Method for improving continuous cropping adaptability of panax notoginseng

Abstract

The invention relates to the technical field of traditional Chinese medicinal material planting, and particularly discloses a method for improving continuous cropping adaptability of panax notoginseng, which comprises the following steps: S1, collecting ageratina adenophora plants, adding arbuscular mycorrhizal fungi, and carrying out decomposition treatment to obtain a plant-derived fungus compound; s2, applying the composted compound to a pseudo-ginseng continuous cropping field, covering a film for several days, uncovering the film, and airing; according to the technical scheme, the composted eupatorium adenophorum and the arbuscular mycorrhizal fungi compound are introduced into the soil, energy is provided for microbial agents in biological prevention and control, the effects of green prevention and control and efficiency improvement are achieved in cooperation with the microbial agents, pollution of pesticides and chemical fertilizers to the environment is effectively reduced, the yield of the panax notoginseng seedlings is increased, and the yield of the panax notoginseng seedlings is increased. The microenvironment and rhizosphere ecology of the continuous cropping soil are optimized, the production site conditions of the panax notoginseng are restored, and the emergence rate of the panax notoginseng in the continuous cropping soil treated by the crofton weed and the arbuscular mycorrhiza fungi is increased by about 258.05%.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese herbal medicine planting, and particularly relates to a method for improving the continuous cropping adaptability of Panax notoginseng. Background Art

[0002] Panax notoginseng (Burk.) F.H.Chen is a traditional precious Chinese herbal medicine in China. The main medicinal part of Panax notoginseng is the root system. The root rot disease caused by the continuous cropping obstacle of Panax notoginseng can lead to the failure of the industry and the reduction of drug efficacy. At present, chemical agents are commonly used in production to prevent and control the occurrence of root rot of Panax notoginseng. Not only the cost is high, the control effect is low, and the pathogenic bacteria are easy to produce resistance, but also it will cause soil acidification and the reduction of organic matter content, and finally lead to soil non-point source pollution, heavy metal and pesticide residues, threatening the balance of the ecosystem. At the same time, these pesticides enter the human body, causing serious carcinogenic, teratogenic and mutagenic risks. Therefore, the green prevention and control technology of continuous cropping obstacles has always been a bottleneck problem for the quality of Chinese herbal medicines and promoting traditional Chinese medicine to the world, and it is also the key point of scientific and technological innovation for green and efficient production in Yunnan Province and the urgent need for ecological civilization construction. Summary of the Invention

[0003] The main object of the present invention is to provide a green prevention and control method for improving the continuous cropping obstacle of Panax notoginseng by using the symbiotic system of Ageratum conyzoides and arbuscular mycorrhizal fungus Rhizophagus irregularis. This technology solves the problems of soil pathogen accumulation, poor root development and yield decline caused by continuous cropping of Panax notoginseng through the synergistic regulation of plant-derived plants and microbial activities, and is applicable to large-scale ecological planting of Chinese herbal medicines.

[0004] The present invention provides a method for improving the continuous cropping adaptability of Panax notoginseng, comprising the following steps:

[0005] S1: Collect Ageratum conyzoides plants, add arbuscular mycorrhizal fungus Rhizophagus irregularis for composting treatment to obtain a plant-derived fungal complex;

[0006] S2: Apply the composted complex to the continuous cropping field of Panax notoginseng, cover the film for several days and then uncover the film for sun drying;

[0007] S3: Transplant Panax notoginseng seedlings to the sun-dried soil for planting.

[0008] In step S1, the composting treatment is to regularly turn the pile under ventilation conditions to ensure that the temperature in the pile is maintained at 40-60 °C for 30 days of composting.

[0009] Further, in step S2, the continuous cropping field of Panax notoginseng is deeply plowed by 20 cm and the plant-derived fungal complex is applied.

[0010] Further, in step S2, the complex is applied to the continuous cropping field of Panax notoginseng at a dosage of 1500 kg / ha. After 45 days of film mulching, the film is removed and the field is sun-dried for 10 days.

[0011] The present invention also provides an application of a plant-derived fungal complex in improving the soil with continuous cropping obstacles of Panax notoginseng. The plant-derived fungal complex is obtained by adding arbuscular mycorrhizal fungus Rhizophagus irregularis to Eupatorium adenophorum plants for composting treatment.

[0012] The present invention also provides an application of a plant-derived fungal complex in improving the soil with continuous cropping obstacles of Panax notoginseng. The plant-derived fungal complex is prepared by culturing Rhizophagus irregularis, an arbuscular mycorrhizal fungus, in the crude methanol extract of Eupatorium adenophorum.

[0013] The present invention also provides an application of the crude methanol extract of Eupatorium adenophorum in promoting the spore germination of arbuscular mycorrhizal fungus Rhizophagus irregularis.

[0014] The technical effects achieved by the present invention:

[0015] The plant-derived "Eupatorium adenophorum" provided by the present invention comes from the natural plant resource treasure house, and has the characteristics of high selectivity, environmental friendliness, safety for non-target organisms, low toxicity, low residue, promoting crop growth and improving crop disease resistance, etc., meeting the requirements of international green prevention and control and the concept of safe and sustainable development. It not only effectively prevents and controls the occurrence of root rot of Panax notoginseng, but also its secondary metabolites have functions such as inhibiting pathogen proliferation, balancing soil microbial communities, controlling soil acidification, aggregating beneficial microorganisms, mobilizing nutrient cycling and promoting growth through competition in the microecological cycle, becoming an effective way to alleviate the continuous cropping obstacles of Panax notoginseng, reduce agricultural residues and restore soil ecology.

[0016] The plant-derived "Eupatorium adenophorum" as a nutrient power bank has a synergistic effect in controlling root rot and alleviating continuous cropping obstacles when compounded with the microbial inoculant "arbuscular mycorrhizal fungus Rhizophagus irregularis". The secondary metabolites of plants can also aggregate specific microbial communities related to soil nutrient cycling, change the nitrogen metabolism and nutrient cycling among plants, convert the difficult-to-absorb nutrient forms into absorbable organic substances, increase nutrient return, and be utilized by symbiotic plants. Therefore, introducing suitable plant-derived substances into the soil can provide energy for microbial inoculants in biological control, and synergistically with microbial inoculants to achieve the effect of improving green prevention and control, making the soil transform into a "bacterial type" of "disease-suppressing and organic nutrient soil", effectively reducing the environmental pollution caused by pesticides and fertilizers, optimizing the microenvironment of continuous cropping soil and rhizosphere ecology, and restoring the production site conditions of Panax notoginseng.

[0017] The arbuscular mycorrhizal fungus Rhizophagus irregularis can significantly promote the absorption of mineral nutrients and water by plants, improve the survival ability of plants in extreme environments, and play a huge role in ecological restoration. Since Rhizophagus irregularis is an obligate biotroph, its application in research and production is greatly limited. Therefore, the in vitro culture of Rhizophagus irregularis and the exploration of growth-promoting substances are particularly important. This invention verified through experiments that the methanol extract of Eupatorium adenophorum can promote the spore germination of Rhizophagus irregularis. The germination rate of AMF spores reached 10.3% at 0.25 mg / mL and increased to 45.7% at 8 mg / mL, showing a dose-dependent response. The extract of Eupatorium adenophorum significantly promoted the growth of hyphae. This invention provides a new scheme for the culture of Rhizophagus irregularis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Survival rate diagram of the control group in the field in June of the second year;

[0019] Figure 2 Survival rate diagram of the Eupatorium adenophorum + Rhizophagus irregularis treatment group in the field in June of the second year. DETAILED DESCRIPTION OF THE INVENTION

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. For purchased commodities in the test methods, if specific conditions are not indicated, they shall be carried out according to conventional conditions or the conditions recommended by the manufacturer. For reagents or instruments whose manufacturers are not indicated, they are all conventional products that can be obtained through commercial purchase.

[0021] Example 1

[0022] Adding Eupatorium adenophorum + Rhizophagus irregularis to continuous cropping soil of Panax notoginseng to promote the adaptability of Panax notoginseng to continuous cropping soil

[0023] 1. Preparation and application of plant-derived fungal complex

[0024] (1) Collection and composting of plant source: Collect Eupatorium adenophorum plants, ensuring the plants are healthy and free from pests and diseases. Add the arbuscular mycorrhizal fungus Rhizophagus irregularis and stack them in a well-ventilated area for composting. During the composting process, regularly turn the pile to ensure ventilation and uniform fermentation, and ensure that the temperature inside the pile is maintained at 30-39 °C. The composting period is 30 days to obtain the plant-source fungal complex. After composting, the plant source has a loose texture, no obvious odor, and a dark brown color.

[0025] (2) Application method: Select the experimental field after the harvest of Panax notoginseng. Before planting Panax notoginseng, deeply plow the continuous cropping experimental field to a depth of 20 cm. Evenly spread the plant-source fungal complex into the soil, with a solid dosage of 1500 kg / ha. Then cover the film for 45 days to promote the activities of microorganisms in the soil and further decomposition of the plant source. After uncovering the film, sun-dry for 10 days to ensure good soil air permeability, and then transplant the Panax notoginseng seedlings.

[0026] 2. Preparation and planting of Panax notoginseng seedlings

[0027] (1) Seedling preparation: Select the mature period of Panax notoginseng (from September to November), pick the fruits from the healthy and pest-free Panax notoginseng plants, and retain the seeds. Use plug tray substrate for seedling raising, and the substrate formula is a mixture of humus soil, perlite and vermiculite in a volume ratio of 2:1:1. After sowing, place it in the greenhouse, keep the temperature at 20-25 °C, humidity at 60%-70%, and regularly spray to keep moist. After 8 months of cultivation, harvest the one-year-old Panax notoginseng seedlings in August of the following year. The seedling height is about 15 cm and the roots are intact.

[0028] (2) Planting method: In the field plots (AP) and control plots (CK) where the plant-source fungal complex has been applied, transplant the one-year-old Panax notoginseng seedlings according to the specifications of a plant spacing of 10 cm and a row spacing of 15 cm. Set up 10 plots for each treatment. Immediately water after planting to ensure the soil is moist. Follow-up management is carried out according to the conventional Panax notoginseng planting techniques, including shading, irrigation, fertilization and pest and disease monitoring.

[0029] 3. Calculation methods for emergence rate, growth rate and survival rate of seedlings

[0030] (1) Calculation method for emergence rate

[0031] The emergence rate refers to the percentage of the actual number of emerged seedlings in the total number of planted seeds within a certain period, which is used to measure the germination ability of seeds and the treatment effect. The calculation formula is as follows:

[0032] Emergence rate (%) = (Actual number of emerged seedlings / Total number of planted seeds) × 100%

[0033] (2) Calculation method for growth rate

[0034] The growth rate is the percentage of the increase in the emergence rate of the Panax notoginseng seedling treatment group compared to the control group, and is used to measure the high or low emergence effect of the treatment group. The calculation formula is as follows:

[0035] Growth rate (%) = [(Emergence rate of treatment group - Emergence rate of control group) / Emergence rate of control group] × 100%

[0036] (3) Calculation method of seedling survival rate

[0037] The seedling survival rate refers to the percentage of the number of surviving seedlings in the total number of planted seedlings during the planting process, and is used to measure the survival situation of the seedlings and the effect of the treatment. The calculation formula is as follows:

[0038] Seedling survival rate (%) = (Number of surviving seedlings / Total number of planted seedlings) × 100%

[0039] 3. Effect

[0040] See Table 1 and Table 2:

[0041] (1) The interaction between Eupatorium adenophorum and arbuscular mycorrhizal fungus Rhizophagus irregularis promoted the emergence rate of Panax notoginseng in continuous cropping soil. Compared with the control group, the growth rate of the emergence rate of Panax notoginseng in the continuous cropping soil treated with Eupatorium adenophorum + arbuscular mycorrhizal fungus Rhizophagus irregularis was about 258.05%.

[0042] (2) The interaction between Eupatorium adenophorum and arbuscular mycorrhizal fungus Rhizophagus irregularis promoted the adaptability of Panax notoginseng to continuous cropping soil. The seedling survival rate of Panax notoginseng in continuous cropping soil (control group) was 0 in July during growth, while the seedling survival rate of Panax notoginseng in the continuous cropping soil treated with Eupatorium adenophorum + arbuscular mycorrhizal fungus Rhizophagus irregularis was 78.32%, and the seedling survival rate remained 67.43% until September.

[0043] In summary, the interspecific interaction effect of Eupatorium adenophorum + arbuscular mycorrhizal fungus Rhizophagus irregularis promoted the adaptability of Panax notoginseng to continuous cropping.

[0044] Table 1 Comparison of emergence rate percentages of Panax notoginseng in continuous cropping soil and treatment

[0045]

[0046] Table 2 Comparison of seedling survival rate percentages of Panax notoginseng in continuous cropping soil and treatment

[0047]

[0048] Example 2

[0049] The methanol extract of Eupatorium adenophorum plants promotes the spore germination of arbuscular mycorrhizal fungus Rhizophagus irregularis

[0050] 1. Preparation of crude methanol extract of Eupatorium adenophorum:

[0051] Collect fresh whole plants of Eupatorium adenophorum, wash, dry and remove impurities, then crush them into fine powder with a pulverizer and pass through a 40-mesh sieve for standby. Next, weigh a certain amount (such as 100 g) of Eupatorium adenophorum powder, put it into a filter paper tube, place it in a Soxhlet extractor, and add an appropriate amount of anhydrous methanol (about 500 mL) to ensure that the solvent can completely immerse the plant powder. Connect the Soxhlet extractor to a reflux condenser, heat and reflux for 6 - 12 hours until the color of the extract becomes lighter, indicating that the active ingredients have been basically extracted completely. Then, transfer the extract to a rotary evaporator and concentrate it under reduced pressure at a water bath temperature of 50 - 60 °C until there is no solvent residue to obtain the crude methanol extract of Eupatorium adenophorum. Finally, place the crude extract in an oven and dry it at 60 °C to constant weight to obtain the crude extract powder, and store it sealed in a dry and light-proof environment for subsequent experiments.

[0052] 2. Experiment on the promotion of Rhizophagus irregularis spore germination by Eupatorium adenophorum:

[0053] Dissolve the prepared crude methanol extract of Eupatorium adenophorum, and dilute each sample to 6 concentrations of 8, 4, 2, 1, 0.5 and 0.25 mg·mL －1 . Use 10 mL of 1.5% water agar medium as the basic medium. Take 10 g of sand samples containing AMF stored at low temperature, separate AMF spores by wet sieving and decantation method, operate under a microscope, pick the sterilized spores with a sterile pipette tip onto different media, 50 spores per dish, and 10 dishes for each medium. After inoculation, place the dishes in a dark box at 26 °C for 10 d, observe the spore germination situation, calculate the spore germination rate, and the spore germination rate = (the number of germinated spores in the treatment ÷ the total number of inoculated spores in the treatment) × 100%. Measure the length of branched hyphae in the image processing system.

[0054] 3. Effect, see Table 3. The arbuscular mycorrhizal fungus Rhizophagus irregularis cannot reproduce and survive under non-symbiotic conditions. However, Eupatorium adenophorum promotes the spore germination of arbuscular mycorrhizal fungus Rhizophagus irregularis, and has a concentration effect. As the concentration increases, the spore germination rate also increases, and the hypha length also increases, indicating that the co-culture of Eupatorium adenophorum and arbuscular mycorrhizal fungus Rhizophagus irregularis can induce the production of pure spores of arbuscular mycorrhizal fungus Rhizophagus irregularis and promote hypha growth.

[0055] Table 3 Pure Spore Germination of Eupatorium adenophorum Inducing Arbuscular Mycorrhizal Fungi

[0056]

[0057]

[0058] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for improving the adaptability of continuous cropping of Panax notoginseng, characterized in that, It includes the following steps: S1: Collect Eupatorium adenophorum plants, add arbuscular mycorrhizal fungus Rhizophagus irregularis for composting treatment to obtain a plant-derived fungal complex; S2: Apply the composted complex to the continuous cropping field of Panax notoginseng, cover with a film, and uncover the film for sun drying after several days; S3: Transplant Panax notoginseng seedlings to the sun-dried soil for planting.

2. The method according to claim 1, wherein In step S1, the composting treatment is to regularly turn the pile under ventilation conditions to ensure that the temperature inside the pile is maintained at 30-39°C and compost for 30 days.

3. The method according to claim 1, wherein In step S2, the continuous cropping field of Panax notoginseng is deeply plowed by 20 cm and the plant-derived fungal complex is applied.

4. The method according to claim 3, wherein In step S2, the complex is applied to the continuous cropping field of Panax notoginseng at a dosage of 1500 kg / ha, the film is uncovered after 45 days of covering, and sun drying is carried out for 10 days.

5. Application of a plant-derived fungal complex in improving the soil with continuous cropping obstacles of Panax notoginseng, characterized in that, The plant-derived fungal complex is obtained by adding arbuscular mycorrhizal fungus Rhizophagus irregularis to Eupatorium adenophorum plants for composting treatment.

6. Use of a plant-derived fungal complex in improving the soil with continuous cropping obstacles of Panax notoginseng, characterized in that, The plant-derived fungal complex is prepared by adding arbuscular mycorrhizal fungus Rhizophagus irregularis to the crude methanol extract of Eupatorium adenophorum for cultivation.

7. Application of the crude methanol extract of Eupatorium adenophorum in promoting the spore germination of arbuscular mycorrhizal fungus Rhizophagus irregularis.

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