A method for interplanting corn and marigold

Abstract

The application provides a method for interplanting corn and marigold, and belongs to the technical field of agricultural planting. Funneliformis mosseae The application proposes inoculating Cylindrocarpon musae and interplanting to jointly regulate the growth of corn and marigold and the soil nutrients in the rhizosphere. Inoculating Cylindrocarpon musae promotes the growth of crops, increases the plant height and biomass of corn, and interplanting increases the leaf number and biomass of marigold. Inoculating Cylindrocarpon musae and interplanting both increase the biomass of crop roots, promote the absorption of nutrients by crops, and improve the soil nutrients and soil enzyme activity, so that the growth of crops and the soil environment are both optimized.

Description

Technical Field

[0001] This application belongs to the field of agricultural planting technology, specifically relating to a method for intercropping corn and marigolds. Background Technology

[0002] marigold( Tagetes erecta Marigolds (L.) are an important economic crop with ornamental, edible, and medicinal value. Their petals are rich in lutein, making them an ideal raw material for lutein production and development. Lutein is widely used in the food, feed, pharmaceutical, and chemical industries. Marigolds have achieved large-scale cultivation in my country, forming a distinctive sunshine industry centered on lutein extraction. Corn (… Zea mays Corn is an important food and feed crop. Silage corn has the characteristics of high biomass yield, good palatability, and high nutritional value. Moreover, because it is covered with a lot of lactic acid bacteria, it is easier to successfully ensilage it, thus becoming one of the important feed resources for animal husbandry in my country.

[0003] Arbuscular mycorrhiza fungi (AMF) are important soil microorganisms that live in symbiosis within plants and are widely involved in the nutrient uptake process. Although AMF inoculation has been reported to have general effects on root expansion and nutrient uptake, soil improvement, carbon sequestration, and ecological stability, there are no reports on its effects on improving agronomic traits and rhizosphere soil nutrients in maize and marigolds. Summary of the Invention

[0004] The purpose of this invention is to provide a *Moses tube-handle cysticercosis* (… Funneliformis mosseae Application in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0005] This invention provides the application of *Mammillaria mosierifolia* in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0006] Preferably, promoting maize growth includes increasing at least one of maize plant height, stem diameter, and biomass; Promoting marigold growth includes increasing the plant height.

[0007] Preferably, the improvement of rhizosphere soil nutrients includes increasing the fertilizer content in the soil and / or improving the activity of extracellular enzymes in the soil.

[0008] Preferably, increasing the fertilizer content in the soil includes increasing the content of total nitrogen, total potassium, available potassium and organic matter in the rhizosphere soil of maize and / or increasing the content of at least one of total nitrogen, total potassium, available potassium and alkaline nitrogen in the rhizosphere soil of marigold; The improvement of soil extracellular enzyme activity includes improving the activity of at least one of the following enzymes in maize rhizosphere soil: urease, nitrate reductase, acid phosphatase, sucrase, and catalase, and / or improving the activity of soil phytase and / or catalase in marigold rhizosphere.

[0009] This invention provides the application of *M. mossiostomata* and intercropping in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0010] Preferably, promoting maize growth includes increasing maize stalk diameter and / or biomass; The promotion of marigold growth includes increasing at least one of the following: number of marigold leaves, stem diameter, and biomass.

[0011] Preferably, the improvement of rhizosphere soil nutrients includes increasing the fertilizer content in the soil and / or improving the activity of extracellular enzymes in the soil; The improvement of fertilizer content in the soil includes increasing the content of at least one of total nitrogen, total phosphorus, total potassium, available nitrogen, and available potassium in the rhizosphere soil of corn and / or the content of at least one of total nitrogen, total potassium, available nitrogen, and available potassium in the rhizosphere soil of marigold.

[0012] Preferably, the improvement of soil extracellular enzyme activity includes improving the activity of urease and / or catalase in the rhizosphere soil of maize and / or marigold.

[0013] This invention provides a method for intercropping corn and marigolds, comprising the following steps: Corn and marigolds were intercropped in a substrate inoculated with *Morchella mosierifolia*.

[0014] Preferably, the *Morchella mosierifolia* is inoculated in the form of an inoculum, with an inoculation amount of 230-270 g / plant. The method for preparing the inoculum involves transplanting sterile seedlings into a cultivation substrate containing *Bacillus mosierifolius* for 30-40 days, removing the above-ground parts and topsoil, collecting the roots and cultivation substrate, and obtaining the inoculum.

[0015] This invention provides the application of *Aureobasidium mosesi* in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients. Experiments showed that *Aureobasidium mosesi* (AMF) successfully colonized the roots of both marigolds and maize using various structures: hyphae, arbuscular mycelia, and hyphal rings. The colonization rate in maize ranged from 61.11% to 83.47%, and in marigolds from 38.31% to 53.54%. Electron microscopy revealed a greater prevalence of hyphae and arbuscular mycelia, while spore and vesicle structures were less abundant (<2%). *Aureobasidium mosesi* successfully colonized and exerted its effects on the roots of both maize and marigolds. Compared to uninoculated treatment, AMF inoculation significantly increased maize stem circumference, significantly increased maize plant biomass, and increased the plant height of both maize and marigolds. In terms of plant nutrients, compared to the uninoculated treatment, AMF inoculation significantly increased the total nitrogen and total potassium content in both maize and marigolds, and significantly reduced the total phosphorus content. Simultaneously, AMF inoculation significantly reduced rhizosphere soil pH in the maize treatment and significantly increased available potassium and organic matter content in the rhizosphere soil of the maize treatment compared to the uninoculated treatment, while significantly increasing available potassium, total potassium, and available nitrogen content in the rhizosphere soil of the marigold treatment. AMF inoculation also significantly increased the activities of urease, nitrate reductase, acid reductase, sucrase, and catalase in the rhizosphere soil of the maize treatment compared to the uninoculated treatment, while significantly increasing the activities of phytase and catalase in the rhizosphere soil of the marigold treatment. AMF inoculation promotes crop growth and improves crop agronomic traits.

[0016] This invention provides the application of *Amanita mossicae* and intercropping in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients. Experiments showed that intercropping had no significant effect on AMF colonization rate. Two-factor results of AMF inoculation and intercropping patterns indicated that the planting pattern significantly affected marigold stem diameter and maize and marigold biomass; AMF inoculation significantly affected maize plant height, stem diameter, and biomass, and marigold leaf number; the interaction between the two significantly affected stem diameter and biomass. Regarding plant nutrients, the planting pattern significantly affected the total nitrogen, total phosphorus, and total potassium content of both maize and marigolds; AMF inoculation significantly affected the total nitrogen, total phosphorus, and total potassium content of both maize and marigolds; the interaction between the two significantly affected the total nitrogen, total phosphorus, and total potassium content of maize, and significantly affected the total nitrogen and total potassium content of marigolds; simultaneously, the planting pattern, AMF inoculation, and the combined effect of both affected the pH, available nitrogen, and available potassium content of the crop root soil. Regarding the effects on extracellular enzymes in rhizosphere soil, two-factor analysis showed that planting patterns significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of maize and marigold treatments; AMF inoculation significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of maize and marigold treatments; and the interaction between the two significantly affected the activities of urease and catalase in the rhizosphere soil of maize and marigold treatments. It is evident that both AMF inoculation and intercropping increased crop root biomass, promoted crop nutrient uptake, and improved soil nutrient and enzyme activity. Attached Figure Description

[0017] Figure 1 Optical micrographs of root segments of plants infected with AMF after 60 days of pot cultivation; (A) is a photograph of arbuscular structure, a represents arbuscular structure; (B) is a photograph of hyphae, b represents hyphae structure; c represents hyphal ring structure; (C) is a photograph of spore structure; d represents spore structure; (D) is a photograph of vesicle structure, e represents vesicle structure; scale bar is 50 μm; Figures 2-4 The results of AMF colonization rates for maize (Y) and marigold (W) in different treatment groups; Figures 5-7 Results of agronomic traits determination for maize (Y) and marigold (W) in different treatment groups; Figure 8 SPAD values ​​for maize (Y) and marigold (W) in different treatment groups; Figure 9 Biomass results of maize (Y) and marigold (W) in different treatment groups Figure 10 Results of plant nutrient content in maize (Y) and marigold (W) under different treatment groups; Figures 11-16Results of rhizosphere soil agrochemical properties of maize (Y) and marigold (W) in different treatment groups; Figure 17 Results of total nitrogen and organic matter content in rhizosphere soil of maize (Y) and marigold (W) in different treatment groups; Figures 18-23 Results of rhizosphere soil enzyme activity in maize (Y) and marigold (W) under different treatment groups. Detailed Implementation

[0018] This invention provides the application of *Mammillaria mosierifolia* in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0019] In this invention, the *Tetranychus moses* comprises hyphae and / or spores. This invention does not impose any particular limitation on the source of the *Tetranychus moses*; any source known in the art is acceptable. The *Tetranychus moses* exists in the form of an inoculum. Preferably, the inoculum is prepared by transplanting sterile seedlings into a cultivation substrate containing *Tetranychus moses*, culturing for 30-40 days, removing the above-ground parts and topsoil, and collecting the roots and cultivation substrate. The sterile seedlings include common crops such as corn, wheat, and sorghum. This invention does not impose any particular limitation on the preparation method of the sterile seedlings; any preparation method known in the art is acceptable. For example, after sterile crop seeds have germinated, they are inoculated with *Tetranychus moses*, transplanted into a sterilized cultivation substrate, and nutrient solution is applied during cultivation. When the colony is at least 90% established, the above-ground parts and topsoil are removed, and the roots and cultivation substrate are collected. The cultivation substrate preferably comprises a mixture of farmland soil and vermiculite. The preferred volume ratio of farmland soil to vermiculite is 3:1. The preferred particle size of the vermiculite is 1-2 mm. In this embodiment of the invention, sterile seedlings are obtained by culturing corn seeds, and the preferred corn variety is Wugu 1790. The preferred nutrient solution is 1 / 2 Hoagland nutrient solution. The culturing time is 30-40 days, and can be 35 days.

[0020] In this invention, promoting maize growth preferably includes increasing at least one of maize plant height, stem diameter, and biomass. Promoting marigold growth preferably includes increasing marigold plant height. To verify the effects of *Tetranychus moses* inoculation on the growth of maize and marigolds, this invention used a cultivation group without *Tetranychus moses* inoculation as a control, and measured agronomic traits (plant height, stem diameter, number of leaves, number of branches, and chlorophyll content). The results showed that *Tetranychus moses* inoculation increased maize stem diameter, biomass, and plant height, while also increasing marigold plant height.

[0021] In this invention, improving rhizosphere soil nutrients preferably includes increasing the fertilizer content in the soil and / or improving the activity of extracellular enzymes in the soil. Increasing the fertilizer content in the soil preferably includes increasing the content of total nitrogen, total potassium, available potassium, and organic matter in the rhizosphere soil of maize and / or increasing the content of at least one of total nitrogen, total potassium, available potassium, total potassium, and alkaline nitrogen in the rhizosphere soil of marigold. Improving the activity of extracellular enzymes in the soil preferably includes reducing the activity of at least one of urease, nitrate reductase, acid phosphatase, sucrase, and catalase in the rhizosphere soil of maize and / or increasing the activity of phytase and nitrate reductase in the rhizosphere soil of marigold and / or decreasing the activity of catalase.

[0022] In another embodiment of the present invention, to verify the effects of AMF inoculation on the nutrient composition of rhizosphere soil in maize and marigolds, the uninoculated AMF group was used as a control, and the content of macronutrients, pH, and extracellular enzymes in the soil were measured. Regarding plant nutrients, compared with the uninoculated AMF treatment, the AMF inoculation treatment significantly increased the total nitrogen and total potassium content in maize and marigolds, and significantly decreased the total phosphorus content. The AMF inoculation treatment significantly reduced the pH of the rhizosphere soil in the maize treatment, significantly increased the available potassium and organic matter content in the rhizosphere soil in the maize treatment, and significantly increased the available potassium, total potassium, and available nitrogen content in the rhizosphere soil in the marigold treatment compared to the uninoculated treatment. Regarding the effects of extracellular enzymes in the soil, the AMF inoculation treatment significantly increased the activities of urease, nitrate reductase, acid reductase, sucrase, and catalase in the rhizosphere soil of the maize treatment compared to the uninoculated treatment, and significantly increased the activities of phytase and catalase in the rhizosphere soil of the marigold treatment.

[0023] The present invention also preferably provides the application of intercropping in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0024] In this invention, intercropping significantly increased the number of leaves and biomass in the marigold treatment compared to monocropping. Compared to monocropping, intercropping significantly increased the total nitrogen content of maize plants, decreased the total phosphorus content, and increased the total nitrogen content of marigold plants. Compared to monocropping, intercropping significantly increased the pH, available phosphorus, total phosphorus, and total nitrogen content in the rhizosphere soil of maize, and significantly increased the total nitrogen and organic matter content in the rhizosphere soil of marigold. The intercropping pattern had a significant impact on the activities of urease, phytase, and catalase in the rhizosphere soils of both maize and marigold treatments.

[0025] This invention provides the application of *M. mossiostomata* and intercropping in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

[0026] In this invention, promoting maize growth preferably includes increasing maize stem diameter and / or biomass. Promoting marigold growth preferably includes increasing at least one of marigold leaf number, stem diameter, and biomass.

[0027] In one embodiment of the present invention, the effects of two factors, namely, *Amygdaloides mosierifolia* inoculation and intercropping, on the growth or agronomic traits of maize and / or marigold were as follows: the planting pattern had a significant effect on marigold stem diameter and on the biomass of both maize and marigold; AMF inoculation had a significant effect on maize plant height, stem diameter, and biomass, and on the number of marigold leaves; the interaction between the two factors had a significant effect on stem diameter and biomass.

[0028] In this invention, the improvement of rhizosphere soil nutrients preferably includes increasing the fertilizer content in the soil and / or improving the activity of soil extracellular enzymes. The improvement of soil fertilizer content preferably includes increasing the content of at least one of total nitrogen, total phosphorus, total potassium, available nitrogen, and available potassium in the rhizosphere soil of maize and / or the content of at least one of total nitrogen, total potassium, available nitrogen, and available potassium in the rhizosphere soil of marigold. The improvement of soil extracellular enzyme activity preferably includes improving the activity of urease and / or catalase in the rhizosphere soil of maize and / or marigold.

[0029] In one embodiment of the present invention, the effects of *Amygdalus mosierifolius* inoculation and intercropping on rhizosphere soil nutrients in maize and / or marigold were as follows: Planting pattern significantly affected the total nitrogen, total phosphorus, and total potassium contents of both maize and marigold; AMF inoculation significantly affected the total nitrogen, total phosphorus, and total potassium contents of both maize and marigold; the interaction between the two significantly affected the total nitrogen, total phosphorus, and total potassium contents of maize and marigold; planting pattern, AMF inoculation, and the combined effect of both affected the pH, available nitrogen, and available potassium contents of the crop root soil. Simultaneously, the effects on soil extracellular enzymes were as follows: planting pattern significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of both maize and marigold treatments; AMF inoculation significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of both maize and marigold treatments; the interaction between the two significantly affected the activities of urease and catalase in the rhizosphere soil of both maize and marigold treatments.

[0030] This invention provides a method for intercropping corn and marigolds, comprising the following steps: Corn and marigolds were intercropped in a substrate inoculated with *Morchella mosierifolia*.

[0031] In this invention, the *Morchella mosierifolia* is preferably inoculated in the form of an inoculum, with an inoculation amount preferably of 230-270 g / plant. The inoculation method is preferably a sandwich inoculation, specifically: first, a cultivation substrate is laid out, then the inoculum is spread evenly on top, corn and marigolds are inoculated on top of the inoculum, and then another layer of inoculum is inoculated.

[0032] This invention does not impose any special restrictions on the varieties of corn and marigolds; varieties well-known in the art can be used. In this embodiment, the corn variety is Wugu 1790, purchased from Gansu Wugu Seed Industry Co., Ltd. The marigold variety is Lidaer Jinju No. 1, purchased from Yunnan Lidaer Biotechnology Co., Ltd. The floating seedling substrate for corn or marigolds is purchased from Dali National Plastics Factory Co., Ltd., with a total porosity of 80-95%, conductivity ≤1000us / cm, particle size 1-5mm, and conforming to standard YC / T310-2024. The floating seedling substrate mainly contains organic matter, perlite, vermiculite, and rooting powder, with an organic matter content of 31%.

[0033] The following detailed description of a method for intercropping corn and marigolds provided by the present invention, with reference to specific embodiments, should not be construed as limiting the scope of protection of the present invention.

[0034] Example 1 A method for jointly regulating the growth of maize and marigold and rhizosphere soil nutrients by AMF and planting patterns 1. Preparation of inoculum 1.1 Preparation of cultivation substrate Large soil clumps were removed by sieving farmland soil through a 5mm sieve. The sieved material was collected and mixed with commercially available (1-2mm) vermiculite at a volume ratio of 3:1. The mixture was then placed in a high-pressure steam sterilizer and sterilized at 121℃ for 2 hours. The sterilization was repeated 3 times after 24 hours. The mixture was then cooled to room temperature to obtain a sterile cultivation substrate for inoculum propagation.

[0035] 1.2 Preparation of sterile maize seedlings Select corn seeds of uniform size, shape and plumpness (e.g., Wugu 1790), disinfect them with 10% sodium hypochlorite for 10 minutes, rinse them 3 times with sterile water, place them in a petri dish with double-layer sterile filter paper, and culture them in the dark until the corn sprouts (temperature: 28℃) to obtain sterile corn seedlings.

[0036] 1.3 Thoroughly wipe the flowerpots (350 mm × 250 mm) with 75% alcohol to sterilize them, let them dry, and label them. Lay the sterile cultivation substrate prepared in step 1.1 at the bottom of each sterilized flowerpot; weigh 500 g of substrate containing AMF (… F. mosseae The rhizosphere soil containing mycelium and spores (purchased from the Institute of Plant Nutrition and Resources, Beijing Agricultural and Forestry College) was spread on top of the cultivation substrate in a sterile flowerpot. The sterile corn seedlings obtained in step 1.2 were then transplanted into the sterile flowerpot substrate and covered with 3-4 cm of sterile cultivation substrate to form a "sandwich" method for propagating AMF.

[0037] Use 1 / 2 Hoagland nutrient solution (Hoagland nutrient solution formula: calcium nitrate 94.5 g / L, ammonium nitrate 8 g / L, potassium nitrate 50.6 g / L, potassium dihydrogen phosphate 13.6 g / L, magnesium sulfate heptahydrate 100.9 g / L, Na2EDTA·2H2O 7.46 g / L, FeSO4·7H2O 5.56 g, MnSO4·4H2O 4.46 g / L, H3BO3 1.24 g / L, ZnSO4·7H2O 1.72 g / L, CuSO4·5H2O 0.005 g / L, CoCl2·6H2O 0.005 g, KI 0.166 g, Na2MoO4·2H2O). Water thoroughly with 0.05g / L nutrient solution until the bottom of the pot seeps out. During the growing season, maintain regular management to ensure normal plant growth. Then, water with 300ml of 1 / 2 Hoagland nutrient solution every 15 days. After about 35 days of cultivation, randomly check the corn roots and observe the colonization of AMF in the corn roots under an optical microscope using the cross-cross method. If typical structures such as AMF hyphae, arbuscular mycelium, vesicles, and hyphal rings are observed, the colonization is considered successful. After culturing for about 60 days until the AMF colonization rate at the corn roots is no less than 90%, the above-ground parts of the corn are cut off and the top 1-2 cm of soil is removed. The roots and culture medium are collected to obtain an inoculum containing AMF.

[0038] 2. Cultivation of corn and marigold seedlings 2.1 The floating seedling substrates for corn and marigolds were sterilized by high-pressure steam (121℃, 2 h; sterilized once every 24 h, for a total of 3 times) to obtain sterile seedling substrates. The silage corn Wugu 1790 and the marigold Lidaer Jinju No. 1 were floated and raised in the special floating seedling substrate that had been sterilized. The marigold seedlings were raised for 45 days and the corn seedlings for 30 days. They were transplanted on the same day.

[0039] 2.2 Commercially available horticultural vermiculite (1-2mm) and farmland soil were used as the basic cultivation substrate for marigolds and corn at a volume ratio of 1:4. The farmland soil was sieved through a 5mm sieve to remove larger impurities. It was then mixed evenly with the commercially available vermiculite and sterilized in a high-pressure steam sterilizer at a temperature of 121℃ for 2 hours. The sterilization process was repeated three times with an interval of more than 24 hours. The mixture was then air-dried naturally to serve as a sterile cultivation substrate.

[0040] The AMF-containing inoculum obtained in step 1.3 was divided into two equal parts. One part was used as the inoculation group, and the other part was sterilized by autoclaving (121℃, sterilization for 2 h, repeated 3 times at 24 h intervals) and air-dried to obtain inactivated AMF as the non-inoculation control group.

[0041] 3. Experiment Implementation To investigate the combined effects of AMF and planting patterns on the growth of maize and marigolds and rhizosphere soil nutrients, two different treatment combinations were set up: AMF inoculation versus no AMF inoculation, and intercropping versus no intercropping. Flower pots (55 cm × 33 cm × 18 cm) were thoroughly cleaned with 75% alcohol, dried, and labeled. A layer of sterile cultivation substrate was laid at the bottom (to 2 / 3 of the pot's height). For the inoculation group, an inoculum containing AMF (250 g / plant, added twice, before and after transplanting) was added above the sterile cultivation substrate. For the control group, an inoculum containing inactivated AMF (250 g / plant) was added above the sterile cultivation substrate.

[0042] Transplant the marigold and corn seedlings from step 2.1. For the marigold monoculture group, plant 2 marigold seedlings per pot, spaced 30cm apart; for the corn monoculture group, plant 2 corn seedlings per pot, spaced 30cm apart; for the marigold-corn intercropping group, plant one marigold and one corn seedling, spaced 30cm apart. Spread the aseptic cultivation substrate evenly over the surface of the inoculum layer up to the top of the cultivation container, forming a "sandwich" method to inoculate *M. moses* onto the roots of both the marigolds and corn.

[0043] After culturing marigold and corn seedlings for 60 days, agronomic traits and SPAD values ​​were measured. Marigold and corn samples were collected at harvest, resulting in 8 sample groups. ① Maize monoculture without AMF: A-YD inoculation; ②Maize monoculture inoculation with AMF: A+YD; ③ Marigolds planted alone without AMF:A-WD inoculation; ④ Marigold monoculture inoculation with AMF: A+WD; ⑤ Intercropping maize with marigolds without AMF inoculation: A-YJ; ⑥ Corn intercropped with marigolds and inoculated with AMF: A+YJ; ⑦ Marigolds intercropped with maize without AMF inoculation: A-WJ; ⑧ Marigolds intercropped with maize and inoculated with AMF: A+WJ.

[0044] 4. Sample collection and determination Samples of maize and marigolds were collected after 60 days of cultivation in a greenhouse. Agronomic traits of the crops were measured before sampling: plant height, stem diameter, and number of leaves for maize; and plant height, stem diameter, number of leaves, and number of branches for marigolds. The chlorophyll content of marigold and maize leaves was measured using a SPAD instrument. When collecting samples, carefully remove the plants, wash the roots, and absorb excess water with filter paper. Weigh the fresh weight of the above-ground and underground parts of the crop separately. Take fresh root samples and use the acid fuchsin staining method and the cross-cross method to detect and count the AMF colonization rate. Collect marigold and corn root segments separately, wash them clean, add 10% KOH solution until the root samples are completely submerged, heat in a 90 ℃ water bath until the root samples are translucent, cool, rinse with tap water 2-3 times, add lactic acid for acidification, and soak for 10-20 minutes. Cut the root samples into root segments of about 1 cm, transfer them to a glass slide, add 0.5% acid fuchsin staining solution, and stain the corn and marigold root segments. During the staining process, the back of the glass slide can be ignited with an alcohol lamp to accelerate the staining until white smoke appears 2-3 times. The stained root segments were decolorized with lactic acid glycerol, and an appropriate amount of root segments were pressed onto a glass slide. The pressed slides were observed under a regular optical microscope, and the colonization rate of *Tetranychus mosierifolius* was statistically analyzed using the cross-multiplication method. At least 300 fields of view were examined for each root sample, and the total colonization rate of *Tetranychus mosierifolius* was calculated according to the following formula I. The remaining samples were divided into aboveground and underground parts, placed in kraft paper envelopes, and placed in an oven at 105℃ for 30 minutes to blanch, then dried at 75℃ to constant weight. The dry weight of the samples was measured. The samples were pulverized using a micro-pulverizer and mixed thoroughly for measuring the content of crop nutrients N, P, and K. According to the method in the Agricultural Industry Standard of the People's Republic of China NY / T2017-2011, three replicate samples were set up for each experimental treatment.

[0045] Colony rate of *M. mossioides* (%) = (Total number of fields of view - Number of blank fields of view) / Total number of fields of view × 100% Formula I.

[0046] Rhizosphere soil samples were taken from marigolds and corn in intercropping patterns for analysis. For monoculture potted plants, rhizosphere soil samples were mixed thoroughly before analysis. During soil collection, the crops were carefully removed from the pots along with their roots and soil. The topsoil was removed, and the loose soil attached to the roots was gently shaken off. Soil tightly bound to the roots was brushed off with a small brush, mixed thoroughly, and used as rhizosphere soil. This mixture was then transferred to a cool, shaded area to air dry and passed through a 60-mesh sieve. Three replicates were selected for each treatment to determine soil physicochemical properties: pH, available nitrogen, available phosphorus, available potassium, total nitrogen, total phosphorus, total potassium, and organic matter. Simultaneously, soil nutrient-related enzyme activities were measured: acid phosphatase, catalase, urease, phytase, sucrase, and nitrate reductase. Soil pH was determined using the water extraction method; soil alkaline nitrogen content was determined using the alkaline diffusion method; soil available phosphorus content was determined using the NaHCO3 extraction-molybdenum antimony colorimetric method; soil available potassium content was determined using the flame photometry method; soil organic matter content was determined using the potassium dichromate titration method; soil total nitrogen content was determined using the flow analysis method; soil total phosphorus content was determined using the molybdenum blue colorimetric method; and soil total potassium content was determined using the flame photometry method. Specific methods are detailed in "Soil Agrochemical Analysis". Soil enzyme activity kits were purchased from Suzhou Grees Biotechnology Co., Ltd.

[0047] 5. Results like Figure 1 and Figures 2-4 As shown: The roots of marigolds and corn inoculated with AMF were successfully infected with AMF (… F. mosseae Multiple structures successfully colonized the mycelium, including hyphae, arbuscular mycelium, and mycelial rings. The colonization rate in maize ranged from 61.11% to 83.47%, while that in marigolds ranged from 38.31% to 53.54%. Electron microscopy revealed a higher prevalence of hyphae and arbuscular structures, while spores and vesicles were less abundant (<2%). AMF successfully colonized and functioned in the roots of both maize and marigolds. Intercropping did not significantly affect the AMF colonization rate.

[0048] like Figures 5-9 As shown, compared with no inoculation, AMF inoculation significantly increased maize stem circumference, significantly increased maize plant biomass, and increased plant height of both maize and marigolds. Compared with monoculture, intercropping significantly increased leaf number and biomass in the marigold treatment. Whether or not AMF was applied and different planting patterns had no significant effect on SPAD values.

[0049] Two-factor results showed that planting pattern significantly affected marigold stem diameter and marigold biomass; AMF inoculation significantly affected marigold plant height, stem diameter, and biomass, and marigold leaf number; the interaction between the two had a significant effect on stem diameter and biomass.

[0050] like Figure 10As shown: From the perspective of plant nutrients, compared with the uninoculated treatment, the AMF inoculation treatment significantly increased the total nitrogen and total potassium content in maize and marigold, and significantly decreased the total phosphorus content. Compared with the monoculture treatment, the intercropping treatment significantly increased the total nitrogen content in maize plants, decreased the total phosphorus content, and increased the total nitrogen content in marigold plants. The two-factor results indicate that: planting pattern has a significant impact on the total nitrogen, total phosphorus, and total potassium content in both maize and marigold; AMF inoculation has a significant impact on the total nitrogen, total phosphorus, and total potassium content in both maize and marigold; the interaction between the two has a significant impact on the total nitrogen, total phosphorus, and total potassium content in maize, and a significant impact on the total nitrogen and total potassium content in marigold.

[0051] like Figures 11-17 As shown, compared with the non-AMF inoculation group, the AMF inoculation treatment significantly reduced the rhizosphere soil pH of the maize treatment and significantly increased the rhizosphere soil contents of available potassium and organic matter in the maize treatment, while significantly increasing the rhizosphere soil contents of available potassium, total potassium, and available nitrogen in the marigold treatment. Compared with monoculture treatment, intercropping treatment significantly increased the rhizosphere soil pH, available phosphorus, total phosphorus, and total nitrogen content of maize, and significantly increased the rhizosphere soil contents of total nitrogen and organic matter in marigold treatment. The two-factor results indicate that planting pattern, AMF inoculation, and the combined effect of both all affect the contents of crop rhizosphere soil, including pH, available nitrogen, and available potassium.

[0052] like Figures 18-23 As shown, AMF inoculation significantly increased the activities of urease, nitrate reductase, acid reductase, sucrase, and catalase in the rhizosphere soil of maize compared to the uninoculated treatment, and significantly increased the activities of phytase and catalase in the rhizosphere soil of marigold. Two-factor analysis indicated that planting pattern significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of both maize and marigold treatments; AMF inoculation significantly affected the activities of urease, phytase, and catalase in the rhizosphere soil of both maize and marigold treatments; and the interaction between the two significantly affected the activities of urease and catalase in the rhizosphere soil of both maize and marigold treatments.

[0053] The experimental results show that AMF inoculation and intercropping jointly regulate the growth of maize and marigolds, as well as rhizosphere soil nutrients. AMF inoculation promotes crop growth, increases maize plant height and biomass, and intercropping increases the number of leaves and biomass in marigolds. Both AMF inoculation and intercropping increase crop root biomass, promote nutrient absorption, and improve soil nutrients and soil enzyme activity.

[0054] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Moses tube-handle cysticercosis ( Funneliformis mosseae Applications in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

2. The application according to claim 1, characterized in that, The promotion of maize growth includes increasing at least one of maize plant height, stem diameter and biomass; Promoting marigold growth includes increasing the plant height.

3. The application according to claim 1, characterized in that, The improvement of rhizosphere soil nutrients includes increasing the fertilizer content in the soil and / or improving the activity of extracellular enzymes in the soil.

4. The application according to claim 3, characterized in that, The improvement of fertilizer content in the soil includes increasing the content of total nitrogen, total potassium, available potassium and organic matter in the rhizosphere soil of corn and / or increasing the content of at least one of total nitrogen, total potassium, available potassium and alkaline nitrogen in the rhizosphere soil of marigolds. The improvement of soil extracellular enzyme activity includes improving the activity of at least one of the following enzymes in maize rhizosphere soil: urease, nitrate reductase, acid phosphatase, sucrase, and catalase, and / or improving the activity of soil phytase and / or catalase in marigold rhizosphere.

5. Application of *M. mossiostomata* and intercropping in promoting the growth of maize and / or marigolds and / or improving rhizosphere soil nutrients.

6. The application according to claim 5, characterized in that, The promotion of maize growth includes increasing maize stalk diameter and / or biomass; The promotion of marigold growth includes increasing at least one of the following: number of marigold leaves, stem diameter, and biomass.

7. The application according to claim 5, characterized in that, The improvement of rhizosphere soil nutrients includes increasing the fertilizer content in the soil and / or improving the activity of extracellular enzymes in the soil; The improvement of fertilizer content in the soil includes increasing the content of at least one of total nitrogen, total phosphorus, total potassium, available nitrogen, and available potassium in the rhizosphere soil of corn and / or the content of at least one of total nitrogen, total potassium, available nitrogen, and available potassium in the rhizosphere soil of marigold.

8. The application according to claim 7, characterized in that, The improvement of soil extracellular enzyme activity includes improving the activity of urease and / or catalase in the rhizosphere soil of maize and / or marigold.

9. A method for intercropping corn and marigolds, characterized in that, Includes the following steps: Corn and marigolds were intercropped in a substrate inoculated with *Morchella mosierifolia*.

10. The method according to claim 9, characterized in that, The *M. mossiosus* was inoculated in the form of an inoculum, with an inoculation amount of 230-270 g / plant. The method for preparing the inoculum involves transplanting sterile seedlings into a cultivation substrate containing *Bacillus mosierifolius* for 30-40 days, removing the above-ground parts and topsoil, collecting the roots and cultivation substrate, and obtaining the inoculum.

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