Desert algae sand burying preservation method

Abstract

The invention relates to a desert algae sand burying preservation method. The desert microalgae are inoculated into the sandy soil culture medium obtained by mixing the algae culture medium and the sandy soil, so that desert algae cells are preserved in the culture medium close to the original living environment, the preservation time of the desert algae cells can be effectively prolonged, the survival rate can be increased, and long-term stable preservation of the desert algae cells with specific forms and physiological characteristics can be ensured. The method is simple and rapid to operate, low in application cost and capable of keeping the original hereditary characteristics of algae species for a long time and improving the preservation effect.

Description

Technical Field

[0001] The invention relates to the technical field of desert algae preservation, in particular to a method for preserving desert algae by burying them in sand. Background Art

[0002] The long-term preservation of algae seed resources is an important foundation for conducting algae theory and applied research. Proper algae seed preservation methods can maintain the growth advantages and genetic traits of microalgae for a long time, are not easy to pollute, are simple and easy to operate, reduce the possibility of mutation, facilitate research work, and are conducive to more excellent microalgae strains.

[0003] The preservation technology of desert microalgae has always restricted the normal supply and preservation of algae species by algae production and scientific research units. Although there are several methods for preserving algae, no preservation method has been found that can meet the long-term, high survival rate of desert microalgae and maintain the growth advantages and genetic traits of microalgae for a long time.

[0004] There are many factors that affect the efficiency of algae preservation, such as the characteristics of the algae species, algae density, protective agents, storage temperature, etc. As an extreme environment algae that survives under strong light and drought conditions, desert microalgae can maintain their genetic characteristics while achieving long-term preservation of desert microalgae species, which is of great significance for the research and development of extreme environment algae resources. Sand burial preservation is a method of desert microalgae preservation invented to address the shortcomings of desert microalgae preservation. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention discloses a method for preserving desert algae by burying them in sand, comprising the following steps: Step 1: Take a sand sample, add an algae culture medium, place it on a light culture rack, perform value-added culture, observe under a microscope, and after algae cells are found, perform separation and treatment to obtain a single species of desert microalgae; Step 2: Select sandy soil, sieve it, sterilize it under high pressure, and mix it with the algae culture medium to obtain a sandy soil culture medium; Step 3: inoculating a single species of desert microalgae into a sandy soil culture medium, and placing it on a light culture rack for value-added culture to obtain an algae species; Step 4: Store the algae species and sandy soil culture medium in the dark at 15-20°C.

[0006] As a preferred embodiment of the present invention, the main components and concentrations of the algae culture medium are: K2HPO4·3H2O 0.04g / L, NaNO31.5g / L, MgSO4·7H2O 0.075g / L, ammonium ferric citrate 0.006g / L, CaCI2·2H2O 0.036g / L, citric acid 0.006g / L, EDTANa20.00lg / L, Na2CO30.02g / L, H3BO32.86g / L, MnC14·4H2O 1.81g / L, ZnSO4·7H2O 0.222g / L, Na2MoO4·2H2O 0.39g / L, CuSO4·5H2O0.079g / L, Co(NO3)2·6H2O 0.049g / L.

[0007] As a preferred embodiment of the present invention, the culture conditions for the proliferation culture are: light intensity of 5000 lux, light-dark cycle of 14h / 10h, temperature of 28-30°C, and culture for 10-14 days.

[0008] As a preferred embodiment of the present invention, the high pressure sterilization in step 2 is performed at 121° C. for 15-20 minutes.

[0009] As a preferred embodiment of the present invention, if the preserved desert microalgae need to be revived, the preserved algae suspension is inoculated into a fresh liquid culture medium or spread on a solid culture medium plate, and cultured and revived under weak light conditions at 28°C.

[0010] As a preferred embodiment of the present invention, the main components and concentrations of the liquid culture medium and the solid culture medium are: NaNO3 1.5 g / L, K2HPO3·3H2O 0.04 g / L, MgSO4·7H2O 0.075 g / L, CaCI2·2H2O 0.036 g / L, ammonium ferric citrate 0.006 g / L, citric acid 0.006 g / L, EDTANa2 0.001g / L, Na2CO30.02g / L.

[0011] Beneficial effects of the present invention: The present invention inoculates desert microalgae into a sandy soil culture medium obtained by mixing an algae culture medium with sandy soil, so that the desert algae cells are stored in a culture medium close to the original living environment, which can effectively extend their preservation time, improve the survival rate, and ensure the long-term stable preservation of their unique morphology and physiological characteristics. The method is simple and fast to operate, has a low application cost, can maintain the original genetic characteristics of the algae species for a long time, and improve the preservation effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a state diagram of the algae culture medium of the present invention;

[0013] Figure 2 It is a state diagram of the sandy soil culture medium of the present invention;

[0014] Figure 3 This is a diagram of the state of algae species being directly preserved in algae culture medium;

[0015] Figure 4 This is a diagram of the state of algae species buried in sandy soil culture medium for preservation;

[0016] Figure 5 The results of microscopic observation after the recovery of algae cells preserved in algae culture medium;

[0017] Figure 6 Microscope observation results after recovery of algae cells preserved in sand. DETAILED DESCRIPTION

[0018] Example 1

[0019] The present invention discloses a method for preserving desert algae by burying them in sand, comprising the following steps: Step 1. Take a beaker and put in pure water, add K2HPO4·3H2O, NaNO3, MgSO4·7H2O, ammonium ferric citrate, CaCI2·2H2O, citric acid, EDTANa2, Na2CO3, H3BO3, MnC14·4H2O, ZnSO4·7H2O, Na2MoO4·2H2O, CuSO4·5H2O, Co(NO3)2·6H2O in proportion to prepare an algae culture medium. If the pH of the algae culture medium is too high or too low, adjust the pH to 7.1 with 1mol / L NaOH or 1mol / L HCl solution. Take a sand sample and add it to the algae culture medium (such as Figure 1 As shown), placed on a light culture rack, cultured for 10 days under the conditions of a light intensity of 5000 lux, a light-dark cycle of 14h / 10h, and a temperature of 28°C, observed under a microscope, and after algal cells were found, they were separated and processed to obtain a single species of desert microalgae; Step 2: Select sand and sieve it, sterilize it under high pressure at 121°C for 15 minutes, and mix it with the algae culture medium to obtain a sand culture medium (such as Figure 2 shown); Step 3: Inoculate a single species of desert microalgae into a sandy soil culture medium (e.g. Figure 4 As shown), and placed on a light culture rack, cultured for 10 days under the conditions of a light intensity of 5000 lux, a light-dark cycle of 14h / 10h, and a temperature of 28°C to obtain algae species; Step 4: Store the algae seeds and the sand culture medium in the dark at 15°C. Generally, transplant them once after 6-8 months of storage. The storage humidity is expressed as relative humidity, usually 50% to 70%.

[0020] The main components and concentrations of the algae culture medium are: K2HPO4·3H2O 0.04g / L, NaNO3 1.5g / L, MgSO4·7H2O 0.075g / L, ammonium ferric citrate 0.006g / L, CaCI2·2H2O 0.036g / L, citric acid 0.006g / L, EDTANa2 0.00lg / L, Na2CO3 0.02g / L, H3BO3 2.86g / L, MnC14·4H2O 1.81g / L, ZnSO4·7H2O 0.222g / L, Na2MoO4·2H2O 0.39g / L, CuSO4·5H2O 0.079g / L, Co(NO3)2·6H2O 0.049g / L.

[0021] If the preserved algae species need to be revived, the preserved algae species suspension is inoculated into fresh liquid culture medium and cultured and revived under weak light conditions at 28°C; the main components and concentrations of the liquid culture medium are: NaNO3 1.5g / L, K2HPO3·3H2O 0.04g / L, MgSO4·7H2O 0.075g / L, CaCI2·2H2O 0.036g / L, ammonium ferric citrate 0.006g / L, citric acid 0.006g / L, EDTANa2 0.001g / L, and Na2CO3 0.02g / L.

[0022] Example 2

[0023] The present invention discloses a method for preserving desert algae by burying them in sand, comprising the following steps: Step 1. Take a beaker and put in pure water, add K2HPO4·3H2O, NaNO3, MgSO4·7H2O, ammonium ferric citrate, CaCI2·2H2O, citric acid, EDTANa2, Na2CO3, H3BO3, MnC14·4H2O, ZnSO4·7H2O, Na2MoO4·2H2O, CuSO4·5H2O, Co(NO3)2·6H2O in proportion to prepare an algae culture medium. If the pH of the algae culture medium is too high or too low, adjust the pH to 7.1 with 1mol / L NaOH or 1mol / L HCl solution. Take a sand sample and add it to the algae culture medium (such as Figure 1 As shown), placed on a light culture rack, cultured for 14 days under the conditions of a light intensity of 5000 lux, a light-dark cycle of 14h / 10h, and a temperature of 30°C, observed under a microscope, and after algal cells were found, they were separated and processed to obtain a single species of desert microalgae; Step 2: Select sand and sieve it, sterilize it under high pressure at 121°C for 20 minutes, and mix it with the algae culture medium to obtain a sand culture medium (such as Figure 2 shown); Step 3: Inoculate a single species of desert microalgae into a sandy soil culture medium (e.g. Figure 4 As shown), and placed on a light culture rack, cultured for 14 days under the conditions of light intensity of 5000 lux, light-dark cycle of 14h / 10h, and temperature of 30°C to obtain algae species; Step 4: Store the algae seeds and the sand culture medium in the dark at 20°C. Generally, transplant them once after 6-8 months of storage. The storage humidity is expressed as relative humidity, usually 50% to 70%.

[0024] The main components and concentrations of the algae culture medium are: K2HPO4·3H2O 0.04g / L, NaNO3 1.5g / L, MgSO4·7H2O 0.075g / L, ammonium ferric citrate 0.006g / L, CaCI2·2H2O 0.036g / L, citric acid 0.006g / L, EDTANa2 0.00lg / L, Na2CO3 0.02g / L, H3BO3 2.86g / L, MnC14·4H2O 1.81g / L, ZnSO4·7H2O 0.222g / L, Na2MoO4·2H2O 0.39g / L, CuSO4·5H2O 0.079g / L, Co(NO3)2·6H2O 0.049g / L.

[0025] If the preserved algae species need to be revived, the preserved algae species suspension is inoculated into fresh liquid culture medium and cultured and revived under weak light conditions at 28°C; the main components and concentrations of the liquid culture medium are: NaNO3 1.5g / L, K2HPO3·3H2O 0.04g / L, MgSO4·7H2O 0.075g / L, CaCI2·2H2O 0.036g / L, ammonium ferric citrate 0.006g / L, citric acid 0.006g / L, EDTANa2 0.001g / L, and Na2CO3 0.02g / L.

[0026] Example 3

[0027] The present invention discloses a method for preserving desert algae by burying them in sand, comprising the following steps: Step 1. Take a beaker and put in pure water, add K2HPO4·3H2O, NaNO3, MgSO4·7H2O, ammonium ferric citrate, CaCI2·2H2O, citric acid, EDTANa2, Na2CO3, H3BO3, MnC14·4H2O, ZnSO4·7H2O, Na2MoO4·2H2O, CuSO4·5H2O, Co(NO3)2·6H2O in proportion to prepare an algae culture medium. If the pH of the algae culture medium is too high or too low, adjust the pH to 7.1 with 1mol / L NaOH or 1mol / L HCl solution. Take a sand sample and add it to the algae culture medium (such as Figure 1 As shown), placed on a light culture rack, cultured for 12 days under the conditions of a light intensity of 5000 lux, a light-dark cycle of 14h / 10h, and a temperature of 29°C, observed under a microscope, and after algal cells were found, they were separated and processed to obtain a single species of desert microalgae; Step 2: Select sand and sieve it, sterilize it under high pressure at 121°C for 18 minutes, and mix it with the algae culture medium to obtain a sand culture medium (such as Figure 2 shown); Step 3: Inoculate a single species of desert microalgae into a sandy soil culture medium (e.g. Figure 4 As shown), and placed on a light culture rack, cultured for 12 days under the conditions of light intensity of 5000 lux, light-dark cycle of 14h / 10h, and temperature of 29°C to obtain algae species; Step 4: Store the algae seeds and the sand culture medium in the dark at 18°C. Generally, transplant them once after 6-8 months of storage. The storage humidity is expressed as relative humidity, usually 50% to 70%.

[0028] The main components and concentrations of the algae culture medium are: K2HPO4·3H2O 0.04g / L, NaNO3 1.5g / L, MgSO4·7H2O 0.075g / L, ammonium ferric citrate 0.006g / L, CaCI2·2H2O 0.036g / L, citric acid 0.006g / L, EDTANa2 0.00lg / L, Na2CO3 0.02g / L, H3BO3 2.86g / L, MnC14·4H2O 1.81g / L, ZnSO4·7H2O 0.222g / L, Na2MoO4·2H2O 0.39g / L, CuSO4·5H2O 0.079g / L, Co(NO3)2·6H2O 0.049g / L.

[0029] If the preserved algae species need to be revived, the preserved algae species suspension is inoculated into fresh liquid culture medium and cultured and revived under weak light conditions at 28°C; the main components and concentrations of the liquid culture medium are: NaNO3 1.5g / L, K2HPO3·3H2O 0.04g / L, MgSO4·7H2O 0.075g / L, CaCI2·2H2O 0.036g / L, ammonium ferric citrate 0.006g / L, citric acid 0.006g / L, EDTANa2 0.001g / L, and Na2CO3 0.02g / L.

[0030] like Figure 3 , Figure 4 As shown in the figure, a comparative experiment was conducted between the preservation method directly in the algae culture medium and the preservation method buried in sand: the algae species buried in sand and the algae species directly preserved in the algae culture medium were preserved under the same conditions for 6 months, and the algae suspension was taken out and inoculated into fresh liquid culture medium or spread on solid culture medium plates, and cultured and revived under weak light conditions at 28°C. After culturing for about a week, the growth was observed, and the cell morphology was observed at 100 times the differential interference microscope. The results of microscopic observation are shown in the figure below. Figure 5 , Figure 6 The results showed that the cells preserved in sand had good morphology and could continue to be preserved using this method.

[0031] Components not described in detail herein are prior art.

[0032] Although the specific embodiments of the present invention are described in detail above, the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in this field without departing from the purpose of the present invention, and modifications or deformations without creative labor are still within the protection scope of the present invention.

Claims

1. A method for preserving desert algae by burying them in sand, characterized in that: The steps include: Step 1: Take a sand sample, add an algae culture medium, place it on a light culture rack, perform value-added culture, observe under a microscope, and after algae cells are found, perform separation and treatment to obtain a single species of desert microalgae; Step 2: Select sandy soil, sieve it, sterilize it under high pressure, and mix it with the algae culture medium to obtain a sandy soil culture medium; Step 3: inoculating a single species of desert microalgae into a sandy soil culture medium, and placing it on a light culture rack for value-added culture to obtain an algae species; Step 4: Store the algae species and sandy soil culture medium in the dark at 15-20°C.

2. A method for preserving desert algae by burying them in sand according to claim 1, characterized in that: The main components and concentrations of the algae culture medium are: K2HPO4·3H2O 0.04g / L, NaNO3 1.5g / L, MgSO4·7H2O 0.075g / L, ammonium ferric citrate 0.006g / L, CaCl2·2H2O 0.036g / L, citric acid 0.006 g / L, EDTANa20.00lg / L, Na2CO30.02g / L, H3BO32.86g / L, MnC14·4H2O 1.81g / L, ZnSO4·7H2O 0.222g / L, Na2MoO4·2H2O 0.39g / L, CuSO4·5H2O 0.079g / L, Co(NO3)2·6H2O 0.049g / L.

3. A method for preserving desert algae buried in sand according to claim 1, characterized in that: The culture conditions for the proliferation culture are: light intensity of 5000 lux, light-dark cycle of 14h / 10h, temperature of 28-30°C, and culture for 10-14 days.

4. The method for preserving desert algae by burying them in sand according to claim 1, characterized in that: The high pressure sterilization in step 2 is carried out at 121° C. for 15-20 minutes.

5. The method for preserving desert algae by burying them in sand according to claim 1, characterized in that: The preserved algae suspension was inoculated into fresh liquid culture medium and cultured and revived under weak light conditions at 28°C.

6. A method for preserving desert algae buried in sand according to claim 5, characterized in that: The main components and concentrations of the liquid culture medium are: NaNO3 1.5 g / L, K2HPO3·3H2O 0.04 g / L, MgSO4·7H2O 0.075 g / L, CaCI2·2H2O 0.036 g / L, ammonium ferric citrate 0.006 g / L, citric acid 0.006 g / L, EDTANa2 0.001 g / L, and Na2CO3 0.02 g / L.

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