A Real-time Dynamic Monitoring Method for the Yield of Mass Cultivation of Desert Cyanobacteria

Abstract

The present invention discloses a method for real-time dynamic monitoring of the yield of large-scale cultivation of desert cyanobacteria, which comprises the following steps: cultivating multiple groups of cyanobacteria under multiple initial underwater light intensity conditions, recording the sample dry weights of the multiple groups of cyanobacteria at different time periods and the average underwater light intensity of the day, and constructing a calibration curve of the average underwater light intensity of the day - cyanobacteria sample dry weight therefrom; measuring the average underwater light intensity of the day under the photoperiod conditions in the target cyanobacteria culture pond, and realizing real-time dynamic monitoring of the cyanobacteria sample dry weight in the target cyanobacteria culture pond according to the average underwater light intensity of the day and the standard curve, so as to infer the cyanobacteria yield. Compared with the prior art, the advantages of the present invention include: 1) realizing real-time dynamic monitoring of the yield of desert cyanobacteria; 2) having a simple usage method and being easy to operate; 3) having low cost and high detection efficiency.

Description

Technical Field

[0001] The present invention relates to a monitoring method, and particularly to a real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria, belonging to the technical field of cyanobacteria cultivation. Background Art

[0002] The artificial biological soil crust desertification land treatment technology is a new green and environmental protection desertification land treatment technology at home and abroad. It utilizes the function of biological soil crust to fix the sand surface and resist wind erosion. The main organisms in the biological soil crust (such as desert cyanobacteria) are artificially cultivated and inoculated on the sand surface. Through maintenance and survival, the biological soil crust is accelerated to form on the ground surface, playing a role in wind prevention and sand fixation and ecological restoration. It is the forefront and hot spot of the current research on the restoration of damaged desertified land. The large-scale cultivation of desert cyanobacteria organisms is an important environment for this technology and an important prerequisite and guarantee for field construction.

[0003] However, there is currently no method for real-time monitoring of the yield during its large-scale cultivation process. Therefore, there is an urgent need to invent a real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria, providing a basis for the monitoring of large-scale cultivation of desert cyanobacteria organisms and laying a foundation for the large-scale promotion of the artificial biological soil crust desertification land treatment technology. Summary of the Invention

[0004] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria.

[0005] To achieve the foregoing invention purpose, the technical solutions adopted by the present invention include:

[0006] A real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria, comprising the following steps:

[0007] Cultivate multiple groups of cyanobacteria under multiple initial underwater light intensity conditions respectively, record the sample dry weights of multiple groups of cyanobacteria at different time periods and the average underwater light intensity on the same day under the photoperiod condition, and construct a calibration curve of the average underwater light intensity - cyanobacteria sample dry weight on the same day;

[0008] Measure the average underwater light intensity on the same day under the photoperiod condition in the target cyanobacteria culture pool, and realize the real-time dynamic monitoring of the cyanobacteria sample dry weight in the target cyanobacteria culture pool according to the average underwater light intensity on the same day and the standard curve, and thereby deduce the cyanobacteria yield.

[0009] Compared with the prior art, the advantages of the present invention include:

[0010] 1) Realize the real-time dynamic monitoring of the yield of desert cyanobacteria;

[0011] 2) The usage method is simple and easy to operate;

[0012] 3) Low cost and high detection efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 It is the functional relationship between the average underwater light intensity I of the day and the dry weight M of the cyanobacteria sample;

[0014] Figure 2 This is a lookup table for the daily average underwater light intensity I and the dry weight M of cyanobacteria samples. DETAILED DESCRIPTION

[0015] In view of the shortcomings of the prior art, the inventors of this case, after long-term research and extensive practice, have proposed the technical solution of the present invention. The following will further explain this technical solution, its implementation process and principles.

[0016] Example 1

[0017] Multiple groups of cyanobacteria were cultured under different initial underwater light intensity conditions. The dry weight of the samples of the multiple groups of cyanobacteria at different time periods and the average underwater light intensity of the day were recorded. Based on this, a calibration curve of the average underwater light intensity of the day - the dry weight of the cyanobacteria samples was constructed.

[0018] The average underwater light intensity of the day under the photoperiod conditions in the target cyanobacteria culture pond was measured, and the real-time dynamic monitoring of the dry weight of the cyanobacteria samples in the target cyanobacteria culture pond was achieved based on the average underwater light intensity of the day and the standard curve, so as to infer the cyanobacteria yield.

[0019] The specific steps are as follows:

[0020] (1) The cyanobacteria are thoroughly mixed with the culture medium and cultured at a specified underwater temperature for more than 24 hours;

[0021] (2) collecting the mixed system containing cyanobacteria and culture medium in step (1) once every specified time period, drying it and weighing it to obtain the dry weight of the cyanobacteria sample, measuring the average underwater light intensity on the day when the dry weight of the cyanobacteria sample is obtained, and obtaining a standard curve of underwater light intensity-dry weight of the cyanobacteria sample;

[0022] (3) The cyanobacteria and the culture solution are fully mixed, and then the average underwater light intensity of the cyanobacteria mixture is measured and compared with the standard curve to determine the dry weight of the cyanobacteria sample in the cyanobacteria mixture.

[0023] It is understandable that at a specified initial underwater light intensity, which is 1500 - 4000 Lux, the cyanobacteria are fully mixed with the culture medium. The cyanobacteria are selected from one or a combination of two or more of Anabaena, Microcoleus vaginatus, Nostoc, Phormidium, Scytonema, Scytonema javanicum, and Tolypothrix. The above cyanobacteria and culture medium mixture system is cultured at a specified underwater temperature for more than 24 hours. The specified underwater temperature in the cyanobacteria culture pond is 25°C - 30°C. At this temperature, the growth condition of the cyanobacteria is the best. When the water temperature exceeds 40°C, cooling measures should be taken in a timely manner. Preferably, the water temperature can be measured and recorded every hour to adjust the water temperature in a timely manner. The cyanobacteria and culture medium mixture system is collected at a specified interval, and the dry weight of the cyanobacteria sample is weighed. The specified interval here can be 12 hours, 24 hours, 48 hours, or 72 hours. At the same time, the average underwater light intensity on the day when the dry weight of the cyanobacteria sample is obtained is measured and obtained. The obtaining method can be to measure the underwater light intensity at least once every hour within 24 hours, and the average value of the measured underwater light intensities is taken to obtain the average underwater light intensity on the day.

[0024] Furthermore, during the culturing process of the mixture system containing cyanobacteria and culture medium, the mixture system is alternately placed under light or darkroom conditions without light. The ratio of the duration of the light time to the darkroom time is 9 / 15 - 12 / 12; preferably, the mass ratio of the initial dry weight of the cyanobacteria to the culture medium in the mixture system is 8 g / t.

[0025] Example 2

[0026] (1) The cyanobacteria are placed in a cyanobacteria culture pond for large-scale culture.

[0027] The cyanobacteria in this group are cultured under the following conditions: at an initial light intensity of 4000 lux in a 12 / 12 light-dark cycle, the water temperature is maintained at 25°C, and the cyanobacteria are inoculated into the culture medium with an initial biomass of 8 g / t. This can be recorded as the 0th hour of culturing.

[0028] (2) After inoculation, after culturing for 24 hours, the underwater light intensity (hereinafter referred to as: underwater light intensity) is recorded every hour, and the underwater light intensity measured every 24 hours after the 0th to 24th hours is recorded and sorted into the average underwater light intensity on the day. At any time after 24 hours, a cyanobacteria sample is collected from the cyanobacteria culture pond, and a cyanobacteria sample is collected at a specified interval.

[0029] (3) The dry weight of the cyanobacteria sample collected from the cyanobacteria culture pond is measured, and a functional relationship is established between the underwater light intensity of the culture pond when the cyanobacteria sample is collected and the dry weight of the cyanobacteria sample.

[0030] In this embodiment, a cyanobacteria culture pool with a specification of 15 m × 1 m × 1 m (length × width × height) is selected. A culture solution with a water depth of 0.5 m is placed in the cyanobacteria culture pool. For every 1 ton of the culture solution, cyanobacteria with a dry weight of 8 g are proportioned, and the cyanobacteria are inoculated into the culture solution for cultivation.

[0031] During the cultivation process, the light-dark cycle is 12 / 12. The initial underwater light intensity is set to 4000 lux, and the water temperature is maintained at 25 °C.

[0032] After culturing for 24 hours, the underwater light intensity is recorded at least once per hour. Starting from the 24th hour, the underwater light intensities within the subsequent 24 hours are sorted out and the average underwater light intensity under the light cycle conditions within these 24 hours (of the same day) is calculated, denoted as I. When cyanobacteria reproduce in the culture solution, as the number of them increases, the underwater light intensity measured underwater will gradually decrease. Preferably, when recording the underwater light intensity, different regions of the cyanobacteria culture pool are selected for measurement. For example, the underwater light intensity is measured at least once at the front end, middle end, and rear end set along the length direction of the cyanobacteria culture pool, and the measured underwater light intensities are sorted out, recorded, and the average value is calculated, which is recorded as the average underwater light intensity within these 24 hours (of the same day), denoted as I.

[0033] After the cyanobacteria are cultured for 24 hours, cyanobacteria samples can be collected at any time.

[0034] The cyanobacteria can be aseptically cultured under laboratory conditions, and then the aseptically cultured cyanobacteria samples are collected and transferred into the cyanobacteria culture pool for large-scale cultivation with an initial dry weight of 8 g / t of the culture solution.

[0035] In this embodiment, cyanobacteria samples in the cyanobacteria culture pool are collected at the 34th hour after cultivation, and their dry weight is measured, denoted as M. When measuring the dry weight, 1 L (1 liter) of cyanobacteria samples are evenly scooped from different positions in the cyanobacteria culture pool. The cyanobacteria samples are poured into a glass bottle and dried in an 80 °C oven until a constant weight is reached. After taking them out, the mass is weighed with a one-thousandth electronic balance. The data measured here is the dry weight M of the cyanobacteria samples.

[0036] Preferably, blue-green algae liquid is scooped from the front end, middle end, and rear end of the blue-green algae culture pond along its length direction, and the blue-green algae liquid is mixed to form the "blue-green algae sample" collected this time. Since blue-green algae are prone to sedimentation during their cultivation process, the contact area between the sedimented blue-green algae and underwater light decreases, which may affect the accuracy of the final measurement data. Therefore, one or more disturbance devices can be set in the blue-green algae culture pond. The disturbance devices are used to disturb, mix, or stir the blue-green algae and the culture solution in the blue-green algae culture pond, so as to increase the contact area between the blue-green algae and light and improve the accuracy of the data. Preferably, the disturbance device can be a submersible pump, and the submersible pump can control the disturbance duration through a time-controlled switch. For example, the disturbance duration can be controlled to be disturbed for 12 hours every 24 hours, and the corresponding disturbance period can also be set. In this embodiment, the disturbance period is set from 7:00 to 19:00 every day. At the same time, when a method for real-time dynamic monitoring of the yield of large-scale cultivation of desert blue-green algae provided in this embodiment is applied to blue-green algae cultivation, the set disturbance device can increase the contact area between the liquid in the blue-green algae culture pond and carbon dioxide, and carbon dioxide helps the reproduction of blue-green algae. Therefore, the yield of blue-green algae per unit time can be increased.

[0037] The "specified period" between blue-green algae collections can be 12 hours, 24 hours, 48 hours, 72 hours, or any other arbitrarily selected period, that is, the period between each blue-green algae collection is fixed. The "specified period" is selected as 48 hours, that is, the time between each blue-green algae sample collection is 48 hours. The dry weight of the blue-green algae sample collected for the first time is recorded as M1, and the dry weight of the blue-green algae sample collected 48 hours after the first collection is recorded as M2, and so on. The dry weights of the subsequent blue-green algae samples collected are recorded as M3, M4, M5. The average underwater light intensity on the day when the dry weight of the blue-green algae sample is M1 for the first time is recorded as I1, and so on. The average underwater light intensity on the day when the subsequent blue-green algae samples are collected is I2, I3, T4, I5. Please refer to Table 1. Table 1 is a record table of the dry weight M of the blue-green algae sample and the average underwater light intensity I for each collection of the blue-green algae sample with a time interval of 48 hours.

[0038]

[0039] Table 1

[0040] Under these conditions, the growth quantity of the blue-green algae yield over time gradually increases, and the measured average underwater light intensity on the day gradually decreases.

[0041] The cultivation conditions are the same as above. The "specified period" is selected as 72 hours, and the above recording method is repeated. Five sets of data are collected, and the following Table 2 is obtained. Table 2 is a record table of the dry weight M of the blue-green algae sample and the average underwater light intensity I for each collection of the blue-green algae sample with a time interval of 72 hours.

[0042]

[0043] Table 2

[0044] Under this condition, the yield of cyanobacteria gradually increases with time, the measured average underwater light intensity on the same day gradually decreases, and when the yield of cyanobacteria reaches a certain value, the dry weight M of the cyanobacteria sample decreases.

[0045] More specifically, during the cultivation of each group of cyanobacteria, two or more cyanobacteria culture ponds can be selected for large-scale cultivation of cyanobacteria. Multiple said cyanobacteria culture ponds are arranged in the same closed chamber, such as multiple said cyanobacteria culture ponds are arranged in the same greenhouse. Let the cyanobacteria culture ponds be irradiated by sunlight, and the light intensities received by multiple said cyanobacteria culture ponds are different. Measure the data of the average underwater light intensity on the same day and the dry weight of the cyanobacteria sample in different cyanobacteria culture ponds at the same time. When collecting, the difference in the average underwater light intensity on the same day obtained from different cyanobacteria culture ponds is 50 - 100 Lux, and the difference in the dry weight of the obtained cyanobacteria sample is 5 - 10 g. Therefore, it can be judged that the growth rate of cyanobacteria in the same closed chamber is roughly the same.

[0046] The culture conditions are the same as above. Select the "designated period" as 48 hours, and repeat the above recording method. Continuously collect data 15 times to obtain the following Table 3. Table 3 is a record table of the dry weight M of the cyanobacteria sample and the average underwater light intensity I on the same day with a time interval of 12 hours for each collection.

[0047]

[0048]

[0049] Table 3

[0050] Under this condition, the yield of cyanobacteria gradually increases with time, the measured average underwater light intensity on the same day gradually decreases, and when the yield of cyanobacteria reaches around the value of 2000 Lux, with the increase of time, the dry weight M of the cyanobacteria sample begins to decrease. Referring to the data recorded in the table, under the 12 / 12 light-dark cycle with an initial light intensity of 4000 lux, the water temperature is maintained at 25 °C, and the cyanobacteria are inoculated into the culture solution with an initial biomass of 8 g / t. The growth of cyanobacteria is in an overall upward trend, and the dry weight M of the sample gradually increases; in the later stage, the cyanobacteria are in the process of decline, and the decline rate of cyanobacteria is greater than the growth rate of cyanobacteria, resulting in the gradual decrease of the dry weight M of the cyanobacteria sample.

[0051] Based on this, the average underwater light intensity on the same day and the dry weight of the cyanobacteria sample are curve-fitted to establish a functional relationship:

[0052] M = 185.57 + 0.058I - 0.0013I 2

[0053] By statistical methods, a functional relationship between the daily average underwater light intensity and the dry weight of the cyanobacteria sample is established, and a calibration curve of the daily average underwater light intensity - cyanobacteria sample dry weight is formulated. As Figure 1 shown, Figure 1 the squares in it represent the actually collected sample data, Figure 1 and the curve in it represents the calibrated curve of the daily average underwater light intensity - cyanobacteria sample dry weight obtained by fitting.

[0054] Furthermore, according to the calibration curve of the daily average underwater light intensity - cyanobacteria sample dry weight, a real-time yield retrieval table of the daily average underwater light intensity - cyanobacteria sample dry weight is established. As Figure 2 shown. When actually culturing and propagating cyanobacteria, under the initial light intensity of 4000 lux in a 12 / 12 light-dark cycle, with the water temperature maintained at 25 °C, the cyanobacteria are inoculated into the culture solution with an initial biomass of 8 g / t. By detecting the average underwater light intensity, the instant dry weight data of the cyanobacteria sample can be obtained through the functional relationship between the daily average underwater light intensity and the dry weight of the cyanobacteria sample. Compared with the conventional method of scooping cyanobacteria and then drying and weighing them, it is not only convenient for timely recording of data, but also has a simple method and is easy to operate, and can quickly obtain the dry weight data of the cyanobacteria sample under the daily average underwater light intensity.

[0055] More specifically, the culture solution is preferably BG11 culture solution. In the BG11 culture solution, the concentration of MgSO4·7H2O is 0.07 g / L, the concentration of K2HPO4 is 0.04 g / L, the concentration of CaCl2·2H2O is 0.036 g / L, the concentration of NaNO3 is 1.5 g / L, the concentration of citric acid is 0.006 g / L, the concentration of ferric ammonium citrate is 0.006 g / L, the concentration of sodium ethylenediaminetetraacetate (EDTA) is 0.001 g / L, and the concentration of CaCO3 is 30.02 g / L; the concentration of trace element A5+Co solution is 1 mL; among them, in each liter of distilled water in the A5+Co solution: the concentration of H3BO3 is 2.86 g / L, the concentration of MnCl2·4H2O is 1.86 g / L, the concentration of ZnSO4·7H2O is 0.22 g / L, the concentration of Na2MoO4·2H2O is 0.39 g / L, the concentration of CuSO4·5H2O is 0.08 g / L, and the concentration of Co(NO3)2·6H2O is 0.05 g / L. That is, cyanobacteria are cultured in the BG11 culture solution with this ratio, and the calibration curve of the daily average underwater light intensity - cyanobacteria sample dry weight is formulated.

[0056] More specifically, the cyanobacteria are selected from one or more of Anabaena, Microcoleus vaginatus, Nostoc, Phormidium, Scytonema, Scytonema javanicum, and Tolypothrix.

[0057] It should be understood that the above embodiments are only for illustrating the technical concept and features of the present invention, and the purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly. It is not intended to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.

Claims

1. A method for real-time dynamic monitoring of the yield of large-scale cultivation of desert cyanobacteria, characterized in that, The steps include: Multiple groups of cyanobacteria were cultured under different initial underwater light intensity conditions. The dry weight of samples of multiple groups of cyanobacteria at different time periods and the average underwater light intensity of the day under the photoperiod conditions were recorded. Based on this, a calibration curve of the average underwater light intensity of the day - dry weight of cyanobacteria samples was constructed. The average underwater light intensity of the day under the photoperiod conditions in the target cyanobacteria culture pond was measured. Based on the average underwater light intensity of the day and the standard curve, the dry weight of the cyanobacteria samples in the target cyanobacteria culture pond was monitored in real time to estimate the cyanobacteria yield. Specifically include: (1) Fully mixing the cyanobacteria with the culture solution to form a mixed system, and placing the mixed system in a dark room with or without light alternately, with the ratio of the light time to the dark room time being 9 / 15 to 12 / 12; and culturing the system underwater at a temperature of 25°C to 30°C for more than 24 hours; (2) collecting the mixed system containing cyanobacteria and culture medium in step (1) once every specified time period, drying it and weighing it to obtain the dry weight of the cyanobacteria sample, wherein the specified time period is 12 hours, 24 hours, 48 hours or 72 hours; measuring the average underwater light intensity on the day when the dry weight of the cyanobacteria sample is obtained, and obtaining a standard curve of underwater light intensity-dry weight of the cyanobacteria sample; (3) The cyanobacteria and the culture solution are fully mixed, and then the average underwater light intensity of the cyanobacteria mixture is measured and compared with the standard curve to measure the dry weight of the cyanobacteria sample in the cyanobacteria mixture.

2. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria according to claim 1, characterized in that: It involves measuring the underwater light intensity multiple times within 24 hours, and taking the average of the measured underwater light intensities to obtain the average underwater light intensity for the day.

3. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria according to claim 2, characterized in that: Specifically include: Measure underwater light intensity at least once every hour for 24 hours.

4. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria according to claim 1, wherein, Also includes: The mixed system containing cyanobacteria and culture solution is disturbed at a frequency of 12 hours every 24 hours.

5. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria according to claim 1, characterized in that: The initial underwater light intensity is 1500-4000 Lux.

6. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert blue-green algae according to claim 1, characterized in that: The mass ratio of the initial dry weight of the cyanobacteria to the culture solution in the mixed system is 8 g / t.

7. The real-time dynamic monitoring method for the yield of large-scale cultivation of desert cyanobacteria according to claim 1, characterized in that: The cyanobacteria is selected from one or a combination of two or more of Anabaena, Microcoleus, Nostoc, Pseudocranium, and Monochamus.

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