the pH value is 6.8-7.2.
5. The method for preparing the composite nutrient solution for the algae and the bacteria as claimed in claim 3, wherein the culture medium special for culturing the pseudomonas is tryptone 10g/L, yeast extract powder 5g/L, sodium chloride 10g/L and distilled water 1L; the culture temperature of the pseudomonas is 26-30 ℃, and the pH value is 6.8-7.0.
6. The method for preparing the composite nutrient solution for algae and bacteria according to claim 2, wherein the special culture medium for culturing the photosynthetic bacteria is KH 2 PO 4 0.25g，MgSO 4 ·7H 2 0.25g of O, 0.5g of acetic acid, 0.2g of yeast extract and 1L of distilled water; the culture temperature of the photosynthetic bacteria is 27-29 ℃, and the pH value is 7.1-7.3.
7. The method for preparing the algae-bacterium composite nutrient solution according to claim 5, wherein the centrifugal conditions of the chlamydomonas delavayi liquid in the step (2) are as follows: 4500-5500 r/min, 4-6 min; the centrifugation conditions of the composite bacillus liquid, the pseudomonas liquid and the photosynthetic bacteria liquid are as follows: 7500-8500 r/min, 4-6 min.
8. The use of the algae-bacteria complex nutrient solution of claim 1 in pepper or tomato cultivation.
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