ZHOU Kaisheng

(1. Experiment Center of Environment Science， Bengbu University， Bengbu 233030， China; 2. School of Geographic Science， Nanjing Normal University， Nanjing 210046， China)

Abstract: Continuous cropping of watermelon has become increasingly common in China. However， yield and quality of watermelons are seriously affected by soil degradation caused by continuous cropping due to soil acidification and proliferation of Fusarium oxysporum f. sp. niveum (FON). In the present study， anaerobic soil disinfestation (ASD) was used to treat soil samples collected from dry farmlands and paddy fields used annually for planting watermelons. Each soil category was divided into 5 groups， namely: (1) controls (H1， S1)， (2) flooded controls (H2， S2)， (3) flooded and incorporated rice straw alone (H3， S3)， (4) flooded and incorporated rice straw with sulfur powder (H4， S4)， and (5) flooded and incorporated rice straw and ammonia water (H5， S5). Each group had 15 replicates. The soil samples were then placed in plastic bags， mixed thoroughly， sealed， and incubated outdoors for 20 days. Pot experiment was then performed to test the effect of ASD by planting watermelon seedlings instead of grafting. Results showed that the -N and contents in soils were reduced significantly by ASD (P<0.05)， while the pH values of the soil were elevated. The EC values of the soil were effectively adjusted， and the soil-borne pathogenic FON was remarkably suppressed (P<0.05). The mortality rate of the control group was 43.3%. The treated groups had better watermelon growth than the control group， and the watermelon yields of all the treated groups were higher than that of the control group. It was concluded that the ASD method could repair and improve degraded soil caused by continuous cropping.

Remediation of watermelon continuous cropping soil by anaerobic soil disinfestation

ZHOU Kaisheng1，2

(1.ExperimentCenterofEnvironmentScience，BengbuUniversity，Bengbu233030，China; 2.SchoolofGeographicScience，NanjingNormalUniversity，Nanjing210046，China)

soil remediation; anaerobic soil disinfestation; soil degradation

Watermelon is one of the most important summer fruits and serves as a major source of income in some rural households in China. However， continuous cropping of watermelon often causes soil deterioration through soil acidification[1-2]， accumulation of allelochemicals[3-4]， nitrate nitrogen， sulfate radicals[5]， and proliferation of soil-borne pathogen[4，6]. In addition， some complications also occur[4]and seriously affect the sustainable development of watermelons. In watermelon production， lime is typically applied to ameliorate acidic soil[1]， and organic fertilizer is also used to improve soil structure and quality[7]. Soil solarization[8]， high-temperature greenhouse[9]， chemical fumigation， and grafting[10-11]are performed to suppressFusariumoxysporumf. sp.niveum(FON)， which can cause watermelon wilts. However， these methods basically fail to overcome the continuous cropping obstacles.

In this study， ASD was applied to treat watermelon continuous cropping soil to test its remediation effect， and possible material was selected to provide references for further application.

1 Materials and methods

1.1 Experimental materials

For laboratory test， soils were collected from paddy fields (used for watermelon and rice rotation annually) and dry farmlands (used for watermelon and other dry crop rotation annually) continuously planted with watermelon for more than 10 years in the village of Zhangxiang， Bengbu City， Anhui Province， China， at the beginning of July， 2013. The rice straws used in this study were also collected from the same village. The total nitrogen (TN) and total carbon (TC) contents in the rice straw were approximately 14.54 and 337.15 g kg-1， respectively. Sulfur powder and ammonia water were bought from the supply station of Huachang Chemical Procurement， Bengbu City， Anhui Province， China. Wasteland soil samples were collected from the campus of Bengbu University near the village of Zhangxiang at the end of April， 2013 and were tested at the beginning of May， 2013.

A pot experiment for watermelons was performed from the beginning of May， 2016 until the middle of August， 2016. The soil samples used in the pot experiment were collected from the watermelon test field of Bengbu University， where watermelons were continuously planted for more than 3 years.

1.2 Experimental design

1.2.1Laboratorytest

The soil samples were divided into 5 groups. Each sample amounted to 3 kg of dry soil weight and was treated as follows: (1) control (non-amended and non-flooded， H1 and S1, H and S referred to dry land and paddy field soils， respectively， the same as below); (2) flooded control (flooded but non-amended， H2 and S2); (3) flooded and incorporated rice straw in soil (H3 and S3, the dosage of rice straw amounted to 1% dry soil weight, the same as below); (4) flooded and incorporated rice straw with sulfur powder in soil (H4 and S4, the dosage of sulfur accounted for 0.1% dry soil weight); and (5) flooded and incorporated rice straw and ammonia water in soil (H5 and S5, the dosage of ammonia water was 300 mg·L-1， which referred to the content of NH3amounted to dry soil weight). Each soil sample group had 15 replicates. Each soil sample was thoroughly mixed with the aforementioned design ratio and then placed in plastic bags. Pond water was added into the plastic bags， which were then sealed and incubated outdoors for 20 days. The wasteland soils were divided into 10 samples， which were then used for direct testing.

1.2.2Potexperiment

The pot experiments were divided into 6 groups of incubated soils， and each group was further separated into 30 parallel samples. The weight of each soil sample was equivalent to approximately 3 kg of dry soil. The materials of each incubated group were incorporated with the following designs: (1) control (CK， non-amended and non-flooded); (2) flooded and incorporated with alfalfa powder in soil (1% Al， the dosage of alfalfa powder amounted to 1% dry soil weight); (3) flooded and incorporated acetic acid in soil (0.25% Ac， the ratio of acetic acid accounted for 0.25% of dry soil weight); (4) flooded and incorporated with ammonia in soil (0.25% AW， the rate of ammonia was equivalent to 0.25% of dry soil weight); (5) flooded and incorporated alfalfa powder with acetic acid in soil (1%Al + 0.25%Ac， the dosage of alfalfa powder and acetic acid amounted to 1% and 0.25% of dry soil weight， respectively); (6) flooded and incorporated alfalfa powder with ammonia in soil (1% Al+0.25% AW， the dosage of alfalfa powder and ammonia amounted to 1% and 0.25% of dry soil weight， respectively). Each soil sample was thoroughly mixed with the aforementioned design ratio and then placed in plastic bags. Water was added into plastic bags， which were sealed and incubated outdoors for 21 d. Ungrafted watermelon seedlings were planted after the soil was dried. The growth conditions of the watermelons were examined throughout the entire growing season to verify the effect of ASD on the soil used for watermelon continuous cropping.

1.3 Determination of soil pH and EC

A S220K pH meter (Mettler-Toledo International Inc.， Shanghai， China) was used to measure soil pH in a 1∶2.5 (m/V) ratio of soil to deionized water. Soil EC value was measured in a 1∶5(m/V) ratio of soil to deionized water through a DDS-320 conductivity meter (Shanghai Great Temple Instrument Co.， Ltd. Shanghai， China). The results were presented as the mean of 3 arbitrary samples from the 15 replicates of each treatment.

1.5 Analysis of soil-culturable microbes

Beef extract peptone agar medium was used for growing soil-culturable bacteria. Gause’s No. 1 medium was used to incubate soil-culturable actinomyces. Rose Bengal medium was used to incubate soil-culturable fungi[31]， and improved Komada’s culture medium was used for the growth ofF.oxysporum[39]. The amount of soil-culturable microbes was counted by smearing-plate method and was incubated in a microbiological incubator at 35 ℃. The number of microbes was determined by counting the number of colony-forming units (CFU). The number of bacteria was counted after 2 d， and the number of actinomyces， fungi， and FON were all counted after 4 d of incubation.

1.6 Data analysis

SPSS 16.0 (SPSS Inc.， Chicago， USA) and Microsoft Excel 2007 were used for statistical analysis. Independent-samplet-test was performed at the end of each assay， and differences withPvalues<0.05 were considered significant.

2 Results and analysis

2.1 Laboratory test

2.1.1ChangesinsoilpHandEC

The pH values in wasteland soil ranged from 6.0 to 6.5， while the EC values ranged from 0.026 to 0.770 mS·cm-1. Meanwhile， the pH values of the controls were all below 6.0， and the pH values of the treated soil samples (including the flooded controls) were all significantly (P<0.05) higher than those of the controls (Fig. 1). The soil pH values of the samples with sulfur (H4 and S4) were lower than 6.5， whereas the soil pH values of other treatments (H2， S2， H3， S3， H5， and S5) were higher than 6.5 after 20 d. The pH values of the soil samples with ammonia water were higher than 7.5 after 20 d.

The EC values of H1 and S1 ranged from 0.131 to 0.195 mS·cm-1， and 0.094 to 0.132 mS·cm-1， respectively， which were higher than those of the flood controls (H2 and S2). The highest EC value was observed in the dry farmland soil samples with ammonia water (H5). After 20 d， the EC values of H3 and H4 were all lower than that of H1， while those of S4 and S5 were higher than those of S1 and other treated soil samples (Fig. 2). Thus， it was shown that compared to wasteland， continuous cropping of watermelon would reduce soil pH and cause minor changes in soil EC value.

Fig.1 Dynamic changes of pH values in soil samples

Fig.2 Dynamic changes of EC values in soil samples

2.1.3Changesinculturablemicrobesinsoil

Fig.3 Dynamic changes in contents in soil samples

Fig.4 Dynamic changes in contents in soil samples

Fig.5 Dynamic changes in contents in soil samples

The number of bacteria and actinomycetes ranged from 2.3×108to 7.0 ×108CFU·g-1and 0.47×107to 2.3×107CFU·g-1， respectively， in wasteland soils. The population of fungi andF.oxysporumranged from 0.5×105to 2.7×105CFU·g-1and 0.83×104to 5.33×104CFU·g-1， respectively, in wasteland soils. In dry farmland and paddy field soils with continuous cropping of watermelon， the number of bacteria was 108CFU·g-1in all soil samples after 20 d (Fig. 6). Meanwhile， all the populations of actinomycetes in all treatments decreased along with time except H1 and S1. The number of fungi was 105CFU·g-1in the controls (H1， S1) throughout the experiment， while the fungus populations in H2 and H3 ranged from 105CFU·g-1to 104CFU·g-1， and the number of fungi in H4 and H5 ranged from 105CFU·g-1to 103CFU·g-1. Similarly， the number of fungi in S2， S3， S4， and S5 ranged from 105CFU·g-1to 104CFU·g-1throughout the experiment. The number of FON in H1 and S1 were 105CFU·g-1and 104CFU·g-1， respectively, during the experiment， yet the population

Fig.6 Dynamic changes of numbers of culturable microorganisms in soil samples

of FON in H2， H3， H4， and H5 decreased from 105CFU·g-1to 103CFU·g-1. Similarly， FON populations in S2， S3， S4 and S5 decreased from 104CFU·g-1to 103CFU·g-1.

2.2 Pot experiment

Compared to control (CK)， the number of dead watermelons in various treated groups was lower， as well as the mortality rate (Fig. 7). Three groups with 0 mortality rate were observed， i.e. 1%Al， 0.25%Ac， and 1%Al+0.25%Ac. The mortality rates of 0.25%AW and 1%Al+0.25%AW groups were 3.3% and 6.6%， respectively. Meanwhile， the mortality rate of CK was 43.3%.

The watermelons in all the treated groups had better growth than those in the control. The water-

Fig.7 Effect of treatment on death of watermelon in pot experiment

melon yields of all the treated groups were higher than those of CK (Fig. 8)， and the differences were significant (P<0.05) for the treatment of 1%Al， 0.25%Ac and 1%Al+0.25%Ac.

Fig.8 Effect of treatment on yield of watermelon in pot experiment

3 Discussion and conclusion

Current research has shown that acetic acid， propionic acid， and other volatile organic acids produced by organic materials decomposition[16-17，28，30，45]， NH3produced by ammoniation of rice straw and H2S produced by sulfur reduction[29]can inhibit soil-borne pathogensF.oxysporum[16，28-30，43]. The Fe2+and Mn2+ions produced under reductive conditions in the soil samples may be the induced factors that inhibit soil-borneF.oxysporum[19]. The suppressing effect on FON was confirmed by the experimental results in this study. The least number of FON was observed in the H5 and S5 treatments， followed by H4， S4， H3， S3 treatments. NH3and H2S can effectively suppress FON， and this finding is consistent with the aforementioned mechanism underlyingF.oxysporuminhibition[16-17，28-29]. Studies have shown that the critical pathogenic concentration ofF.oxysporumf. sp.cubenseis 103CFU·g-1[46]， and the FON contents after ASD treatment decreased from 105CFU·g-1to 103CFU·g-1in dry farmland soil samples and from 104CFU·g-1to 103CFU·g-1in paddy field soil samples in the present study (Fig. 6). In addition， significant negative correlation between soil pH and number of FON was observed. The absolute values of the correlation coefficient between pH value and number of FON exceeded 0.80， and thus the elevated pH values were also observed to have an inhibitory effect on FON. Based on these results， the researchers found that ASD could effectively inhibit FON， whereas the content of soil-culturable bacteria showed nearly no change and maintained at the magnitudes of 108CFU·g-1in the soil samples throughout the experiment.

The pot experiment showed that the watermelon seedlings in various treated groups were more exuberant than those in the controls， and the yields of the watermelons in various treated groups were also higher than those in controls. These results were consistent with those of the laboratory test. Therefore， ASD method could be used to prevent and control complication of watermelon continuous cropping.

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(責(zé)任編輯 高 峻)

2017-02-22

蚌埠學(xué)院優(yōu)秀人才項(xiàng)目([2014]182)；安徽省級(jí)質(zhì)量工程項(xiàng)目(2015zy068)；安徽省振興計(jì)劃項(xiàng)目(2014zdjy137)；安徽省級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)計(jì)劃項(xiàng)目(201611305073)

周開勝(1970—), 男, 安徽鳳陽(yáng)人，博士研究生，副教授，研究方向?yàn)橥寥栏牧?。E-mail: zks606@sina.com

土壤修復(fù)；厭氧土壤滅菌法；土壤退化

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1004-1524(2017)07-1179-10

http://www.zjnyxb.cn

10.3969/j.issn.1004-1524.2017.07.17

S471 Document: A

1004-1524(2017)07-1179-10

浙江農(nóng)業(yè)學(xué)報(bào)ActaAgriculturaeZhejiangensis， 2017，29(7): 1179-1188

周開勝. 厭氧土壤滅菌修復(fù)西瓜連作退化土壤(英文)[J]. 浙江農(nóng)業(yè)學(xué)報(bào)，2017，29(7)： 1179-1188.

厭氧土壤滅菌修復(fù)西瓜連作退化土壤：周開勝1，2(1.蚌埠學(xué)院 環(huán)境科學(xué)實(shí)驗(yàn)中心，安徽 蚌埠 233030； 2.南京師范大學(xué) 地理科學(xué)學(xué)院，江蘇 南京 210046)