Method for rapidly estimating species composition in phycomycete symbiont

Abstract

The invention belongs to the field of species analysis, and particularly relates to a method for rapidly estimating species composition in phycomycete symbionts. The invention provides the dual-wavelength spectrophotometric method capable of simultaneously and rapidly estimating the biomass of the algae and the biomass of the bacteria in the phycomycete symbiont for the first time. When the biomass is small, a simple and convenient linear model can be directly used for estimation. When the biomass is large, the estimation effect of a linear model is not ideal, a nonlinear model is further constructed, the application range of the method is expanded, and a good prediction effect can also be achieved for the biomass composition of species of a co-culture system of high-concentration biomass.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of species analysis, and particularly relates to a method for rapidly estimating the species composition in an algal-bacterial symbiotic body. BACKGROUND

[0002] In an algal-bacterial symbiotic body, algae fix carbon dioxide and provide organic matter through photosynthesis, and bacteria participate in material circulation (such as nitrogen fixation and decomposition of organic matter). Under the background of rapid development of synthetic biology, the metabolic interaction characteristics of the algal-bacterial symbiotic body have attracted widespread attention for creating an efficient green production microbial system, and therefore, rapid and non-invasive detection of the biomass of algae and bacteria in the algal-bacterial symbiotic body is of great significance for online detection of the state of the green production system. In addition, the algal-bacterial symbiotic body may also appear in ecological environment related application scenarios such as sewage treatment systems and natural water bodies, and separately measuring the biomass of the two is helpful for quantifying the ecological contribution of each.

[0003] At present, the methods and limitations for measuring the biomass of different species in an algal-bacterial symbiotic body mainly include: (1) direct counting method by microscope. This method is time-consuming and laborious, and has a large error; especially when the symbiotic body has a high biomass concentration and the morphology of the internal species is complex, the measurement result is very inaccurate. (2) single-wavelength spectrophotometry combined with chlorophyll content determination method. This method is complex to operate, and needs centrifugation, filtration, extraction and other multiple steps of processing; and only the high and low of the chlorophyll content under a specific optical density can be judged based on experience, and it is difficult to accurately obtain the index of quantifying the abundance of bacteria. (3) plate counting method. This method needs to wait for the growth of microbial colonies / algal colonies, and it is difficult to obtain the result in a short time. (4) quantitative PCR method. This method is complex to operate, and has high professional requirements, and needs precise instruments such as fluorescence quantitative PCR instrument, and has high measurement cost. (5) flow cytometry. This method needs expensive equipment, and has low popularization rate. And it needs to be pre-calibrated, and has high requirements for sample pretreatment. (6) quantitative technology based on marker gene sequencing. This method has high cost and high professional requirements, and the copy number of the commonly used marker gene is different in different organisms, and the detection result cannot completely correspond to the biomass concentration.

[0004] Spectrophotometry has become one of the key methods for detecting microbial biomass due to its advantages such as simplicity, rapidness and compatibility with non-invasive detection. Based on the absorbance additivity of Lambert-Beer law, through constructing a species-specific absorbance model, it is expected to realize independent quantification of the biomass of each species in a two-species co-culture system. However, for a two-species algal-bacterial symbiotic body, the light absorption values representing the biomass of algae and bacteria interfere with each other, and if the ordinary spectrophotometry is used without processing the data, the measurement data will inevitably have large differences from the actual situation.

[0005] Therefore, if a method can be provided based on the basic principles of spectrophotometry that can relatively accurately and quickly measure the biomass of algae and bacteria in an algal-bacterial symbiosis, it will have significant research and application value. Summary of the Invention

[0006] The purpose of this invention is to provide a method for rapidly estimating the species composition in algal-bacterial symbionts.

[0007] To achieve the above-mentioned objectives, the technical solution adopted by this invention is: a method for rapidly estimating the biomass of algae and bacteria in an algal-bacterial symbiosis, the method comprising the following steps:

[0008] (1) The total absorbance of the algal-bacterial symbiotic organism is expressed as Equation (1):

[0009]

[0010] A 600 The total absorbance of the algae-bacterial symbiotic organism at 600 nm, A 680 The total absorbance of the algae-bacterial symbiotic organism at 680 nm, A 600，菌 A refers to the absorbance of bacteria in a symbiotic organism at 600 nm. 600，藻 A refers to the absorbance of algae in a symbiotic organism at 600 nm. 680，菌 A refers to the absorbance of bacteria in a symbiotic organism at 680 nm. 680，藻 This refers to the absorbance of algae in a symbiotic organism at 680 nm.

[0011] (2) Establish the absorbance A of the bacterial cells at 680 nm. 680,菌 Absorbance A at 600 nm 600,菌 Mathematical relation A 680,菌 =f(A 600,菌 ), and the absorbance of the algae at 600 nm (A 600,藻 ) and absorbance at 680 nm (A 680,藻 Mathematical relation A 600,藻 =g(A 680,藻 Then equation (1) is transformed into equation (2):

[0012]

[0013] (3) Wavelength conversion equation for fungi A 680,菌 =f(A 600,菌 And the wavelength conversion equation A for algae. 600,藻 =g(A 680,藻 Both can be represented by general linear models, namely equations (3) and (4):

[0014] A 680,菌 =w×A 600,菌+v (3)

[0015] A 600,藻 =w′×A 680,藻 +v′ (4)

[0016] (4) Substituting equations (3) and (4) into equation (1), we obtain equation (5):

[0017]

[0018] Solve equation (5) to obtain the biomass of fungi and algae.

[0019] Accordingly, a method for rapidly estimating the biomass of algae and bacteria in an algal-bacterial symbiosis includes the following steps: constructing formula (13):

[0020]

[0021] Among them, A′ 600 A′ represents the measured absorbance of the diluted algal-bacterial symbiont at 600 nm. 680 A represents the measured absorbance of the diluted algae-bacterial symbiont at 680 nm, where DR is the dilution ratio of the algae-bacterial symbiont; A 600，菌 A refers to the absorbance of bacteria in a symbiotic organism at 600 nm. 600，藻 A refers to the absorbance of algae in a symbiotic organism at 600 nm. 680，菌 A refers to the absorbance of bacteria in a symbiotic organism at 680 nm. 680，藻 This refers to the absorbance of algae in a symbiotic organism at 680 nm.

[0022] Solve equation (13) to obtain the biomass of fungi and algae.

[0023] Accordingly, a computer device / apparatus / system includes a memory, a processor, and a computer program stored in the memory, wherein the processor executes the computer program to implement the steps of the method.

[0024] Accordingly, a computer-readable storage medium stores a computer program / instructions thereon, which, when executed by a processor, implement the steps of the method.

[0025] Accordingly, a computer program product includes a computer program / instructions that, when executed by a processor, implement the steps of the method.

[0026] This invention offers the following advantages: It provides, for the first time, a dual-wavelength spectrophotometric method for simultaneously and rapidly estimating the biomass of both algae and bacteria in an algal-bacterial symbiosis. When the biomass is small, a simple linear model can be used directly for estimation. However, when the biomass is large, the linear model's estimation effect is not ideal. This invention further constructs a nonlinear model, expanding the method's applicability and achieving better prediction results even for species biomass composition in co-culture systems with high biomass concentrations. Attached Figure Description

[0027] Figure 1 The cell density of YliSX and its A 600 The mathematical relationship between them;

[0028] Figure 2 The cell density of CsrLP and its A 680 Mathematical relationships between them (low concentration range);

[0029] Figure 3 The cell density of CsrLP and its A 680 Mathematical relationships between them (complete intervals);

[0030] Figure 4 Schematic diagrams of the wavelength conversion equations for YliSX and CsrLP, respectively;

[0031] Figure 5 A schematic diagram showing the comparison between the estimated CsrLP and YliSX biomass based on equation (5) and the measured values;

[0032] Figure 6 This is a schematic diagram of the ICC test results based on the estimation results of equation (5);

[0033] Figure 7 A schematic diagram showing the comparison between the estimated CsrLP and YliSX biomass based on equation (13) and the measured values;

[0034] Figure 8 This is a schematic diagram of the ICC test results based on the estimation results of equation (13). Detailed Implementation

[0035] This invention provides a method for rapidly estimating the biomass of algae and bacteria in an algal-bacterial symbiosis, specifically comprising the following steps:

[0036] 1. For algae and bacteria, their biomass can be represented by absorbance at 680 nm and 600 nm, respectively. Therefore, within the range where the relationship between absorbance and biomass is linear, the total absorbance of the algae-bacterial symbiosis can be expressed as equation (1):

[0037]

[0038] A 600 The total absorbance of the algae-bacterial symbiotic organism at 600 nm, A 680 The total absorbance of the algae-bacterial symbiosis at 680 nm. A 600，菌 A refers to the absorbance of bacteria in a symbiotic organism at 600 nm. 600，藻 This refers to the absorbance of algae in a symbiotic organism at 600 nm. A 680，菌 A refers to the absorbance of bacteria in a symbiotic organism at 680 nm. 680，藻 This refers to the absorbance of algae in a symbiotic organism at 680 nm.

[0039] 2. Establish the absorbance A of the bacterial cells at 680 nm. 680,菌 Absorbance A at 600 nm 600,菌 Mathematical relation A 680,菌 =f(A 600,菌 ), and the absorbance of the algae at 600 nm (A 600,藻 ) and absorbance at 680 nm (A 680,藻 Mathematical relation A 600,藻 =g(A 680,藻 Then equation (1) is transformed into equation (2):

[0040]

[0041] Because the overall absorbance of the algal-bacterial symbiosis at 680nm is A 680 and absorbance A at 600 nm 600 It can be measured by instruments, therefore theoretically A can be calculated by solving the above system of equations. 680,藻 and A 600,菌 This allows for the estimation of the biomass of algae and bacteria within the symbiotic organism.

[0042] 3. When biomass is low, i.e., absorbance is low, the wavelength conversion equation A for fungi is... 680,菌 =f(A 600,菌 And the wavelength conversion equation A for algae. 600,藻 =g(A 680,藻 Both can be expressed as general linear models, namely equations (3) and (4):

[0043] A 680,菌 =w×A 600,菌 +v (3)

[0044] A 600,藻 =w′×A 680,藻 +v′ (4)

[0045] Among them, A 680，菌 A refers to the absorbance of fungi at 680 nm. 600，菌 A refers to the absorbance of fungi at 600 nm.680，藻 A refers to the absorbance of algae at 680 nm. 600，藻 This refers to the absorbance of algae at 600 nm. w, v, w', and v' are parameters from a linear regression equation, which can be obtained by least-squares fitting (e.g., using the basic lm function in R). An alternative approach is to substitute the absorbance data of pure cultured bacteria or algae at 600 nm and 680 nm, respectively, and then fit the data using least-squares fitting.

[0046] 4. Substituting equations (3) and (4) into equation (1), we obtain equation (5):

[0047]

[0048] 5. By solving the system of linear equations in equation (5), A can be calculated. 600，菌 and A 680，藻 This indirectly characterizes the biomass of fungi and algae. Using absorbance to obtain the corresponding biomass is a well-known technique in the industry and will not be elaborated upon here.

[0049] 6. To further overcome the limitation that "the measured absorbance of the co-culture system is not the sum of the absorbance of the algae and bacteria at the corresponding wavelength" when the biomass is high, a theoretical absorbance (tA) is constructed, which is made to satisfy the linear change of tA with the change of biomass, specifically expressed by equation (6):

[0050]

[0051] Among them, tA 680 tA represents the total theoretical absorbance of the algae-bacterial symbiosis at 680 nm. 600 tA represents the total theoretical absorbance of the algae-bacterial symbiosis at 600 nm. 680，菌 The theoretical absorbance of fungi at 680 nm, tA 600，菌 The theoretical absorbance of fungi at 600 nm, tA 680，藻 The theoretical absorbance of algae at 680 nm, tA 600，藻 This refers to the theoretical absorbance of algae at 600 nm.

[0052] 7. For each item in equation (6), when the biomass is low, the measured absorbance can maintain a good linear relationship with the biomass. Therefore, the theoretical absorbance of the algal-bacterial symbiosis can be expressed as the product of the measured absorbance of the diluted algal-bacterial symbiosis and the dilution ratio, i.e., equations (7) and (8):

[0053] tA 600 =A′ 600 ×DR (7)

[0054] tA 680 =A′ 680×DR (8)

[0055] Among them, A′ 600 A′ represents the measured absorbance of the diluted algal-bacterial symbiont at 600 nm. 680 The absorbance of the diluted algae-bacterial symbiotic at 680 nm is denoted as DR, where DR represents the dilution ratio of the algae-bacterial symbiotic system (the dilution factor of the algae-bacterial symbiotic system; a 1-fold dilution results in a dilution ratio of 1). Preferably, the dilution principle for the algae-bacterial symbiotic is as follows: A = (Absorbance A of the diluted co-culture system) / (Absorbance A of the diluted system) 600 The wavelength does not exceed or slightly exceeds the linear range of the wavelength transformation equation when the bacteria are cultured alone, and A 680 The wavelength conversion equation does not exceed or slightly exceeds the linear range of the wavelength conversion equation when algae are cultured alone.

[0056] Extensive experiments have shown that the absorbance of fungi exhibits a linear relationship with cell density and absorbance at two wavelengths. Since cell density can represent the absolute value of biomass, the measured absorbance of fungi can be directly used as an approximation of its theoretical absorbance, i.e., equations (9) and (10):

[0057] tA 600,菌 ≈A 600,菌 (9)

[0058] tA 680,菌 ≈A 680,菌 (10)

[0059] Among them, tA 600，菌 The theoretical absorbance of fungi at 600 nm, tA 680，藻 A refers to the theoretical absorbance of algae at 680 nm. 600，菌 A refers to the measured absorbance of fungi at 600 nm. 680，菌 This refers to the measured absorbance of fungi at 680 nm.

[0060] Numerous experiments have shown that the absorbance of algae is related to cell density at higher absorbance levels (e.g., A). 680 When the absorbance exceeds 1.5), a linear relationship is no longer observed. The relationship between the theoretical absorbance tA and the measured absorbance A of the algae is constructed, where tA is obtained by substituting the cell density of sampling points in the pure culture system exceeding the low concentration range into the linear equation of absorbance-cell density in the low concentration range. The relationship between the theoretical absorbance and the measured absorbance can be obtained by fitting a quadratic polynomial, namely equations (11) and (12):

[0061] tA 600藻 =α×A 600,藻 2 +β×A 600,藻 +γ (11)

[0062] tA 680,藻 =α′×A680,藻 2 +β′×A 680,藻 +γ′ (12)

[0063] Among them, tA 680，藻 The theoretical absorbance of algae at 680 nm, tA 600，藻 A refers to the theoretical absorbance of algae at 600 nm. 680，藻 A refers to the actual absorbance of algae at 680 nm. 600，藻 This refers to the actual absorbance of algae at 600 nm. α, β, γ, α', β', and γ' are all conventional parameters for quadratic polynomial nonlinear fitting, which can be obtained through nonlinear least squares fitting (e.g., the nls function in R). An alternative approach is to substitute the absorbance data of pure cultured bacteria or algae at 600 nm and 680 nm respectively, and then fit the data using least squares fitting.

[0064] 8. In summary, we obtain equation (13):

[0065]

[0066] The biomass of algae and bacteria in the symbiont can be calculated by solving equation (13).

[0067] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art, and the data obtained are all average values ​​obtained after at least three repetitions, and each repetition yields valid data.

[0068] Example

[0069] Chlorella sorokiniana LP was selected as the experimental algal species (labeled CsrLP), and Yarrowia lipolitica SX was selected as the experimental bacterial species (labeled YliSX). Both were co-cultured on AYX-2 medium, the formulation of which is shown in Table 1.

[0070] Table 1 AYX-2 Culture Medium Formulation Table

[0071]

[0072] 1. Detection of absorbance and cell density in pure algae and bacteria culture systems (algae and bacteria cultured separately)

[0073] CsrLP (cultured in TAP medium) and YliSX (cultured in SD medium) at the late logarithmic growth stage were inoculated at a 1% (v / v) inoculation rate into 120mL PC tissue culture flasks containing 50mL of AYX-2 medium. The flasks were placed in a light-shaking incubator (using a 6000K DC LED lamp as the light source) and cultured under conditions of 2000lx light intensity, 24h continuous illumination, 30℃, and 120rpm until the plateau phase. At regular intervals, 1mL of well-mixed culture medium was added to 1.5mL EP tubes, with 200μL used for A detection using a microplate reader. 680 and A 600 The remaining culture medium was used to detect cell density. Data from multiple batches of replicate experiments were recorded, and the results of each batch were finally combined. The results are shown in Tables 2 and 3. "-" indicates that the test was not performed. There is a negative value in Table 3 because the absorbance value needs to be subtracted from the blank value. When the absorbance is low, the result may be negative due to large random errors, and it can be regarded as 0.

[0074] Table 2 Comparison of absorbance and cell density data for CsrLP cultured alone (pure culture)

[0075]

[0076] Table 3 Comparison of absorbance and cell density data for YliSX cultured alone (pure culture)

[0077]

[0078] Based on the data in Tables 2 and 3, the cell density of YliSX and its correlation with A were established. 600 Mathematical relationships between them (e.g.) Figure 1 As shown), and the cell density of CsrLP and its A 680 The mathematical relationship between them (relationship in low concentration range, such as...) Figure 2 ; complete interval such as Figure 3 As shown in the figure, it is used to evaluate the calculation results of the dual-wavelength spectrophotometry in subsequent experiments.

[0079] The results showed that within the lower absorbance range, both YliSX and CsrLP exhibited a good linear relationship between cell density and corresponding absorbance. However, as biomass increased, the relationship between CsrLP cell density and corresponding absorbance needed to be expressed as a non-linear relationship.

[0080] 2. Detection of absorbance and cell density in the algae-bacteria co-culture system

[0081] CsrLP and YliSX were simultaneously inoculated into 120mL PC tissue culture flasks containing 50mL AYX-2 medium at different initial inoculation ratios (CsrLP:YliSX = 1:1, 1:3, 3:1, 3:3). The flasks were then placed in a light-shaking incubator (using a 6000K DC LED lamp as the light source) and co-cultured under conditions of 2000 lx light intensity, 24h continuous illumination, 30℃, and 120 rpm. Every so often, 1mL of the well-mixed culture medium was added to a 1.5mL EP tube, with 200μL used for detecting the A value of the co-culture system using a microplate reader. 600 and A 680 The remaining culture medium was used to detect cell density. The results are shown in Table 4.

[0082] Table 4. Comparison of absorbance and cell density in the CsrLP and YliSX co-culture system.

[0083]

[0084]

[0085] While measuring the data in Table 4, the culture medium was diluted several times using AYX-2 medium, and the A content of the diluted co-culture system was then measured. 600 A 680 Cell density. The results are shown in Table 5.

[0086] Table 5. Absorbance and cell density comparison of CsrLP and YliSX co-culture systems (after dilution)

[0087]

[0088] 3. Use linear formulas to estimate the biomass of CsrLP and YliSX respectively.

[0089] (1) A measured using Table 3 680 and A 600 The data were linearly regressed using equation (3) as the model to determine the wavelength conversion equation coefficients w and v of YliSX. Figure 4 a) A measured using Table 2 680 and A 600 The data (scatter plot) shows a linear trend within a certain range, i.e., range A. 680 <1.75), using equation (4) as the model, linear regression was performed to determine the wavelength conversion equation coefficients w' and v' of CsrLP. Figure 4 (b) Wherein, the specific values ​​of w, v, w' and v' are 0.952598, -0.000103, 0.71593 and -0.00559, respectively.

[0090] (2) The A measured in Table 4 680 and A 600 Substituting the wavelength conversion equation coefficients w, v, w', and v' of YliSX and CsrLP into equation (5), the absorbance A of YliSX and CsrLP in the co-culture system is calculated respectively. 600,菌 and A 680,藻 And substitute them into their respective absorbance-cell density conversion equations (YliSX uses) Figure 1 Corresponding equation; CsrLP uses Figure 2 and Figure 3 Corresponding equation, where: A 680,藻 <1.5 usage Figure 2 A 680,藻 ≥1.5 use Figure 3 ), calculate the corresponding estimated cell density value, and compare it with the measured cell density value corresponding to each sampling point in Table 4.

[0091] Growth curves showing the measured and estimated cell densities of CsrLP and YliSX co-culture systems based on different initial inoculum amounts are plotted as follows: Figure 5 As shown. Figure 5 In the diagram, dots represent estimated values, lines represent measured values, and the horizontal axis represents the relative time of sampling for different inoculum levels. An intraclass correlation coefficient (ICC) test was performed between the measured and estimated values, and the results are shown below. Figure 6 As shown, Figure 6 The horizontal axis represents the estimated cell density, and the vertical axis represents the measured cell density.

[0092] The results show that relying solely on the linear model (5) can effectively estimate the biomass of YliSX and CsrLP when their biomass is relatively balanced (i.e., as shown in equation (5)). Figure 5 The experimental groups with Y:C = 1:1 and Y:C = 3:1. However, when the biomass of a certain microorganism is high and the overall biomass of the system is high (e.g., Figure 5 In the experimental groups with Y:C = 1:3 and Y:C = 3:3, the initial inoculum of CsrLP was high, resulting in a very high CsrLP cell density in the system after 20 hours of culture. At this point, a large error occurred between the estimated and measured values.

[0093] 4. Use nonlinear formulas to estimate the biomass of CsrLP and YliSX respectively.

[0094] (1) To estimate the biomass of YliSX and CsrLP using the nonlinear model (13), the relationship between the theoretical absorbance tA and the measured absorbance A of the algae was first constructed: the cell density of each sampling point measured in Table 2 was substituted into the linear equation of cell density-absorbance in the low concentration range of CsrLP ( Figure 2The theoretical absorbance of each sampling point is calculated. Further, within the range where the wavelength conversion equation shows a good linear relationship, the absorbance and theoretical absorbance are fitted using quadratic polynomials according to equations (11) and (12) to determine the parameters α, β, γ, α', β', and γ'. The values ​​of α, β, γ, α', β', and γ' are 0.62541, 0.44035, 0.06367, 0.4292, 0.4740, and 0.0835, respectively.

[0095] (2) According to equations (7) and (8), if an attempt is made to extend the detection range of the method using a nonlinear model, the product of the absorbance of the diluted culture medium and the dilution ratio must be used for calculation. Therefore, the A measured in Table 5 (after dilution) 680 and A 600 Substituting the data, the wavelength conversion equation of YliSX, the wavelength conversion equation of CsrLP, and the quadratic polynomial model of absorbance and theoretical absorbance into equation (13), solve for A. 600,菌 and A 680,藻 The results were then substituted into the mathematical relationship between absorbance and cell density in YliSX and CsrLP (YliSX uses...). Figure 1 Corresponding equation; CsrLP uses Figure 2 and Figure 3 Corresponding equation, where: A 680,藻 <1.5 usage Figure 2 A 680,藻 ≥1.5 use Figure 3 The data were converted to cell density and then compared with the measured cell density. The results are as follows: Figure 7 As shown. Figure 7 In the diagram, dots represent estimated values, lines represent measured values, and the horizontal axis represents the relative time of sampling for different inoculum levels. An ICC test was performed on the measured and estimated values, and the results are shown below. Figure 8 As shown. Figure 8 In the diagram, the horizontal axis represents the estimated cell density, and the vertical axis represents the measured cell density.

[0096] Results based on nonlinear models and Figure 5 and Figure 6 In comparison, it is evident that the biomass estimation method based on a nonlinear model can better meet the needs of higher biomass conditions, thus expanding the applicability of the method to some extent. The results show that, compared to the dual-wavelength spectrophotometric method based on a linear model, the introduction of a nonlinear model expands the applicability of this method and has better performance in predicting the species biomass composition of co-culture systems with relatively high biomass concentrations.

[0097] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Any modifications, alterations, substitutions, or variations made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention shall fall within the protection scope defined by the claims of the present invention.

Claims

1. A method for rapidly estimating the biomass of algae and bacteria in an algal-bacterial symbiosis, characterized in that: The method includes the following steps: (1) The total absorbance of the algal-bacterial symbiotic organism is expressed as Equation (1): A 600 The total absorbance of the algae-bacterial symbiotic organism at 600 nm, A 680 The total absorbance of the algae-bacterial symbiotic organism at 680 nm, A 600，菌 A refers to the absorbance of bacteria in a symbiotic organism at 600 nm. 600，藻 A refers to the absorbance of algae in a symbiotic organism at 600 nm. 680，菌 A refers to the absorbance of bacteria in a symbiotic organism at 680 nm. 680，藻 This refers to the absorbance of algae in a symbiotic organism at 680 nm. (2) Establish the absorbance A of the bacterial cells at 680 nm. 680,菌 Absorbance A at 600 nm 600,菌 Mathematical relation A 680,菌 =f(A 600,菌 ), and the absorbance of the algae at 600 nm (A 600,藻 ) and absorbance at 680 nm (A 680,藻 Mathematical relation A 600,藻 =g(A 680,藻 Then equation (1) is transformed into equation (2): (3) Wavelength conversion equation for fungi A 680,菌 =f(A 600,菌 And the wavelength conversion equation A for algae. 600,藻 =g(A 680,藻 Both can be represented by general linear models, namely equations (3) and (4): And 680,菌 =w×A 600,菌 +in (3) A 600,藻 =w′×A 680,藻 +v′ (4) Among them, w, v, w' and v' are all equation parameters obtained based on linear regression, which can be fitted by the least squares method; (4) Substituting equations (3) and (4) into equation (1), we obtain equation (5): Solve equation (5) to obtain the biomass of fungi and algae.

2. A method for rapidly estimating the biomass of algae and bacteria in an algal-bacterial symbiosis, characterized in that: The method includes the following steps: Constructing formula (13): Among them, A′ 600 A′ represents the measured absorbance of the diluted algal-bacterial symbiont at 600 nm. 680 A represents the measured absorbance of the diluted algae-bacterial symbiont at 680 nm, where DR is the dilution ratio of the algae-bacterial symbiont; A 600，菌 A refers to the absorbance of bacteria in a symbiotic organism at 600 nm. 600，藻 A refers to the absorbance of algae in a symbiotic organism at 600 nm. 680，菌 A refers to the absorbance of bacteria in a symbiotic organism at 680 nm. 680，藻 The absorbance of algae in the symbiotic organism at 680 nm; w, v, w', v', α, β, γ, α', β' and γ' are all conventional parameters for quadratic polynomial nonlinear fitting, which can be obtained by fitting using the nonlinear least squares method; Solve equation (13) to obtain the biomass of fungi and algae.

3. A computer device / equipment / system, comprising a memory, a processor, and a computer program stored in the memory, characterized in that, The processor executes the computer program to implement the steps of the method of claim 1 or 2.

4. A computer-readable storage medium having a computer program / instructions stored thereon, characterized in that, When the computer program / instructions are executed by the processor, they implement the steps of the method described in claim 1 or 2.

5. A computer program product, comprising a computer program / instructions, characterized in that, When the computer program / instructions are executed by the processor, they implement the steps of the method described in claim 1 or 2.

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