CN113773848A - Improvement method of saline soil in yellow river delta region - Google Patents
Improvement method of saline soil in yellow river delta region Download PDFInfo
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Abstract
The invention relates to the technical field of improvement of saline soil, in particular to a method for improving the saline soil in the yellow river delta area. The salt-tolerant trichoderma can further improve the salt tolerance of the peppermint, and increase the yield of the peppermint in the saline soil of the yellow river delta. And the salt-tolerant trichoderma and the salt-tolerant peppermint have synergistic effect to ensure that the soil desalination rate reaches 26.1-33.19%, and meanwhile, the content of soil organic matters, effective nitrogen, quick-acting phosphorus and quick-acting potassium can be increased, the soil fertility is enhanced, the soil enzyme activity and microbial community diversity are increased, the soil micro-ecological environment is improved, and a good effect is achieved on the restoration of the saline soil.
Description
Technical Field
The invention relates to the technical field of improvement of saline soil, in particular to a method for improving saline soil in yellow river delta areas.
Background
Soil salinization seriously restricts the sustainable development of agriculture and becomes a worldwide resource and ecological problem. The underground water level in the yellow river delta area of China is shallow in buried depth, the mineralization degree is high, natural vegetation is simple, the salinization area of soil reaches 44.29 ten thousand hectares, and the salinization area occupies more than 50% of the total area. Comprehensive treatment and utilization of the regional salinized land are related to a series of important practical problems of national farmland reserve resources, regional grain safety, economic development and the like. At present, the saline soil improvement technology mainly comprises physical improvement measures for adjusting soil structures such as field lifting, deep ploughing, straw mulching and soil dressing improvement, water conservancy improvement projects for irrigating salt, draining salt from underground concealed pipes and draining water from deep wells of ditches, chemical improvement methods for adjusting pH and salinity, and biological improvement methods mainly for planting saline-alkali resistant crops. Physical improvement and hydraulic improvement engineering costs are high, and the excessive use of chemical improvement mode can cause secondary pollution of soil. The biological improvement measures are considered to be the most effective improvement technology due to the advantages of low cost, environmental protection and the like, and the vegetation planting can promote the virtuous circle of the plant-soil and plays a vital role in the sustainable development of the ecological environment. Therefore, in recent years, the saline soil improvement technology taking salt-tolerant plant planting as a core is widely applied, but the special soil property of the saline soil causes the survival rate and the preservation rate of plants to be lower, so that the technology becomes a bottleneck for limiting the development of the technology, the salt tolerance and the survival rate of the plants under the salt stress condition are promoted, the comprehensive utilization of the saline soil in the yellow river delta area is enhanced, and the technology is an important embodiment for meeting the national strategic demands.
Peppermint (Mentha piperita L.) is an economic crop using peppermint essential oil as a main product, has cooling property and analgesic effect, and is widely applied to the industries of food, medicine and daily cosmetics. The 'Keyao No. 1' is a salt-tolerant peppermint, can be widely planted in typical saline soil of yellow river delta, and has a good desalting effect. But earlier researches find that the peppermint has good growth in saline soil of less than 0.4 percent of yellow river delta and has limited growth in saline soil of 0.4 percent of yellow river delta. Aiming at the problems that the survival rate and the preservation rate of plants in the yellow river delta area are low and the growth of the salt-tolerant peppermint in the high-salinity soil is limited, a salt-tolerant trichoderma strain ST02 is obtained by early screening, and the spore liquid of the strain is found to be capable of promoting the germination of tomato seeds, the growth of seedlings and the physiological and biochemical indexes of salt tolerance of the seedlings under the condition of salt stress and improving the saline soil of tomato roots. On the basis, the invention combines the research and development and the use method of the salt-tolerant trichoderma agent and the planting of the salt-tolerant peppermint to research and develop an improvement method of the saline soil in the yellow river delta area, and can solve the technical bottleneck in the plant repair of the saline soil.
Disclosure of Invention
Aiming at the problems of serious soil salinization and low plant survival rate and preservation rate in the yellow river delta area, the invention provides the method for improving the salinized soil in the yellow river delta area, which promotes the yield of the peppermint in the east-Ying yellow river delta area, lightens the salinization degree of the soil, improves the physicochemical property of the soil, enhances the fertility and the activity of the soil enzyme, improves the diversity of microorganisms in the soil and improves the salinized soil through the synergistic action of a microbial inoculum of salt-tolerant trichoderma harzianum and salt-tolerant peppermint 'Coacademy 1'.
The technical scheme of the invention is as follows:
a method for improving saline soil in yellow river delta areas adopts a salt-tolerant trichoderma agent and salt-tolerant peppermint to jointly restore the saline soil.
Furthermore, the salt-tolerant trichoderma is trichoderma harzianum ST02 (patent application: 201811629417.9, preservation number: CGMCC No.16964), and the salt-tolerant peppermint is 'Coacademy 1' (variety registration number: 038).
Further, the method comprises: the prepared salt-tolerant trichoderma viride agent and organic fertilizer are mixed and spread in a field, the land is leveled after the mixture is uniform, the salt-tolerant peppermint is planted in the field according to the plant-row spacing of 20cm multiplied by 30cm, and the subsequent field management is carried out.
Furthermore, the dosage of the salt-tolerant trichoderma agent is 5-10kg per mu, and the dosage of the organic fertilizer is 50-150kg per mu.
Further, the salt-tolerant trichoderma agent is wettable powder, and a liquid-solid two-phase fermentation method is adopted, wherein the fermentation process comprises the following steps: the method comprises the steps of seed preparation, liquid fermentation, solid fermentation, spore suspension preparation and carrier adsorption.
Further, the fermentation process of the liquid-solid two-phase fermentation method comprises the following steps: the seed preparation comprises culturing ST02 seed preserved on slant with PDA culture medium at 28 deg.C for 3-4 days, wherein hypha and conidium are abundant and free of contamination of bacteria; the liquid fermentation is to inoculate 1-2 fungus cakes with diameter of 0.5cm into 100mL PDW culture medium, to shake culture at 30 deg.C and 200rpm for 36h, and the mycelia should be branched more during microscopic examination, without pollution and mixed fungus; the solid fermentation is to mix liquid seeds of the liquid fermentation with a solid fermentation culture medium according to the proportion of 1:50(W: W), subpackage the mixture into culture bags and culture the mixture in a solid culture chamber; the spore suspension is prepared by soaking fermented solid fermentation culture medium into sterile water at a ratio of 1:1 (W: W), stirring to disperse conidia in water, filtering to remove solid fermentation culture medium and large mycelia to obtain spore suspension; the carrier adsorption is to centrifuge the prepared spore suspension, precipitate the spores, remove the supernatant, and utilize the carrier for adsorption to form the solid microbial inoculum.
Further, the solid fermentation medium is a wheat bran solid medium, and the fermentation conditions are that the temperature of a culture room is 25-30 ℃ and the air humidity is more than 90%.
Further, the wheat bran solid fermentation medium comprises: boiling semen Tritici Aestivi in water for about 1 hr, soaking overnight to make semen Tritici Aestivi water-swelling and water content about 70%, sterilizing at 121 deg.C under high temperature and high pressure for 30min, and cooling with circulating water.
Further, the carrier is diatomite or medical stone.
Furthermore, the final viable count of the microbial inoculum is 2.0 multiplied by 109cfu/g。
Further, the PDA medium (potato dextrose agar medium) formula (1L): 200g of potato, 20g of glucose, 12g of agar powder and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min.
Further, the PDW medium (potato dextrose water medium) formulation (1L): 200g of potato, 20g of glucose and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min.
The invention has the following effects:
the microbial inoculum of the salt-tolerant trichoderma ST02 can relieve the stress of NaCl on the growth and photosynthesis of the peppermint, relieve the oxidative damage and osmotic imbalance caused by the stress of NaCl, and promote the salt tolerance of the peppermint by improving the antioxidant defense reaction of the microbial inoculum.
In typical saline soil of Dongying yellow river delta, by utilizing the preferred embodiment of the invention, the saline soil is repaired by utilizing the combination of the salt-tolerant trichoderma agent and the salt-tolerant peppermint, so that the yield of the peppermint can be improved, and the yield of the saline soil of the yellow river delta is increased; the salinization degree of the soil is reduced, and the desalinization rates of a soil layer of 0-20cm and a soil layer of 20-40cm reach 26.1% -27.68% and 31.1% -33.19%; the physical and chemical properties of the soil are improved, the contents of organic matters, available nitrogen, available phosphorus and available potassium in the soil are increased, and the activities of urease, sucrase, phosphatase and catalase in the soil are improved; the method has the advantages of increasing the content of bacteria and fungi in the soil, improving the richness and diversity of microbial communities, improving the micro-ecological environment of the soil and having good improvement effect on the saline soil.
Drawings
FIG. 1 shows a potting experiment.
Figure 2 is a graph of the effect of different treatments on peppermint growth and photosynthesis.
A2: the plant height of peppermint; b2: fresh weight of aerial parts of peppermint; c2: neat photosynthetic rate of peppermint (Pn); d2: the chlorophyll content in peppermint leaves.
CK: control, + NaCl: NaCl treatment experimental group, + ST 02: trichoderma harzianum ST02 treatment experimental group, + NaCl + ST 02: the experimental groups were treated simultaneously with NaCl and ST02 and the values were mean ± standard deviation (n ═ 5), different lower case letters indicate ANOVA analysis of variance between groups, and P < 0.05 indicates significant differences.
Figure 3 is a graph of the effect of different treatments on oxidative damage to peppermint and the content of osmoregulation related substances.
A3: malondialdehyde (MDA) content of Mentha piperita leaf; b3: peppermint leaf Soluble Protein (SP) content; c3: proline (Pro) content of Mentha piperita leaf.
CK: control, + NaCl: NaCl treatment experimental group, + ST 02: trichoderma harzianum ST02 treatment experimental group, + NaCl + ST 02: the experimental groups were treated simultaneously with NaCl and ST02 and the values were mean ± standard deviation (n ═ 5), different lower case letters indicate ANOVA analysis of variance between groups, and P < 0.05 indicates significant differences.
FIG. 4 is a graph of the effect of different treatments on peppermint antioxidant enzyme activity.
A4: peppermint leaf superoxide dismutase (SOD) activity; b4: peppermint leaf Peroxidase (POD) activity; c4: catalase (CAT) activity of peppermint leaves.
FIG. 5 shows the effect of different treatments of halotolerant Trichoderma in field on soil enzyme activity.
A5: soil urease (S-UE) activity; b5: soil sucrase (S-SC) activity; c5: soil phosphatase (S-ACP) activity; d5: soil catalase (S-CAT) activity.
FIG. 6 shows the number of bacteria and fungi in soil of different experimental groups of field experiments.
FIG. 7 is a graph showing the relative abundance of bacterial populations at the phylum level in soil from different test groups of a field trial.
FIG. 8 is a graph of the relative abundance of fungal populations at the phylum level in soil from different test groups of a field trial.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 preparation of halotolerant Trichoderma ST02 inoculum
The preparation method of the microbial inoculum comprises the following steps: the halotolerant trichoderma ST02 microbial inoculum is prepared according to a liquid-solid two-phase fermentation method. The fermentation process comprises the following steps: the method comprises the steps of seed preparation, liquid fermentation, solid fermentation, spore suspension preparation and carrier adsorption.
(1) Seed preparation: culturing ST02 seeds preserved on the inclined plane on a PDA culture medium at 28 ℃ for 3-4 days, wherein hypha and conidium are required to be plump without infectious microbe pollution;
PDA culture medium (potato dextrose agar culture medium) formula (1L):
200g of potato, 20g of glucose, 12g of agar powder and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min.
(2) Liquid fermentation: inoculating 1-2 fungus cakes with the diameter of 0.5cm into 100mL of PDW culture medium, performing shake culture at 30 ℃ and 200rpm for 36h, wherein the hyphae are more branched during microscopic examination and have no pollution and mixed fungi;
formula (1L) of PDW culture medium (potato dextrose water culture medium):
200g of potato, 20g of glucose and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min.
(3) Solid fermentation: fully mixing liquid seeds subjected to liquid fermentation with a solid culture medium according to the proportion of 1:50(W: W), subpackaging the mixture into culture bags, and culturing in a solid culture chamber; the temperature of the culture room is 25-30 ℃, and the air humidity is more than 90%;
solid medium: a wheat bran culture medium, wherein wheat is boiled by adding water for about 1h, soaked overnight to ensure that the water absorption expansion water content of the wheat is about 70 percent, and is cooled by circulating water after being sterilized at the high temperature of 121 ℃ for 30min under high pressure;
(4) preparing spore suspension: soaking the fermented solid culture medium into sterile water according to a ratio of 1:1 (W: W), stirring to disperse conidia into water, and filtering to remove the solid culture medium and large mycelia to obtain spore suspension;
(5) carrier adsorption: centrifuging the prepared spore suspension, precipitating spores, removing supernatant, obtaining the number of spores per gram of spore mud by platelet counting method, adsorbing with diatomaceous earth or Maifanitum to form solid microbial inoculum, adjusting the number of bacteria to 2.0 × 109cfu/g。
Example 2 salt tolerant Trichoderma preparation to promote salt tolerance of Mentha piperita
(1) Experiment design: soil was taken from the experimental site of the institute of ecology of academy of sciences of Shandong province, and potting experiments were carried out using a pot 40cm in height and 9cm in diameter (FIG. 1). The pot experiment was performed with 5 replicates of NaCl treatment, ST02 treatment, combined NaCl and ST02 treatment, and control. Adding soil 2.5kg into each culture container, adding 200mL water, cutting herba Menthae Rotundifoliae with uniform growth for 7 days, mixing ST02 microbial inoculum with water at ratio of 1:100(V: V) (viable count 2 × 10)7cfu/g) was prepared as a bacterial solution, and the solution was irrigated with 200mL of bacterial solution, as compared to direct irrigation with water. After ST02 treatment for 7d, 200mL of 600mM NaCl solution was used for irrigationAnd carrying out NaCl treatment 1 time every 3 days, continuously irrigating for 3 times, then irrigating with normal water, and after NaCl treatment for 21 days, detecting the growth and salt tolerance physiological and biochemical indexes of the peppermint.
(2) As a result: NaCl stress influences the growth and photosynthesis intensity of the peppermint, and the plant height (SL), the overground Fresh Weight (FW), the photosynthesis intensity (Pn) and the chlorophyll content (SPAD) of leaves of the peppermint are obviously smaller than those of a control under the NaCl stress condition; trichoderma harzianum ST02 promoted the growth of peppermint, ST02 treated peppermint SL was significantly higher than control, FW, Pn and SPAD were not significantly changed from control; under the conditions of simultaneous NaCl and ST02 treatment, SL, FW, Pn and SPAD of the peppermint have no obvious difference compared with the control, but are all obviously higher than the SL, FW, Pn and SPAD values of the peppermint under the NaCl treatment, which shows that Trichoderma harzianum ST02 can relieve the stress of NaCl on the growth and photosynthesis of the peppermint and promote the growth and photosynthesis intensity of the peppermint under the salt stress condition, as shown in the attached figure 2 and figure 2, CK: control, + NaCl: NaCl treatment experimental group, + ST 02: trichoderma harzianum ST02 treatment experimental group, + NaCl + ST 02: the experimental groups were treated simultaneously with NaCl and ST02 and the values were mean ± standard deviation (n ═ 5), different lower case letters indicate ANOVA analysis of variance between groups, and P < 0.05 indicates significant differences.
NaCl stress caused the generation of Reactive Oxygen Species (ROS) in the cells, causing oxidative damage to the cells, and lipid peroxidation in the cell membranes to produce MDA, so that the amount of MDA in NaCl-treated peppermint leaves was significantly increased (A3 in FIG. 3). The MDA content in the peppermint leaves treated with trichoderma harzianum ST02 did not change significantly, but under NaCl stress conditions, the MDA content in the ST02 treated peppermint leaves was significantly increased compared to the control and significantly decreased compared to NaCl treatment, indicating that ST02 can alleviate oxidative damage caused by NaCl stress (A3 in fig. 3). Soluble Protein (SP) and proline (Pro) are important osmoregulation substances in cells, the SP and the Pro are accumulated in the cells in a large amount under NaCl stress to maintain the osmotic balance inside and outside the cells, the SP and the Pro content in NaCl-treated peppermint leaves is obviously increased, and Trichoderma harzianum ST02 can reduce the increase of the SP and the Pro content caused by NaCl stress (B3 and C3 in figure 3).
The antioxidant enzyme can eliminate ROS generated by NaCl stress in cells, maintain ROS balance and promote the tolerance of the cells to the NaCl stress. The results showed that the antioxidant enzyme (SOD, POD and CAT) activity in NaCl-treated peppermint leaves was significantly lower than that of the control, and that the SOD, POD and CAT activity in Trichoderma harzianum ST 02-treated peppermint leaves was higher than that of the control (FIG. 4). In leaf discs of peppermint treated with both NaCl and ST02, SOD activity was lower than control and higher than NaCl-stressed peppermint, POD activity was higher than control and lower than ST 02-treated peppermint, and CAT activity was similar to control (FIG. 4). The trichoderma harzianum ST02 can improve the antioxidant enzyme activity of the peppermint under NaCl stress and promote the salt tolerance of the peppermint by improving the antioxidant defense reaction.
Example 3 field test
(1) Test site: a test base of eastern academy of forestry science research, Shandong province, is coastal saline soil with salt content of about 0.5, pH value of the soil of about 7.6, organic matter content of the soil of 5.23g/kg, alkaline hydrolysis nitrogen content of 55.65mg/kg, quick-acting phosphorus of 7.88mg/kg and quick-acting potassium of 62.33 mg/kg. According to different application methods of the salt-tolerant trichoderma viride agent and the salt-tolerant peppermint planting method, different test groups and control groups are arranged, subsequent field management is carried out, and the peppermint yield, the salt content in soil, the soil physicochemical property, the enzyme activity and the microbial diversity are detected after the peppermint is harvested.
Test group 1 (T1): before planting, uniformly mixing an organic fertilizer (5-150 kg/mu) and a salt-tolerant trichoderma viride agent (2 kg/mu), spreading the mixture into a field, leveling the field after uniformly mixing, planting salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Test group 2 (T2): before planting, uniformly mixing an organic fertilizer (5-150 kg/mu) and a salt-tolerant trichoderma viride agent (5 kg/mu), spreading the mixture into a field, leveling the field after uniformly mixing, planting salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Test group 3 (T3): before planting, uniformly mixing an organic fertilizer (5-150 kg/mu) and a salt-tolerant trichoderma viride agent (10 kg/mu), spreading the mixture into a field, leveling the field after uniformly mixing, planting salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Test group 4 (T4): before planting, uniformly mixing an organic fertilizer (5-150 kg/mu) and a salt-tolerant trichoderma viride agent (15 kg/mu), spreading the mixture into a field, leveling the field after uniformly mixing, planting salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Test group 5 (T5): before planting, spreading an organic fertilizer (5-150 kg/mu) in a field, uniformly mixing, leveling the field, mixing trichoderma agent and water into agent suspension according to a ratio of 1:100, soaking root systems of peppermint for 20-30min, planting the peppermint in the field according to a plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Test group 6 (T6): before planting, spreading an organic fertilizer (5-150 kg/mu) in a field, uniformly mixing, leveling the land, planting the salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, irrigating the ST02 microbial inoculum to the field according to the dosage of 2 kg/mu, and performing subsequent field management.
Test group 7 (T7): before planting, spreading an organic fertilizer (5-150 kg/mu) in a field, uniformly mixing, leveling the land, planting the salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, irrigating the ST02 microbial inoculum to the field according to the dosage of 5 kg/mu, and performing subsequent field management.
Test group 8 (T8): before planting, spreading an organic fertilizer (5-150 kg/mu) in a field, uniformly mixing, leveling the land, planting the salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, irrigating the ST02 microbial inoculum to the field according to the dosage of 10 kg/mu, and performing subsequent field management.
Test group 9 (T9): before planting, spreading an organic fertilizer (5-150 kg/mu) in a field, uniformly mixing, leveling the land, planting the salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, irrigating the ST02 microbial inoculum to the field according to the use amount of 15 kg/mu, and performing subsequent field management.
Control 1(CK 1): control group of peppermint was planted without trichoderma preparation.
Control 2(CK 2): uniformly mixing the organic fertilizer (5-150 kg/mu) and the salt-tolerant trichoderma agent (10 kg/mu), spreading the mixture to a field, and leveling the field after uniformly mixing without planting peppermint.
Control group 3(CK 3): blank control group without planting peppermint and applying trichoderma agent
TABLE 1 field test peppermint yield and soil salt content for different test groups and control group
(2) As a result: the method comprises the steps of detecting the yields of peppermint from different test groups and control groups in typical saline soil of yellow river delta, applying CK1 without applying trichoderma agents to the control group to increase the yield of peppermint by 932 kg/mu, and applying salt-tolerant trichoderma agents to increase the yield of peppermint, wherein the test groups with the highest yield are T3, T2 and T5 test groups, and increase by 26.93%, 24.14% and 23.82% respectively (Table 1); the salt content of a typical saline soil used in the test is 0.52-0.58% in a soil layer with the salt content of 0-20cm and 0.41-0.47% in a soil layer with the salt content of 20-40cm, the salt content of a blank control CK3 soil layer is increased, and the salt content of the soil layers with the salt content of 0-20cm and 20-40cm is respectively increased by 8.11% and 7.36%; the salt rates of soil layers of CK1 and CK2 control groups are reduced to a certain degree, and the salt rejection rates of 0-20cm soil layers and 20-40cm soil layers of different test groups are obviously higher than that of CK1 and CK2 control groups and also higher than the sum of the salt rejection rates of CK1 and CK2 control groups, so that the desalination effect on the game soil can be achieved by planting peppermint and applying a salt-tolerant trichoderma agent, and the desalination effect on the salt soil is obviously enhanced through the synergistic effect between the peppermint and the salt-tolerant trichoderma (Table 1); the test groups with the best desalting effect on 0-20cm soil layers are T2, T3 and T4 which are respectively 26.1%, 28.68% and 26.62%, the test groups with the best desalting effect on 0-20cm soil layers are T2 and T3 which are respectively 32.94% and 33.19% (Table 1), and the test groups with the best desalting effect on the soil are T2 and T3 due to the conditions of yield increase of the peppermint and the desalting rate of the soil.
TABLE 2 field test soil physicochemical Properties for different test groups
The physicochemical properties of the peppermint root soil of each test group are detected, and the results show that: the peppermint is planted and the salt-tolerant trichoderma agent is applied, so that the pH value of the soil is not obviously influenced; because the planting of the peppermint and the application of the halotolerant trichoderma have desalting effects, the conductivity of the soil of CK1 and CK2 control groups is obviously reduced compared with that of CK3, and the peppermint and halotolerant trichoderma inoculants in each test group have synergistic effect, so that the reduction amplitude of the conductivity in the soil is obviously increased; the contents of organic matters, alkaline hydrolysis nitrogen, quick-acting phosphorus and quick-acting potassium in soil are increased by 53.32%, 55.68%, 37.13% and 21.82% respectively by planting peppermint, and salt-tolerant trichoderma agents are applied, so that the contents of the organic matters, the effective nitrogen, the quick-acting phosphorus and the quick-acting potassium in the soil are increased by the synergistic effect of the salt-tolerant trichoderma agents and the peppermint, the soil is obviously higher than CK1 without the trichoderma agents and also higher than CK2 without the peppermint, the spreading effect is better than the irrigation and root dipping effects, and the comprehensive T2 and T3 effects are optimal (Table 2).
The activity of urease, sucrase, phosphatase and catalase in the soil of different test groups is detected, and the result shows that: the enzyme activity in the soil can be improved by planting peppermint and haloxyfop and applying trichoderma agent, and the peppermint and haloxyfop are synergistic, so that the enzyme activity of the soil of each test group is obviously higher than that of a control group, the application effect of broadcasting and irrigation is similar, the enzyme activity tends to rise first and then fall along with the rise of the usage amount, the optimal usage amount is 5-10 kg/mu, as shown in figure 5, wherein the numerical value is the average value plus or minus standard deviation (n is 3).
The number of bacteria and fungi in soil of each test group is detected by utilizing a qRT-PCR method, the content of the bacteria and fungi in the soil can be increased by planting peppermint (CK1) and applying trichoderma (CK2), and the content of the bacteria and fungi in the soil is greatly increased by the interaction of the peppermint and the trichoderma of each test group; the number of bacteria in the soil is not obviously affected by the use amount of different trichoderma agents, because the trichoderma agents survive and propagate in the soil, the number of fungi in the soil is obviously increased, and the increasing trend of the number is positively correlated with the use amount, as shown in figure 6, wherein the numerical value is the average value +/-standard deviation (n-3).
Analyzing the community diversity of bacteria and fungi in the soil of different test groups by a high-throughput sequencing technology and a bioinformatics analysis method, wherein the Shannon index and the Simpson index are used for evaluating the community diversity of microorganisms; the Chao index is an index that estimates the number of OTUs measured for a sample and the Coverage index is the Coverage measured for the sample. The results in Table 3 show that the bacterial and fungal diversity analysis Coverage indexes of each test group respectively reach more than 97% and 99%, which indicates that the sequencing depth of the sample is enough to carry out subsequent data analysis; the Simpson indexes of bacterial communities and fungal communities in soil of the control group CK1 only planted with the peppermint and the control group CK2 only applied with the trichoderma are smaller than those of the empty control group CK3, and the Shannon index and the Chao index are larger than those of the blank control group CK3, so that the richness and diversity of soil microbial communities can be increased by planting the peppermint and applying the trichoderma; and the Simpson index of each test group under the time combined action of the peppermint planting and the trichoderma agent is smaller than that of any control group, and the Shannon index and the Chao index are larger than those of the control group, so that the synergistic promotion between the salt-tolerant trichoderma agent and the peppermint is realized, and the richness and diversity of soil microbial communities are increased (table 3).
TABLE 3-field test soil bacteria and fungi community diversity for different test groups
The abundance of bacterial and fungal communities in soil of different test groups at the phylum level is shown in fig. 7 and fig. 8, and the bacterial and fungal communities of each test group are similar in composition and mainly comprise: proteobacteria (Proteobacteria), actinomycetemcomita (Actinobacteria), Bacteroidetes (Bacteroidetes), Acidobacteria (Acidobacteria), Cyanobacteria (Cyanobacteria _ chlorest), planktonic bacteria (Planctomycetes), Firmicutes (Firmicutes), Verrucomicrobia (Verrucomicrobia), clocurvatia (Chloroflexi), gemmatamustinates (gemmatiadetes), saccharobacter bacteria (saccharomyces), nitrospiramium (nitrospiraee), Proteobacteria and actinomyces are dominant phyla, and planting peppermint and applying trichoderma agent increase the abundance of Proteobacteria; the fungi mainly comprise: ascomycota (Ascomycota), Basidiomycota (Basidiomycota), Trichosporon (Mortierella) and Chytridiomycota, wherein the active state of Trichoderma is Ascomycota, and Trichoderma is applied to increase the abundance of Ascomycota.
By combining the analysis, in typical saline soil of east-Ying Huanghe Delta, the salt-tolerant peppermint and the salt-tolerant trichoderma fungicide synergistically promote the yield of the peppermint, reduce the salinization degree of the soil, improve the physical and chemical properties of the soil, enhance the soil fertility and the soil enzyme activity, improve the microbial diversity in the soil and improve the soil micro-ecological environment, and the most preferable planting mode and trichoderma application mode of the peppermint are as follows: before planting, uniformly mixing the organic fertilizer (5-150 kg/mu) and the salt-tolerant trichoderma viride agent (5-10 kg/mu), spreading the mixture into a field, leveling the field after uniformly mixing, planting the salt-tolerant peppermint in the field according to the plant-row spacing of 20cm multiplied by 30cm, and performing subsequent field management.
Although the present invention has been described in detail by referring to the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. A method for improving saline soil in yellow river delta areas is characterized in that salt-tolerant trichoderma agents and salt-tolerant peppermint are combined to restore the saline soil.
2. The method for improving saline soil in yellow river delta area according to claim 1, wherein trichoderma halodurans is trichoderma harzianum ST02, and mentha halodurans is "kechow 1".
3. The method for improving saline soil in yellow river delta area according to claim 2, wherein the method comprises the following steps: the prepared salt-tolerant trichoderma viride agent and organic fertilizer are mixed and spread in a field, the land is leveled after the mixture is uniform, the salt-tolerant peppermint is planted in the field according to the plant-row spacing of 20cm multiplied by 30cm, and the subsequent field management is carried out.
4. The method for improving the saline soil in the yellow river delta area as claimed in claim 3, wherein the dosage of the salt-tolerant trichoderma agent is 5-10kg per mu, and the dosage of the organic fertilizer is 50-150kg per mu.
5. The method for improving the saline soil in the yellow river delta area according to claim 3, wherein the halotolerant trichoderma agent is wettable powder, a liquid-solid two-phase fermentation method is adopted, and the fermentation process comprises the following steps: the method comprises the steps of seed preparation, liquid fermentation, solid fermentation, spore suspension preparation and carrier adsorption.
6. The method for improving the saline soil in the yellow river delta area according to claim 5, wherein the fermentation process of the liquid-solid two-phase fermentation method comprises the following steps: the seed preparation comprises culturing ST02 seed preserved on slant with PDA culture medium at 28 deg.C for 3-4 days, wherein hypha and conidium are abundant and free of contamination of bacteria; the liquid fermentation is to inoculate 1-2 fungus cakes with diameter of 0.5cm into 100mL PDW culture medium, to shake culture at 30 deg.C and 200rpm for 36h, and the mycelia should be branched more during microscopic examination, without pollution and mixed fungus; the solid fermentation is to mix liquid seeds of the liquid fermentation with a solid fermentation culture medium according to the proportion of 1:50, subpackage the mixture into culture bags and culture the mixture in a solid culture chamber; the preparation of spore suspension comprises soaking fermented solid fermentation culture medium in sterile water at a ratio of 1:1, stirring to disperse conidia in water, and filtering to remove solid fermentation culture medium and large mycelia to obtain spore suspension; the carrier adsorption is to centrifuge the prepared spore suspension, precipitate the spores, remove the supernatant, and utilize the carrier for adsorption to form the solid microbial inoculum.
7. The method for improving saline soil in yellow river delta area according to claim 6, wherein the solid fermentation medium is wheat bran solid medium, the fermentation condition is that the temperature of the culture room is 25-30 ℃, and the air humidity is more than 90%.
8. The method for improving saline soil in yellow river delta area according to claim 7, wherein the wheat bran solid fermentation culture medium comprises the following components: boiling semen Tritici Aestivi in water for 1h, soaking overnight to make semen Tritici Aestivi water swelling and water content 70%, sterilizing at 121 deg.C under high temperature and high pressure for 30min, and cooling with circulating water.
9. The method for improving the saline soil in the yellow river delta area as claimed in claim 6, wherein the carrier is diatomite or maifanite; the final viable count of the microbial inoculum is 2.0 multiplied by 109cfu/g。
10. The method for improving saline soil in yellow river delta area as claimed in claim 6, wherein the formula of 1L PDA culture medium is as follows: 200g of potatoes, 20g of glucose, 12g of agar powder and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min; 1L PDW culture medium formula: 200g of potato, 20g of glucose and distilled water with the constant volume of 1L, and sterilizing at the high temperature and the high pressure of 110 ℃ for 30 min.
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