Dephenolization pyrolysis fluid improved physicochemical properties and microbial community structure of saline-alkali soils

doi:10.21203/rs.3.rs-1516314/v1

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Dephenolization pyrolysis fluid improved physicochemical properties and microbial community structure of saline-alkali soils

https://doi.org/10.21203/rs.3.rs-1516314/v1

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Because of its acidity, biomass pyrolytic fluid (BPF) had been paid attention in the amelioration of saline soil. However, BPF contained biotoxic phenolic compounds and must be refined before use. Most of the existing refining methods were cumbersome and uneconomical. A simple one-step salting-out extraction system of was developed. BPF was successfully divided into upper and lower phases, and most phenolic substances were concentrated in the upper phase. The lower phase diluted 400 times at the pyrolysis temperature of 500℃ was added into saline-alkali soil, which greatly increased the content of soil available nutrients. Under the action of organic acids, soil pH and total salt content could be reduced effectively, and soil enzyme activities can be increased. Microbial community analysis showed that the addition of LP could increase the proportion of Actinomycetes, which played a beneficial role in improving soil fertility, and then improved the growth of Chinese cabbage.

Pyrolysis fluids

Saline-alkali soil

Salting-out extraction

Microorganisms

Enzyme activity

Soil remediation

Soil salinization and alkalization were global problem restraining the improvement of land-use capability and the development of agricultural production (Na et al. 2015). It has been reported that over 1.1 billion hectares of land, involving more than 100 countries and regions, are affected by soil salinity (Xia et al. 2019). On the one hand, soil salinity affected the growth of crops and exacerbates the crisis of food shortages; on the other hand, it caused an annual economic loss of at least 27.2 billion dollars in agriculture production all over the world (Kou et al. 2019). Thus, the improvement and utilization of salinized land have gradually attracted people's extensive attention.

The restoration of saline soils has been extensively studied by many researchers in terms of theory, measures and evaluation methods (Soldo et al. 2020). Physical, chemical and biological methods have also been extensively studied, for example, artificial irrigation (Lei et al. 2019), salt-tolerant plants (Jin et al. 2017), compost and manure. However, these measures were costly and difficult to apply in practice on a large scale.

It was well known that biomass (i.e. wood, straw, etc.) was a natural widespread renewable resource (Laëtitia et al. 2019). Biomass, under conditions of oxygen isolation, was pyrolyzed at high temperatures to produce a liquid called pyrolysis fluid (BPF) (Ys et al. 2020). Due to its high content of organic acids (Mamaeva et al. 2016), it has been used by some researchers for the study of saline soil improvement. It was reported that the diluted BPF could reduce the pH and soil salt in salinized land, improve soil organic matter mass fraction, significantly increase the number of soil microorganisms and promote plant growth (Xinyan et al. 2018). BPF can not only improve the physical and chemical properties of the soil, but also have varying degrees of influence on the enzyme activity in the soil (Wang et al. 2010). Sirimuji studied the effects of different concentrations of BPF on soil enzyme activity through pot experiments, and the results showed that BPF promoted the activity of soil amylase, protease, phosphatase, urease, and catalase (Sirimuji et al. 2018). Despite the positive effect of BPF on saline land improvement, a large number of phenolic substances exist in BPF. Phenol and its derivatives are aromatic compounds and a kind of protoplasmic poison, which can produce toxicity to all biologically active bodies (Avnish et al. 2021). Therefore, the refinement of BPF becomes an important condition for its better utilization.

Currently, conventional distillation (Wang et al. 2021), molecular distillation (Liu et al. 2020) and graded condensation (Moutsoglou et al. 2018) have been investigated as the main BPF refining methods. However, these methods were expensive and required additional energy consumption. Salting-out extraction (SOE) was a separation method used to extract hydrophilic product from its aqueous solution with the aid of inorganic salt as the salting-out reagent and organic solvent as the extractant (Wang et al. 2014). As a novel, easy-to-use, low-cost and low-energy method, it has been used by several researchers in the field of extraction and purification of organic substances. Fu et al. demonstrated that salting-out extraction system composed of monosodium phosphate and ethanol exhibited excellent extraction efficiency for both butyric acid (∼99%) and acetic acid (∼90%) (Fu et al. 2017). Jia et al. developed an effective method for extracting total phenolic (TP)。 from grape seed using enzyme and microwave co-assisted salting-out extraction (Jia et al. 2021)

However, SOE had not been used in the removal of phenolic substances in BPF. Therefore, in this study, we established the NH₄SO₄-BPF-ethanol extraction system. BPF was successfully divided into upper phases (UP) and lower phases (LP), and the biotoxic phenolics were mainly concentrated in UP. The physicochemical properties of original BPF, UP and LP were studied respectively. LP at different dilution rates was added to saline-alkaline soil. Not only was the pH of the soil reduced, but the nutrient content of the soil increased. In addition, the activities of some enzymes and microorganisms in soil were increased, and the microbial community structure was improved obviously. The changes of these indexes significantly improved the growth of Chinese cabbages. The method provided in this study had the advantages of low cost, simple process and obvious effect. It also provided a new ideas for the possibility of large-scale saline-alkaline soil improvement.

2.1 Material Preparation

The cotton stalk (CS) was picked from the agricultural experimental farm of Shihezi University and air-dried in the ventilation area. The CS was cut into small pieces with a particle size of 2 cm, dried in an oven at 105°C for 4 h, cooled and put into sealing bags, and then placed in a desiccator for standby use.

In this experiment, the saline-alkali soil (white alkali saline-alkaline soil) was taken from the vicinity of Mushroom Lake Reservoir in Shihezi City (85°9′39″E, 44°4′75″N), and sampling points were set in the sampling area in the S-shaped. The soil mining depth was 0‒20 cm, and a total of about 100 kg saline-alkali soil was collected. Brought it back to the laboratory, spread the soil on a plastic cloth for air drying, picked out impurities such as gravel grass root residues, passed a 3 mm sieve and put it into a sack for use. The basic physicochemical properties of soils were shown in Table 1

Table 1

The Basic Soil Properties
Indicators	pH	Full salt content Wt.%	Moisture content %	Alkaline nitrogen mg/kg	Organic matter g/kg	Fast-acting phosphorusmg/kg	Fast-acting potassium mg/kg
Data	7.98 ± 0.09	1.36 ± 0.14	4.09 ± 0.21	44.26 ± 1.06	7.24 ± 0.11	4.31 ± 0.25	85.01 ± 1.24

2.2 Pyrolysis Process of Biomass

In each experiment, 100 ± 0.5 g CS was loaded into the furnace and heated from room temperature to five final sample temperatures (300, 400, 500, 600 and 700 ℃) separately at a constant heating rate of 10 ℃/min, during which ultra-high purity helium (N₂) was flown inside and outside the glass tube as the purging and protective gas at the rate of 0.3 L/ min. After each experiment, the pyrolysis fluids (BPF) taken out and weighed when the furnace temperature was the same as room temperature. Put it into a brown bottle and put the bottle in a refrigerator at 4℃ for later use. Each treatment was conducted three times and all of BPF were subjected to subsequent analyses.

2.3 Salting-out Extraction

The saturated ammonium sulfate solution was allocated and added to the centrifuge tube, and ethanol was added to mix with BPF (The mass ratio of ammonium sulfate solution, BPF and alcohol was 6:6:1). After shaking, ultrasonic processing was carried out for 15 min, followed by centrifugation in a frozen centrifuge at 8000 r/min at room temperature for 15 min. After standing for a period of time, the upper phases (UP) and lower phases (LP) were separated. The above experiment was repeated three times.

2.4 Pot Experiment

In this experiment, Chinese cabbage was used as a planting plant for pot cultivation. LP was applied into the soil, and 20 groups of treatments were performed. LP prepared at different temperatures were diluted 50, 100, 200 and 400 times. Then they were added to 20 groups of potted plants respectively. At the same time, the blank (CK) control of water irrigation was conducted. Three parallel experiments were performed for each of the above treatments.

250 g of saline soil was put and 12 Chinese cabbage seeds were planted in each pot. The pots were put in the greenhouse and the numbers of sprouting of Chinese cabbage in each pot were recorded after sowing for 3 days. The two worst seedlings were plucked out every 7 days and the plant height and weight were measured. The test period was 35 days.The test period was 35 days. The plants were irrigated every 3 days, 30 mL of diluted LP was irrigated in the first 10 days, and 50 mL of diluted LP was irrigated in the next 25 days. The thiobarbituric acid method was used to determine the content of malondialdehyde (MDA) in plants (Zhang et al. 2019a).

2.5 Methods of Analysis and Characterization

2.5.1 Analysis and Characterization of BPF

The density of the BPF was determined by the pycnometer method. The pH of the BPF was determined by pH meter. The determination of total polyphenols (TP) used a slightly modified Folin-phenol (Luque-Rodríguez et al. 2007) reagent method, using gallic acid as the standard, and drew a standard curve. The determination of the total acid (TA) referred to the national standard GB/T 12456-90 and it was determined by titration, and TA was calculated as acetic acid. The water content was determined using a Karl Fischer moisture meter (787KF, Metrohm, Swiss). The organic composition of BPF was analyzed by GC-MS (GCMS-QP2020, Shimadzu, Japan).

2.5.2 Soil nutrient and enzyme activity analysis

Urease, Sucrase, catalase and alkaline phosphatase activity were measured by sodium phenol-sodium hypochlorite colorimetric method, 3,5-dinitrosalicylic acid colorimetric method, potassium titanate titration method (Tapia et al. 2016) and colorimetric method of benzyl phosphate (Mishra et al. 2014), respectively. The alkaline diffusion method (Leng et al. 2020), 0.5 mol/L NaHCO₃ method, NH₄OAc and flame photometry and volumetric method of potassium dichromate - dilution calorimetry (Musadji et al. 2020) were used to measure alkaline nitrogen, fast-acting phosphorus, fast-acting potassium and organic matter content in soil separately.

2.5.3 High-throughput sequencing

The sequences were initially screened according to the following criteria and eliminated from further analysis: requiring sequence length ≥ 160 bp and not allowing ambiguous base N. We will exclude: 1) sequences with primer mismatch base number > 1 at the 5' end; 2) sequences containing consecutive identical base numbers > 8 (Tanja et al. 2011). After chimera checking using USEARCH, UCLUST was used to cluster 97% sequence identity into an operable taxonomic unit (OUT) from which the alpha diversity indices (Simpson index) were determined. Principal component analysis (PCA) was used to assess the inter-relationships between different acclimatized microbial communities (Ramette 2007).

3.1 Total Phenol and Total Acid Content in BPF, UP and LP

The pH, density, yield, moisture content and heating value of BPF were measured in the study period (See supplementary materials). When the pyrolysis temperature was low (300–400℃), the aliphatic hydroxyl groups on the branched chain of lignin structure were broke, and the cellulose would depolymerize and dehydrate. The main products of their reactions were acetic acid, formic acid and other acid compounds, a small amount of tar substances and phenolic compounds (Ren et al. 2017). Therefore, as shown in Fig. 1a, the acid content was higher at a lower temperature, up to 139.23mg/g. With the increase of pyrolysis temperature, the secondary pyrolysis of BPF was strengthened and the organic acids decomposed. For example, levoglucose was decomposed into small molecular gas products through molecular reforming, decarboxylation and other reactions, thus reducing the acid content. On the other hand, the increase of the total amount of BPF also led to the decrease of the proportion of acid compounds in the total organic matter. The higher the pyrolysis temperature was, the more intense the pyrolysis reaction of lignin was. Guaiacol (also known as o-methoxy-phenol), one of the main products of pyrolysis, and tar-like substances produced at low temperature had secondary cracking. The GC-MS analysis of BPF showed that the content of guaiacol did decrease with the increase of pyrolysis temperature (See supplementary materials) (Chen et al. 2020). This led to an increase in the number of phenols and a rapid increase in TP content. In addition, the chemical bonds of oxygen-containing compounds in BPF broke in the order of C-O, C-H and C-C bonds from weak to strong, and decarboxylation, decarbonylation and dehydrogenation occurred respectively (Hoyong et al. 2018), so oxygen-containing compounds converted into more stable phenolic compounds. Therefore, as shown in Fig. 1a, TP reached the minimum 67.27mg /g at 300℃ and the maximum 76.79mg /g at 700℃.

From Fig. 1b and supplementary materials, it can be seen that the two-phase product obtained by salting-out extraction, UP was much smaller than LP product, and the yield of LP product was first increased and then decreased, the highest yield was 96.29% at 500 ℃. TP were mainly enriched in UP. The average TP content in UP was 193.27 mg/g, while that in LP was 64.52 mg/g (Fig. 1c). Because when inorganic salt solutions were added, the cations produced by ionization attract more water molecules to form a hydration layer, causing more organic molecules to be repelled into the organic phase (Xiao et al. 2009) (See supplementary materials). As shown in Fig. 1d, the effect of salting-out extraction on the TA of UP and LP in each temperature segment was the same as that of BPF, and the TA content in UP was slightly higher than LP after the salting-out extraction of BPF at each temperature.

3.2 Potted plant experiment

Firstly, BPF was diluted to a factor of 0,200,400,600 and was added to saline-alkali soil. The effects of BPF on the growth of Chinese cabbage were investigated by pot experiment. As we can see (see supplementary material), only a 600-fold dilution of BPF resulted in very few seeds germinating, with a germination rate of only 13.33%. This was because BPF contained more phenols and acids, which have a great impact on the germination rate of plants (Muscolo et al. 2001).

Next, LP with different dilution ratio (50,100,200,400) was added into saline-alkali soil for pot experiment. Compared with CK, LP had a significant effect on the germination rate (3 d), plant height (35 d) and plant weight (35 d) of Chinese cabbage (Fig. 2a-c). In particular, a 400-fold dilution of LP increased the germination rate by 25.76% on average, while a 50-fold dilution of LP only increased the germination rate by 2.4%. The plant average height and plant weight of Chinese cabbage, which reached a maximum height of 6.7 cm and a maximum weight of 0.9661 g, both of which were higher than CK, by 67.51% and 93.22%, respectively. As the dilution increases the plants grew better (see supplementary material). When the dilution was low, the phenolic concentration in LP was higher, and the excessive phenolic compounds have an inhibitory effect on seed germination and plant growth. On the other hand, the addition of sulfate, which increases the soluble salt content in the soil, will change the osmotic pressure in the plant, thus causing some hindrance to the plant root system for nutrient and water uptake. It even inhibits plant growth, while salt stress can cause ion toxicity in the plant body (Shahzad et al. 2021). This also results in severe plant senescence and high MDA content (Fig. 2d). With 400-fold LP diluted in each temperature segment having a good effect on the plant's MDA, with an average decrease of 42.35% compared with the blank control group. Through pot experiments, we found that BPF inhibited plant growth, while LP promoted plant growth to varying degrees. In order to further investigate the specific mechanism of LP, soil nutrients, enzyme activities and microbial community structure were analyzed.

3.3 Effects of LP on soil nutrients and enzyme activities

The change in soil pH was inversely proportional to the TA content in LP (Fig. 3a). The salt used in the test was (NH₄)₂SO₄, which was a strong acid and weak base salt. Its solution was acidic, so it made a more significant impact on soil pH value. In addition, the addition of (NH₄)₂SO₄ had a certain influence on soil total salt content (Fig. 3b) and alkali-hydrolyzable nitrogen content (Fig. 3d). Also, the addition of high concentration of LP promoted the formation of macromolecule complexation in the soil, but it was not discharged in a short time (Shang et al. 2021). With the increase of test period, small molecule salt was regenerated, leading to the increase of salt content. When using the appropriate dilution ratio (200 and 400 times), it could effectively complex salt ions and reduce the total salt content in the soil. LP was rich in organic materials, so when it was added to the soil, the larger the dilution ratio of LP was, the less the content of organic matter was in the soil. When LP was diluted by 50, 100, 200 and 400 times, the average organic matter content of soil increased by 66.75%, 47.16%, 29.39% and 24.76% respectively (Fig. 3c).

As shown in Fig. 3d-f, LP can significantly increase the effective soil nutrient content. LP contained a certain amount of acidic organic matter. On the one hand, the acid dissolution, chelation dissolution and desorption of these acidic substances had an important and direct effect on the increase of soil nitrogen, phosphorus and potassium elements. And small molecular acids could inhibit the mineralization of these elements and promote the release of available elements in soil (Li et al. 2020). The migration of colloidal ions in soil was promoted by acidic ions and organic active molecules in LP. This resulted in the release of the original fixed nutrients in the soil and at the same time increased the activity of the original nutrients (Zhang et al. 2019b). On the other hand, the acidic components of LP could interact with materials in the soil at the right concentrations. This could promote enzyme and microbial activity, thus further improve soil activity (Zhang et al., 2019a). Therefore, with the addition of LP, the contents of basic nitrogen, available phosphorus and available potassium significantly increased, up to 139.56 mg/kg, 14.82mg/kg and 71.53mg/g respectively.

Soil colloids and minerals had adsorption effects on enzymes, this can reduce soil enzyme activity(Li et al. 2021). LP contained a large amount of acidic substances such as acetic acid, which can compete with soil colloids and minerals for adsorption sites on soil enzymes, thus reducing adsorption (Kabiri et al. 2016). From Fig. 4, it can be seen that LP had a significant promotion effect on soil enzyme activity. The urease and alkaline phosphatase activity increased with the increase of the dilution ratio of LP. Especially when the dilution ratio of LP was 200 and 400 times, the promotion of both enzymes was greater. Compared with CK, urease activity increased on average 3.13-fold (Fig. 4a). The trend graph of alkaline phosphatase activity (Fig. 4c) was consistent with the change in the trend graph of fast-acting phosphorus (Fig. 3e). Sucrase activity was related to the metabolism of carbon (Bastida et al. 2016). At lower dilutions, the LP contained more carbon sources, which led to increased microbial metabolism. So the sucrase activity was higher (Fig. 4b). The activity of catalase was negatively correlated with pH. Therefore, a small dilution of LP with high organic acid content was more favorable to promote the activity of catalase (Fig. 4d) in the soil.

To further represent more visually the effects of pyrolysis temperature and dilution times on soil nutrients, physicochemical properties and enzyme activity, we did a correlation analysis, as shown in Fig. 4e. The similarity of the color level indicates that the effect was close. As can be seen from the figure, the dark red color is mainly concentrated in the area of dilution multiples 50 and 400. Low dilutions have a positive effect on organic matter, total salinity, alkaline nitrogen, sucrase and catalase activity in the soil. Higher dilutions had positive effects on fast-acting phosphorus, fast-acting potassium, plant growth, alkaline phosphatase and urease in the soil. In order to investigate more clearly which microorganisms are affected by LP at high and low dilutions, high throughput sequencing was performed in this study.

3.4 Changes of microbial communities in acclimatized soil

We made gene sequencing on 16SRNA to evaluate the richness and diversity under various suitable conditions. Figure 5a displayed the Shannon-Wiener curve of bacterial communities in this research. As the increase of measured sequence, the Shannon-Wiener curve leveled off. LP diluted 50 times and 400 times at different temperatures were added to LP. Generally, The Shannon and Simpson indices reflected the diversity and uniformity of samples, and Chao 1 and ACE reflected the richness of communities (Pd et al. 2021). As can be seen from the Table 2, Shannon values of samples were all higher than CK, indicating that the addition of LP was beneficial to enrich microbial community. Simpson value in low dilution ratio samples decreased, while that in high dilution ratio samples increased. This was due to the fact that LP at low dilutions contained more phenolics, which affected the biological activity of some microorganisms. And the appropriate concentration of LP had positive effect on the growth of microorganisms. Compared with the values of Chao 1 and ACE, CK increased, indicating that the richness of microorganisms in the samples raised. It can also be seen from the abundance class curves (Fig. 5b). PCA results of multiple samples (Fig. 5c) showed that samples at different pyrolysis temperatures and in the same dilution multiple did not appear aggregation. The results showed that the microbiota of each sample changed in different degrees during the adaptation process.

Table 2

Microbial diversity index of the flora
	Simpson	Chao1	ACE	Shannon
CK	0.99826	2366	2366	10.32
L-3-50	0.994174	3064.3	3299.12	9.54
L-4-50	0.993492	3749.14	4141.72	9.74
L-5-50	0.986293	2639.5	2827.96	9.16
L-6-50	0.989704	3195.91	3365	9.11
L-7-50	0.990269	2873.36	3093.39	9.16
L-3-400	0.998486	4186.03	4588.54	10.72
L-4-400	0.998096	3956.81	4276.93	10.57
L-5-400	0.998278	4296.08	4674.51	10.69
L-6-400	0.998256	3484.17	3709.49	10.48
L-7-400	0.997807	4542.93	4971.09	10.69

Multiple studies have shown, in phylum level, Proteobacteria had an absolute advantage in the alkali-saline soils (Bai et al. 2019). As shown in Fig. 5d, there are also distributions of Acidobacteria, Gemmatimonadetes, Chloroflexi, Firmicutes and Acidobacteris. When LP (L-50) diluted 50 times was added, the abundance of Proteobacteria raised to varying degrees, while the abundance of Acidobacteria reduced and the abundance of Firmicutes greatly increased. The reason was that LP contains more (NH₄)₂SO₄ at lower dilution ratio, resulting in higher overall salt content than CK. The abundance of Proteobacteria, as the dominant bacteria, increased. From the analysis of order level (Fig. 5e), Clostridiales had strong resistance under high salt condition, and became the dominant bacteria under the condition of abundant organic matter (Du et al. 2022). From the analysis of genus level (Fig. 5f), it showed that Pseudomonas mostly appeared in poor environment and releases catalase while consuming organic matter in soil. This was also one of the reasons for the high catalase content in soil at low dilution ratio (Fig. 4d). Soil organic matter content was too high, causing C: N was higher than 70:1. Under this condition, nitrogen-fixing bacteria start to fix nitrogen, which was the main reason why Azoarcus became the dominant genus (Du et al. 2021). Therefore, the content of alkali-hydrolyzed nitrogen was higher at low dilution ratios. Desulfosporosinus (Pester et al. 2012), a reductive microorganism often found in environments of high sulfate concentrations, restored sulfate to stable sulfides. At low dilution ratio, the abundance of dominant bacteria in saline-soil environment increased and the diversity of bacteria was low due to the addition of sulfate. Soil fertility was poor which leaded to poor growth of, but it also improved to a certain extent compared with CK.

LP diluted 400 times added, the richness of Proteobacteria slightly reduced, while Acidobacteria and Gemmatimonadetes significantly increased, sequenced in phylum level. Studies have shown that Actinomycetes mainly undertook the decomposition of organic matter, played a beneficial role in improving soil fertility, and could provide certain protection against abiotic stress(Shi et al. 2022). The increase in the abundance of Gemmatimonadetes was attributed to the raising available potassium, available phosphorus and organic matter content (Mk et al. 2020). From the analysis of order level, Micrococales and Rhizobiales (Aliashkevich et al.) were the main phospho-solubilizing bacteria orders, and the abundance of these two orders was higher when the dilution ratio was higher, which was also one of the reasons why the content of available phosphorus in soil was higher. Nitrosomonadales becoming the dominant bacterium mainly because the soil was rich in NH₄⁺, under the action of which, NH₄⁺ can be converted into nitrate that can be directly used by the Chinese Cabbage (Boden et al. 2017). From genus level analysis, Sphingomonas and Pseudarthrobacter can be used for degradating aromatic compounds (Lha et al.). Because LP contained part of phenolic substances, the emergence of these two bacteria also meant the degradation of phenolic substances. Therefore, under the high dilution ratios, Chinese Cabbage grew better.

The addition of LP increased the contents of soil organic matter and available nutrients. LP diluted 400 times at pyrolysis temperature of 500℃ had a suitable concentration of organic acids. They could neutralize soil alkalinity and reduce soil salt stress through effective complexation of salt ions. Additionally, acidic organic substances could compete with enzymes for adsorption sites on soil colloid and minerals, reduce enzyme adsorption and increase soil enzyme activity. The increase of Actinomycetes ratio optimized soil population structure and improved soil quality. The positive amelioration effect of LP on saline soil significantly improved the growth index of Chinese cabbage.

Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

Authors contributions

Shuai Wang: Conceptualization, Methodology, Software, Validation, Formal analysis, Investigation, Writing-Original Draft, Data Curation, Visualization. Hanyu Chang: Data Curation, Writing-Review & Editing. Zhenfei Dong: Software, Methodology, Writing-Review & Editing. Yansen Ren: Validation, Writing-Review & Editing. Tianle Tan: Visualization.

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Funding

This work was supported by the National Natural Science Foundation of China under No. 51768061 and Key Scientific and Technological project for Double firstclass discipline construction of Shihezi University under No. SHYL-ZD201803.

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Table 1 The Basic Soil Properties

Indicators

Full salt content

Wt.%

Moisture content

Alkaline nitrogen

mg/kg

Organic matter

g/kg

Fast-acting phosphorusmg/kg

Fast-acting potassium

mg/kg

Data

7.98±0.09

1.36±0.14

4.09±0.21

44.26±1.06

7.24±0.11

4.31±0.25

85.01±1.24

Table 2 Microbial diversity index of the flora

	Simpson	Chao1	ACE	Shannon
CK	0.99826	2366	2366	10.32
L-3-50	0.994174	3064.3	3299.12	9.54
L-4-50	0.993492	3749.14	4141.72	9.74
L-5-50	0.986293	2639.5	2827.96	9.16
L-6-50	0.989704	3195.91	3365	9.11
L-7-50	0.990269	2873.36	3093.39	9.16
L-3-400	0.998486	4186.03	4588.54	10.72
L-4-400	0.998096	3956.81	4276.93	10.57
L-5-400	0.998278	4296.08	4674.51	10.69
L-6-400	0.998256	3484.17	3709.49	10.48
L-7-400	0.997807	4542.93	4971.09	10.69

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Editorial decision: Major Revision
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Editor invited by journal
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You are reading this latest preprint version

Dephenolization pyrolysis fluid improved physicochemical properties and microbial community structure of saline-alkali soils

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Abstract

Figures

1 Introduction

2 Material And Method

2.1 Material Preparation

2.2 Pyrolysis Process of Biomass

2.3 Salting-out Extraction

2.4 Pot Experiment

2.5 Methods of Analysis and Characterization

2.5.1 Analysis and Characterization of BPF

2.5.2 Soil nutrient and enzyme activity analysis

2.5.3 High-throughput sequencing

3 Results And Discussion

3.1 Total Phenol and Total Acid Content in BPF, UP and LP

3.2 Potted plant experiment

3.3 Effects of LP on soil nutrients and enzyme activities

3.4 Changes of microbial communities in acclimatized soil

4 Conclusions

Declarations

References

Tables

Supplementary Files

Status:

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