Use of ethoxylated chlortetracycline for soil fumigation and its application as fumigant

Abstract

The present invention relates to the use of ethyl garlicin for soil fumigation and its application as a fumigant, and specifically relates to a use of ethyl garlicin for fumigating the soil of crops to be planted before crop planting, wherein the amount of ethyl garlicin used for soil fumigation is 10 - 60 g / m<supgt;2< / supgt;. By studying the effects of soil fumigation with ethyl garlicin on soil physical and chemical properties, the taxonomic composition of soil bacteria and fungi, crop plant growth, and crop yield, the present invention discovers that ethyl garlicin can be used as an efficient soil fumigant, and the growth of crops planted on the soil after fumigation with ethyl garlicin is promoted, and diseases and root-knot nematode pests are reduced.

Description

Technical Field

[0001] The invention belongs to the field of agricultural plant protection, in particular to the field of prevention and control of soil-borne diseases of crops and root-knot nematodes, and specifically to the application of ethiocarbamide and a composition thereof for soil fumigation and as a fumigant. Background Art

[0002] Soil-borne plant diseases and root-knot nematodes are seriously affecting the yield and quality of my country's crops, and have become important diseases that threaten the safe production of my country's grain and cash crops. With the rapid development of large-scale planting, cross-regional services, and high added value of crops in my country, coupled with global warming, soil-borne diseases are showing a serious trend, causing serious economic losses to society.

[0003] Although agricultural production has increased and farmers' income has increased through plant protection technologies, such as the use of pesticides and green control methods, this benefit is often offset by the increase in soil-borne diseases, resulting in a decline in crop quality and yield. For example, in the process of tomato cultivation, soil-borne diseases caused by late blight, tomato bacterial wilt, root nematode disease, etc. significantly reduce the quality and yield of tomatoes. If soil pathogens are not controlled in time, the yield can be reduced by more than 60.0%, and even if the soil-borne diseases cannot be effectively controlled and the crops are seriously infected by the diseases, the tomato yield will be completely lost.

[0004] Soil fumigation before planting is the most effective method for controlling soil-borne diseases. Methyl bromide (MB) was once a widely used and effective soil fumigant in the world, but it was banned due to its ozone depleting properties. Currently, other commonly used fumigants include chloropicrin (Pic), imidazole (DZ), 1,3-dichloropropylene (1,3-D), allyl isothiocyanate (AITC) and methyl sodium (MS). Although these fumigants have broad spectrum and can effectively kill soil pathogens and improve crop quality and yield, in order to expand the types of diseases controlled and improve the operational convenience of control, the field of soil health is in great need of new, green and efficient soil fumigants.

[0005] Soil fumigants have multiple effects on soil microorganisms, for example, fumigation significantly reduced bacterial community diversity and altered soil bacterial and fungal community structure. Soil nitrogen cycling was affected by 1,3-dichloropropene fumigation. Imidazole reduced soil bacterial taxonomic diversity. Allyl isothiocyanate initially reduced soil bacterial taxonomic diversity, but later stimulated soil fungal taxonomic diversity and significantly altered fungal community structure.

[0006] Ethylalicin is an organic sulfur fungicide independently developed in China. Its molecular formula is C4H 10O2S2, boiling point is 283.71℃, flash point is 125.38℃, density is 1.191g / cm3, it is a colorless or slightly yellow oily liquid. Ethyl garlic is characterized by high efficiency, broad spectrum and no pollution. Ethyl garlic is highly volatile, has a garlic smell, is effective against a variety of pathogenic fungi and bacteria, can reduce the growth rate of pathogenic populations and reduce plant diseases. Currently registered preparations of ethyl garlic include emulsifiable concentrates and wettable powders. Ethyl garlic is usually used as a spray to control diseases of rice, cotton and watermelon. Ethyl garlic is used as a conventional fungicide to control 76%-79% of bacterial horn rot of cucumbers, and inhibit more than 50% of gray mold, smut fungus, Rhizoctonia solani and sclerotinia. In addition, ethyl garlic is an easily degradable pesticide that is quickly digested in vegetables and soil, and will not cause residue accumulation and pollution in the soil when used at the recommended dose.

[0007] Although ethyl allicin can be used to control a variety of plant diseases, it is currently mainly used to control pathogenic pathogens suffered by plants during the growth stage, especially to inhibit the growth and reproduction of pathogenic bacteria during the growth stage of plants. Ethylicin has not been used for soil fumigation, and there is no report on how to use ethyl allicin for soil fumigation and simultaneously achieve the control of root-knot nematodes in the soil. Summary of the invention

[0008] In the present invention, the application studies the use of ethionine for soil fumigation to treat soil, reduce the number of pathogenic bacteria in the soil and can reduce the number of root-knot nematodes in the soil, control crop root-knot nematodes, for example, by fumigating the soil in a greenhouse where tomatoes are grown all year round to achieve the control of tomato soil-borne diseases and root-knot nematodes and promote tomato growth. Different concentrations of ethionine and different concentrations of ethionine and different concentrations of sulfur are used in soil fumigation to study the optimal ethionine concentration for ethionine fumigation to achieve soil-borne disease control, and the optimal combined use concentration of ethionine and sulfur. By regularly collecting soil samples after fumigation, the effects of Fusarium spp, Phytophthora spp pathogens and root-knot nematode abundance in the soil of different treatment schemes, the effects of soil physicochemical properties, soil enzyme activity, crop growth and crop yield are observed and analyzed. High-throughput gene sequencing technology is used to study the effects of different fumigation treatments on bacterial and fungal communities in the soil. The present invention has found that fumigating the soil before planting crops with ethionine and a composition comprising ethionine and sulfur can reduce soil pathogens and root-knot nematodes after planting crops in the fumigated soil, improve soil ecology, promote crop growth and increase crop yields.

[0009] Specifically, the present invention provides a use of ethionine for fumigating soil where crops are to be planted before planting the crops.

[0010] The fumigation is to mix ethoxylated and soil and then seal them, and then release the seal after a period of time to reduce the number of pathogenic microorganisms in the soil, so as to prevent the crops planted in the later stage from suffering from the adverse effects of soil-borne diseases.

[0011] Preferably, in the above use, the amount of ethoxylated used for soil fumigation is 10-60 g / m 2 It is particularly preferred that the amount of ethoxylated used for soil fumigation is 10-50 g / m 2 Particularly preferably, the amount of ethoxylated used for soil fumigation is 20-40 g / m 2 .

[0012] Preferably, in the above use, the amount of ethionine used for soil fumigation is 6000-40000 g / mu, and particularly preferably, the amount of ethionine used for soil fumigation is 10000-35000 g / mu.

[0013] The invention provides a composition containing ethionine for fumigating soil for planting crops before planting the crops. The content of ethionine in the composition is 5-90%.

[0014] The fumigation is to mix the composition containing ethionine with the soil and then seal it, and then release the seal after a period of time to reduce the number of pathogenic microorganisms in the soil, so as to prevent the crops planted in the later stage from suffering from the adverse effects of soil-borne diseases.

[0015] Preferably, in the above-mentioned use, the composition used for soil fumigation is calculated as ethyl allicin, and the amount of ethyl allicin is 10-60 g / m 2 , particularly preferably, the amount of ethionine is 10-50 g / m 2 Particularly preferably, the amount of the active ingredient ethionine is 20-40 g / m 2 .

[0016] Preferably, in the above use, the composition contains sulfur, wherein the mass ratio of ethionine to sulfur is 10:0.1-5.0, and the weight content of sulfur in the composition is 1-45%.

[0017] Preferably, in the above use, the amount of ethoxylated used for soil fumigation is 10-60 g / m 2 The amount of sulfur used is 0.1-30g / m 2 Preferably, the amount of ethoxylated used for soil fumigation is 10-50 g / m 2 The amount of sulfur used is 0.3-25g / m 2 , particularly preferably, the amount of sulfur used is 0.3-5g / m 2 Particularly preferably, the amount of ethoxylated used for soil fumigation is 10-40 g / m2 The amount of sulfur used is 0.9-20g / m 2 In particular, the amount of sulfur used is 0.9-1.8 g / m 2 ,

[0018] Preferably, in the above-mentioned use, the amount of ethionine used for soil fumigation is 6000-40000 g / mu, and the amount of sulfur used is 60-20000 g / mu. Particularly preferably, the amount of ethionine used for soil fumigation is 6000-30000 g / mu. Preferably, the amount of sulfur used is 600-15000 g / mu. Particularly preferably, the amount of sulfur used is 600-1500 g / mu.

[0019] The present invention also provides a method for fumigating soil for crops to be planted before planting the crops, the method comprising the following steps:

[0020] Step 1: Before planting crops, ethionine or a composition containing ethionine is mixed with the soil where the crops are to be planted, and then the surface of the soil is sealed; or the surface of the soil where the crops are to be planted is sealed, and then ethionine or a composition containing ethionine is mixed with the soil where the crops are to be planted;

[0021] Step 2: Lift the blockade after 3-60 days, and plant crops within 3-60 days after lifting the blockade.

[0022] Preferably, in the above method, the mixing method is to mix ethionine or a composition comprising ethionine with water and drip or sprinkle it into the soil, or to mix ethionine or a composition comprising ethionine with soil.

[0023] Preferably, in the above method, the sealing method is to cover the soil surface with a film, wherein the film is a film made of organic matter or a film made of a mixture of organic matter and inorganic matter.

[0024] Preferably, in the above method, the amount of ethionine or the composition containing ethionine applied in the soil is calculated based on the area of ​​the soil, and the amount of the active ingredient ethionine is 10-60 g / m 2 , particularly preferably, the amount of the active ingredient ethionine is 10-50 g / m 2 Particularly preferably, the amount of the active ingredient ethionine is 20-40 g / m 2 .

[0025] Preferably, in the above method, the mass content of ethionine in the composition containing ethionine is 5-90%.

[0026] Preferably, in the above method, in step 2, the sealing is lifted after 20-50 days, and the crops are planted again within 5-50 days after lifting the sealing.

[0027] The present invention also provides the use of ethionine or a composition containing ethionine for soil fumigation before crop planting to reduce the CFU value of pathogenic bacteria in the fumigated and crop-planted soil and increase the fungal Shannon index and the fungal super index.

[0028] The present invention also provides the use of ethionine or a composition containing ethionine for soil fumigation before crop planting to reduce the CFU value of pathogenic bacteria in the fumigated soil where crops have been planted, increase the fungal Shannon index and the fungal super index, and reduce the number of root-knot nematodes.

[0029] The CFU (colony forming unit) means colony forming unit.

[0030] Preferably, in the above use, the amount of ethoxylated used for soil fumigation is 10-60 g / m 2 It is particularly preferred that the amount of ethoxylated used for soil fumigation is 10-50 g / m 2 Particularly preferably, the amount of ethoxylated used for soil fumigation is 20-40 g / m 2 .

[0031] Preferably, in the above use, the amount of ethionine used for soil fumigation is 6000-40000 g / mu, and particularly preferably, the amount of ethionine used for soil fumigation is 10000-35000 g / mu.

[0032] Preferably, in the above use, the content of ethionine in the composition is 5-90%.

[0033] Preferably, in the above-mentioned use, the composition used for soil fumigation is calculated as ethyl allicin, and the amount of ethyl allicin is 10-60 g / m 2 , particularly preferably, the amount of ethionine is 10-50 g / m 2 Particularly preferably, the amount of the active ingredient ethionine is 20-40 g / m 2 .

[0034] Preferably, in the above use, the composition contains sulfur, wherein the mass ratio of ethionine to sulfur is 10:0.1-5.0, and the weight content of sulfur in the composition is 1-45%.

[0035] Preferably, in the above use, the amount of ethoxylated used for soil fumigation is 10-60 g / m 2 The amount of sulfur used is 0.1-30g / m 2 Preferably, the amount of ethoxylated used for soil fumigation is 10-50 g / m 2 The amount of sulfur used is 0.3-25g / m 2Preferably, the amount of sulfur used is 0.3-5 g / m 2 Particularly preferably, the amount of ethoxylated used for soil fumigation is 10-40 g / m 2 The amount of sulfur used is 0.9-20g / m 2 The amount of sulfur used is 0.9-1.8g / m 2 .

[0036] Preferably, in the above-mentioned use, the amount of ethionine used for soil fumigation is 6000-40000 g / mu, and the amount of sulfur used is 60-20000 g / mu. Particularly preferably, the amount of ethionine used for soil fumigation is 6000-15000 g / mu, and particularly preferably, the amount of sulfur used is 600-1500 g / mu.

[0037] The invention also provides the use of ethionine in preparing a fumigant for fumigating soil to prevent and control soil-borne diseases and root-knot nematodes of crops planted on the fumigated soil.

[0038] In the above uses and methods, the crops are selected from food crops, cash crops, fruits and vegetables, and lawn and garden crops.

[0039] Ethylalicin or a composition containing ethylalicin significantly reduced the relative prevalence of bacteria, reduced the number and species diversity of bacteria in the soil, and increased the number of fungi through soil fumigation. Ethylalicin or a composition containing ethylalicin changed the physical and chemical properties of the soil and the taxonomic dominance of microorganisms through soil fumigation, thereby controlling pathogens and increasing crop yields.

[0040] Ethylion or a composition comprising ethylion significantly reduces the number of Phytophthora, Fusarium and Root-knot Nematode by soil fumigation. Ethylion or a composition comprising ethylion, such as a composition of ethylion and sulfur, significantly increases the mortality of these pathogenic bacteria and root-knot nematodes in the soil, promotes plant growth, and increases crop yield. Ethylion or a composition comprising ethylion changes the physicochemical properties of the soil by soil fumigation, thereby changing the relative abundance and classification composition of soil microorganisms. Ethylion or a composition comprising ethylion reduces the number of bacteria and species diversity in the soil by soil fumigation, increases the number of fungi, and causes complex interactions between soil microorganisms.

[0041] Beneficial Effects

[0042] The present invention shows that ethionine or a composition containing ethionine as a soil fumigant has obvious inhibitory and killing effects on plant pathogens (such as Fusarium and Phytophthora), and also has excellent effects on plant root-knot nematodes.

[0043] The present invention studies the influence of ethyl allicin on soil physical and chemical properties, soil bacteria and fungi classification composition, enzyme activity, tomato plant growth and tomato yield through soil fumigation, and obtains that ethyl allicin can be used as a soil fumigant. Through ethyl allicin soil fumigation, the growth of crops planted on the fumigated soil is promoted and diseases and root-knot nematode pests are reduced.

[0044] Soil fumigation with ethylalicin significantly reduced the abundance of Fusarium and Phytophthora by 67.7%-84.0% and 53.8%-81.0%, respectively, and ethylalicin reduced root-knot nematodes by 67.2%-83.6%. Soil fumigation with ethylalicin significantly increased the growth of tomato plants and increased tomato yield by 18.3%-42.0%. Due to the effect of ethylalicin, soil ammonium nitrogen concentration increased significantly, while nitrate nitrogen concentration and soil urease activity decreased significantly. High-throughput gene sequencing showed that soil fumigation with ethylalicin reduced soil bacterial diversity and bacterial abundance, but increased the taxonomic diversity of soil fungi. The increase in some microbial genera, such as Firmicutes, Steroidobacteria and Chytridiomycetes, may be due to changes in the physical and chemical properties of the soil, and the soil is more conducive to the reproduction of beneficial microorganisms, so ethylalicin is effective as a soil fumigant, and it will be a useful supplement to the currently limited varieties of soil fumigants.

[0045] The results of the study on the fumigation of cowpea with allicin in soil with continuous cropping showed that increasing the concentration of allicin fumigation and shortening the fumigation time would not cause crop damage, but would increase the emergence rate of crops and promote the growth of crops in the seedling stage. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 Fuscarium spp. in soil samples collected during the first, second, and third fumigation treatments ( Figure 1 A), Phytophthora spp. Figure 1 B) and Meloidogyne spp. ( Figure 1 C) influence diagram.

[0047] Mean values ​​(N=3) and the same letter in the same bar graph were not statistically different (P=0.05) according to Duncan's new multiple range test.

[0048] Figure 1 The meaning of CFU (colony forming unit) is colony forming unit.

[0049] Figure 2 is the effect of fumigation treatment on stem diameter (A), leaf chlorophyll (B), fruit number (C) and tomato yield (D).

[0050] Figure 2 The English title of the vertical axis means:

[0051] Stem diameter: stem diameter; Leaf Cholorophyll: leaf cholorophyll; No. of Fruit: number of fruits; Tomato yield: tomato yield.

[0052] Figure 3 The effects of different fumigation treatments on soil bacteria ( Figure 3 A, Figure 3 C) and fungi ( Figure 3 B, Figure 3 D) Impact diagram of alpha diversity.

[0053] Figure 3 The English title of the vertical axis means:

[0054] Shannon index of OUT level: Shannon index of classification operation unit level (or Shannon index); Chao index of OUT level: Chao index of classification operation unit level (or Chao index).

[0055] Translation of the category names in the picture: Bacteria: bacteria; Fungi: fungi.

[0056] The t-test was used to determine the differences in alpha diversity between soil fumigation treatments. Statistical differences between two groups are indicated by 1-3 asterisks (P < 0.05): *0.01 < P ≤ 0.05; **0.001 < P ≤ 0.01; ***P ≤ 0.001.

[0057] Figure 4 The effects of different fumigation treatments on soil bacteria ( Figure 4 A) and fungi ( Figure 4 B) PCoA influence plot of the community.

[0058] English translation of the figure: PCoA on OUT level: PCoA on the operational classification unit level.

[0059] Figure 5 The effects of different fumigation treatments on bacteria ( Figure 5 A) and fungi ( Figure 5 B) Analysis of changes in taxonomic composition.

[0060] in Figure 5 The Chinese meaning of the English names of the microorganisms on the vertical axis in A:

[0061] Actinobacteriota: Actinobacteria; Firmicutes: Firmicutes; Patescibacteri a: Patellar bacteria; Planctomycetota: Planctomycetes; Deinococcota: Deinococcus; Des ulfobacterota: Desulfobacteria;

[0062] in Figure 5 The Chinese meaning of the English names of the microorganisms on the vertical axis in B:

[0063] Ascomycota: Ascomycota; Unclassified_k_Fungi: Unclassified fungi; Chytridiomycota: Chytridiomycota.

[0064] Figure 5 The Chinese meaning of the horizontal axis mean proportion is: average proportion.

[0065] The number of asterisks indicates significant differences (P < 0.05) between treatments according to one-way ANOVA and FDR (false discovery rate) adjustment: *0.01 < P ≤ 0.05; **0.001 < P ≤ 0.01; ***P ≤ 0.001.

[0066] Figure 6 Figure 5 is the LEfSe cladogram analysis of the effects of different doses of ethionine or the combined fumigation of ethionine and sulfur (P=0.05) on the abundance of different species in the soil bacterial (LDA=3.5) community.

[0067] The five rings in the cladogram represent phylum, class, order, family, and genus, respectively. Different colored nodes on the rings represent significant changes in taxonomic composition due to different treatments.

[0068] Figure 6 The names of Chinese fungi and some of their English names are translated as follows, where "no Chinese translation yet" means that the fungus currently has no Chinese translation.

[0069] Nitrosomonadaceae, Pseudomonadales, Pseudomonadaceae, Pseudomonas, Xanthomonadaceae, Lysobacter, Cellvibrionales, Cellvibrionaceae, Cellvibrio, Steroidobacterales, Steroidobacteraceae The genus is also known as the family of bacteria, including Steroidobacteria, Steroidobacter, Alphaprotcobacteria, Rhizobiales, Beijerinckiaceae, Microvirga, Devosiaceae, Dongiales, Dongiaceae, Sphingomonadales, Sphingomonadaceae, and Sphingomonadales. gomonas, Alphaproteobacteria, Firmicutes, Bacilli, Bacillales, Bacillaceae, Bacillus, Virgibacillus, Paenibacillales, Chryseolinea, Rhodothermia, Rhodothermales, Rhodoth ermaceae, Patescibacteria, Dehalococcoidia, Ardenticatenales, Ardenticatenaceae, Ardenticatenaceae, Actinobacteriota, Actinomarinales, Actinomarinales, Actinomarinales, Actinomarinales,Nocardioides, Streptomycetales, Streptomycetaceae, Streptomyces, Gaiellales, Longimicrobia, Longimicrobiales, Longimicrobiaceae, terrestrial, group, norank.

[0070] Figure 7 Figure 5. LEfSe cladogram analysis of different differentially abundant species in soil fungal (LDA=4.0) communities after fumigation with different doses of ethionine or combination of ethionine and sulfur (P=0.05).

[0071] Figure 7 The names of Chinese fungi and some of their English names are translated as follows, where "no Chinese translation yet" means that the fungus currently has no Chinese translation.

[0072] Ascomycota, Pezizomycetes, Pezizales, Ascobolaceae, Ascobolus, Pezizaceae, Iodophanus, Sordariomycetes, Chaetomiaceae, Humicola, Myceliophthora, Sordariales, unclassified, Incertae_sedis, Microascales, Microascomycetes, croasouceae, Hypocreales, Hypocreales, Acremonium, Glomerellales, Plectosphaerellaceae, Sodiomyces, Sordariomycetes, Chytridiomycota, Olpidiomycota, Olpidiomycetes, Olpidiales, Olpidiaceae, Olpidium, Fungi.

[0073] Figure 8 For soil bacteria ( Figure 8 A) and fungi ( Figure 8 B) Heat map analysis of the correlation between the taxonomic composition of phyla, environmental factors, and tomato yield. Correlation analysis was performed using the Spearman method. The legend shows the color intervals for different R values. The number of asterisks indicates the degree of correlation (P < 0.05): * 0.01 < P ≤ 0.05; ** 0.001 < P ≤ 0.01; *** P ≤ 0.001.

[0074] Figure 8 The full name corresponding to the code in the horizontal axis of A is the name in brackets after the following code.

[0075] OM (organic matter), EC (electrical conductivity), NN (nitrate ammonium), S-UE (soil urease), Mel (root-knot nematode), Fus (Fusarium), Phy (Phytophthora), K (available potassium), S-SC (soil sucrase), pH (pH), P (available phosphorus), S-CAT (soil catalase), AN (ammonium nitrogen), TY (tomato yield).

[0076] Figure 8 The translation of the Latin name in the vertical axis of A is as follows, where "no Chinese translation yet" means that the fungus currently has no Chinese translation name.

[0077] The main phyla included Chloroflexi, Patescibacteria, Nitrospirota, Armatimonadota, Halanaerobiaeota, Verrucomicrobiota, Sumerlaeota, Acidobacteriota, Cyanobacteria, Elusimicrobiota, Fibrobacterota, Bdellovibrionota, Deinococcota, Proteobacteriota, and Cyanococcus. ria, Gemmatimonadota, Bacteroidota, Firmicutes, Myxococcota, Actinobacteriota, group, cladeMarine, Desulfobacterota, Dependentiae, Planctomycetota, Latescibacterota, Methylomirabilota, unclassified, norank, Bacteria.

[0078] Figure 8 The full name corresponding to the code in the horizontal axis of B is in the brackets after the following code.

[0079] pH (pH), S-SC (soil sucrase), P (available phosphorus), S-CAT (soil catalase), AN (ammonium nitrogen), TY (tomato yield), EC (electrical conductivity), Mel (root-knot nematode), Fus (Fusarium), Phy (Phytophthora), NN (nitrate nitrogen), S-UE (soil urease), K (available potassium), OM (organic matter).

[0080] Figure 8 The translation of the Latin name in the vertical axis B is as follows, where "no Chinese translation yet" means that the fungus currently has no Chinese translation name.

[0081] Ascomycota, Glomeromycota, Chytridiomycota, unclassified_k_Fungi, Blastocladiomycota, Basidiobolomycota, Basidiomycota, Neocallimastigomycota, Olpidiomycota, Aphelidiomycota, Entomophthoromycota, Zoopagomycota, Rozellomycota, Mortierellomycota, Calcarisporiellomycota, Kickxellomycota.

[0082] Fig. 9 This is a picture of the situation before and after the emergence of cowpea after watering with 1L / hole at 500 times the concentration of 80% ethidium bromide and sealing for 3 days.

[0083] Fig.10 This is a picture of the situation before and after the emergence of cowpea after watering with 500mL / hole with 80% ethyl allicin 500 times and sealing for 3 days.

[0084] Fig.11 This is a picture of the situation before and after the emergence of cowpea after watering with 500mL / hole and sealing for 3 days.

[0085] Fig.12 This is a picture of the situation before and after the emergence of cowpea after watering with 1L / hole at 500 times the concentration of 80% ethidium bromide and sealing for 7 days.

[0086] Fig.13 This is a picture of the situation before and after the emergence of cowpea after watering with 500mL / hole with 80% ethyl allicin 500 times and sealing for 7 days.

[0087] Fig.14 The picture shows the situation before and after the emergence of cowpea after watering with 500mL / hole with 250 times 80% allicin and sealing for 7 days.

[0088] Fig.15 This is a picture of the situation before and after the emergence of cowpea after watering with 500mL / hole and sealing for 7 days. DETAILED DESCRIPTION

[0089] The experiment was conducted in a greenhouse located in Fangshan District, Beijing, which has been used for continuous tomato production for more than 20 years. The greenhouse soil is characterized as "sandy loam" (average soil composition: sand: 35.12%, silt: 51.78%, clay: 13.10%). The fumigation experiment was conducted in this greenhouse.

[0090] The agents used for fumigation are ethionine EC (80%), ethionine (1.5%), and sulfur AS (3%), and the agents are provided by Hainan Zhengye Zhongnong High-tech Co., Ltd.

[0091] The greenhouse fumigation experiment set up six treatments, which were:

[0092] (1) Blank treatment (CK);

[0093] (2) Low concentration of ethoxylated 10.0 g / m 2 Fumigation (E10);

[0094] (3) Medium concentration of ethoxylated 20.0 g / m 2 Fumigation (E20);

[0095] (4) High concentration of ethoxylated 40.0 g / m 2 Fumigation (E40);

[0096] (5) Low concentration of ethoxylated 10.0 g / m 2 + Sulfur 0.9g / m 2 Fumigation (ES0.9);

[0097] (6) High concentration of ethoxylated 40.0 g / m 2 + Sulfur 1.8g / m 2 Fumigation (ES1.8).

[0098] The fumigation agent was mixed evenly with the soil using soil drip irrigation technology. The application process was as follows: Step 1: Cover the soil surface with a 0.04 mm polyethylene film ("PE film", from Shandong Shouguang Longxing Technology Co., Ltd., China); Step 2: Wet the soil using water drip irrigation; Step 3: Mix the fumigation agent and water evenly, and then apply them to the soil through drip irrigation; Step 4: Use water drip irrigation again to fully mix the fumigation agent into the soil; Step 5: Remove the PE film after 4 weeks.

[0099] Fifteen days after the PE film was removed, tomato seedlings were transplanted into the fumigated soil. Tomatoes were planted in a 10.0 cm high and 120.0 cm wide seedbed with a spacing of 80.0 cm between seedlings and a row spacing of 1.5 m. Each treatment was replicated three times, with a total of 30 tomato plants in each treatment. All treatments adopted the same field management practices.

[0100] The soil was sampled three times in the greenhouse, 2 months (first time), 3 months (second time), and 4 months (third time) after transplanting the tomato seedlings. In each treated plot, three points were randomly selected and soil samples were collected at 5.0-20.0 cm below the soil surface. Each soil sample was sieved through a sieve with a 2.0 mm hole to remove gravel and plant residues. Each soil sample was divided into three parts for testing different indicators. The first part was refrigerated at 4°C to detect soil inorganic nitrogen content, soil pathogen counts, and root-knot nematode counts. The second part was air-dried at 4°C to measure changes in soil physicochemical properties and soil enzyme activity, and the third part was refrigerated at -80°C to analyze changes in microbial classification.

[0101] Soil pathogens Fusarium, Phytophthora and Root-knot Nematodes were isolated and the colony forming units per gram of soil (CFU / g soil) and the number of root-knot nematodes in each treatment were recorded. Figure 1 .

[0102] from Figure 1 It can be seen that compared with the blank treatment, the fumigation treatment of ethyl allicin greatly reduced the abundance of Fusarium, Phytophthora and Root-knot Nematode. Over time, the number of pathogens and root-knot nematodes in the blank soil increased rapidly, but the ethyl allicin fumigation treatment still maintained effective control over pathogens and root-knot nematodes. Before the third survey, the effect of each ethyl allicin fumigation treatment on reducing the CFU (colony forming units) of Fusarium and Phytophthora in the soil was still 67.7-84.0% ( Figure 1 A) and 53.8-81.0% (see Figure 1 B). The number of soil root-knot nematodes decreased by 67.2-83.6% ( Figure 1 C). E40 treatment with increasing concentrations of ethoxylated chloranil significantly reduced the number of Fusarium and Phytophthora in the soil and greatly reduced the CFU of Phytophthora ( Figure 1 B). The control effect of ES1.8 was similar to that of E10 and better than that of ES0.9. Moreover, the control effect of ES1.9 on Phytophthora was significantly higher than that of ES0.8. These treatments were effective in controlling root-knot nematodes ( Figure 1 C).

[0103] The stem diameter of tomato plants was measured with a digital vernier caliper. The chlorophyll content in tomato leaves was measured by a handheld non-destructive SPAD-502plus chlorophyll meter (Konica Minolta, Japan). The harvest date, yield per plant and its weight were recorded. The total tomato yield (TY) of each treatment was calculated from the ripening of tomato fruits to the completion of harvest. For specific results, see Figure 2 .

[0104] from Figure 2It can be seen that all fumigation treatments significantly increased stem diameter, tomato fruit number and yield compared to the blank treatment. In the first and second surveys, E40 significantly increased stem diameter by 16.5% and 18.9%, respectively ( Figure 2 All fumigation treatments increased the leaf chlorophyll content of tomato plants by 2.1-9.3% ( Figure 2 B). The results of the first chlorophyll survey showed that each fumigation treatment significantly increased the chlorophyll content of tomatoes compared with the blank treatment, and E40 was significantly higher than ES1.8. In the second and third surveys, E40 significantly increased leaf chlorophyll compared with all other fumigation treatments except E20.

[0105] Ethylalicin fumigation treatment significantly increased the fruit yield by 13.8-38.6% ( Figure 2 C), tomato yield increased by 18.3-42.0% ( Figure 2 D). Except for E20 and ES1.8, there were significant differences in the number of fruits and tomato yield among the other fumigation treatments. Although the number of fruits and tomato yield increased significantly with the increase in the dose of ethoxylated chloranil, ES0.9 and ES1.8 both significantly increased tomato yield compared with the blank treatment.

[0106] The concentrations of ammonium nitrogen and nitrate nitrogen in the soil were determined using the FuturaTM continuous flow analysis system. The effective potassium concentration in each treatment was measured using an FP6410 flame photometer. The effective phosphorus concentration in each treatment was measured using a UV2012-PC spectrometer. The content of organic matter was measured. The soil conductivity and pH value (soil: water = 1:2.5) were measured using an MP513 conductivity meter and an MP512-02 precision water meter (Shanghai Sanxin Instrument Co., Ltd.), respectively. The specific results are shown in Table 1 below.

[0107] Table 1 Effects of fumigation treatment on nitrogen, P, K, organic matter, pH and conductivity in soil

[0108]

[0109] As can be seen from Table 1, ethyl allicin fumigation significantly increased soil ammonium nitrogen (NH4 + -N), reducing soil nitrate nitrogen (NO3 - -N), compared with other fumigation treatments, E40 significantly increased soil ammonium nitrogen and significantly reduced soil nitrate nitrogen. Compared with the blank treatment, the allicin fumigation treatment increased the available phosphorus concentration, among which E10 and E40 significantly increased soil available phosphorus. E40 was significantly higher than other fumigation treatments. The allicin fumigation treatment did not significantly change the soil organic matter content. However, compared with the blank treatment, allicin soil fumigation increased the soil pH and significantly reduced the soil electrical conductivity.

[0110] The activities of soil sucrase (S-SC), soil catalase (S-CAT) and soil urease (S-UE) were detected based on the corresponding enzyme activity detection kits. 3 The multimode microplate measures the concentration of each substance based on absorbance, as shown in Table 2 below.

[0111] Table 2 Changes in soil enzyme activities

[0112]

[0113] As can be seen from Table 2, soil sucrase activity decreased in all fumigation processes compared with the control in the first survey, except for the treatment ES1.8. In the second survey, the enzyme activity decreased significantly in all fumigations. However, soil sucrase activity increased in the third survey compared with the control. Soil sucrase content decreased over time, but the rate of decrease was slow in the ethyl allicin fumigation treatment. Soil catalase activity was not affected by ethyl allicin fumigation in the first survey compared with the control: but it increased in E40 in the second and third surveys. Ethylicin fumigation treatment significantly reduced soil urease activity compared with the control. This ratio decreased significantly with the increase in the amount of ethyl allicin in the second and third surveys. Soil urease activity decreased significantly by 55.6-73.5% in the third survey compared with the control. There were significant differences in soil urease activity in all treatments except E20 and ES1.8.

[0114] according to Soil total genomic DNA was extracted following the instructions provided in the DNA isolation kit. The extracted DNA was confirmed using 1% (W / V) agarose gel electrophoresis. DNA concentration was determined using a NanoDrop ND-1000 spectrophotometer.

[0115] Total DNA extracted from soil was used as template for PCR amplification and MiSeq sequencing in the bacterial V3-V4 region of 16S rRNA and the fungal ITS1 region, respectively. 338F and 806R were used as universal primers for bacteria. ITS1F and ITS2R were used as universal primers for fungi. PCR amplification and MiSeq sequencing were performed on Illumina's MiSeq PE300 platform.

[0116] In order to obtain high-quality and complete sequence data for subsequent information analysis, Flash (V.1.2.11) software was used to splice the sequences obtained by MiSeq sequencing from the raw data. The UPARSE (V.11) clustering of optimized sequences with 97% similarity was used as classification operational units (OTUs). In order to obtain the species classification information corresponding to each classification operational unit, the RDP classifier (V.2.13) Bayesian algorithm was used to classify and analyze the representative sequences of the classification operational units with a similarity level of 97%, and compared with the bacterial 16SrRNA database Silva (V.138) and the fungal ITS database Unite (V.8.0). In order to calculate α diversity and evaluate the differences in taxonomic diversity between different treatments, the Mothur bioinformatics method was used. Qiime (V.1.9.1) software was used for β diversity analysis in hierarchical cluster analysis and PCoA analysis. One-way analysis of variance (ANOVA) was used to describe the statistical significance between treatments and species.

[0117] After optimizing the original sequences, we obtained a total of 1,420,786 bacterial and 1,183,371 fungal sequences from all samples, and clustered 7,740 bacterial OTUs and 864 fungal OTUs based on 97.0% similarity, respectively. Figure 3 .from Figure 3 It can be seen that, except for E40, all fumigation treatments significantly reduced the Shannon diversity index of bacteria. ES1.8 significantly reduced the Shannon diversity index and Chao diversity index of bacteria (P = 3.1 × 10 -4 , P = 6.7 × 10 -3 , ES1.8CK). Except for E40 and ES0.9, the Chao diversity index of other treatments was significantly lower than that of the blank treatment. All fumigation treatments significantly increased the Shannon diversity index of fungi. Compared with other treatments, E40 significantly increased the Shannon diversity index of fungi. ES1.8 significantly increased the Shannon diversity index of fungi higher than ES0.9 (P = 3.9 × 10 -2 , ES0.8 and ES1.9, respectively (t-test). E20 and E40 significantly increased the fungal diversity index. Compared with other treatments, E40 significantly increased the Chao diversity index of fungi.

[0118] The results of hierarchical clustering and principal coordinates analysis can be found in Figure 4Hierarchical cluster analysis based on β-diversity POW markers demonstrated that the three replicates of each treatment clustered together in bacterial and fungal communities, indicating good repeatability between samples. In general, allicin fumigation had a relatively high contribution to soil bacterial and fungal diversity. The results showed that PC1 and PC2 contributed 20.9% and 9.9% (16S), 23.1% and 13.2% (ITS) to the differences in species composition between treatments, respectively.

[0119] The results of the analysis of changes in bacterial and fungal taxonomic composition can be found in Figure 5 ,from Figure 5 It can be seen that Actinobacteria, Firmicutes and Patella are the dominant phyla in the bacterial community ( Figure 5 A). Fumigation treatments with ethionine or a combination of ethionine and sulfur significantly reduced the relative abundance of Actinobacteria by 18.3%-32.1% compared to the blank treatment. E40 reduced the abundance of Actinobacteria by 32.1%. All fumigation treatments significantly increased the relative prevalence of Firmicutes by 11.4%-34.0%. E40 increased the relative abundance of Firmicutes by 11.4%. Fumigation treatments greatly increased the relative prevalence of Patellar bacteria by 25.3%-83.7%. E10 significantly increased the relative prevalence of Patellar bacteria by 83.7%. Ascomycota was the dominant phylum in the fungal community. Fumigation treatments significantly reduced the relative prevalence of Ascomycota by 8.5%-55.5%. E40 and ES1.8 significantly reduced the relative prevalence of Ascomycota by 51.9% and 55.5%.

[0120] Bacillus, Steroidobacterium, and Sphingomonas were the dominant genera in the bacterial community. Figure 6 It can be seen that E10 and E40 significantly reduced the relative prevalence of Bacillus by 40.3% and 36.3%, respectively. Fumigation increased the relative prevalence of Bacillus steroidogenes by 25.2-106.8%. E10 and E40 significantly reduced the relative prevalence of Sphingomonas by 20.6% and 28.1%, respectively. Figure 6 In the fungal community, Sodiomyces and Iodophanus were observed to be the dominant genera ( Figure 7 Fumigation significantly increased the relative prevalence of Soxera and Iodonophores, ranging from 1793.5% to 15546.0% and 325.9% to 18797.1%, respectively.

[0121] LEfSe software was used to detect significant differences in bacterial and fungal abundance at the phylum and genus levels. LEfSe analysis showed that at the bacterial phylum level, CK (Actinobacteria), E10 (Patellarium), and ES0.9 (Firmicutes) each obtained one biomarker ( Figure 6). At the fungal phylum level, one marker was obtained for each of E40 (Chytridiomycota), ES0.9 (Oleophycota), and ES1.8 (Unclassified_k_fungi), and at the bacterial genus level, 5, 3, 2, 2, 3, and 3 biomarkers were found in CK, E10, E20, E40, ES0.9, and ES1.8, respectively. At the fungal genus level, 2, 2, 1, 3, 2, and 1 biomarker were found in CK, E10, E20, E40, ES0.9, and ES1.8, respectively ( Figure 7 ). The results showed that bacterial colonies were more sensitive to ethoxylated fumigation than fungal colonies.

[0122] Effects of environmental factors on soil microbial diversity Figure 8 , Figure 8 Heat map analysis showed the relationships between the prevalence of the 30 most common bacteria and fungi and soil physicochemical properties (N, P, K, organic matter, pH, electrical conductivity), soil pathogenic fungi and root-knot nematodes, tomato yield, and soil sucrase, urease, and catalase activities.

[0123] In general, bacteria are more often associated with environmental factors than fungi ( Figure 8 ). In the bacterial community at the phylum level, the relative prevalence of Fibrobacter was positively correlated with tomato yield ( Figure 8 A). Firmicutes were positively correlated with soil pH. Late bacilli were negatively correlated with available potassium. Fibrobacterium were negatively correlated with soil pathogenic fungi and root-knot nematodes. Actinomycetes were significantly positively correlated with soil pathogenic fungi, root-knot nematodes, soil urease and soil nitrate nitrogen, respectively.

[0124] At the fungal level, Ascomycota was positively correlated with electrical conductivity, organic matter, and nitrate nitrogen ( Figure 8 A). Cladosporium was positively correlated with available phosphorus, ammonium nitrogen, catalase activity, and tomato yield. Cladosporium was negatively correlated with electrical conductivity, soil pathogenic fungi, root-knot nematodes, nitrate nitrogen, and urease activity. Chytridiomycetes was positively correlated with soil sucrase activity, available phosphorus, ammonium nitrogen, and tomato yield. Chytridiomycetes was negatively correlated with root-knot nematodes, soil pathogenic fungi, nitrate nitrogen, and urease activity. Chytridiomycetes was positively correlated with soil sucrase activity, available phosphorus, ammonium nitrogen, and tomato yield. Glomeromycetes was negatively correlated with root-knot nematodes, soil pathogenic fungi, nitrate nitrogen, and urease activity ( Figure 8 B).

[0125] In the bacterial community, genus-level association analysis showed that Sphingomonas was negatively correlated with soil sucrase, catalase, and available phosphorus ( Figure 8 A). Bacillus spp. was negatively correlated with soil available phosphorus and catalase activity.

[0126] In the fungal community, genus-level association analysis showed that Plectosphaerella was positively correlated with soil catalase activity, soil pH, ammonium nitrogen, and tomato yield. Plectosphaerella was negatively correlated with soil electrical conductivity, nitrate nitrogen, urease activity, soil pathogenic fungi, and root-knot nematodes. Figure 8 B).

[0127] Heat map correlation analysis showed that soil microbial communities were closely associated with the genus Meloidogyne. The bacterial Shannon index and super index were positively correlated with the abundance of Meloidogyne. In contrast, the fungal Shannon index and super index were negatively correlated with the genus Meloidogyne.

[0128] In order to investigate the safety of soil fumigation with ethyl allicin, short-term fumigation and seed germination and growth tests were conducted to verify the safety. Specifically, cowpea was sown after fumigating the soil with high concentrations of ethyl allicin to observe the emergence of cowpea seedlings to verify the safety of ethyl allicin fumigation.

[0129] Test varieties: cowpea, Bohong white golden horse bean seeds, purchased

[0130] Experimental agent: 80% ethoxylated emulsifiable concentrate, provided by Hainan Zhengye Zhongnong Hi-Tech Co., Ltd.

[0131] The test was conducted at the test base of Hainan Zhengye Laocheng Factory on a ridge about 10 cm high and 60 cm wide. Cowpea planting holes were set after the previous crop of cowpea seedlings were cleaned. After the planting holes were determined, the planting holes were irrigated with a solution diluted with 80% ethionyl acetate emulsifiable concentrate. The irrigating and applying treatments were carried out as shown in Table 3. After the agent was irrigated, the clear water control treatment and the ethionyl acetate treatment were irrigated at the same time. After irrigating, the film was covered and sealed within 1 hour. After sealing, a hole was dug in the film directly above the planting hole. Cowpea seeds were sown within 1 day after digging the hole, and covered with soil at the location of the planting hole. Three seeds were sown in each planting hole. The management after sowing was the same, and no other agents and fertilizers were applied.

[0132] Table 3 Treatment table of ethyl allicin fumigation test

[0133] Serial number deal with dose Closure days (days) 1 80% Ethyl Allicin 500 times 1L / hole 3 2 80% Ethyl Allicin 500 times 500mL / hole 3 3 Clear water control 500mL / hole 3 4 80% Ethyl Allicin 500 times 1L / hole 7 5 80% Ethyl Allicin 500 times 500mL / hole 7 6 80% Ethyl Allicin 250 times 500mL / hole 7 7 Clear water control 500mL / hole 7

[0134] When the cowpeas germinate and grow to the 2-3 leaf stage, the emergence of cowpeas in each treatment was observed and counted. The emergence rate is shown in Table 4.

[0135] Table 4 Summary of cowpea seedling emergence rate

[0136] Serial number Seedling rate (%) 1 92.97aA 2 94.53aA 3 80.47bAB 4 84.38abA 5 90.63abA 6 87.50abA 7 70.31cB

[0137] For observation of the growth of cowpea after emergence, see Figures 9 to 15, Specific statistical data can be seen from Table 4. The application of 80% ethyl allicin in holes before sowing had no effect on the emergence of cowpea seedlings. The emergence of cowpea seedlings sealed with ethyl allicin for 3 days was slightly better than that sealed with ethyl allicin for 7 days. The ethyl allicin soil fumigation had no symptoms of stem burning on cowpea stems.

[0138] The cowpea seedling emergence experiment verified that when the closure time is relatively short during the fumigation of soil with ethyl allicin, it will not cause phytotoxicity to the germination and seedling growth of cowpeas that are sown immediately, because the seed germination and seedling emergence stages are easily affected by phytotoxicity, and high-concentration ethyl allicin fumigation, even if it is closed for a short time (the closure stage is actually the fumigation stage), will not cause phytotoxicity to seed germination and seedling growth, but can increase the germination rate. It can also be seen from the growth of cowpea that ethyl allicin soil fumigation can promote the growth of cowpeas in the seedling stage, and the cowpea seedlings grow better.

Claims

1. A composition comprising ethionine for fumigating soil for crops to be planted before planting, wherein the content of ethionine in the composition is 5-90%, the composition comprises sulfur, wherein the mass ratio of ethionine to sulfur is 10:0.1-5.0, the weight content of sulfur in the composition is 1-45%, and the amount of ethionine in the composition used for soil fumigation is 10-60 g / m 2 .

2. The use according to claim 1, characterized in that The amount of ethoxylated used in soil fumigation is 10-60 g / m 2 The amount of sulfur used is 0.1-30g / m 2 .

3. The use according to claim 2, characterized in that The amount of ethoxylated used for soil fumigation is 10-50g / m 2 The amount of sulfur used is 0.3-25g / m 2 .

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