Methods for promoting growth of ananas comosus roots

Abstract

The application provides a method for promoting the growth of an anthurium andraeanum root system. The method for promoting the growth of the anthurium andraeanum root system is characterized in that the anthurium andraeanum plant is subjected to root irrigation with an anthurium andraeanum root growth-promoting liquid to soak the roots of the anthurium andraeanum plant, and the anthurium andraeanum root growth-promoting liquid is an endophytic fungal culture solution of an orange-yellow calanthe. The method for promoting the growth of the anthurium andraeanum root system can effectively promote the growth of the anthurium andraeanum root system, and further promote the rapid growth of the anthurium andraeanum plant.

Description

Technical Field

[0001] This invention relates to the field of plant cultivation technology, and in particular to a method for promoting the root growth of Anoectochilus roxburghii. Background Technology

[0002] The entire plant of *Anoectochilus roxburghii* can be used medicinally. It has a neutral and sweet taste and possesses properties such as clearing heat and cooling the blood, dispelling wind and dampness, detoxifying, relieving pain, and suppressing cough. It is primarily used to treat hemoptysis, bronchitis, nephritis, cystitis, diabetes, hematuria, rheumatoid arthritis, tumors, and other difficult-to-treat diseases. This has led to a continuous increase in domestic and international market demand, resulting in a sharp decline in wild resources. To alleviate the shortage of wild *Anoectochilus roxburghii* resources, people have begun to focus on and study its ecological habits, growth conditions, and artificial propagation techniques, hoping to protect and propagate *Anoectochilus roxburghii* through scientific methods.

[0003] Currently, the artificial cultivation of Anoectochilus roxburghii has achieved initial success. The promotion of this method can not only protect wild Anoectochilus roxburghii from the threat of over-harvesting, but also provide people with a sustainable and stable source of Anoectochilus roxburghii. For example, Chinese invention patent applications with application number 201210442593.8 and 201510488134.7 have achieved rapid seedling cultivation of Anoectochilus roxburghii through tissue culture. However, due to the underdeveloped root system of Anoectochilus roxburghii, the plants grow slowly and it is still difficult to meet market demand. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for promoting the root growth of Anoectochilus roxburghii, thereby promoting the rapid growth of the Anoectochilus roxburghii plant.

[0005] The objective of this invention is achieved through the following technical solution:

[0006] A method for promoting the root growth of Anoectochilus roxburghii involves applying a root growth promoter to the Anoectochilus roxburghii plant through root irrigation, so that the root growth promoter soaks the roots of the Anoectochilus roxburghii plant.

[0007] The root growth promoter solution of *Anoectochilus roxburghii* is a culture solution of endophytic fungi of *Anoectochilus roxburghii*.

[0008] In one embodiment, the root irrigation of *Anoectochilus roxburghii* plants with a root growth promoter solution includes the following steps:

[0009] Obtain the aforementioned Anoectochilus roxburghii plant;

[0010] The *Anoectochilus roxburghii* plant is subjected to cultivation treatment so that it is planted in a cultivation substrate;

[0011] The cultivation substrate is perforated to form multiple holes, each hole being positioned adjacent to the root system of the Anoectochilus roxburghii plant.

[0012] The borehole is irrigated with the root growth promoting solution of *Anoectochilus roxburghii*, so that the root growth promoting solution is poured back into the borehole.

[0013] In one embodiment, after the step of irrigating the drilled hole with the *Anoectochilus roxburghii* root growth-promoting solution, the method for promoting *Anoectochilus roxburghii* root growth further includes the following steps:

[0014] The drilled hole is filled so that the cultivation substrate fills the drilled hole.

[0015] In one embodiment, the cultivation substrate is peat moss.

[0016] In one embodiment, the root growth promoter of *Anoectochilus roxburghii* is obtained through the following steps:

[0017] Picking wild orange-yellow jade phoenix flowers;

[0018] The rootstock of the orange-yellow jade phoenix flower is treated to clean the surface of the rootstock and obtain new nutrient roots that form mycorrhizae.

[0019] The newly formed nutrient roots are scraped to extract the fungal colonies from them, thus obtaining the orange-yellow jade phoenix flower fungal community.

[0020] The orange-yellow jade phoenix flower fungus was cultured to obtain the root growth promoter of Anoectochilus roxburghii.

[0021] In one embodiment, the root and stem treatment of the orange-yellow jade phoenix flower specifically includes the following steps:

[0022] The rhizomes of the orange-yellow jade phoenix flower are washed and cut to remove particulate matter from the surface of the rhizomes, resulting in rhizome segments of the orange-yellow jade phoenix flower.

[0023] The root and stem segments of the orange-yellow jade phoenix flower are subjected to deep cleaning treatment to remove bacteria and harmful substances from the surface of the root and stem segments;

[0024] The orange-yellow jade phoenix flower root and stem segments, after deep cleaning, are further cleaned to remove the liquid from their surface, thus obtaining new nutrient roots.

[0025] In one embodiment, the rhizomes of the orange-yellow jade phoenix flower are washed and cut, specifically as follows:

[0026] Rinse the rhizomes of the orange-yellow jade phoenix flower under tap water, then cut the rhizomes into sections to obtain orange-yellow jade phoenix flower rhizome sections.

[0027] In one embodiment, the length of the orange-yellow jade phoenix flower rhizome segment is 1cm to 2cm.

[0028] In one embodiment, the root and stem segments of the orange-yellow jade phoenix flower are subjected to deep cleaning treatment, specifically as follows:

[0029] The root and stem segments of the orange-yellow jade phoenix flower were soaked in sodium hypochlorite solution, then taken out and soaked in ethanol solution.

[0030] In one embodiment, the root and stem segments of the orange-yellow jade phoenix flower, after deep cleaning, are subjected to a cleaning process, specifically as follows:

[0031] The root and stem segments of the orange-yellow jade phoenix flower were rinsed with sterile water at least three times, and then the liquid on the surface of the root and stem segments was dried with sterile absorbent paper.

[0032] Compared with the prior art, the present invention has at least the following advantages:

[0033] The method for promoting root growth of *Anoectochilus roxburghii* of this invention uses an endophytic fungal culture solution of *Anoectochilus roxburghii* as a root growth promoter. This solution is then applied via root irrigation to the roots of the *Anoectochilus roxburghii* plant, allowing the roots to be thoroughly immersed in the root growth promoter. This ensures the root growth promoter acts directly on the roots, promoting symbiosis between the endophytic fungi of *Anoectochilus roxburghii* and the roots of *Anoectochilus roxburghii*, thereby effectively promoting root growth. This increases the number and intensity of the roots, which in turn facilitates nutrient uptake by the *Anoectochilus roxburghii* plant, resulting in faster growth. Detailed Implementation

[0034] To facilitate understanding of the present invention, a more comprehensive description will be provided below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of the present invention.

[0035] It should be noted that when an element is referred to as being "fixed to" another element, it can be directly attached to the other element or there may be an intervening element. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or there may be an intervening element. The terms "vertical," "horizontal," "left," "right," and similar expressions used herein are for illustrative purposes only and do not represent the only possible implementation.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0037] This application provides a method for promoting the root growth of *Anoectochilus roxburghii*. To better understand this method, the following further explanation is provided:

[0038] One embodiment of the method for promoting the root growth of Anoectochilus roxburghii involves applying a root growth promoter to the Anoectochilus roxburghii plant through root irrigation, so that the root growth promoter soaks into the roots of the Anoectochilus roxburghii plant; wherein, the root growth promoter is an endophytic fungal culture solution of Anoectochilus roxburghii.

[0039] The above-mentioned method for promoting the root growth of *Anoectochilus roxburghii* uses the endophytic fungal culture solution of *Anoectochilus roxburghii* as the root growth promoter. This solution is then applied via root irrigation to the roots of the *Anoectochilus roxburghii* plant, allowing the roots to be thoroughly soaked in the growth promoter. This ensures the growth promoter acts directly on the roots, promoting symbiosis between the endophytic fungi of *Anoectochilus roxburghii* and the roots of *Anoectochilus roxburghii*. This effectively promotes root growth, increasing the number and intensity of the roots, which in turn facilitates nutrient uptake by the plant, resulting in rapid growth.

[0040] In one embodiment, the orange-yellow jade phoenix flower was collected from the Xiangtoushan Nature Reserve. Further, the orange-yellow jade phoenix flower was collected from a location at 114°E, 23°N, and an altitude of 460m.

[0041] In one embodiment, the root irrigation of *Anoectochilus roxburghii* plants with a root growth promoter solution includes the following steps:

[0042] S110. Obtain *Anoectochilus roxburghii* plants. Further, the *Anoectochilus roxburghii* plants were tissue-cultured seedlings derived from wild *Anoectochilus roxburghii* species native to Longmen County, provided by Longmen County Hujinyuan Ecological Agriculture Development Co., Ltd.

[0043] S120. Cultivate the *Anoectochilus roxburghii* plants so that they are planted in the cultivation substrate.

[0044] S130. Drill holes in the cultivation substrate to create multiple holes, with each hole positioned close to the root system of the Anoectochilus roxburghii plant.

[0045] S140. Use Anoectochilus roxburghii root growth promoter to irrigate the boreholes so that the Anoectochilus roxburghii root growth promoter is poured back into the boreholes.

[0046] It's understandable that planting the obtained *Anoectochilus roxburghii* (golden thread orchid) plants in a cultivation substrate, and then pouring the root growth promoter into the drilled holes, allows the promoter to effectively contact and saturate the plant's roots. This promotes a symbiotic relationship between the endophytic fungi of *Anoectochilus roxburghii* and the *Anoectochilus roxburghii* roots. Alternatively, directly spraying or watering the plants would result in the promoter remaining largely in the soil. Since soil is not suitable for the survival and growth of all fungi, much of the promoter would be lost, hindering root growth. Conversely, increasing the amount of promoter would significantly increase the prevalence of *Anoectochilus roxburghii* endophytic fungi in the soil, potentially leading to pathogenic dominance and negatively impacting the plant's growth.

[0047] In one embodiment, after the step of irrigating the borehole with the root growth promoter of Anoectochilus roxburghii, the method for promoting the root growth of Anoectochilus roxburghii further includes the following step: filling the borehole so that the cultivation substrate fills the borehole, thereby further reducing the loss of the root growth promoter of Anoectochilus roxburghii.

[0048] In one embodiment, the cultivation substrate is peat moss, which is beneficial to the growth of Anoectochilus roxburghii plants.

[0049] In one embodiment, the amount of root growth promoter used is 5ml to 15ml. It is understood that when the root growth promoter is applied via root irrigation to fully saturate the roots of the *Anoectochilus roxburghii* plant, a dosage of 5ml to 15ml promotes root growth more effectively. However, if the dosage is too low, the endophytic fungi of *Anoectochilus roxburghii* are less dominant and less likely to form a symbiotic relationship with the *Anoectochilus roxburghii* roots, thus hindering root growth. Conversely, if the dosage is too high, the endophytic fungi of *Anoectochilus roxburghii* are more dominant, which can negatively impact the dominance of the native fungi in the *Anoectochilus roxburghii* roots, ultimately hindering root growth.

[0050] In one embodiment, the root growth promoter solution for Anoectochilus roxburghii is obtained through the following steps:

[0051] S210, Picking wild orange-yellow jade phoenix flowers;

[0052] S220. Treat the rhizomes of the orange-yellow jade phoenix flower to clean the surface of the rhizomes and obtain new nutrient roots that form mycorrhizae.

[0053] S230. Perform a scraping operation on the newly grown nutrient roots to extract the fungal clusters on the newly grown nutrient roots and obtain the Orange-Yellow Jade Phoenix Flower Fungal Community.

[0054] S240. The orange-yellow jade flower fungus was propagated and cultured to obtain a root growth promoter for Anoectochilus roxburghii.

[0055] It is understood that the endophytic fungi of *Anoectochilus roxburghii* are cultured together to form a root growth promoter for *Anoectochilus roxburghii*. At least one endophytic fungus from *Anoectochilus roxburghii* can effectively promote the root growth of *Anoectochilus roxburghii* plants. This application does not purify or isolate the specific species of endophytic fungi; it only aims to protect the endophytic fungi of *Anoectochilus roxburghii* so that they are cultured together to form a root growth promoter for *Anoectochilus roxburghii*. When applied to the roots of *Anoectochilus roxburghii* plants, it has a good effect on promoting root growth. It is worth mentioning that although the root growth promoter for *Anoectochilus roxburghii* contains multiple endophytic fungi, under controlled dosage, it will not affect the root growth of *Anoectochilus roxburghii* plants.

[0056] In one embodiment, the rhizome treatment of the orange-yellow jade phoenix flower specifically includes the following steps:

[0057] S310. The rhizomes of the orange-yellow jade phoenix flower are rinsed and cut to remove particulate matter from the surface of the rhizomes, resulting in multiple rhizome segments.

[0058] S320. Deeply clean the root and stem segments of the orange-yellow jade phoenix flower to remove bacteria and harmful substances from the surface of the root and stem segments.

[0059] S330. The root and stem segments of the orange-yellow jade phoenix flower after deep cleaning are cleaned to remove the liquid on the surface of the root and stem segments and obtain new nutrient roots.

[0060] It is understandable that by cleaning the surface of the roots and stems of the Orange-Yellow Jade Phoenix Flower separately to remove particulate matter, bacteria, and harmful substances, it is easier for the newly formed nutrient roots to obtain a high purity of the Orange-Yellow Jade Phoenix Flower's endophytic fungal community. This helps to reduce the impact of miscellaneous bacteria or other substances on the growth of the fungal community at the newly formed nutrient roots during the propagation and culture process.

[0061] In one embodiment, the rhizomes of the orange-yellow jade phoenix flower are rinsed and cut. Specifically, the rhizomes of the orange-yellow jade phoenix flower are rinsed under tap water, and then cut into segments to obtain rhizome segments. This effectively ensures the removal of particulate matter on the surface of the rhizomes of the orange-yellow jade phoenix flower, and the formation of rhizome segments is beneficial for scraping the mycelial masses of newly formed nutrient roots later.

[0062] In one embodiment, the length of the rootstock segment of the orange-yellow jade phoenix flower is 1cm to 2cm, which is beneficial for scraping out the mycelial mass of newly formed nutrient roots in the later stage.

[0063] In one embodiment, the root and stem segments of the orange-yellow jade phoenix flower were subjected to deep cleaning treatment. Specifically, the root and stem segments of the orange-yellow jade phoenix flower were soaked in sodium hypochlorite solution, and then taken out and soaked in ethanol solution, which effectively removed bacteria and harmful substances from the surface of the root and stem segments of the orange-yellow jade phoenix flower.

[0064] In one embodiment, the sodium hypochlorite solution has a mass percentage of 2.5%. Furthermore, soaking the rhizome segments of *Echeveria elegans* in the sodium hypochlorite solution for 0.5 to 1 minute further effectively removes bacteria and harmful substances from the surface of the rhizome segments.

[0065] In one embodiment, the ethanol solution has a mass percentage of 70%. Furthermore, immersing the rhizome segments of *Echeveria elegans* in the ethanol solution for 0.8 min to 1 min further effectively removes bacteria and harmful substances from the surface of the rhizome segments.

[0066] In one embodiment, the root and stem segments of the orange-yellow jade phoenix flower after deep cleaning are further cleaned by rinsing the root and stem segments of the orange-yellow jade phoenix flower with sterile water at least three times, and then using sterile absorbent paper to dry the liquid on the surface of the root and stem segments of the orange-yellow jade phoenix flower, which further reduces the residue of various reagents or bacteria on the surface of the orange-yellow jade phoenix flower.

[0067] In one embodiment, a mycelial clump scraping operation is performed on the newly formed nutrient roots. Specifically, the newly formed nutrient roots are placed in a culture dish with sterile water, and a single mycelial clump of the newly formed nutrient roots is scraped off using a sterilized inoculation needle, which effectively achieves the acquisition of endophytic fungi on the newly formed nutrient roots.

[0068] In one embodiment, before the step of propagating and culturing the *Anoectochilus roxburghii* microbial community and after the step of scraping the microbial community from the newly formed nutrient roots, the *Anoectochilus roxburghii* root growth promoter solution further includes the following step: selectively culturing the *Anoectochilus roxburghii* microbial community to inhibit the excessive growth of miscellaneous bacteria in the *Anoectochilus roxburghii* microbial community.

[0069] In one embodiment, selective culture of the *Anoectochilus roxburghii* fungal community was performed by adding caspofungin to the PDB medium. Further, the amount of caspofungin used was 0.05 g / L to 0.3 g / L. It is understood that since there are many types of endophytic fungi in *Anoectochilus roxburghii*, and at least one of these endophytic fungi promotes root growth in *Anoectochilus roxburghii*, caspofungin was used for selective proliferation culture of the endophytic fungi to better promote root growth in *Anoectochilus roxburghii*. Thus, the resulting *Anoectochilus roxburghii* root growth promoter had a better promoting effect on the root growth of *Anoectochilus roxburghii* plants.

[0070] In one embodiment, the preparation of PDB culture medium is specifically carried out as follows: 200g of peeled potatoes are boiled in boiling water for 30 minutes, filtered through gauze, 20g of glucose is added, water is added to make up to 1L, and sterilized at 121℃ for 30 minutes.

[0071] In one embodiment, the *Anoectochilus roxburghii* microbial community was proliferated and cultured. Specifically, the microbial community was inoculated into PDB medium, and then the PDB medium was placed in a shaker at a temperature of 25°C–28°C for cultivation. Further, the shaker speed was 110–120 rpm. Further, the PDB medium volume was 100–150 ml. Further, the PDB medium was cultured in a shaker at 25°C–28°C for at least 3 days. This effectively achieved rapid proliferation of the *Anoectochilus roxburghii* microbial community, reduced mutual interference among the microbial communities, and thus ensured the promoting effect of the *Anoectochilus roxburghii* root growth promoter solution formed after the *Anoectochilus roxburghii* microbial community proliferation culture on the root growth of *Anoectochilus roxburghii* plants. It is understandable that if the *Anoectochilus roxburghii* fungal community overgrows, it will affect the mutual interference among the fungal communities, thus affecting the effect of the *Anoectochilus roxburghii* root growth promoter on the root growth of the *Anoectochilus roxburghii* plant. If the *Anoectochilus roxburghii* fungal community does not grow sufficiently, the concentration of endophytic fungi of *Anoectochilus roxburghii* in the *Anoectochilus roxburghii* root growth promoter will be low, making it difficult to meet the root growth promotion needs of the *Anoectochilus roxburghii* plant.

[0072] In one embodiment, the PDB medium is placed in a shaker at a temperature of 25°C to 28°C for cultivation, and then the PDB medium is broken up and mixed so that the clumps of mycelium in the PDB medium are evenly dispersed in the PDB medium to obtain a root growth promoter for Anoectochilus roxburghii.

[0073] Compared with the prior art, the present invention has at least the following advantages:

[0074] The method for promoting root growth of *Anoectochilus roxburghii* of this invention uses an endophytic fungal culture solution of *Anoectochilus roxburghii* as a root growth promoter. This solution is then applied via root irrigation to the roots of the *Anoectochilus roxburghii* plant, allowing the roots to be thoroughly immersed in the root growth promoter. This ensures the root growth promoter acts directly on the roots, promoting symbiosis between the endophytic fungi of *Anoectochilus roxburghii* and the roots of *Anoectochilus roxburghii*, thereby effectively promoting root growth. This increases the number and intensity of the roots, which in turn facilitates nutrient uptake by the *Anoectochilus roxburghii* plant, resulting in faster growth.

[0075] The following are some specific examples. Where %, it refers to a percentage by weight. It should be noted that the following examples do not exhaustively cover all possible scenarios, and unless otherwise specified, the materials used in the following examples are commercially available.

[0076] Example 1

[0077] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0078] Preparation of PDB medium: Boil 200g of potatoes (peeled) in boiling water for 30 minutes, filter with gauze, add 20g of glucose, add water to make up to 1L, put it in an electric pressure steam sterilizer at 121℃ for 30 minutes, divide into Erlenmeyer flasks and set aside. The capacity of the Erlenmeyer flasks is 100ml.

[0079] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 0.5min. Then soak each sample plant in 70% ethanol solution for 0.8min. Wash with sterile water (rinse 3 times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 5 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0080] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0081] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up with sterilized pipettes and bamboo sticks and placed into PDB medium. They are then cultured for 3 days on a constant temperature shaker at 25°C and 110 r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors, and the mycelium and mycelium solution are evenly mixed with the liquid using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0082] Example 2

[0083] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0084] Preparation of PDB medium: Boil 200g of potatoes (peeled) in boiling water for 30 minutes, filter with gauze, add 20g of glucose, add water to make up to 1L, put it in an electric pressure steam sterilizer at 121℃ for 30 minutes, divide into Erlenmeyer flasks and set aside. The capacity of the Erlenmeyer flasks is 150ml.

[0085] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstocks of the orange-yellow jade phoenix flower, cut them with scissors, place them in a fixed cup, and transfer them to a clean bench. Cut the rootstocks of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 0.8min. Then soak each sample plant in 70% ethanol solution for 0.9min. Wash them with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse them with sterile water 6 times. Place them on sterile absorbent paper to dry, and obtain new nutrient roots.

[0086] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0087] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 4 days on a constant temperature shaker at 26℃ and 116r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors, and the mycelium and mycelium solution are evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0088] Example 3

[0089] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0090] Preparation of PDB medium: Boil 200g of potatoes (peeled) in boiling water for 30 minutes, filter with gauze, add 20g of glucose, add water to make up to 1L, put it in an electric pressure steam sterilizer at 121℃ for 30 minutes, divide into Erlenmeyer flasks and set aside. The capacity of the Erlenmeyer flasks is 150ml.

[0091] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 1 minute. Then soak each sample plant in 70% ethanol solution for 1 minute. Wash with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 6 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0092] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0093] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 5 days on a constant temperature shaker at 28℃ and 120r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors. The mycelium and mycelium solution are then evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0094] Example 4

[0095] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0096] Preparation of PDB medium: 200g of potato (peeled) was boiled in boiling water for 30 minutes, filtered through gauze, 20g of glucose was added, and 0.05g / L of caspofungin was added. Water was added to make up to 1L, and the mixture was placed in an electric pressure steam sterilizer at 121℃ for 30 minutes. The mixture was then divided into 150ml Erlenmeyer flasks for later use.

[0097] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 1 minute. Then soak each sample plant in 70% ethanol solution for 1 minute. Wash with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 6 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0098] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0099] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 5 days on a constant temperature shaker at 28℃ and 120r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors. The mycelium and mycelium solution are then evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0100] Example 5

[0101] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0102] Preparation of PDB medium: 200g of potatoes (peeled) were boiled in boiling water for 30 minutes, filtered through gauze, 20g of glucose and 0.1g / L of caspofungin were added, and water was added to make up to 1L. The mixture was then placed in an electric pressure steam sterilizer and sterilized at 121℃ for 30 minutes. The mixture was then divided into 150ml Erlenmeyer flasks for later use.

[0103] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 1 minute. Then soak each sample plant in 70% ethanol solution for 1 minute. Wash with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 6 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0104] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0105] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 5 days on a constant temperature shaker at 28℃ and 120r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors. The mycelium and mycelium solution are then evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0106] Example 6

[0107] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0108] Preparation of PDB medium: 200g of potatoes (peeled) were boiled in boiling water for 30 minutes, filtered through gauze, 20g of glucose and 0.2g / L of caspofungin were added, and water was added to make up to 1L. The mixture was then placed in an electric pressure steam sterilizer at 121℃ for 30 minutes and sterilized. The mixture was then divided into Erlenmeyer flasks with a capacity of 150ml for later use.

[0109] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 1 minute. Then soak each sample plant in 70% ethanol solution for 1 minute. Wash with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 6 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0110] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0111] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 5 days on a constant temperature shaker at 28℃ and 120r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors. The mycelium and mycelium solution are then evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0112] Example 7

[0113] Sampling: Orange-yellow jade phoenix flower, collected from Xiangtoushan Nature Reserve (114°E, 23°N, 460m altitude);

[0114] Preparation of PDB medium: 200g of potatoes (peeled) were boiled in boiling water for 30 minutes, filtered through gauze, 20g of glucose and 0.3g / L of caspofungin were added, and water was added to make up to 1L. The mixture was then placed in an electric pressure steam sterilizer and sterilized at 121℃ for 30 minutes. The mixture was then divided into 150ml Erlenmeyer flasks for later use.

[0115] Rinse the collected roots of the orange-yellow jade phoenix flower with running water to remove sand and other particles from the surface of the rootstock. Select the rootstock of the orange-yellow jade phoenix flower, cut it with scissors, place it in a fixed cup, and transfer it to an ultra-clean workbench. Cut the rootstock of the orange-yellow jade phoenix flower into small sections of 1.5±0.5cm each. Then soak them in 2.5% sodium hypochlorite solution for 1 minute. Then soak each sample plant in 70% ethanol solution for 1 minute. Wash with sterile water (rinse three times). Gently scrape the surface of the rootstock with a sterile inoculation needle. Rinse with sterile water 6 times. Place it on sterile absorbent paper to dry, and obtain the new nutrient roots.

[0116] Place the newly formed nutrient roots in a sterile water petri dish and scrape off a single mycelial mass from the newly formed nutrient roots using a sterilized inoculation needle.

[0117] On a clean bench, single mycelial clusters of newly formed nutrient roots are picked up into PDB medium using sterilized pipettes and bamboo sticks. The culture is carried out for 5 days on a constant temperature shaker at 28℃ and 120r / min. The mycelial balls in the PDB medium are then cut into pieces with sterilized pliers and scissors. The mycelium and mycelium solution are then evenly mixed using a vortex shaker to prepare a root growth promoter for Anoectochilus roxburghii.

[0118] The root growth promoters obtained in Examples 1 to 7 were used for the cultivation of Anoectochilus roxburghii plants.

[0119] 1. Materials:

[0120] 1.1 The Anoectochilus roxburghii plants with the same growth rate were tissue culture seedlings from wild Anoectochilus roxburghii in Longmen, provided by Longmen County Hujinyuan Ecological Agriculture Development Co., Ltd.

[0121] 1.2 Cultivation substrate: peat moss;

[0122] 2. Cultivation conditions:

[0123] The cultivation experiment of *Anoectochilus roxburghii* was conducted in a shaded greenhouse at the Hujin Garden cultivation base in Longmen County, Huizhou City. Located in a tropical monsoon climate zone, the area is situated at a low latitude and near the South China Sea, exhibiting typical maritime characteristics due to its marine influence. The average annual temperature is stable at 23℃, with almost no frost throughout the year for more than 350 days. The annual rainfall is approximately 1770 mm, concentrated between April and September.

[0124] 3. Test methods:

[0125] 3.1 A completely randomized design was adopted, with 10 tissue culture seedlings of Anoectochilus roxburghii forming one small group, and 3 small groups of tissue culture seedlings forming one large group, which were numbered in total, with the 8th large group serving as the blank control group;

[0126] 3.2 The dosage of the root growth promoter for Anoectochilus roxburghii was as follows:

[0127] The tissue culture seedlings of Anoectochilus roxburghii in groups 1 to 7 were respectively treated with the root growth promoting solution of Anoectochilus roxburghii in Examples 1 to 7;

[0128] For each of the 1st to 7th groups: the amount of root growth promoter used for the tissue culture seedlings of Anoectochilus roxburghii in the 1st group was 5ml; the amount of root growth promoter used for the tissue culture seedlings of Anoectochilus roxburghii in the 2nd group was 10ml; and the amount of root growth promoter used for the tissue culture seedlings of Anoectochilus roxburghii in the 3rd group was 15ml.

[0129] Group 8: Each group of Anoectochilus roxburghii tissue culture seedlings uses the same amount of sterile water as each group of Anoectochilus roxburghii tissue culture seedlings in the other groups.

[0130] 3.3 Make three holes in the cultivation substrate of each Anoectochilus roxburghii tissue culture seedling. All three holes are located around the root system of the Anoectochilus roxburghii tissue culture seedling. Pour the corresponding amount of Anoectochilus roxburghii root growth promoter for each Anoectochilus roxburghii tissue culture seedling into the corresponding hole. Then fill the hole. Collect the Anoectochilus roxburghii spider after it has grown for 150 cm.

[0131] 4. Measurement method:

[0132] The root length of each group of *Anoectochilus roxburghii* tissue culture seedlings was measured and recorded. The biomass of each group of *Anoectochilus roxburghii* plants was measured and recorded. The drying method was used, and the plants were dried at 105℃ for 2 hours until constant weight. After drying, the dry weight of the *Anoectochilus roxburghii* spiders was weighed, and the biomass and water content of the *Anoectochilus roxburghii* plants were calculated using formulas.

[0133] 5. Measurement Results:

[0134] 5.1 Table 1: Root length of *Anoectochilus roxburghii* tissue culture seedlings (unit: cm)

[0135]

[0136]

[0137] 5.2 Table 2: Root water content of *Anoectochilus roxburghii* tissue culture seedlings (unit: %)

[0138]

[0139]

[0140] Table 3.3: Content of anoectochilin in tissue culture seedlings of *Anoectochilus roxburghii* (unit: %); average value

[0141]

[0142]

[0143] Table 1 shows that the root length of *Anoectochilus roxburghii* tissue culture seedlings treated with the root growth promoter was significantly longer than that of seedlings not treated with the same promoter, indicating that the promoter had a positive effect on root growth. Table 2 shows that there was no significant difference in root water content between the seedlings treated with the promoter and those not treated, indicating that the promoter improved root water absorption and... There was no effect, and therefore no impact on the overall growth of *Anoectochilus roxburghii* tissue culture seedlings. As shown in Table 3, the content of *Anoectochilus roxburghii* glycosides in the tissue culture seedlings treated with the *Anoectochilus roxburghii* root growth promoter was significantly higher than that in the tissue culture seedlings not treated with the *Anoectochilus roxburghii* root growth promoter, indicating that the *Anoectochilus roxburghii* root growth promoter had a positive promoting effect on the content of *Anoectochilus roxburghii* glycosides in the tissue culture seedlings. Furthermore, as shown in Tables 1 and 2, the *Anoectochilus roxburghii* root growth promoters in Examples 4 to 7 had a more significant positive promoting effect on root growth and the increase in *Anoectochilus roxburghii* glycoside content.

[0144] The above embodiments merely illustrate several implementation methods of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A method for promoting the root growth of *Anoectochilus roxburghii*, characterized in that, The roots of *Anoectochilus roxburghii* plants were irrigated with a root growth promoter solution to allow the solution to soak the roots of the plants. The root growth promoter of Anoectochilus roxburghii is a culture solution of endophytic fungi of Anoectochilus roxburghii. The root growth promoter solution for Anoectochilus roxburghii is obtained through the following steps: Picking wild orange-yellow jade phoenix flowers; The rootstock of the orange-yellow jade phoenix flower is treated to clean the surface of the rootstock and obtain new nutrient roots that form mycorrhizae. The newly formed nutrient roots are scraped to extract the fungal colonies from them, thus obtaining the orange-yellow jade phoenix flower fungal community. The orange-yellow jade phoenix flower fungus was cultured to obtain the root growth promoter of the golden thread lotus; Before the step of propagating and culturing the *Anoectochilus roxburghii* fungal community, and after the step of scraping the fungal clusters from the newly formed nutrient roots, the *Anoectochilus roxburghii* root growth promoter solution also includes the following steps: selectively culturing the *Anoectochilus roxburghii* fungal community to inhibit the excessive growth of miscellaneous bacteria in the *Anoectochilus roxburghii* fungal community. Selective culture of the orange-yellow jade flower fungus was carried out by adding caspofungin to PDB medium at a concentration of 0.05 g / L to 0.3 g / L.

2. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 1, characterized in that, The root irrigation of *Anoectochilus roxburghii* plants with a root growth-promoting solution includes the following steps: Obtain the aforementioned Anoectochilus roxburghii plant; The *Anoectochilus roxburghii* plant is subjected to cultivation treatment so that it is planted in a cultivation substrate; The cultivation substrate is perforated to form multiple holes, each hole being positioned adjacent to the root system of the Anoectochilus roxburghii plant. The borehole is irrigated with the root growth promoting solution of *Anoectochilus roxburghii*, so that the root growth promoting solution is poured back into the borehole.

3. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 2, characterized in that, After the step of irrigating the drilled holes with the root growth-promoting solution of *Anoectochilus roxburghii*, the method for promoting the root growth of *Anoectochilus roxburghii* further includes the following steps: The drilled hole is filled so that the cultivation substrate fills the drilled hole.

4. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 2, characterized in that, The cultivation substrate is peat moss.

5. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 4, characterized in that, The root and stem treatment of the orange-yellow jade phoenix flower specifically includes the following steps: The rhizomes of the orange-yellow jade phoenix flower are washed and cut to remove particulate matter from the surface of the rhizomes, resulting in rhizome segments of the orange-yellow jade phoenix flower. The root and stem segments of the orange-yellow jade phoenix flower are subjected to deep cleaning treatment to remove bacteria and harmful substances from the surface of the root and stem segments; The orange-yellow jade phoenix flower root and stem segments, after deep cleaning, are further cleaned to remove the liquid from their surface, thus obtaining new nutrient roots.

6. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 5, characterized in that, The rhizomes of the orange-yellow jade phoenix flower are washed and cut. The specific operation is as follows: Rinse the rhizomes of the orange-yellow jade phoenix flower under tap water, then cut the rhizomes into sections to obtain orange-yellow jade phoenix flower rhizome sections.

7. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 5 or 6, characterized in that, The length of the orange-yellow jade phoenix flower rootstock segment is 1cm to 2cm.

8. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 5, characterized in that, The root and stem segments of the orange-yellow jade phoenix flower were subjected to deep cleaning treatment, specifically as follows: The root and stem segments of the orange-yellow jade phoenix flower were soaked in sodium hypochlorite solution, then taken out and soaked in ethanol solution.

9. The method for promoting root growth of *Anoectochilus roxburghii* according to claim 5, characterized in that, The orange-yellow jade phoenix flower root and stem segments after deep cleaning are cleaned by rinsing them with sterile water at least three times, and then using sterile absorbent paper to dry the liquid on the surface of the orange-yellow jade phoenix flower root and stem segments.

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