CN111011382B - Application of beta-glucan and succinoglycan in prevention and control of soil-borne fungal diseases - Google Patents
Application of beta-glucan and succinoglycan in prevention and control of soil-borne fungal diseases Download PDFInfo
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Abstract
The invention discloses application of beta-glucan and succinyl glycan in preventing and controlling soil-borne fungal diseases. According to the invention, the mixture of beta-glucan and succinyl polysaccharide is added into soil, so that the increase of the number of streptomycete in the soil can be induced, the increase of the activity of hydrolases such as beta-glucanase, chitinase and cellulase in the soil can be induced, and the effective prevention and control of various soil-borne pathogenic fungal diseases can be realized through the combined action of the streptomycete and various hydrolases. The invention has the advantages of wide source of required raw materials, simple operation, better effect than that of singly using one polysaccharide and good environmental compatibility.
Description
Technical Field
The invention belongs to the technical field of biological control, and relates to application of beta-glucan and succinyl glycan in controlling soil-borne fungal diseases.
Background
The soil-borne diseases refer to diseases that pathogens come from soil and attack plants from roots or stems of the plants, and have the characteristics of wide distribution, serious harm, wide host range, multiple propagation ways, long survival time and the like, and have great harm to the plants. The prevention and control of soil-borne diseases by a biological method is a technology with less environmental risk. Wherein, the biological prevention and control by adding an environment-friendly inducer from an external source mainly depends on the effects of competitive growth, antagonism, heavy parasitism and the like of soil microbial populations, has little damage to the ecological environment and has higher feasibility. Reasonable inducer is selected, the microorganism with antagonistic action on pathogenic fungi in the soil is enriched through the induction action, and simultaneously the activity of hydrolase system capable of degrading the cell wall of the pathogenic fungi is induced to be increased, so that the method has very important significance for reducing soil-borne diseases.
The actinomycetes are the microorganisms which are discovered to have biological control effect at the earliest time, the actinomycetes are various in species and have different metabolic functions, and Streptomyces (Streptomyces) has obvious effect on controlling plant diseases. For example, agricultural streptomycin can be used for preventing and controlling various plant pathogenic bacteria such as crucifer soft rot, cucumber angular leaf spot, phytophthora phaseoli, tomato canker and the like. The actinomycete living preparation Mycostop is mainly used for preventing and treating common soil-borne pathogens such as pythium, fusarium, phytophthora and rhizoctonia. However, the above preparations are produced on a large scale and then added to soil, and have limited adaptability to the soil environment.
Beta-glucans are a class of naturally synthesized non-starch polysaccharides which are primarily characterized by a polysaccharide backbone containing a relatively high proportion of beta-1, 3-glucosides. Beta-glucan is an important component of cell walls, widely exists in the kingdoms of microorganisms, plants and animals, and part of microorganisms also have the capacity of secreting extracellular beta-glucan. Succinoglycan is an extracellular polysaccharide secreted by nitrogen-fixing microorganisms such as rhizobium and is mainly characterized in that each polysaccharide repeating unit contains 1 or 2 succinyl modification groups. The succinoglycan plays an important role in the symbiotic process of rhizobia and plant roots. Researches show that the beta-glucan and the succinyl glycan both have higher physiological activity and are environment-friendly. At present, certain reports are made on the prevention and treatment of soil-borne diseases by using beta-glucan and succinoglycan, and researches show that the plant growth is promoted by using the beta-glucan or the succinoglycan alone. But due to the difference in the mechanism of action between the two.
Disclosure of Invention
The invention aims to provide application of beta-glucan and succinoglycan in preventing and controlling soil-borne fungal diseases. The inventor firstly finds that the combination of beta-glucan and succinyl glycan with different action mechanisms can further enrich streptomycete in soil, improve the activity of soil hydrolase and prevent and treat soil-borne pathogenic fungi by controlling the proportional relation of the beta-glucan and the succinyl glycan.
The technical scheme for realizing the purpose of the invention is as follows:
the application of the beta-glucan and the succinyl glycan in preventing and controlling soil-borne fungal diseases comprises the following specific steps:
adding a mixture of beta-glucan and succinyl glycan into soil according to the mass ratio of the beta-glucan to the succinyl glycan of 1: 0.1-1: 2.0.
In the present invention, the beta-glucan is a beta-glucan conventionally used in the art, and may be yeast glucan, laminarin, lentinan or schizomycete polysaccharide, and is preferably yeast glucan or laminarin.
In the invention, the structural general formula of the succinoglycan is as follows:
in the present invention, the mass ratio of the beta-glucan to the succinoglycan is preferably 1:0.5 to 1: 2.0.
In the invention, the addition amount of the mixture of the beta-glucan and the succinyl glycan is 0.01-0.5% of the mass of the soil, and preferably 0.05-0.25% of the mass of the soil.
In the invention, the addition mode of the mixture of the beta-glucan and the succinyl glycan can be directly adding solid powder or preparing mixture solution for spraying.
Compared with the prior biological control technology, the invention has the following advantages:
(1) the mixture of the beta-glucan and the succinyl glycan has better selective enrichment effect on streptomyces, can induce the hydrolase activities such as beta-glucanase, chitinase, cellulase and the like in soil to be obviously increased, and the beta-glucan, the chitinase, the cellulase and the like are synergistically enhanced, so that the effect of preventing and controlling soil-borne fungal diseases of the mixture is obviously better than that of singly adding the beta-glucan or the succinyl glycan.
(2) The effect of the mixture for preventing and controlling soil-borne fungal diseases can be further promoted by regulating the proportional relation of the beta-glucan and the succinyl glycan.
(3) The method has the advantages of low economic cost, high feasibility and environmental friendliness.
Drawings
FIG. 1 is a graph showing the trend of the relative abundance of streptomyces in soil in original forest land of Enshi over time, in soil in alternate tillage of Nanjing city over time, in soil in potato continuous cropping land of Tengzhou city over time, and in soil in tobacco continuous cropping land of Kunming city over time, after different concentrations of mixed polysaccharides were added in example 1.
FIG. 2 is a graph of the change in soil beta-glucanase activity over time with the addition of different concentrations of mixed polysaccharides as in example 2.
FIG. 3 is a graph of the change in soil cellulase activity over time after the addition of different concentrations of mixed polysaccharides in example 2.
FIG. 4 is a graph of the time course of chitinase activity in soil after the addition of various concentrations of mixed polysaccharides as in example 2.
FIG. 5 is a statistical graph of the number of culturable fungi in soil after the addition of different concentrations of mixed polysaccharides as described in example 3.
FIG. 6 is a graph showing the relative content of Phytophthora infestans in soil after different concentrations of mixed polysaccharides were added in example 3.
FIG. 7 is a graph showing the inhibitory effect of Streptomyces melaleukii on Fusarium oxysporum in example 4.
Detailed Description
The present invention will be described in more detail with reference to the following examples and the accompanying drawings.
The succinoglycan employed in the examples below has the general structural formula:
Example 1
Determination result of relative abundance of streptomyces in typical soil
The original forest land of Enshi province of Hubei province, the rotary tillage land of Nanjing city of Jiangsu province, the potato continuous cropping land of Tengzhou city of Shandong province and the tobacco continuous cropping land of Kunming city of Yunnan province are respectively selected as test areas. Powder of a mixture of β -glucan and succinoglycan (laminarin: succinoglycan ═ 1:1w/w) was added to the selected soil surface layer in amounts of 0%, 0.05%, and 0.25%, respectively. Then, the sampling is performed every 15d, and the process is ended at 60 d. The total DNA of the soil is extracted from the sampled soil by adopting an Ezup column type soil DNA extraction kit (living creatures (Shanghai)) and the soil metagenome sequencing is carried out. And analyzing the relative abundance of the streptomyces sequences in the soil microbial region system according to the sequencing result. FIG. 1 shows the variation trend of the relative abundance of Streptomyces in the above 4 soils with time. As can be seen from the figure, the relative abundance of Streptomyces increases significantly with time in 4 soils after addition of the mixture of β -glucan and succinoglycan. The content of the streptomyces in the soil can be further improved by increasing the addition amount of the mixture. The results show that the mixture of beta-glucan and succinyl glycan has the effect of enriching streptomyces in soil.
Example 2
Activity changes of beta-glucanase, cellulase and chitinase in soil
Selecting alternate cultivated land of Nanjing city of Jiangsu province as a test soil sample, and adding mixture powder of beta-glucan and succinoglycan (yeast glucan: succinoglycan ═ 1:10w/w) to the surface layer of the selected soil, wherein the usage amounts are 0%, 0.05% and 0.25% respectively. Then, the sampling is performed every 15d, and the process is ended at 60 d. Taking laminarin, sodium carboxymethylcellulose and chitin colloid as substrates respectively, and determining the activity of the hydrolase of the soil leachate at pH5.5, wherein the reaction temperature is 35 ℃ and the reaction time is 24 h. FIGS. 2 to 4 show the time-dependent changes of the activities of 3 hydrolases in soil. As can be seen from the figure, after the mixture of beta-glucan and succinoglycan is added, the activity of 3 enzymes is obviously improved along with the time and the adding amount of the mixture. The results show that the addition of the mixture of beta-glucan and succinyl glycan contributes to improving the activity of the hydrolase in the soil.
Example 3
Soil fungus content determination results
A potato continuous cropping land in Tengzhou, Shandong province is selected as a soil sample, a mixture of beta-glucan and succinoglycan (lentinan: succinoglycan ═ 1:0.2w/w) is prepared into a solution with the concentration of 25g/L, and then the solution is sprayed on the surface layer of the selected soil, and the using amounts are 0%, 0.05% and 0.25% respectively.Then, the sampling is performed every 15d, and the process is ended at 60 d. Then, measuring the abundance change process of the culturable fungi in the soil by adopting a dilution plating counting method: diluting 1g of soil sample into 10mL of sterile phosphate buffer, violently shaking for 30min to uniformly disperse the soil in the buffer, then carrying out gradient dilution, finally selecting a proper concentration to coat the soil sample on a PDA (personal digital assistant) plate, culturing at 30 ℃ for 48h, and counting. FIG. 5 shows that the fungal content in the soil gradually decreased with increasing time of addition and polysaccharide concentration. The number of culturable fungi in the soil with the content of the mixed polysaccharide of 0.25 percent is reduced to 1.1 to 1.2 multiplied by 10 in 60 days5CFU/g。
In addition, the analysis of the relative content of the phytophthora infestans in the soil shows that the relative content of the phytophthora infestans is obviously reduced after the mixed polysaccharide is added (fig. 6), and the relative amount of the phytophthora infestans in the soil with the addition amount of the mixed polysaccharide of 0.25% is reduced from 0.97 to 0.63 compared with the control group at 60 days. The results show that the addition of the mixture of beta-glucan and succinoglycan can effectively reduce the relative abundance of the soil-borne fungi.
Example 4
Inhibition of streptomycete on fusarium oxysporum
Selecting a tobacco continuous cropping land of Kunming city in Yunnan province as a test soil sample, preparing a solution with the concentration of 25g/L by using a mixture of beta-glucan and succinoglycan (schizomycete polysaccharide: succinoglycan: 1:10w/w), spraying the solution on the surface layer of the selected soil, and sampling when the usage amount is 0.5% to 60 days. Streptomyces is separated from a soil sample by adopting a Chao' S medium to obtain a purified strain, and the strain is Streptomyces pactum (Streptomyces tenebrio) after 16S rDNA sequence alignment. Then the bacterial strain is used for carrying out bacteriostasis test on fusarium oxysporum. As shown in FIG. 7, the zones of inhibition of Fusarium oxysporum were consistently present in the petri dishes with Streptomyces pictum patches after the co-cultivation for 4 days. The result shows that the streptomycete enriched by the mixture of the beta-glucan and the succinyl glycan has the effect of inhibiting the growth of fusarium oxysporum.
Example 5
The effect of the addition of the mixture of beta-glucan (laminarin) and succinoglycan was compared to the effect of the addition of either beta-glucan (laminarin) or succinoglycan alone
Soil samples of the alternative cultivated land of Nanjing city, Jiangsu province, described in example 1 were selected and processed according to the method described in example 1. The relative abundance of streptomyces in the soil, beta-glucanase activity and fusarium oxysporum were measured at 30d and 60d, respectively, and the results are shown in tables 1 and 2.
TABLE 1 comparison of measurement results when the mixed polysaccharide is added at a concentration of 0.05%
TABLE 2 comparison of measurement results when the mixed polysaccharide is added at a concentration of 0.25%
The comparison shows that when the beta-glucan and the succinoglycan are used together, the key data index is better than the effect of the beta-glucan or the succinoglycan when used alone. Meanwhile, when the ratio of the beta-glucan to the succinyl glycan is in the range of 1: 0.5-1: 2, the effect is optimal.
Claims (7)
1. The application of the mixture of beta-glucan and succinyl glycan in preventing and controlling soil-borne fungal diseases is characterized in that in the mixture of beta-glucan and succinyl glycan, the mass ratio of beta-glucan to succinyl glycan is 1: 0.1-1: 2.0.
2. The application of claim 1, comprising the following steps:
adding a mixture of beta-glucan and succinyl glycan into soil according to the mass ratio of the beta-glucan to the succinyl glycan of 1: 0.1-1: 2.0.
3. Use according to claim 1 or 2, wherein the β -glucan is selected from yeast glucan, laminarin, lentinan or schizophyllan.
4. The use according to claim 1 or 2, wherein the succinoglycan has the general structural formula:
5. the use according to claim 1 or 2, wherein the mass ratio of β -glucan to succinoglycan is 1:0.5 to 1: 2.0.
6. The use according to claim 1 or 2, wherein the amount of the mixture of β -glucan and succinoglycan added is 0.05% to 0.25% of the mass of the soil.
7. The use according to claim 1 or 2, wherein the mixture of β -glucan and succinoglycan is added directly to the solid powder or is prepared as a solution of the mixture and sprayed.
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| WO2011065831A2 (en) * | 2009-11-27 | 2011-06-03 | Ceradis B.V. | Improved formulations comprising heteropolysaccharides |
| CN104012535A (en) * | 2014-06-11 | 2014-09-03 | 山东棉花研究中心 | Bactericide composition containing fluoride ether bacteria amide and laminarin |
| CN106135230A (en) * | 2015-04-13 | 2016-11-23 | 深圳诺普信农化股份有限公司 | A kind of nematicide disease-resistant composition and application thereof |
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| WO2011065831A2 (en) * | 2009-11-27 | 2011-06-03 | Ceradis B.V. | Improved formulations comprising heteropolysaccharides |
| CN104012535A (en) * | 2014-06-11 | 2014-09-03 | 山东棉花研究中心 | Bactericide composition containing fluoride ether bacteria amide and laminarin |
| CN106135230A (en) * | 2015-04-13 | 2016-11-23 | 深圳诺普信农化股份有限公司 | A kind of nematicide disease-resistant composition and application thereof |
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| 防治小麦纹枯病和根腐病高效药剂的筛选及其与香菇多糖联用的防效评价;姜莉莉 等;《麦类作物学报》;20180131;第38卷(第1期);第113-118页 * |
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