CN113593642B - Identification method of microorganism influence mechanism by slope tillage ridge ditch layout - Google Patents

Identification method of microorganism influence mechanism by slope tillage ridge ditch layout Download PDF

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CN113593642B
CN113593642B CN202111041681.2A CN202111041681A CN113593642B CN 113593642 B CN113593642 B CN 113593642B CN 202111041681 A CN202111041681 A CN 202111041681A CN 113593642 B CN113593642 B CN 113593642B
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樊晶晶
刘姗姗
秦天玲
赵爽
严登华
林帅
董碧琼
冯贱明
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China Institute of Water Resources and Hydropower Research
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Abstract

本发明提供了一种坡耕地垄沟布局对微生物影响机理的识别方法，属于土壤微生物学技术领域，包括：确定坡耕地垄沟布局参数；布设实验小区和对照小区；进行土壤微生物高通量测序，得到土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据；确定对所述土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据产生影响的坡耕地垄沟布局参数的主次顺序；构建线性回归方程；对所述线性回归方程进行显著性检验，完成对坡耕地垄沟布局对微生物影响机理的识别。本发明选择四种具有代表性的垄沟布局参数，通过高通量测序技术对不同垄沟布局下土壤微生物群落多样性进行分析，识别坡耕地垄沟布局对土壤微生物影响机理，为制定合适的坡耕地垄沟布局方式提供参考。

The invention provides a method for identifying the mechanism of the impact of ridge and furrow layout on sloping farmland on microorganisms. It belongs to the technical field of soil microbiology and includes: determining the ridge and furrow layout parameters of sloping farmland; laying out experimental plots and control plots; performing high-throughput sequencing of soil microorganisms to obtain Soil microbial community structure diversity related index and soil carbon and nitrogen cycle process functional gene abundance data; determine the ridge and furrow layout of sloping farmland that affects the soil microbial community structure diversity related index and soil carbon and nitrogen cycle process functional gene abundance data. The primary and secondary order of parameters; constructing a linear regression equation; conducting a significance test on the linear regression equation to complete the identification of the impact mechanism of the ridge and furrow layout of sloping farmland on microorganisms. The present invention selects four representative ridge and furrow layout parameters, uses high-throughput sequencing technology to analyze the diversity of soil microbial communities under different ridge and furrow layouts, and identifies the impact mechanism of ridge and furrow layout on sloping farmland on soil microorganisms, in order to formulate appropriate ridge and furrow layout for sloping farmland. The layout method provides reference.

Description

Identification method of microorganism influence mechanism by slope tillage ridge ditch layout
Technical Field
The invention belongs to the technical field of soil microbiology, and particularly relates to a method for identifying a microorganism influence mechanism by slope cultivation land ridge ditch layout.
Background
The hillside fields are used as a common agricultural soil resource, and the special landform types severely restrict the agricultural production of dry lands, and the furrow cultivation is used as a common protective cultivation measure, so that the problems of water running, fertilizer running and the like caused by the hillside fields can be effectively solved under certain conditions. Microorganisms in soil participate in material circulation and energy flow among soil ecosystems, a promoting effect is provided for growth and development of crop root systems, at present, research on influence of ridge and ditch cultivation on soil microorganisms in hillside cultivation is mainly focused on the influence of ridge and ditch cultivation combined with film covering and straw covering on the microorganisms, and whether the mode of ridge and ditch cultivation affects the soil microorganisms cannot be known simply due to restriction of multiple factors of film covering and straw.
Disclosure of Invention
Aiming at the defects in the prior art, the identification method for the microbial influence mechanism of the slope cultivation ridge ditch layout solves the problem that whether the ridge ditch cultivation mode affects soil microorganisms cannot be known simply.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the scheme provides a method for identifying a microorganism influence mechanism by slope cultivation ridge ditch layout, which comprises the following steps:
s1, selecting a research area, and determining a planting type and a slope cultivation ridge ditch layout parameter, wherein the slope cultivation ridge ditch layout parameter comprises the following steps: ridge gradient, ridge height, furrow width ratio and furrow trend;
s2, arranging an experimental cell and a control cell according to a four-factor three-level orthogonal table by using a ridge gradient, a ridge height, a furrow width ratio and a furrow trend, and planting selected crops;
s3, respectively collecting soil samples of each experimental cell and each control cell before crop planting, during the middle growing period and during the harvesting period, and carrying out high-throughput sequencing on the soil microorganisms to obtain the diversity related indexes of soil microbial communities of different experimental cells and control cells and the functional gene abundance data of the soil carbon nitrogen circulation process;
s4, checking the influence of the ridge gradient, the ridge height, the furrow width ratio and the furrow trend on the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen cycle process by using an analysis of variance method, and determining the major and minor sequences of four factors affecting the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen cycle process;
s5, constructing a linear regression equation by the primary and secondary sequences and the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen circulation process respectively;
s6, performing significance test on the linear regression equation, and taking standard coefficients of ridge gradient, ridge height, furrow width ratio and furrow trend of the linear regression equation as evaluation slope furrow layout to influence soil microorganisms according to test results, so as to complete identification of a microorganism influence mechanism of slope tillage ridge furrow layout.
The beneficial effects of the invention are as follows: according to the invention, the experimental design is carried out on different furrow layout parameters by selecting orthogonal experiments, so that the experimental times are greatly reduced; the analysis of variance method is utilized to test and screen out furrow layout parameters with the greatest influence on soil microorganisms, the importance degree of each furrow layout parameter on experimental results can be accurately estimated, a linear regression equation is constructed between each furrow layout parameter estimated by the analysis of variance method and soil microorganism community structure and functional diversity, and a standard coefficient is used for evaluating the influence of furrow layout on soil microorganisms, so that theoretical reference can be provided for furrow layout of hillside lands, and the purpose of agricultural high yield is achieved.
Further, the step S3 includes the steps of:
s301, selecting two sampling areas of a ridge and a ditch in the upper, middle and lower sections of an experimental cell and a control cell before crop planting, in the middle of the growing period and in the harvesting period, removing surface layer floating soil, collecting soil samples, removing impurities of a soil sample mixture of the same cell, and filling the soil samples into a sterilizing packaging bag for preservation;
s302, performing high-throughput sequencing on soil microorganisms on a soil sample to obtain the sequence number of microorganism DNA or RNA, OTU abundance information and species classification information of the soil sample in each experimental cell and each control cell;
s303, calculating and obtaining the diversity related indexes of the soil microbial community structures of different experimental cells and control cells and the functional gene abundance data of the soil carbon and nitrogen circulation process according to the sequence number of the soil sample microbial DNA or RNA, the OTU abundance information and the species classification information.
The beneficial effects of the above-mentioned further scheme are: by sampling soil samples of the experimental plot in three periods, the influence on experimental errors on a time scale is avoided; the high-throughput sequencing is carried out on the soil sample, so that the abundance and the diversity of the microbial community structure in the soil sample can be judged more quickly and accurately, and the soil microbial community structure diversity and the functional gene abundance data in the soil carbon nitrogen circulation process are used as evaluation indexes for the influence of soil microorganisms, thereby being beneficial to comprehensively analyzing the influence of the furrow layout on the soil microbial community in structural and functional aspects.
Still further, the step S4 includes the steps of:
s401, calculating the related indexes of the microbial community structure diversity in soil samples of each experimental cell and each control cell, and the average value of the functional gene abundance data in the soil carbon nitrogen circulation process;
s402, calculating to obtain the dispersion square sum and the degree of freedom of the microbial community structure diversity related indexes and the soil carbon nitrogen circulation process functional gene abundance data in the soil samples of each experimental cell and the control cell according to the average value;
s403, according to the dispersion square sum and the degree of freedom, using an analysis of variance method to test the influence of different ridge gradients alpha, ridge heights h, furrow width ratios beta and furrow trend d on the diversity related indexes of the soil microbial community structure and the functional gene abundance data of the soil carbon nitrogen circulation process;
s404, performing significance test on the influence result obtained in the step S403;
s405, sorting the influence results obtained in the step S403 from large to small according to the significance test result to obtain a major-minor sequence of four factors of ridge gradient, ridge height, furrow width ratio and furrow trend, which influence the structural diversity related index of the soil microbial community and the functional gene abundance data in the soil carbon nitrogen circulation process.
The beneficial effects of the above-mentioned further scheme are: the influence of different furrow layout parameters on the soil microbial community structure diversity related index and the soil carbon nitrogen circulation process functional gene abundance is tested by calculating the average value, the dispersion square sum and the degree of freedom of the microbial community structure diversity related index and the soil carbon nitrogen circulation process functional gene abundance data in the soil samples of each experimental cell and the control cell, so that the importance degree of the influence of the experimental result of each factor can be accurately estimated.
Still further, the expression of the average value in step S401 is as follows:
j＝1,2,…,p；k＝1,2,…,r；i＝1,2,…,N
wherein ,the average value of the microbial community structure diversity related index and the soil carbon nitrogen circulation process functional gene abundance data of the soil samples of each experimental cell and the control cell is represented, p represents the number of the experimental cells, r represents the measurement times, j represents the jThe k represents the kth measurement of the experimental cell, x jk The index related to the structural diversity of the microbial community measured in the jth experimental community at the kth time is represented, N represents the number of functional genes related to the soil carbon nitrogen cycle process, i represents the number of functional genes related to the ith soil carbon nitrogen cycle process, and FCN ijk The functional gene abundance value of the ith soil carbon nitrogen circulation process measured in the kth experimental community is shown.
The beneficial effects of the above-mentioned further scheme are: and respectively calculating the related indexes of the structural diversity of the soil microbial community and the average value of the functional gene abundance data of the soil carbon nitrogen circulation process, and preparing for the subsequent analysis of variance.
Still further, the expression of the sum of squares of the dispersion and the degree of freedom in the step S402 is as follows:
f T ＝p-1
f A ＝q-1
A＝{α，β，h，d}
wherein ,ST The total dispersion square sum of the microbial community structure diversity related index and the soil carbon nitrogen cycle process functional gene abundance data in the soil samples of each experimental cell and the control cell is represented, S A Representing the sum of squares of dispersion caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, S e Representing error dispersionSum of squares, f T 、f A 、f e All represent S T 、S A and Se Corresponding degrees of freedom, y j The related index of the structural diversity of the microbial community in the soil sample of the j-th experimental plot and the functional gene abundance data of the soil carbon-nitrogen circulation process are expressed,the average value of the microbial community structure diversity related index and the functional gene abundance data in the soil carbon nitrogen circulation process in each experimental cell and control cell is represented, q represents the horizontal number of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, m represents the column number of an orthogonal table, u represents the u th column and k th column of the orthogonal table o Represents the arithmetic mean of the experimental results obtained when the four factors on any column were taken at level o.
The beneficial effects of the above-mentioned further scheme are: and respectively calculating the deviation square sum and the degree of freedom of the experimental and control district soil microbial community structure diversity related indexes and the soil carbon nitrogen circulation process functional gene abundance data caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, which are beneficial to comprehensively analyzing the soil microbial community structure and functional influence from different furrow layout parameters and preparing for analysis of variance calculation.
Still further, in the step S403, the expression for checking the influence of different ridge gradients α, ridge heights h, furrow width ratios β, furrow directions d on the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen cycle process is as follows:
wherein ,FA Soil microorganisms caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d are represented by the mean square of soil microbial community structure diversity related indexes and functional gene abundance data in the soil carbon-nitrogen circulation process, wherein the soil microbial community structure diversity related indexes are caused by ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend dThe ratio of the mean square of the data error of the functional gene abundance in the community structure diversity related index and the soil carbon nitrogen cycle process.
The beneficial effects of the above-mentioned further scheme are: the analysis of variance is carried out on the functional gene abundance data of the soil microorganism community structure diversity related index and the soil carbon nitrogen circulation process, which are affected by the ridge gradient alpha, the ridge height h, the furrow width ratio beta and the furrow trend d, respectively, the difference significance of the results of the influence of different furrow layout parameters on soil microorganisms is checked, the importance degree of the influence of each furrow layout parameter on the experimental results can be accurately estimated, and then the furrow layout parameters with the greatest influence on the soil microorganisms are screened out.
Still further, the expression for performing the significance test in step S404 is as follows:
wherein phi represents F of each ridge gradient alpha, ridge height h, ridge width ratio beta and ridge trend d α 、F h 、F β 、F d Ratio to F value at gamma=0.05 significance level, F A The degree of freedom of each experimental cell and the control cell caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d is shown as (f) A ，f e ) The ratio of the mean square of the soil microbial community structure diversity related index and the functional gene abundance data of the soil carbon nitrogen circulation process to the mean square of experimental errors caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, F 0.05 The degree of freedom of each experimental cell and the control cell, which is caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d, is expressed as (f A ，f e ) The ratio of the mean square of the soil microbial community structure diversity related index and the soil carbon nitrogen circulation process functional gene abundance data to the mean square of experimental errors caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d at the significance level of gamma=0.05.
The beneficial effects of the above-mentioned further scheme are: whether the furrow layout has obvious influence on the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen circulation process is checked from four influencing factors of the furrow gradient alpha, the furrow height h, the furrow width ratio beta and the furrow trend d, and the process analysis result is further improved.
Still further, the expression of the linear regression equation in the step S5 is as follows:
or (b)
wherein ,b0 、b 1 、b 2 、b 3 、b 4 Coefficients, alpha, respectively representing linear regression equations j 、h j 、β j 、d j Respectively representing the data of the ridge gradient alpha, the ridge height h, the furrow width ratio beta and the furrow trend d, y of the jth experimental cell j The data of the related index of the structural diversity of the soil microbial community and the functional gene abundance in the soil carbon nitrogen circulation process of the jth experimental plot are represented, p represents the number of the experimental plot, and x jk The index related to the structural diversity of the microbial community, which represents the kth measurement of the jth experimental cell, FCN ijk The functional gene abundance value of the ith soil carbon nitrogen circulation process measured in the kth experimental community is shown.
The beneficial effects of the above-mentioned further scheme are: by constructing a linear regression equation, the linear relation between the furrow layout parameters and the structural diversity related indexes of the soil microbial communities and the functional gene abundance data in the carbon-nitrogen circulation process of the soil is more clearly expressed, and the statistical rule hidden behind the randomness is found.
Still further, the expression for performing the significance test on the linear regression equation in the step S6 is as follows:
wherein ,RJ 、R G Representing complex correlation coefficients of each linear regression equation, S RJ Regression square sum of linear regression equation representing ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and soil microbial community structure diversity correlation index of each experimental cell and control cell, S TJ Representing the total sum of squares of linear regression equations of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and soil microbial community structure diversity correlation index of each experimental cell and control cell, S RG Expressing regression square sum of linear regression equation of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and functional gene abundance data of soil carbon nitrogen circulation process of each experimental cell and control cell, and S TG And the total sum of squares of linear regression equations of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and functional gene abundance data in the soil carbon nitrogen circulation process of each experimental cell and the control cell are represented.
The beneficial effects of the above-mentioned further scheme are: the reliability of the equation is checked by performing a significance test on the solved linear regression equation.
Drawings
FIG. 1 is a flow chart of the method of the present invention.
Fig. 2 is a layout diagram of furrow parameters of each experimental cell in this embodiment.
Fig. 3 is a cross-sectional view of the distribution of the furrow parameters of each experimental cell in this embodiment.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and all the inventions which make use of the inventive concept are protected by the spirit and scope of the present invention as defined and defined in the appended claims to those skilled in the art.
Examples
As shown in fig. 1, the invention provides a method for identifying the microorganism influence mechanism of a slope cultivation ridge ditch layout, which comprises the following steps:
s1, selecting a research area, and determining a planting type and a slope cultivation ridge ditch layout parameter, wherein the slope cultivation ridge ditch layout parameter comprises the following steps: ridge gradient, ridge height, furrow width ratio and furrow trend;
in the embodiment, a research area is selected, and the experimental area furrow arrangement control parameters, the parameter variation range and the typical crop are determined by combining local cultivation habits and typical crop through field investigation.
S2, arranging experimental cells and comparison cells according to a four-factor three-level orthogonal table by using ridge gradients, ridge heights, furrow width ratios and furrow directions, and planting selected crops, wherein the experimental cells and the comparison cells are arranged as shown in fig. 2 and 3, alpha in fig. 3 represents the ridge gradients, h represents the ridge heights, c represents the furrow widths, e represents the furrow widths, beta represents the furrow width ratios (beta=c/e), and d represents the furrow directions;
in this embodiment, the orthogonal table selects four-factor three level L without consideration of interaction 9 (3 4 ) The orthogonal table is designed as shown in table 1.
TABLE 1
In the orthogonal table, alpha 1 、α 2 、α 3 Three levels representing the slope of the ridge; h is a 1 、h 2 、h 3 Three levels representing ridge heights; beta 1 、β 2 、β 3 Three levels representing furrow-to-width ratios; d, d 1 、d 2 、d 3 Three levels representing the trend of the furrows are distributed among the levels according to an arithmetic progression within the variation range of furrow control parameters in a study area.
S3, respectively collecting soil samples of each experimental cell and each control cell before crop planting, during the middle growing period and during the harvesting period, and carrying out high-throughput sequencing on the soil microorganisms to obtain the diversity related indexes of soil microbial communities of different experimental cells and control cells and the functional gene abundance data of the soil carbon nitrogen circulation process, wherein the implementation method comprises the following steps:
s301, selecting two sampling areas of a ridge and a ditch in the upper, middle and lower sections of an experimental cell and a control cell before crop planting, in the middle of the growing period and in the harvesting period, removing surface layer floating soil, collecting soil samples, removing impurities of a soil sample mixture of the same cell, and filling the soil samples into a sterilizing packaging bag for preservation;
s302, performing high-throughput sequencing on soil microorganisms on a soil sample to obtain the sequence number of microorganism DNA or RNA, OTU abundance information and species classification information of the soil sample in each experimental cell and each control cell;
s303, calculating and obtaining the diversity related indexes of the soil microbial community structures of different experimental cells and control cells and the functional gene abundance data of the soil carbon and nitrogen circulation process according to the sequence number of the soil sample microbial DNA or RNA, the OTU abundance information and the species classification information.
In the embodiment, two sampling areas of ridges and furrows (sampling areas at the same positions are selected in a flat comparison cell) are selected in an experiment cell and a comparison cell of 3 stages of a period of time of growing, a period of growing and a period of harvesting before crop planting, the period of growing and the period of harvesting, floating soil with 0-5cm on the surface layer is removed, 10g of soil sample is collected, and after the soil sample of the same cell is mixed, impurities such as animal and plant residues are removed, the mixture is filled into a sterilizing self-sealing bag and stored at the temperature of minus 80 ℃.
In this embodiment, soil samples are subjected to high-throughput sequencing of soil microorganisms, and the high-throughput sequencing platform is an Illumina Miseq platform. Clustering and flattening OTUs at a similar level of 97% on the data after high-throughput sequencing to obtain the number of sequences of microorganism DNA or RNA, the abundance information of the OTUs and the species classification information of samples of each experimental cell and the control cell.
In this example, the microbial community structure diversity related index Chao, shannon, PD and the functional gene abundance data of the soil carbon nitrogen cycle process in each soil sample were calculated and respectively recorded as CH jk ，S jk ，PD jk ，FCN ijk Where i=1, 2, …, N represents the number of functional genes involved in carbon nitrogen cycling, as determined based on actual experimental procedures; j=1, 2, …,10, j denotes the number of test cells, k denotes the kth measurement, k=1, 2,3.
S4, checking the influence of the ridge gradient, the ridge height, the furrow width ratio and the furrow trend on the diversity related index of the soil microbial community structure and the functional gene abundance data of the soil carbon nitrogen cycle process by using an analysis of variance method, and determining the major and minor sequences of the four factors influencing the diversity related index of the soil microbial community structure and the functional gene abundance data of the soil carbon nitrogen cycle process, wherein the implementation method is as follows:
s401, calculating the related indexes of the microbial community structure diversity in soil samples of each experimental cell and each control cell, and the average value of the functional gene abundance data in the soil carbon nitrogen circulation process:
j＝1,2,…,p；k＝1,2,…,r；i＝1,2,…,N
wherein ,the average value of the microbial community structure diversity related index and the soil carbon nitrogen circulation process functional gene abundance data of the soil samples of each experimental cell and the control cell is represented, p represents the number of the experimental cells, r represents the measurement times, j represents the j-th experimental cell, k represents the k-th measurement of the experimental cell, and x jk The index related to the structural diversity of the microbial community measured in the kth experimental community is represented, N represents the number of functional genes related to the carbon-nitrogen circulation of soil,i represents the number of functional genes related to the carbon nitrogen cycle of the ith soil, FCN ijk Expressing the functional gene abundance value of the ith soil carbon nitrogen circulation process measured by the kth experimental plot;
s402, calculating to obtain the dispersion square sum and the degree of freedom of the microbial community structure diversity related indexes and the soil carbon nitrogen circulation process functional gene abundance data in the soil samples of each experimental cell and the control cell according to the average value:
f T ＝p-1
f A ＝q-1
A＝{α，β，h，d}
wherein ,ST The total dispersion square sum of the microbial community structure diversity related index and the soil carbon nitrogen cycle process functional gene abundance data in the soil samples of each experimental cell and the control cell is represented, S A Representing the sum of squares of dispersion caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, S e Represents the sum of squares of error dispersion, f T 、f A 、f e All represent S T 、S A and Se Corresponding degrees of freedom, y j The related index of the structural diversity of the microbial community in the soil sample of the j-th experimental plot and the functional gene abundance data of the soil carbon-nitrogen circulation process are expressed,the average value of the microbial community structure diversity related index and the functional gene abundance data in the soil carbon nitrogen circulation process in each experimental cell and control cell is represented, q represents the horizontal number of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, m represents the column number of an orthogonal table, u represents the u th column and k th column of the orthogonal table o An arithmetic average value representing experimental results obtained when the four factors on any column are taken as level o;
s403, according to the dispersion square sum and the degree of freedom, using an analysis of variance method to test the influence of different ridge gradients alpha, ridge heights h, furrow width ratios beta and furrow trend d on the diversity related indexes of the soil microbial community structure and the functional gene abundance data of the soil carbon nitrogen circulation process:
wherein ,FA The method is characterized by comprising the steps of expressing the ratio of the mean square of the structural diversity related index of the soil microbial community and the functional gene abundance data in the carbon-nitrogen circulation process, which are caused by ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, to the mean square of the structural diversity related index of the soil microbial community and the functional gene abundance data error in the carbon-nitrogen circulation process, which are caused by the ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d;
s404, performing significance test on the influence result obtained in the step S403:
wherein phi represents F of each ridge gradient alpha, ridge height h, ridge width ratio beta and ridge trend d α 、F h 、F β 、F d Ratio to F value at gamma=0.05 significance level, F A The degree of freedom of each experimental cell and the control cell caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d is shownIs (f) A ，f e ) The ratio of the mean square of the soil microbial community structure diversity related index and the functional gene abundance data of the soil carbon nitrogen circulation process to the mean square of experimental errors caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d, F 0.05 The degree of freedom of each experimental cell and the control cell, which is caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d, is expressed as (f A ，f e ) The ratio of the mean square of the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon and nitrogen circulation process to the mean square of experimental errors caused by four factors of ridge gradient alpha, ridge height h, furrow width ratio beta and furrow trend d under the significance level of gamma=0.05;
s405, sorting the influence results obtained in the step S403 from large to small according to the significance test result to obtain a major-minor sequence of four factors of ridge gradient, ridge height, furrow width ratio and furrow trend, which influence the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen circulation process;
s5, constructing a linear regression equation by the primary and secondary sequences and the structural diversity related index of the soil microbial community and the functional gene abundance data of the soil carbon nitrogen circulation process respectively;
in this embodiment, it is assumed that four factors of the ridge gradient α, the ridge height h, the furrow width ratio β, and the furrow trend d are F according to the arrangement order affecting the primary and secondary α >F h >F β >F d Let equation y=b 0 +αb 1 +hb 2 +βb 3 +db 4 The system of linear regression equations is:
or (b)
wherein ,b0 、b 1 、b 2 、b 3 、b 4 Coefficients, alpha, respectively representing linear regression equations j 、h j 、β j 、d j Respectively representing the data of the ridge gradient alpha, the ridge height h, the furrow width ratio beta and the furrow trend d, y of the jth experimental cell j The data of the related index of the structural diversity of the soil microbial community and the functional gene abundance in the soil carbon nitrogen circulation process of the jth experimental plot are represented, p represents the number of the experimental plot, and x jk The index related to the structural diversity of the microbial community, which represents the kth measurement of the jth experimental cell, FCN ijk The functional gene abundance value of the ith soil carbon nitrogen circulation process measured in the kth experimental community is shown.
S6, performing significance test on the linear regression equation, and taking standard coefficients of ridge gradient, ridge height, furrow width ratio and furrow trend of the linear regression equation as evaluation slope furrow layout to influence soil microorganisms according to test results, so as to complete identification of a microorganism influence mechanism of slope tillage ridge furrow layout.
In this embodiment, the linear regression equation is subjected to significance test, and α of each cell is determined j 、h j 、β j 、d j Substituting the values into the obtained linear regression equation, and calculating the substitution valueAnd then respectively calculating the regression square sum, the total square sum and the degree of freedom of the linear regression equation, and checking the significance of the regression equation, wherein the checking method is a correlation coefficient checking method, and the calculation formula is as follows:
wherein ,RJ 、R G Representing complex correlation coefficients of each linear regression equation, S RJ Representing the ridge gradient alpha and ridge of each experimental cell and the comparison cellRegression square sum of linear regression equation of high h, furrow width ratio beta, furrow trend d and soil microbial community structure diversity correlation index, S TJ Representing the total sum of squares of linear regression equations of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and soil microbial community structure diversity correlation index of each experimental cell and control cell, S RG Expressing regression square sum of linear regression equation of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and functional gene abundance data of soil carbon nitrogen circulation process of each experimental cell and control cell, and S TG And the total sum of squares of linear regression equations of ridge gradient alpha, ridge height h, furrow width ratio beta, furrow trend d and functional gene abundance data in the soil carbon nitrogen circulation process of each experimental cell and the control cell are represented.
In the present embodiment, the complex correlation coefficient R for each linear regression equation J 、R G If the calculation result is between 0 and 1 or equal to 1, the linear regression equation passes the significance test, and if the calculation result is equal to 0, the linear regression equation has no significance.

Claims (2)

1.一种坡耕地垄沟布局对微生物影响机理的识别方法，其特征在于，包括以下步骤：1. A method for identifying the mechanism of the impact of ridge and furrow layout on sloping farmland on microorganisms, which is characterized by including the following steps: S1、选取研究区，并确定种植作类型和坡耕地垄沟布局参数，所述坡耕地垄沟布局参数包括：垄坡度、垄高、垄沟宽比以及垄沟走向；S1. Select the research area, and determine the planting type and ridge and furrow layout parameters of sloping farmland. The ridge and furrow layout parameters of sloping farmland include: ridge slope, ridge height, ridge and furrow width ratio, and ridge and furrow direction; S2、以垄坡度、垄高、垄沟宽比以及垄沟走向按四因素三水平正交表布设实验小区和对照小区，并种植所选作物；S2. Set up the experimental plot and the control plot according to the four-factor three-level orthogonal table based on the ridge slope, ridge height, ridge-furrow width ratio and ridge-furrow direction, and plant the selected crops; S3、分别于作物种植前、生育中期、收获期采集各实验小区及对照小区土壤样品进行土壤微生物高通量测序，得到不同实验小区及对照小区土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据；S3. Collect soil samples from each experimental plot and control plot before crop planting, mid-growth period, and harvest period for high-throughput sequencing of soil microorganisms, and obtain soil microbial community structure diversity-related indexes and soil carbon and nitrogen cycles in different experimental plots and control plots. Process functional gene abundance data; S4、利用方差分析法检验垄坡度、垄高、垄沟宽比以及垄沟走向对所述土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的影响，并确定对所述土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据产生影响的垄坡度、垄高、垄沟宽比以及垄沟走向四因素的主次顺序；S4. Use the variance analysis method to test the effects of ridge slope, ridge height, ridge-furrow width ratio, and ridge-furrow direction on the soil microbial community structure diversity-related index and soil carbon and nitrogen cycle process functional gene abundance data, and determine the impact on the soil The primary and secondary order of the four factors affecting the ridge slope, ridge height, ridge and furrow width ratio, and ridge and furrow direction influenced by the microbial community structure diversity correlation index and soil carbon and nitrogen cycle process functional gene abundance data; 所述步骤S4包括以下步骤：The step S4 includes the following steps: S401、计算各实验小区和对照小区土壤样品中微生物群落结构多样性相关指数及土壤碳氮循环过程功能基因丰度数据的平均值；S401. Calculate the average value of the diversity-related index of the microbial community structure and the functional gene abundance data of the soil carbon and nitrogen cycle process in the soil samples of each experimental plot and the control plot; S402、根据所述平均值，计算得到各实验小区和对照小区土壤样品中微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的离差平方和及自由度；S402. Based on the average value, calculate the sum of squared deviations and degrees of freedom of the microbial community structure diversity correlation index and functional gene abundance data of the soil carbon and nitrogen cycle process in the soil samples of each experimental plot and the control plot; S403、根据所述离差平方和及自由度，利用方差分析法检验不同垄坡度、垄高/>、垄沟宽比/>、垄沟走向/>对土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的影响；S403. Use the variance analysis method to test different ridge slopes based on the sum of squares of deviations and degrees of freedom. , ridge height/> , furrow width ratio/> , furrow direction/> Impact on soil microbial community structure diversity-related indexes and functional gene abundance data of soil carbon and nitrogen cycle processes; S404、对步骤S403得到的影响结果进行显著性检验；S404. Perform a significance test on the influence results obtained in step S403; S405、根据显著性检验结果，按从大到小对步骤S403得到的影响结果进行排序，得到对所述土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据产生影响的垄坡度、垄高、垄沟宽比以及垄沟走向四因素的主次顺序；S405. According to the significance test results, sort the impact results obtained in step S403 from large to small to obtain the ridges that have an impact on the soil microbial community structure diversity related index and soil carbon and nitrogen cycle process functional gene abundance data. The order of priority of the four factors: slope, ridge height, ridge-furrow width ratio and ridge-furrow direction; 所述步骤S401中平均值的表达式如下：The expression of the average value in step S401 is as follows: 其中，表示各实验小区和对照小区土壤样品的微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的平均值，/>表示实验小区的个数，/>表示测量次数，/>表示第/>个实验小区，/>表示对实验小区的第/>次测量，/>表示第/>个实验小区第/>次测量的微生物群落结构多样性相关指数，/>表示土壤碳氮循环相关功能基因数目，/>表示第/>个土壤碳氮循环相关功能基因数目，/>表示第/>个实验小区第/>次测量的第/>个土壤碳氮循环过程功能基因丰度值；in, Represents the average value of the diversity-related index of microbial community structure and functional gene abundance data of soil carbon and nitrogen cycle processes in soil samples from each experimental plot and control plot,/> Indicates the number of experimental plots,/> Indicates the number of measurements,/> Indicates the first/> An experimental community,/> Indicates the No./> of the experimental community measurements,/> Indicates the first/> Experimental community No./> The measured diversity correlation index of microbial community structure,/> Represents the number of functional genes related to soil carbon and nitrogen cycle,/> Indicates the first/> The number of functional genes related to soil carbon and nitrogen cycle,/> Indicates the first/> Experimental community No./> Measurement No./> The functional gene abundance value of each soil carbon and nitrogen cycle process; 所述步骤S402中离差平方和及自由度的表达式如下：The expressions of the sum of squares of deviations and degrees of freedom in step S402 are as follows: 其中，表示各实验小区和对照小区土壤样品中微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的总离差平方和，/>表示垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素引起的离差平方和，/>表示误差离差平方和，/>均表示/>、/>以及/>相应的自由度，/>表示第/>个实验小区土壤样品中微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据，/>表示各实验小区和对照小区土壤样品中微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的平均值，/>表示垄坡度、垄高/>、垄沟宽比/>、垄沟走向/>的水平数，/>表示正交表的列数，/>表示正交表的第u列，/>表示任意列上四因素取水平/>时所得到的实验结果的算数平均值；in, Represents the total sum of squared deviations of the microbial community structure diversity correlation index and functional gene abundance data of soil carbon and nitrogen cycle processes in soil samples from each experimental plot and control plot,/> Indicates ridge slope/> , ridge height/> , furrow width ratio/> , furrow direction/> The sum of squared deviations caused by the four factors,/> Represents the sum of squared error deviations,/> All indicate/> ,/> and/> The corresponding degrees of freedom,/> Indicates the first/> Microbial community structure diversity correlation index and soil carbon and nitrogen cycle process functional gene abundance data in soil samples from each experimental plot,/> Represents the average value of the microbial community structure diversity correlation index and functional gene abundance data of soil carbon and nitrogen cycle processes in soil samples from each experimental plot and control plot,/> Indicates ridge slope , ridge height/> , furrow width ratio/> , furrow direction/> The number of levels,/> Indicates the number of columns of the orthogonal table,/> Represents the u-th column of the orthogonal table,/> Indicates the level of the four factors in any column/> The arithmetic mean of the experimental results obtained; 所述步骤S403中检验不同垄坡度、垄高/>、垄沟宽比/>、垄沟走向/>对土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的影响的表达式如下：In the step S403, different ridge slopes are tested , ridge height/> , furrow width ratio/> , furrow direction/> The expressions of the impact on soil microbial community structure diversity-related indexes and functional gene abundance data of soil carbon and nitrogen cycle processes are as follows: 其中，表示垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>引起的土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的均方与各垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素共同引起的土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据误差的均方的比值；in, Indicates ridge slope/> , ridge height/> , furrow width ratio/> , furrow direction/> The mean square of the correlation index of soil microbial community structure diversity and the functional gene abundance data of soil carbon and nitrogen cycle processes and the slope of each ridge/> , ridge height/> , furrow width ratio/> , furrow direction/> The ratio of the mean square error of the correlation index of soil microbial community structure diversity and the functional gene abundance data of soil carbon and nitrogen cycle processes caused by the four factors; 所述步骤S404中进行显著性检验的表达式如下：The expression for significance testing in step S404 is as follows: 其中，表示各垄坡度/>、垄高/>、垄沟宽比/>和垄沟走向/>的/>、/>、/>、/>分别与/>值在/>显著性水平下的比值，/>表示各实验小区和对照小区由垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素引起的自由度为/>的土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的均方与各垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素共同引起的实验误差均方的比值，/>表示表示各实验小区和对照小区由垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素引起的自由度为/>在/>显著性水平下的土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据的均方与各垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>四因素共同引起的实验误差均方的比值；in, Indicates the slope of each ridge/> , ridge height/> , furrow width ratio/> And the direction of the furrow/> of/> ,/> ,/> ,/> Respectively with/> The value is/> Ratio at significance level,/> Indicates the ridge slope of each experimental plot and control plot/> , ridge height/> , furrow width ratio/> , furrow direction/> The degrees of freedom caused by the four factors are/> The mean square of the correlation index of soil microbial community structure diversity and the functional gene abundance data of soil carbon and nitrogen cycle processes are related to the slope of each ridge/> , ridge height/> , furrow width ratio/> , furrow direction/> The ratio of the mean squares of the experimental errors caused by the four factors,/> Indicates that each experimental plot and control plot are represented by the ridge slope/> , ridge height/> , furrow width ratio/> , furrow direction/> The degrees of freedom caused by the four factors are/> in/> The mean square of soil microbial community structure diversity correlation index and soil carbon and nitrogen cycle process functional gene abundance data at the significance level and the slope of each ridge/> , ridge height/> , furrow width ratio/> , furrow direction/> The ratio of the mean squares of experimental errors caused by the four factors; S5、将所述主次顺序分别与土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据构建线性回归方程；S5. Construct a linear regression equation by combining the primary and secondary order with the soil microbial community structure diversity correlation index and soil carbon and nitrogen cycle process functional gene abundance data; 所述步骤S5中线性回归方程的表达式如下：The expression of the linear regression equation in step S5 is as follows: 或 or 其中，分别表示线性回归方程的系数，/>分别表示第/>个实验小区的垄坡度/>、垄高/>、垄沟宽比/>和垄沟走向/>数据；in, Respectively represent the coefficients of the linear regression equation,/> Respectively represent the first/> Ridge slope of each experimental plot/> , ridge height/> , furrow width ratio/> And the direction of the furrow/> data; S6、对所述线性回归方程进行显著性检验，并根据检验结果将其垄坡度、垄高、垄沟宽比以及垄沟走向的标准系数作为评价坡垄沟布局对土壤微生物的影响，完成对坡耕地垄沟布局对微生物影响机理的识别；S6. Conduct a significance test on the linear regression equation, and use the standard coefficients of ridge slope, ridge height, ridge-furrow width ratio and ridge-furrow direction according to the test results to evaluate the impact of slope-ridge-furrow layout on soil microorganisms, and complete the evaluation of slope-ridge-furrow layout. Identification of the mechanism of influence of layout on microorganisms; 所述步骤S6中对线性回归方程进行显著性检验的表达式如下：The expression for significance testing of the linear regression equation in step S6 is as follows: 其中，、/>表示各线性回归方程的复相关系数，/>表示各实验小区和对照小区的垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>与土壤微生物群落结构多样性相关指数的线性回归方程的回归平方和，/>表示各实验小区和对照小区的垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>与土壤微生物群落结构多样性相关指数的线性回归方程的总平方和，/>表示各实验小区和对照小区的垄坡度/>、垄高/>、垄沟宽比/>、垄沟走向/>与土壤碳氮循环过程功能基因丰度数据的线性回归方程的回归平方和，/>表示各实验小区和对照小区的垄坡度、垄高/>、垄沟宽比/>、垄沟走向/>与土壤碳氮循环过程功能基因丰度数据的线性回归方程的总平方和。in, ,/> Represents the complex correlation coefficient of each linear regression equation,/> Indicates the ridge slope of each experimental plot and control plot/> , ridge height/> , furrow width ratio/> , furrow direction/> The regression sum of squares of the linear regression equation related to the diversity index of soil microbial community structure,/> Indicates the ridge slope of each experimental plot and control plot/> , ridge height/> , furrow width ratio/> , furrow direction/> The total sum of squares of the linear regression equation related to the diversity index of soil microbial community structure,/> Indicates the ridge slope of each experimental plot and control plot/> , ridge height/> , furrow width ratio/> , furrow direction/> Regression sum of squares of linear regression equation with soil carbon and nitrogen cycle process functional gene abundance data,/> Indicates the ridge slope of each experimental plot and control plot. , ridge height/> , furrow width ratio/> , furrow direction/> Total sum of squares of linear regression equation with soil carbon and nitrogen cycle process functional gene abundance data. 2.根据权利要求1所述的坡耕地垄沟布局对微生物影响机理的识别方法，其特征在于，所述步骤S3包括以下步骤：2. The method for identifying the mechanism of influence of ridge and furrow layout on sloping farmland on microorganisms according to claim 1, characterized in that step S3 includes the following steps: S301、在作物种植前、生育期中期以及收获期三个阶段，以及在实验小区和对照小区内上、中、下三段选择田垄和田沟两个取样区域，除去表层浮土，采集土壤样品，除去同一小区土样混合物的杂质，并装入灭菌封装袋下保存；S301. Select two sampling areas, ridges and ditches, in the three stages before crop planting, in the middle of the growth period and in the harvest period, as well as in the upper, middle and lower sections of the experimental plot and the control plot, remove the surface top soil, collect soil samples, and remove Impurities in the soil sample mixture from the same plot are stored in sterilized bags; S302、对土壤样品进行土壤微生物高通量测序，得到各实验小区和对照小区的土壤样品微生物DNA或RNA序列数、OTU丰度信息以及物种分类信息；S302. Perform high-throughput sequencing of soil microorganisms on soil samples to obtain the number of microbial DNA or RNA sequences, OTU abundance information and species classification information of soil samples in each experimental plot and control plot; S303、根据所述土壤样品微生物DNA或RNA序列数、OTU丰度信息以及物种分类信息，计算得到不同实验小区及对照小区土壤微生物群落结构多样性相关指数和土壤碳氮循环过程功能基因丰度数据。S303. According to the soil sample microbial DNA or RNA sequence number, OTU abundance information and species classification information, calculate the soil microbial community structure diversity related index and soil carbon and nitrogen cycle process functional gene abundance data in different experimental plots and control plots. .
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105525025A (en) * 2016-02-17 2016-04-27 南京大学 Method for detecting rhizosphere soil prokaryotic microorganisms of various soybeans based on 16SrDNA deep sequencing
CN108663014A (en) * 2018-03-30 2018-10-16 中国水利水电科学研究院 A kind of furrow farming sloping upland field relative elevation method for computing data
CN109034496A (en) * 2018-08-29 2018-12-18 中国水利水电科学研究院 A kind of slope furrow optimized parameter slowing down slope surface agricultural non-point source pollution determines method
CN109190827A (en) * 2018-09-07 2019-01-11 中国水利水电科学研究院 A kind of furrow is laid out the Influencing Mechanism recognition methods to Rainfall-runoff
CN114145190A (en) * 2021-09-27 2022-03-08 云南良道农业科技有限公司 A high-yield planting method of organic vegetables on sloping land

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105525025A (en) * 2016-02-17 2016-04-27 南京大学 Method for detecting rhizosphere soil prokaryotic microorganisms of various soybeans based on 16SrDNA deep sequencing
CN108663014A (en) * 2018-03-30 2018-10-16 中国水利水电科学研究院 A kind of furrow farming sloping upland field relative elevation method for computing data
CN109034496A (en) * 2018-08-29 2018-12-18 中国水利水电科学研究院 A kind of slope furrow optimized parameter slowing down slope surface agricultural non-point source pollution determines method
CN109190827A (en) * 2018-09-07 2019-01-11 中国水利水电科学研究院 A kind of furrow is laid out the Influencing Mechanism recognition methods to Rainfall-runoff
CN114145190A (en) * 2021-09-27 2022-03-08 云南良道农业科技有限公司 A high-yield planting method of organic vegetables on sloping land

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Studies on the epidemiology of malaria in New Guinea I. Holoendemic malaria [long dash]The clinical picture;PETERS, W.等;TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE;第54卷(第3期);第242-249页 *
典型黑土垄作区耕地沟蚀对土壤养分的影响研究;杨子;刘晓光;宁静;董芳辰;于杰;张鹏;王赛;;土壤(第02期);第174-180页 *
辽西地区坡耕地垄膜沟种对土壤侵蚀和作物产量的影响;肖继兵;孙占祥;蒋春光;郑家明;刘洋;杨宁;冯良山;白伟;;中国农业科学(第20期);第48-61页 *

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