A treatment method for mimicking dry reverse stress and micro-stress interaction metabolism induction of roots of a medicinal plant of the skin class

Abstract

The present application relates to a kind of skin class woody medicinal plant root simulation dry reverse environment and micro stress interaction metabolic induction processing method, comprising the following steps: 1) selecting the skin class woody medicinal plant of root bark medicinal component content difference;2) root simulation dry environment treatment;3) short-term root bark strong stimulation simulation amplification effect treatment.The present application explores the measures of skin class woody medicinal plant to wild and environment cultivation in reverse simulation dry environment cultivation, provides the short-term and effective new cultivation method of stimulating medicinal component content increase for the guidance of skin class woody medicinal plant to wild and environment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of agricultural planting production, and particularly relates to a treatment method of root simulation dry reverse environment and micro-stress interaction metabolism induction for bark wood medicinal plants. BACKGROUND

[0002] In bark medicinal plants, different barks (branch bark, stem bark and root bark) are natural connected bodies with similar morphological structures but distributed in different parts. However, the Pharmacopoeia stipulates that different bark medicinal materials can only be taken from different parts. For example, Eucommia ulmoides Oliv. and Cinnamomum cassia Presl can only be taken from stem bark and branch bark, but root bark cannot be taken; Morus alba L. and Acanthopanax senticosus Harms can only be taken from root bark, but stem bark and branch bark cannot be taken; and Magnolia officinalis Rehd. et Wils. can be taken from stem bark, branch bark and root bark. For the first two types of bark medicinal materials, great waste of non-medicinal bark resources often occurs.

[0003] Eucommia ulmoides Oliv. and Magnolia officinalis Rehd. et Wils. are important bark wood medicinal plants in China. The main medicinal component of Eucommia ulmoides Oliv. is pinoresinol diglucoside (PDG), and the Pharmacopoeia stipulates that Eucommia ulmoides Oliv. can only be taken from stem bark, but root bark cannot be taken. It is found through determination that PDG is mainly accumulated in stem bark, and the content of PDG in root bark is extremely low. The main medicinal components of Magnolia officinalis Rehd. et Wils. are magnolol and honokiol (MN and HN), and the Pharmacopoeia stipulates that Magnolia officinalis Rehd. et Wils. can be taken from root bark, stem bark and branch bark, but the contents of medicinal components in different barks are different. It is known through determination that the content of medicinal components in root bark is higher than that in stem bark. In production, people generally select Eucommia ulmoides Oliv. or Magnolia officinalis Rehd. et Wils. of 10-15 years old for bark collection.

[0004] Traditional Chinese medicinal materials mainly take secondary metabolites as medicinal components, and the synthesis of secondary metabolites is mostly affected by environmental stress. The artificial cultivation of traditional Chinese medicinal materials follows the high-yield cultivation technical system of economic crops, excessively emphasizes economic yield index, and is mostly well managed in terms of fertilizer and water, and has low stress degree of "unfavorable environment", which often leads to the decrease of medicinal quality of medicinal materials or even failure to meet the standard. If the "wild cultivation" mode is completely adopted, or even "unfavorable environment" such as lack of fertilizer, drought, waterlogging, ice disaster and disease and insect pests is intentionally encountered in cultivation, the unit area yield is low, which cannot meet the market demand and is not in line with the mindset of maximizing benefits of planters.

[0005] In recent years, with the concept of "imitation environment cultivation" of Chinese herbal medicines, people have gradually explored various "imitation environment cultivation" measures for different Chinese herbal medicines, aiming to change the traditional cultivation techniques and measures through "imitation environment cultivation" to artificially create high-frequency low-intensity "micro-stress", so as to achieve the purpose of ensuring the medicinal quality of Chinese herbal medicines and taking into account the economic output. Yi Shanyong compared and analyzed the three planting modes of Dendrobium huoshanense, i.e. facility cultivation, under-forest cultivation and imitation environment cultivation, and found that the quality of imitation environment cultivation was better than that of facility and under-forest cultivation, with high yield and significant economic benefits (Yi Shanyong, Kang Chuanzhi, Wang Wei, Song Xiangwen, Xu Tao, Lu Haibo, Luo Shulan, Liu Dong, Guo Lanping, Han Bangxing. Comparison of Dendrobium huoshanense planting modes and advantage analysis of imitation environment cultivation [J]. Chinese Journal of Chinese Materia Medica, 2021, 46(08): 1864-1868. DOI: 10.19540 / j.cnki.cjcmm.20210225.101.). Liao Xianliang, Zhiyun, et al. found that the in vitro biological activity of n-butanol extract of wild-imitated D. officinale was significantly stronger than that of greenhouse cultivation (Liao Xianliang, Liang Zhiyun, Hu Li, et al. Comparison of in vitro biological activity of n-butanol extract of Dendrobium officinale cultivated in greenhouse and wild-imitated [J]. Chinese Pharmaceutical Industry, 2022, 31(24): 43-46.). Many studies have shown that "imitation environment cultivation" of Chinese herbal medicines can improve the quality of Chinese herbal medicines and increase economic output, which is the inevitable development direction of future artificial cultivation of Chinese herbal medicines.

[0006] However, most of the current research and invention patents on "imitation environment cultivation" are focused on the wild-imitated cultivation of herbaceous medicinal plants, such as the wild-imitated cultivation method of Phellinus baumii (Wu Guifen, Peng Jinyun, Lu Qihuang. Wild-imitated cultivation method of Phellinus baumii [P]. CN113016503B, 2023-04-18.), the planting method of wild-imitated Scutellaria baicalensis (Chen Caixia, He Chao, Li Xian'en, Wang Wenzhou. Planting method of wild-imitated Scutellaria baicalensis [P]. CN112715305B, 2022-12-20), and the wild-imitated cultivation technology of D. officinale under forest (Shu Jie. Application value of D. officinale and wild-imitated cultivation technology under forest [J]. Modern Agricultural Science and Technology, 2021(24): 61+69.). However, there is a lack of research and invention on the wild-imitated cultivation of bark wood medicinal plants, which are also important roles in Chinese herbal medicines. The exploration of "imitation environment cultivation" measures for bark wood medicinal plants has great economic effect, and does not need to go through the process of nutrient growth or reproductive growth like herbaceous medicinal plants, which can be directly treated on the basis of existing wood plants. Therefore, it is an important choice for exploring the "imitation environment cultivation" of Chinese herbal medicines. SUMMARY

[0007] The purpose of the present application is to explore whether the root simulation dry reverse habitat cultivation measures in the wild and the simulated environment of the bark of woody medicinal materials can induce the bark content to converge to the simulation site; and whether there is a new "micro stress" induced by the induction measures during the induction process, which has an interactive metabolic induction on the content change of the main and lateral root bark medicinal ingredients, and provides experimental theory for subsequent root dry reverse habitat cultivation of other bark woody medicinal plants. Furthermore, by artificially creating a low-frequency high-intensity "micro stress" which is a strong stimulation of the root bark, it is explored whether the short-term strong stimulation of the root bark can simulate the induction effect of the long-term root simulation dry reverse habitat cultivation measures, so as to provide a short-term and effective new cultivation method for stimulating the increase of medicinal ingredient content in the bark of woody medicinal plants.

[0008] The present application provides a treatment method for root simulation dry reverse habitat and micro stress interactive metabolic induction of bark woody medicinal plants, characterized in that it comprises the following steps:

[0009] 1) Selecting bark woody medicinal plants with significant differences in the content of medicinal ingredients in the root bark and the dry bark

[0010] For example, Eucommia ulmoides Oliv. with significantly lower content of medicinal ingredients in the root bark than in the dry bark, or Magnolia officinalis with significantly higher content of medicinal ingredients in the root bark than in the dry bark; preferably 10-20 year old Eucommia ulmoides Oliv. and Magnolia officinalis plants as treatment plants;

[0011] 2) Root simulation dry habitat treatment

[0012] Selecting the main and lateral roots (preferably the main roots or the developed lateral roots connected to the main roots) below the root stem of the treatment plant as the treatment range (preferably the main and lateral roots within 50 cm below the root stem surface and within 100 cm connected to the root stem), and selecting 30-60 cm long bark at any position in the treatment range for root simulation dry treatment;

[0013] The root simulation dry treatment method is as follows: the main and lateral roots within 50 cm below the root stem surface are dug open in a radial manner on the sunny side, exposing 30-100 cm of the obvious root part, the epidermis is required to be undamaged and connected to the main root, and the residual soil around the root part is brushed clean with a soft brush, and then the soil covering the main and lateral roots and the surrounding area due to wind, rain, etc. is removed regularly;

[0014] If necessary, select the adjacent untreated plants of the treatment plant as controls, and the root bark and dry bark are used as positive and blank controls, respectively;

[0015] Optionally, it further comprises:

[0016] 3) Short-term root bark strong stimulation simulation amplification effect treatment

[0017] The roots of the untreated plants of the same batch adjacent to the root simulation dry treatment plants are selected as the treatment range (preferably the main lateral roots within 50 cm below the root surface of the plant and within 100 cm connected to the rootstock) ; preferably, the treatment measures include the following steps: using root cutting and kinking treatment as short-term root bark strong stimulation treatment;

[0018] The root cutting and kinking treatment refers to the root membrane simulation dry treatment operation of step 2), and further strong stimulation treatment is performed on the exposed 30-100 cm obvious roots, respectively;

[0019] Further root cutting measures are as follows: the tail root is cut off completely or partially at the end of the treatment root away from the rootstock with a saw, a knife or a hoe, and a 5-10 cm long tail root end can be preferably retained and buried in the soil to prevent root rot, and the exposed part of the front end in the soil can be used for root bark strong stimulation sampling;

[0020] Further kinking treatment is as follows: a thin iron wire is wound around the exposed part of the treatment root in the soil, and the degree of winding is to press tightly without damaging the bark, and the root bark near the rootstock above the winding position is selected for kinking strong stimulation sampling.

[0021] Preferably, the 15-20 year old Eucommia ulmoides or Magnolia officinalis selected in step 1) is selected.

[0022] More preferably, the main lateral roots selected for treatment in step 2) need to be completely exposed to the outside during the entire induction process to simulate the growth environment of the main stem.

[0023] In the specific embodiment, the dry bark and root bark of the treatment plants and the control plants are sampled and determined for the content of the medicinal components at the appropriate sampling period (such as April to June every year for Eucommia ulmoides and Magnolia officinalis) and the specific treatment period in step 2).

[0024] More specifically, the main medicinal components specified in the Pharmacopoeia are determined in step 2), for example, the main medicinal components of Eucommia ulmoides are pinoresinol diglucoside (PDG), and the main medicinal components of Magnolia officinalis are magnolol and honokiol (MN and HN).

[0025] Preferably, the sampling length of the root cutting and kinking treatment in step 3) is 30 cm, and the width is 1 / 2 of the root circumference.

[0026] Further preferably, the untreated plants adjacent to the treated plants in step 3) are selected as blank controls, and the corresponding root simulation dry environment treatment is selected to evaluate the short-term root bark strong stimulation simulation amplification effect.

[0027] The application provides the use of the treatment method for guiding the wild-like and environment simulation cultivation method of bark-type woody medicinal plants.

[0028] Specifically, it is used for the cultivation method for improving the content quality of medicinal ingredients of bark-type woody medicinal plants.

[0029] Advantages and beneficial effects of the present application:

[0030] The purpose of the present application is to explore whether the root bark content converges to the simulated part in the process of root simulation dry reverse habitat cultivation in bark-type woody medicinal plants; and to explore whether there is a new "micro stress" induced by the induction measures during the induction process, which has an interactive metabolic induction on the content change of the medicinal ingredients of the main and lateral root bark, and to provide experimental theory for subsequent root dry reverse habitat cultivation of other bark-type woody medicinal plants. Furthermore, by artificially creating a low-frequency high-intensity "micro stress" (root bark strong stimulation), it is explored whether short-term root bark strong stimulation can simulate the induction effect of long-term root simulation dry reverse habitat cultivation measures, so as to provide a short-term and effective new cultivation method for stimulating the increase of medicinal ingredient content in bark-type woody medicinal plants. BRIEF DESCRIPTION OF DRAWINGS

[0031] The present application will be further described below in conjunction with the drawings and examples.

[0032] Figure 1 Schematic diagram of dry treatment of Eucommia ulmoides root;

[0033] Figure 2 Schematic diagram of dry treatment of Magnolia officinalis root;

[0034] Figure 3 Schematic diagram of strong stimulation treatment of Magnolia officinalis root;

[0035] Figure 4 Schematic diagram of strong stimulation treatment of Magnolia officinalis root. DETAILED DESCRIPTION

[0036] In order to facilitate the understanding of the present application, the following embodiments of the present application are given. However, the present application can be realized in many different forms, and is not limited to the embodiments described herein. On the contrary, the purpose of providing the embodiments is to make the disclosure of the present application more thorough and comprehensive.

[0037] 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 application belongs. The terminology used in the specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application.

[0038] Example 1:

[0039] In the longan base of Hunan Agricultural University in Hunan Province, 15-year-old, consistent habitat, and the same growth of many eucommia plants were selected as the same batch of treatment plants for root simulation dry treatment. In December of the previous year, the main lateral roots within 50 cm below the surface of the eucommia plant rhizome were dug up to the sun side soil in a radial pattern, exposing the obvious root part 60 cm, the requirement was that the epidermis was not damaged and connected with the main root, and the residual soil around the root part was brushed clean with a soft brush (such as Figure 1 );

[0040] During the whole year growth period (from December of the previous year to December of the next year), the treated eucommia plant samples were selected for treatment period (treatment 0 months), germination period (treatment 3 months), suitable harvesting period (treatment 5 months), high temperature period (treatment 8 months), and cold period (treatment 12 months). The exposed root bark of the treated plants, the root bark of the untreated plants (control group), and the blank group root bark (blank treatment root bark) were harvested. The dynamic determination of the content of the medicinal ingredients was carried out by high performance liquid chromatography.

[0041] The results of the eucommia root simulation dry treatment are as follows: in the germination period, the contents were 282.58% of the blank group and 356.74% of the control group, respectively; in the suitable harvesting period, the contents were 518.07% of the blank group and 268.63% of the control group, respectively; in the high temperature period, the contents were 180.37% of the blank group and 123.81% of the control group, respectively; and in the cold period, the contents were 606.77% of the blank group and 161.08% of the control group, respectively.

[0042] The results of this example show that eucommia root simulation dry has the effect of inducing changes in the content of root bark medicinal ingredients.

[0043] Example 2

[0044] In the longan base of Hunan Agricultural University in Hunan Province, 15-year-old, consistent habitat, and the same growth of many eucommia plants were selected as the same batch of treatment plants for root simulation dry treatment. In December of the previous year, the main lateral roots within 50 cm below the surface of the eucommia plant rhizome were dug up to the sun side soil in a radial pattern, exposing the obvious root part 60 cm, the requirement was that the epidermis was not damaged and connected with the main root, and the residual soil around the root part was brushed clean with a soft brush (such as Figure 2 );

[0045] During the whole year growth period (from December of the previous year to December of the next year), the treated eucommia plant samples were selected for treatment period (treatment 0 months), germination period (treatment 3 months), suitable harvesting period (treatment 5 months), high temperature period (treatment 8 months), and cold period (treatment 12 months). The exposed root bark of the treated plants, the root bark of the untreated plants (control group), and the blank group root bark (blank treatment root bark) were harvested. The dynamic determination of the content of the medicinal ingredients was carried out by high performance liquid chromatography.

[0046] The MN and HN contents of the root bark of the control were 12.83%, which were 1.83 and 2.27 times of the MN and HN contents of the 0-0.5 m dry bark and 1-1.5 m dry bark, respectively. The MN and HN contents of the root bark of the root simulation dry treatment were 14.37%, which were 1.54% higher than the MN and HN contents of the control. The MN and HN contents of the root bark of the root simulation dry treatment were 2.05 and 2.55 times of the MN and HN contents of the 0-0.5 m dry bark and 1-1.5 m dry bark, respectively. Since the medicinal ingredient content of Magnolia officinalis is higher in the root bark than in the dry bark, theoretically, the root simulation dry treatment should induce a decrease in the content of the root bark. However, the actual result was an increase in the content of the root bark, which indicated that a "micro stress" caused by the induction measure existed in the induction process, and the "micro stress" induced the metabolism of the root bark, resulting in an increase in the content of the root bark.

[0047] The results of this example show that the root simulation dry treatment of Magnolia officinalis has an effect of inducing changes in the content of the medicinal ingredient of the root bark, and a new "micro stress" caused by the induction measure exists in the induction process, which induces the metabolism of the medicinal ingredient of the root bark.

[0048] Example 3

[0049] In the Magnolia officinalis forest in Chang'an base of Hunan Agricultural University in Hunan Province, 15-year-old Magnolia officinalis plants with consistent habitats and the same growth vigor were selected as the same batch of treatment plants, and the following two kinds of root bark strong stimulation treatments were performed in April of the same year.

[0050] The soil of the main lateral roots of Magnolia officinalis plants within 50 cm below the ground surface was dug open in a radial manner, and the obvious root part of 60 cm was exposed. The tail roots 30 cm away from the root stem end were completely cut off with a saw, and the 5 cm long tail root end was buried in the soil. The front end exposed to the soil part could be used for strong stimulation sampling (such as Figure 3 ) after the root was cut off.

[0051] The soil of the main lateral roots of Magnolia officinalis plants within 50 cm below the ground surface was dug open in a radial manner, and the obvious root part of 100 cm was exposed. The root part exposed to the soil was tightly pressed and not damaged by winding with fine iron wire to the extent of the treatment. The root bark close to the root stem above the winding position was selected for strong stimulation sampling (such as Figure 4 ) after the treatment.

[0052] The root bark of the treatment plants after 1 month (the appropriate harvest period), 2 months, and 3 months, the root bark of the untreated adjacent plants, and the dry bark (blank root bark, dry bark at 0-0.5 m above the ground surface of the root stem) were selected for dynamic determination of the content of the medicinal ingredient by high performance liquid chromatography. The results are as follows:

[0053] The total amount of MN and HN of the root bark of the broken root treatment plant was 13.97%, 12.86%, and 11.21% respectively; the total amount of MN and HN of the root bark of the twisted root treatment plant was 14.01%, 13.34%, and 11.05% respectively; the total amount of MN and HN of the root bark of the untreated adjacent plant was 11.73%, 12.40%, and 12.83% respectively, and the total amount of MN and HN of the root bark of the untreated adjacent plant stem was 5.54%, 6.70%, and 7.01% respectively;

[0054] It is found by comparison that the total amount of MN and HN of the root bark of the broken root and twisted root treatment groups is significantly increased by 2.24% and 2.28% respectively compared with the control root bark after one month of treatment; and the total amount of MN and HN of the root bark of the root simulation stem treatment plant is increased by 1.54% compared with the untreated plant after 11 months of treatment;

[0055] This example shows that short-term root bark strong stimulation treatment such as broken root and twisted root for one month can induce the content of the medicinal components of the main and lateral root bark to change to the main stem bark of the simulation part, that is, short-term stem bark strong stimulation can simulate the induction effect of the long-term stem simulation root reverse situation cultivation measures.

[0056] The embodiments described in the present application are only used to describe the embodiments of the present application, and do not limit the concept and scope of the present application. Without departing from the design idea of the present application, various modifications and improvements of the technical solutions of the present application made by the engineering technicians in the field shall fall within the protection scope of the present application. The technical content of the present application which is requested to be protected has been entirely recorded in the claims.

Claims

1. A processing method of barks of medicinal plants of the dicotyledonous woody plants under simulated dry stress and reverse environment and interaction of metabolic induction of micro-stress, characterized in that, Includes the following steps: 1) Select woody medicinal plants with differences in the content of medicinal components in their roots, trunks, and bark. Eucommia ulmoides with significantly lower content of medicinal components in root bark than in dried bark, or Magnolia officinalis with significantly higher content of medicinal components in root bark than in dried bark; Eucommia ulmoides and Magnolia officinalis plants aged 10-20 years were selected as treatment plants. 2) Root simulation of the environment Select the main and lateral roots below the ground surface of the plant's rhizome for treatment. Specifically, select the main and lateral roots within 50cm below the ground surface of the plant's rhizome and within 100cm of the main and lateral roots connected to the rhizome as the treatment range. Within the treatment range, select a 30-60cm long section of the bark at any location for root-simulated stem treatment. The root simulated stem treatment method is as follows: Dig out the soil in a radial pattern within 50cm below the surface of the plant's rootstock on the sunny side of the main and lateral roots, exposing 30-100cm of the obvious root. The epidermis should not be damaged and should be connected to the main root. Then, clean the remaining soil around the roots with a soft brush. Afterward, regularly remove the soil covering the main and lateral roots and the surrounding soil due to wind and rain. Untreated plants adjacent to the treated plants were selected as controls, with their root bark and bark serving as positive and blank controls, respectively. In step 2), the selected primary and lateral roots must be fully exposed to the outside world throughout the entire induction process to simulate the growth environment of the main stem. Also includes: 3) Treatment to simulate the amplification effect of short-term strong root bark stimulation The treatment area is selected from the main and lateral roots below the ground surface of the rootstock of untreated plants from the same batch adjacent to the rootstock of the simulated stem treatment plant. The main and lateral roots connected to the main root are selected within 50cm below the ground surface and within 100cm of the rootstock. The treatment measures include the following steps: root cutting and strangulation are used as short-term strong root bark stimulation treatment. The root pruning and strangulation treatments refer to the root simulated trunk treatment operation in step 2), except that strong stimulation treatment is further applied to the exposed 30-100cm obvious roots. Further root pruning measures are as follows: Cut off the tail root completely or partially with a saw, machete or hoe at the end of the root far from the rhizome, leaving 5-10 cm of the tail root end buried in the soil to prevent root rot, and use the part of the front end exposed in the soil for strong root pruning stimulation sampling. Further strangulation treatment is as follows: Use thin iron wire to wrap around the exposed soil part of the treated root, wrapping tightly without damaging the bark. After treatment, select the root bark above the wrapping position near the rhizome for strong strangulation sampling. In step 3), the root interruption and strangulation treatment sampling length is 30cm and the width is 1 / 2 of the root circumference.

2. The method for inducing metabolic interaction between root simulated stem and micro-stress in bark-type woody medicinal plants according to claim 1, characterized in that, In step 1), select 15-20 year old Eucommia ulmoides or Magnolia officinalis plants.

3. The method for inducing metabolic interaction between root simulated stem and micro-stress in the roots of bark-type woody medicinal plants according to claim 1, characterized in that, In step 3), untreated plants adjacent to the treated plants are selected as blank controls, and corresponding root simulated dry environment treatments are selected to evaluate the short-term root bark strong stimulation simulation amplification effect.

4. The application of the processing method as described in any one of claims 1 to 3, which is used to guide the simulated wild and simulated environment cultivation of bark-type woody medicinal plants.

5. The use according to claim 4, wherein the compound is ###0002### Cultivation methods for improving the content and quality of medicinal components in bark-type woody medicinal plants.

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