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Journal of Resources and Ecology >

2024 , Vol. 15 >Issue 4: 918 - 924

DOI: https://doi.org/10.5814/j.issn.1674-764x.2024.04.013

Impact of Human Activities on Ecosystem

Impact of Anaerobic Soil Disinfestation Treatment on the Properties of Tomato Continuous Cropping Soil

  • ZHOU Kaisheng , *
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  • Center of Environment Science Experiment, Bengbu University, Bengbu, Anhui 233030, China
* ZHOU Kaisheng, E-mail:

Received date: 2023-04-16

  Accepted date: 2023-09-28

  Online published: 2024-07-25

Supported by

The Academic Support Project for Top Talents of Higher Education Subject (Major) in Anhui Province(gxbjZD2022077)

The Agricultural Ecological Conservation and Pollution Control Key Laboratory of Anhui Province(FECPP201901)

The High-level Talent Launch Foundation of the Applied Science Research Project of Bengbu University in 2024(2024YYX54QD)

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Abstract

To investigate the impact of anaerobic soil disinfestation (ASD) treatment with different processing duration on tomato continuous cropping soil, a total of eight treatments were designed, which included two treatment temperatures of 10 ℃ and 30 ℃, and four treatment times of 3, 4, 5 and 7 weeks (w). The results showed that whether the temperature was 10 ℃ or 30 ℃, the pH values, TOC and available K contents in the soil treated by the ASD method were significantly increased (P<0.05), while the EC values and NO3- contents in soils treated by ASD were significantly reduced compared with the untreated group (P<0.05). The Fusarium oxysporum population in soils treated by ASD decreased with the extension of the treatment time. When the treatment temperature was 30 ℃, the effect on the removal of the NO3- that had accumulated in soils treated by ASD was significantly better than the treatment temperature of 10 ℃ (P<0.05). When soil was treated by ASD for more than 5 w at 30 ℃, the NH4+ content in soil was significantly increased (P<0.05) compared with the untreated soil. In conclusion, NO3- accumulation in soils could not be completely eliminated with treatment by ASD, even when the treatment time at 10 ℃ was prolonged. Whether the treatment temperature was 10 ℃ or 30 ℃, the F. oxysporum population in soils treated by ASD decreased significantly with the extension of the treatment time to more than 5 w compared with treatment only for only 3 w.

Key words： anaerobic soil disinfestation; tomato; continuous cropping soil; treatment time

Cite this article

ZHOU Kaisheng . Impact of Anaerobic Soil Disinfestation Treatment on the Properties of Tomato Continuous Cropping Soil[J]. Journal of Resources and Ecology, 2024 , 15(4) : 918 -924 . DOI: 10.5814/j.issn.1674-764x.2024.04.013

1 Introduction

For creating a strong anaerobic soil environment, anaerobic soil disinfestation (ASD) can be used as a soil treatment method. Specifically, perishable organic materials (such as rice bran, wheat bran, and straw powder) are added to the soil before planting the crops, and then water is added to the soil and it is covered with plastic film for 2-15 weeks to create a strong anaerobic soil environment. Under anaerobic conditions, the NO3- and SO42- in soils are reduced (Chen et al., 2019), the pH of acidic soil increases, and the electrical conductivity (EC) of the soil decreases (Wen et al., 2015). Volatile organic acids (such as acetic acid, propionic acid, and butyric acid) are produced by the decomposition of organic materials, H2S is produced by SO42- reduction, and NH3 is produced by NH4+ dehydrogenation, and so on. All of these changes have fumigation effects on soil-borne pathogens (Momma et al., 2006; Huang et al., 2015a).
The reduced iron and manganese ions (Fe2+, Mn2+) in soil (Momma et al., 2011; Bruggen and Blok, 2014), NH4+ produced by the mineralization of organic materials and NO3- dissimilating reduction, and NO2- produced by NO3- reduction, can inhibit soil-borne pathogens (Zhang et al., 2013; Sun et al., 2015). The contents of soil organic matter can be increased by the humification of organic materials, and the content of soil available K can be increased by K-containing minerals that are dissolved by the organic acids. Thus, the soil physical and chemical properties can be adjusted, while disease-causing soil organisms are inhibited, soil microecol ogy is regulated, and soil fertility is improved. In many crop production systems, the ASD method is often used to control obstacles to continuous cropping. To achieve this goal, the length of the treatment time may not be the same, and different organic materials and auxiliary additives can be used to treat the degraded soil during continuous cropping. According to the results published by Momma et al. (Momma et al., 2005; Momma et al., 2010), reducing the chlamydospores of Fusarium oxysporum to below the level of pathogen non-detection in soils required at least 9 days of treatment by ASD with wheat bran as the organic material; if the soils were treated by ASD with 2% (v/v), 1% (v/v) or 0.5% (v/v) alcohol, they required at least 3, 6 and 9 days, respectively. When soil is treated by ASD, different soil temperatures require different treatment times. The treatment of continuous cropping soils based on ASD has been applied in Japan and the Netherlands. However, due to the lower latitude of Japan compared with the Netherlands, the time needed was shorter (about 3 weeks) (Blok et al., 2000; Momma et al., 2006).
Different crops have different fallow periods (i.e., the window of time for the ASD treatment) because of the vast territory and complex climatic conditions in China. In the actual production of many crops, the window of time available for ASD treatment often occurs in the cooler seasons. Therefore, it is of great theoretical and practical significance to examine the effectiveness of continuous cropping soils treated by ASD by extending the treatment time in low- temperature seasons.

2 Materials and methods

2.1 Test materials

The soil treated by ASD indoors was collected from a greenhouse in which tomatoes were continuously planted for 3 years, in Qianli Village, Bengbu City, Anhui Province, China, on July 13, 2019. The basic properties of the untreated soil are shown in Table 1. The rice straw (C:N= 46.19:1) was provided by the School of Geographical Sciences, Nanjing Normal University.
Table 1 The basic properties of the tomato continuous cropping soil before treated by ASD methods
Variable Moisture
content (%)		 pH EC
(µS cm-1)		 NH4+
(mg kg-1)		 NO3-
(mg kg-1)		 TOC
(g kg-1)		 Available K
(mg kg-1)		 TN
(g kg-1)		 lg gene copies g-1 dry soil
Bacteria Fungi F. oxysporum
Value 11.6 4.75 461 4.28 257 10.7 171 1.34 9.38 8.11 6.77

2.2 Method

2.2.1 Experimental design

Soil from tomato continuous cropping for 3 years was selected as the research object in this study. The experiments were designed with two treatment temperatures of 10 ℃ and 30 ℃, and four treatment times of 3, 4, 5 and 7 w, so a total of eight ASD treatments were performed (Table 2). The soil samples (equivalent to dry soil weight of 0.73 kg per soil sample) were weighed and placed in self-sealing plastic bags. Straw powder equivalent to 1% of the dry soil weight was added into each soil sample, and the straw powder was all sifted through a 2 mm sieve. Water was added to achieve 100% soil moisture saturation after the soil samples were mixed with the straw powder, and then the soil samples were sealed before being placed in a biochemical constant temperature incubator at either 10 ℃ or 30 ℃. All the above soil samples were incubated in the biochemical con-stant temperature incubator for 3, 4, 5 and 7 w according to the above experimental design, and three replicates were carried out for each group of treated soil samples.
Table 2 The experimental design of treatments by anaerobic soil disinfestation
Serial number Sample ID Time (w) Temperature (℃) Seria number Sample ID Time (w) Temperature (℃)
1 3w-10℃ 3 10 5 3w-30℃ 3 30
2 4w-10℃ 4 10 6 4w-30℃ 4 30
3 5w-10℃ 5 10 7 5w-30℃ 5 30
4 7w-10℃ 7 10 8 7w-30℃ 7 30

2.2.2 Sample analysis

After weighing the soil samples (equivalent to dry soil weight) and mixing them with deionized water according to 1:2.5 (m/v) and 1:5 (m/v) soil-water ratios, an S220K pH meter (Mettler-Toledo International Inc., Shanghai, China) and a DDS-320 conductivity meter (Dapu Instrument Co., Ltd., Shanghai, China) were used to determine the pH and EC values of the soil, respectively. The contents of total organic carbon (TOC) and total nitrogen (TN) in the soils were determined with the wet burning method of H2SO4- K2Cr2O7 and Kelvin deboiling method in the literature (Lu, 2000), respectively. Soil samples were mixed with 2 mol L-1 KCl solution at a 1:5 (m/v) ratio, oscillated for 1 h at 200 r min-1 in a constant temperature oscillator, filtered, and then a continuous flow analyzer (San++, Skalar Analytical B.V., Breda, the Netherlands) was used to detected the contents of NH4+ and NO3- in the soils. The 5.00 g air-dried soil samples were sifted through a 2 mm sieve and weighed, then placed into 200 mL plastic bottles, and 50 mL of CH3COONH4 solution was added. The samples were then plugged tightly with rubber plugs, and placed on a reciprocating oscillator at a constant temperature of 25 ℃, at 120 cycles min-1. After shaking for 30 min, the suspension was filtered with dry filter paper, and the content of available K was directly measured in the filtrate with a flame spectrophotometer. The microbial DNA was extracted from the soil by referring to the method of the Power Soil TMDNA Isolation Kit (MO BIO Laboratories Inc., USA), and the numbers of soil bacteria, fungi and F. oxysporum were analyzed by referring to the literature (Huang et al., 2015b).

2.3 Data analysis

The significant differences among different treatments were analyzed with univariate variance analysis and Duncan’s test in SPSS 20.0. The experimental data were analyzed with Excel 2010, and illustrations were drawn with Origin 8.0. The log10 transformation of quantitative microbial data was performed before data analysis.

3 Results

After the soil treatment using the ASD method, the general linear model multivariate analysis method was used to analyze the significance of differences in the effects due to the different treatment factors (temperature and time) on soil properties (P), and the results are listed in Table 3.
Table 3 Effects on soil properties of tomato continuous cropping soils treated by ASD with different times, temperatures and their interactions
Treatment factor pH EC NH4+ NO3- TOC K Bacteria Fungi F. oxysporum
Temperature 0.089 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000
Time 0.071 0.000 0.000 0.188 0.001 0.000 0.013 0.000 0.000
Temperature × time 0.298 0.015 0.000 0.170 0.002 0.000 0.008 0.000 0.425

Note: The data in the table are significance level values (P) obtained by two-factor variance analysis of the effects of soil treatment temperature and duration on the indicated soil properties.

3.1 Changes in pH and EC

The data in Table 3 show that the treatment temperature, time and the interaction between temperature and time had no significant effects on soil pH (P>0.05), but they had significant effects on soil EC values (P<0.05). Whether the treated temperature was 10 ℃ or 30 ℃, the values of pH were significantly increased (P<0.05) in soils treated by ASD for 3 w to 7 w compared with the untreated soil, and there were no obvious regular changes in the pH values with the extension of treatment time (Fig. 1a). The EC values of the soils were significantly reduced compared with the untreated soil (P<0.05). When the treatment temperature was 30 ℃, the EC values of soils treated by ASD were significantly lower than those of soils treated by ASD at 10 ℃ for the corresponding treatment times (P<0.05), but there were no significant differences between the EC values at 30 ℃ (P>0.05) (Fig. 1b).
Fig. 1 Changes in pH (a) and EC (b) in continuous cropping soil from facility tomatoes that were field treated by anaerobic soil disinfestation

Note: OS in the figure refers to the original soil sample before treatment. Different letters on the bars represent significant differences after treatment (P<0.05) according to Duncan’s test, and the same letter means no significant difference (P>0.05). This legend also applies to the following figures.

3.2 Changes in the NH4+ and NO3- contents

The treatment temperature, time and the interaction between temperature and time had significant effects on the soil NH4+ content (P<0.001). Treatment temperature had a significant effect on soil NO3- content (P<0.001), but treatment time and the interaction between temperature and time did not have significant effects on soil NO3- content (P>0.05) (Table 3). Whether the treatment temperature was 10 ℃ or 30 ℃, the NH4+ contents in soil samples treated by ASD for 3 w were not significantly different from the corresponding untreated group (P>0.05). The NH4+ contents in soils treated by ASD from 4 w to 7 w were significantly higher than the untreated soils (P<0.05). Except for those treated at 30 ℃, the NH4+ contents in soil samples treated by ASD for 5 w and 7 w were significantly higher than those at 10 ℃, and there were no significant differences between the other ASD treatments at 30 ℃ and the corresponding treatment times at 10 ℃(P>0.05) (Fig. 2a). Whether the treatment temperature was 10 ℃ or 30 ℃, the NO3- contents in soil samples treated by ASD from 3 w to 7 w were significantly reduced compared with those in the untreated (raw) soil (P<0.05), and there were no obvious regular changes in the NO3- contents of soil samples treated by ASD with the extension of treatment time. It should be noted that the accumulated NO3- in soils could not be completely removed from the soils treated by ASD at 10 ℃, and the NO3- contents in the soils were still as high as 77.8 mg kg-1 even when the treatment time was extended to 7 w (Fig. 2b).
Fig. 2 Changes of the NH4+ (a) and NO3- (b) contents in continuous cropping soil from facility tomatoes field that were treated by anaerobic soil disinfestation

3.3 Changes in the TOC and available K contents

The treatment temperature, time and the interaction between temperature and time had significant effects on the TOC and available K contents in soils (P<0.01) (Table 1). Whether the temperature was 10 ℃ or 30 ℃, the TOC contents in the soil treated by ASD were significantly higher than those of the untreated soil (P<0.05). While the TOC contents in soils treated by ASD for 3 w were not significantly different from those treated by ASD during the corresponding treatment time at 10 ℃ (P<0.05), the TOC contents in the other soils treated by ASD at 30 ℃ were significantly lower than those treated by ASD at 10 ℃ (P<0.05) (Fig. 3a). The available K contents in soils treated by ASD were significantly increased compared with those in untreated soil (P<0.05), but the available K contents in all treatments had no obvious regularity in their changes with the extension of treatment time. While the contents of available K in soils treated by ASD for 3, 4 and 7 w at 30 ℃ were significantly lower than those at 10 ℃ for each corresponding treatment time (P<0.05), there were no significant differences between the contents of available K in the other soils treated by ASD at 30 ℃ and those treated by ASD at 10 ℃ (P>0.05) (Fig. 3b).
Fig. 3 Changes of the TOC (a) and K (b) contents in continuous cropping soil from facility tomatoes fields that were treated by anaerobic soil disinfestation

3.4 Changes in soil microbial quantity

3.4.1 Changes in soil bacteria and fungi

The treatment temperature, time and the interaction between temperature and time had significant effects on the numbers of soil bacteria and fungi (P<0.05) (Table 3). The number of bacteria in each treatment was significantly higher than in the untreated soil whether the temperature was 10 ℃ or 30 ℃ (P<0.05). The number of bacteria in soils treated by ASD at 10 ℃ was significantly lower than that at 30 ℃ for each corresponding treatment time (P<0.05) (Fig. 4a). While the number of fungi in soils treated by ASD for 3 w and 7 w at 30 ℃ was not significant different from that in the corresponding treatment time at 10 ℃ (P>0.05), the numbers of fungi in other soils treated by ASD at 30 ℃ were significantly higher than those during the corresponding treatment times at 10 ℃ (P<0.05) (Fig. 4b).
Fig. 4 Changes in the number of bacteria (a) and fungi (b) in continuous cropping soil from facility tomatoes field that were treated by anaerobic soil disinfestation

3.4.2 Changes in F. oxysporum

The population of F. oxysporum in soils treated by ASD was significantly affected by temperature and time (P<0.001), while the interaction between temperature and time had no significant effect on the population of F. oxysporum in soils (P>0.05) (Table 3). Compared with untreated soils (original soils), the number of F. oxysporum in soils treated by ASD for 3 w to 7 w at 10 ℃ did not decrease significantly (P> 0.05). However, when the treatment temperature was 30 ℃, the number of F. oxysporum in each soil sample treated by ASD for 3 w to 7 w was reduced compared with that in untreated soil; especially when the treatment temperature was 30 ℃, the population of F. oxysporum in each soil sample treated by ASD for more than 5 w was significantly reduced compared with that in the untreated soil (P<0.05) (Fig. 5).
Fig. 5 Changes in the number of F. oxysporum in continuous cropping soil from facility tomatoes field that were treated by anaerobic soil disinfestation

4 Discussion

Due to the different fallow periods of various crops, treatment temperature and time are important factors that influence the treatment effect of the ASD method (Blok et al., 2000; Momma et al., 2005; Momma et al., 2006; Momma et al., 2010). Regarding the influence of treatment time on the effect of ASD, Huang et al. (2014a; 2014b) reported that the pH of banana continuous cropping soils treated by ASD with straw as the organic material for 5 days was significantly higher than that of the control, which was consistent with the fact that the pH of soil treated by ASD for 3 w was significantly higher than that of the untreated soil in this study. Consistent with another literature report (Zhu et al., 2013), the EC values of soils treated by ASD at either 10 ℃ or 30 ℃ could be significantly reduced (P<0.05), and the changes in EC values showed a decreasing trend with the extension of treatment time (Fig. 1b). Whether the treatment temperature was 10 ℃ or 30 ℃, the EC values of soils treated by ASD for 3 w to 7 w were lower, under 0.340 mS cm-1, which all fell into the non-salinized soil range (He et al., 2010). It is worth noting that when the treatment temperature was 10 ℃, even the prolonged treatment time could significantly reduce the EC values of soil, and the accumulated NO3- in soils could not be completely removed. Thus, these results show that nitrate was a major contributor to the EC values (Fig. 6), but the EC values of soils were not entirely due to the nitrate.
Fig. 6 The relationships between NO3- contents and soil EC values in tomato continuous cropping soils
The degraded continuous cropping soils treated by ASD can be improved by the killing of soil-borne pathogens and reconstructing the soil microbial community (Huang et al., 2014a; Liu et al., 2016). The treatment time not only affects the physicochemical processes of soil treated by ASD, but it also affects the production and accumulation of H2S, NH3, Mn2+, Fe2+, fatty acids and other bactericidal substances in soils (Tenuta et al., 2002; Momma et al., 2011; Cao et al., 2014; Huang et al., 2015a). The ammonia produced by the dehydrogenation of ammonium ions during soil treatment by ASD has a fumigation effect on F. oxysporum, and a high concentration of NH3 can inhibit F. oxysporum by up to 100% (Huang et al., 2016). The NH4+ (Zhang et al., 2013) and NO2- (Sun et al., 2015) produced by NO3- reduction and H2S (Huang et al., 2016) produced by SO42- reduction can all inhibit F. oxysporum. Consistent with literature reports, the number of F. oxysporum in soils treated by ASD also showed a decreasing trend with an increase in the NH4+ concentration in this study (Fig. 7). Volatile organic acids (such as acetic acid, propionic acid and butyric acid) produced by the anaerobic fermentation of organic materials have toxic effects on soil-borne pathogens (Momma et al., 2006). When the treatment temperature was 30 ℃, microbial activity was enhanced and the reduction reaction was stronger in soils treated by ASD. Many antibacterial substances were produced in a short time, which enhanced the inhibitory effect on F. oxysporum in soil treated by ASD. On the contrary, when the treatment temperature was 10 ℃, the soil microbial activity was low and the reduction reaction was weak. However, extending the treatment time was conducive to the accumulation of bacteriostatic substances. When the accumulation of bacteriostatic substances reached a sufficient concentration, the inhibitory effect on F. oxysporum in soils treated by ASD was also enhanced. The data in Fig. 5 show that F. oxysporum in soils treated by ASD at 10 ℃ were mostly higher than or close to that in the untreated (original) soil, while F. oxysporum in soils treated by ASD at 30 ℃ were lower than in the untreated soil. However, the number of F. oxysporum in soils treated by ASD at 10 ℃ showed a decreasing trend with the extension of the treatment time. Thus, in order to improve the inhibitory effect on F. oxysporum in soils treated by ASD, increasing the treatment temperature and extending the treatment time should be adopted. Especially for the continuous cropping soil with a treatment window period in winter, a scheme with an extending treatment time should be adopted.
Fig. 7 The relationships between NH4+ contents and the number of F. oxysporum in tomato continuous cropping soils

5 Conclusions

Whether the treatment temperature was 10 ℃ or 30 ℃, the treatment time was extended for 3 w to 7 w, which had significant effects on most of the measured indexes, but they did not show regular changes with the increase or decrease of the ASD treatment duration. When the treatment temperature was 10 ℃, the accumulated NO3- could not be completely removed even when the duration of ASD treatment was prolonged. Whether the treatment temperature was 10 ℃ or 30 ℃, all populations of F. oxysporum could be reduced significantly when the treatment time was prolonged (≥5 w) compared with the 3 w treatment. If the goal is to kill pathogenic bacteria, the treatment effect was improved when the treatment time was prolonged at a low temperature.

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