阅读说明：本技术 一种两面针组织培养快速繁殖方法 (Rapid propagation method for radix zanthoxyli tissue culture ) 是由 王继华 蔡时可 梅瑜 徐世强 周芳 顾艳 孙铭阳 李静宇 于 2020-11-23 设计创作，主要内容包括：本发明属于植物快速繁殖技术领域,具体涉及一种两面针组织培养快速繁殖方法,包括以下步骤：(1)外植体选取与消毒：(2)培养基的制备：制备改良MS培养基,然后在改良MS培养基的基础上分别制备不定芽诱导培养基、继代增殖培养基和无菌生根培养基；(3)不定芽诱导培养：将消毒外植体的芽接种于不定芽诱导培养基中进行诱导培养,得到两面针不定芽；(4)继代增殖；(5)无菌生根。本发明两面针组织培养快速繁殖方法,可以快速培养出两面针种苗,能保持母本的优良性状；能有效提高繁殖系数和成苗率,且培养周期短,并且能定向培育单一性别植株,便于标准化种植,种植效益大大提高。(The invention belongs to the technical field of plant rapid propagation, and particularly relates to a method for culturing and rapidly propagating zanthoxylum nitidum tissues, which comprises the following steps: (1) selecting and disinfecting explants: (2) preparation of a culture medium: preparing an improved MS culture medium, and then respectively preparing an adventitious bud induction culture medium, a subculture multiplication culture medium and a sterile rooting culture medium on the basis of the improved MS culture medium; (3) adventitious bud induction culture: carrying out induction culture on the bud of the sterilized explant in an adventitious bud induction culture medium to obtain adventitious buds of the Zanthoxylum nitidum; (4) subculturing and proliferating; (5) and (5) sterile rooting. The method for rapid propagation of zanthoxylum nitidum tissue culture can rapidly culture zanthoxylum nitidum seedlings and can keep the excellent characters of female parents; the method can effectively improve the propagation coefficient and the seedling rate, has short culture period, can directionally culture single-sex plants, is convenient for standardized planting, and greatly improves the planting benefit.)

1. A method for culturing and rapidly propagating zanthoxylum nitidum tissues comprises the following steps:

(1) selecting and disinfecting explants: selecting tender stems of Zanthoxylum nitidum as explants to carry out disinfection treatment to obtain disinfected explants;

(2) preparation of a culture medium: preparing an improved MS culture medium, and then respectively preparing an adventitious bud induction culture medium, a subculture multiplication culture medium and a sterile rooting culture medium on the basis of the improved MS culture medium; the formula of the improved MS culture medium is that the dosage of ammonium nitrate in the MS culture medium is reduced to 550-825mg/L, and other components are unchanged;

(3) adventitious bud induction culture: carrying out induction culture on the bud of the sterilized explant in an adventitious bud induction culture medium to obtain adventitious buds of the Zanthoxylum nitidum;

(4) subculture proliferation: inoculating adventitious buds of Zanthoxylum nitidum into a subculture multiplication medium for subculture multiplication to obtain Zanthoxylum nitidum seedlings;

(5) sterile rooting: inoculating the Zanthoxylum nitidum seedlings into a sterile rooting culture medium for rooting culture to obtain Zanthoxylum nitidum seedlings, and then hardening and transplanting.

2. The nitidissimilis tissue culture rapid propagation method according to claim 1, wherein the adventitious bud induction medium is prepared by adding 6-BA and IAA on the modified MS medium; the concentration of 6-BA in the adventitious bud induction culture medium is 1-3mg/L, and the concentration of IAA is 0.1-0.5 mg/L.

3. The method for rapid propagation in nitidissimilis tissue culture according to claim 2, wherein the concentration of 6-BA in the culture medium for adventitious bud induction is 2mg/L and the concentration of IAA is 0.5 mg/L.

4. The method for rapid propagation of Zanthoxylum nitidum tissue culture according to claim 2, wherein the inducing culture conditions are temperature of 28 ± 2 ℃, relative humidity of 85 ± 10%, illumination time of 12 hours/day, illumination intensity of 600-; the culture time is 4-6 weeks.

5. The nitidissimilis tissue culture rapid propagation method according to claim 1 or 2, wherein the secondary multiplication culture medium is prepared by adding 6-BA and IAA on the modified MS culture medium; the concentration of 6-BA in the subculture multiplication medium is 1-3mg/L, and the concentration of IAA is 0.1-0.2 mg/L.

6. The method for rapid propagation in the culture of Zanthoxylum nitidum tissue according to claim 5, wherein the concentration of 6-BA in the culture medium for subculture propagation is 2mg/L and the concentration of IAA is 0.2 mg/L.

7. The method for rapid propagation of Zanthoxylum nitidum tissue culture according to claim 5, wherein the subculture condition is temperature of 28 ± 2 ℃, humidity of 85 ± 10%, illumination time of 12 hours/day, illumination intensity of 600-; the incubation time was 4 weeks.

8. The Zanthoxylum nitidum tissue culture rapid propagation method according to claim 1 or 7, wherein the sterile rooting medium is prepared by adding NAA and IBA on the basis of the modified MS medium, the concentration of NAA in the rooting medium is 0.5-1mg/L, and the concentration of IBA in the rooting medium is 0.2-0.5 mg/L.

9. The method for rapid propagation of Zanthoxylum nitidum in tissue culture according to claim 8, wherein the concentration of NAA in the culture medium for sterile rooting is 1mg/L and the concentration of IBA is 0.5 mg/L.

10. The method for rapid propagation of Zanthoxylum nitidum tissue culture according to claim 8, wherein the rooting culture conditions are temperature of 28 ± 2 ℃, humidity of 85 ± 10%, illumination time of 10-15 hours/day, illumination intensity of 600-; the culture time is 30-50 days.

Technical Field

The invention belongs to the technical field of plant rapid propagation, and particularly relates to a rapid propagation method for radix zanthoxyli tissue culture.

Background

Radix zanthoxyli (Zanthoxylum nitidum (Roxb.) DC.) belongs to xylem vine plants in the genus of Zanthoxylum of Rutaceae, which are called Acanthopanax trifoliatus, radix zanthoxyli, and the like, and the radix zanthoxyli is a traditional Chinese medicinal material in China, has the effects of promoting blood circulation, removing blood stasis, dispelling wind, relieving swelling and pain, and the like, and is clinically used for treating traumatic injury, rheumatic arthralgia, stomachache, toothache, venomous snake bite, and the like; it can be used for treating scald due to hot liquid or fire for external application. It is mainly distributed in south China, Guangxi, Guangdong, Yunnan, Fujian and Taiwan, and mostly grows in shrubs in mountain and sloping fields. Through the examination of the materia medica of the Zanthoxylum nitidum by scholars at home and abroad, the important Chinese herbal medicine works and the legal standards are both recorded in Lingnan medicine-taking records, Shennong materia medica Jing, Chinese pharmacopoeia and the like. The radix zanthoxyli has complex chemical components, mainly contains compounds such as alkaloids, lignans, flavonoids, sterols and the like, wherein the monomer effective components such as the radix zanthoxyli have the activities of spasmolysis, analgesia, anti-inflammation, antibiosis, anticancer and the like, are used for spasmolysis, analgesia, and develop and produce medicaments such as bony spur tablets, traumatic kaleidoscope oil and the like and daily necessities such as radix zanthoxyli traditional Chinese medicine toothpaste and the like. The market has great demand on the zanthoxylum nitidum, natural excavation is mainly used for a long time, great pressure and damage are brought to wild zanthoxylum nitidum resources, and artificial domestication planting of the zanthoxylum nitidum is urgently needed. However, breeding and seedling breeding of zanthoxylum nitidum are relatively backward at present, wild seeds are mostly directly collected for seedling cultivation and planting in production, and due to the fact that wild zanthoxylum nitidum germplasm resources are high in genetic diversity and large in variation coefficient in a population, offspring are often uneven. The male plant of radix zanthoxyli is rather loved by growers in production, but the sex can not be judged in the seedling stage in seed propagation, and the later-stage standardized planting and quality guarantee are seriously influenced.

The plant tissue culture technology can utilize the asexual rapid propagation of lateral buds, stem segments and the like, retain the excellent characters of female parents, cultivate a large number of seedlings with consistent growth in scale, and relieve the problems of low seedling propagation rate and incapability of directionally cultivating male plants in the traditional production.

Disclosure of Invention

The invention aims to provide a rapid propagation method for the tissue culture of Zanthoxylum nitidum, which aims to solve the defects of low propagation coefficient and long culture period of the existing Zanthoxylum nitidum planting technology.

The purpose of the invention is realized by the following technical scheme:

a method for culturing and rapidly propagating zanthoxylum nitidum tissues comprises the following steps:

(1) selecting and disinfecting explants: selecting tender stems of Zanthoxylum nitidum as explants to carry out disinfection treatment to obtain disinfected explants;

(2) preparation of a culture medium: preparing an improved MS culture medium, and then respectively preparing an adventitious bud induction culture medium, a subculture multiplication culture medium and a sterile rooting culture medium on the basis of the improved MS culture medium; the formula of the improved MS culture medium is that the dosage of ammonium nitrate in the MS culture medium is reduced to 550-825mg/L, and other components are unchanged;

(3) adventitious bud induction culture: carrying out induction culture on the bud of the sterilized explant in an adventitious bud induction culture medium to obtain adventitious buds of the Zanthoxylum nitidum;

(4) subculture proliferation: inoculating adventitious buds of Zanthoxylum nitidum into a subculture multiplication medium for subculture multiplication to obtain Zanthoxylum nitidum seedlings;

(5) sterile rooting: inoculating the Zanthoxylum nitidum seedlings into a sterile rooting culture medium for rooting culture to obtain Zanthoxylum nitidum seedlings, and then hardening and transplanting.

The invention relates to a radix zanthoxyli tissue culture rapid propagation method, which adopts a system to breed excellent single plants of the radix zanthoxyli, and utilizes a tissue culture technology to carry out industrialized seedling culture, thereby not only providing a large amount of high-quality seedlings in a short time, relieving the damage of the current market to wild resources by transitional excavation, but also providing uniform seedlings to planters and ensuring annual market supply of the seedlings.

In the invention, the disinfection treatment is to soak the materials in alcohol and then put mercury bichloride solution containing Tween-20 for disinfection.

Further, the alcohol soaking is to soak for 10 to 30 seconds by adopting 70 percent alcohol, and then to wash by using sterile water; the mercuric chloride solution is sterilized by 0.1-0.3 percent of mercuric chloride solution containing 0.01 percent of Tween-20 in percentage by mass for 15min and then washed by sterile water.

The invention can be improved as follows, the prescription of the adventitious bud induction culture medium is that 6-BA and IAA are added on the improved MS culture medium; the concentration of 6-BA in the adventitious bud induction culture medium is 1-3mg/L, and the concentration of IAA is 0.1-0.5 mg/L.

Preferably, the concentration of 6-BA in the medium for adventitious bud induction is 2mg/L and the concentration of IAA is 0.5 mg/L.

In the present invention, the bud of the sterilized explant is an axillary bud of a bisurface stem segment.

In the invention, the induction culture conditions are that the temperature is 28 +/-2 ℃, the relative humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 600-; the culture time is 4-6 weeks.

The invention can be further improved in the following way, the formulation of the subculture multiplication medium is that 6-BA and IAA are added on the improved MS medium; the concentration of 6-BA in the subculture multiplication medium is 1-3mg/L, and the concentration of IAA is 0.1-0.2 mg/L.

Preferably, the concentration of 6-BA in the medium for subculture proliferation is 2mg/L and the concentration of IAA is 0.2 mg/L.

In the invention, the subculture proliferation conditions comprise a temperature of 28 +/-2 ℃, a humidity of 85 +/-10%, an illumination time of 12 hours/day and an illumination intensity of 600-10001 x; the incubation time was 4 weeks.

The invention can be further improved, the formulation of the sterile rooting medium is that NAA and IBA are added on the basis of the improved MS medium, the concentration of the NAA in the rooting medium is 0.5-1mg/L, and the concentration of the IBA in the rooting medium is 0.2-0.5 mg/L.

Preferably, the concentration of NAA in the medium for sterile rooting is 1mg/L and the concentration of IBA is 0.5 mg/L.

In the invention, the rooting culture conditions are that the temperature is 28 +/-2 ℃, the humidity is 85 +/-10%, the illumination time is 10-15 hours/day, and the illumination intensity is 600-; the culture time is 30-50 days.

Compared with the prior art, the invention has the following beneficial effects:

(1) the method for rapid propagation of zanthoxylum nitidum tissue culture can rapidly culture zanthoxylum nitidum seedlings and can keep the excellent characters of female parents; the method can effectively improve the propagation coefficient and the seedling rate, has short culture period, can directionally culture single-sex plants, is convenient for standardized planting, greatly improves the planting benefit, and can effectively solve the problem of serious insufficient supply of the current Zanthoxylum nitidum.

(2) According to the breeding and growth characteristics of the Zanthoxylum nitidum seedlings, a culture medium for adventitious bud induction, a culture medium for subculture multiplication and a culture medium for sterile rooting which are suitable for Zanthoxylum nitidum tissue culture are prepared on the basis of an MS culture medium.

(3) According to the method for culturing and rapidly propagating the Zanthoxylum nitidum, the produced male Zanthoxylum nitidum seedlings grow neatly and consistently, later-period management is facilitated, the production cost is reduced, the consistency of the seedlings is ensured, the income of planting and sellers is increased, and the development of the Zanthoxylum nitidum industry is promoted.

Drawings

FIG. 1 shows a Zanthoxylum nitidum tissue culture seedling cultured in example 1 of the present invention;

FIG. 2 is a graph showing the effect of different concentrations of 6-BA on the growth factor of Zanthoxylum nitidum in the present invention;

FIG. 3 is a graph comparing the effect of MS medium and modified MS culture of the present invention on plant growth.

Detailed Description

The present invention is further described below in conjunction with specific examples to better understand and implement the technical solutions of the present invention for those skilled in the art.

Example 1

A method for culturing and rapidly propagating zanthoxylum nitidum tissues comprises the following steps:

(1) selecting and disinfecting explants:

selecting young branches of male Zanthoxylum nitidum which grows healthily and has no plant diseases and insect pests as explants;

removing redundant leaves and thorns, cutting into 1-2cm stem sections, washing dirt on the surfaces of the branches with tap water, and then sucking with a paper towel;

thirdly, soaking the workpiece on a super-clean workbench for 15s by using 70 percent alcohol, and then washing the workpiece for 5min by using sterile water;

fourthly, sterilizing the explant for 10min by using 0.2 mass percent mercuric chloride solution containing 0.01 mass percent of Tween-20, and finally washing the sterilized explant for 3 times with sterile water, wherein each time lasts for 5min to obtain a sterilized explant;

(2) preparation of improved MS culture medium:

the formula of the improved MS culture medium is as follows: the dosage of ammonium nitrate is reduced to 825mg/L, and other components are unchanged.

(3) Obtaining a sterile explant and inducing adventitious buds:

cutting the disinfected explant obtained in the step (1) on a superclean workbench by using a sterile scalpel blade to cut the part of the two ends of the disinfected explant, wherein the part of the two ends of the disinfected explant is contacted with a mercuric chloride solution, inserting the lower biological end of the axillary bud of the stem into a culture medium for inducing the adventitious bud according to the direction that the top biological end is upward, placing the axillary bud in an artificial climate box for culturing for 6 weeks, and inducing to obtain adventitious buds of the Zanthoxylum nitidum; the induction culture conditions were as follows: the temperature is 28 +/-2 ℃, the relative humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 7001 x; the formula of the culture medium for adventitious bud induction is as follows: adding 6-BA and IAA on the modified MS culture medium; wherein the concentration of 6-BA is 2mg/L, and the concentration of IAA is 0.5 mg/L;

(4) subculture proliferation:

cutting the adventitious buds of the Zanthoxylum nitidum obtained in the step (3) from the explant, cutting off leaves, inoculating a subculture medium, and performing subculture for 4 weeks to obtain cluster buds of the Zanthoxylum nitidum; the subculture conditions were as follows: the temperature is 28 +/-2 ℃, the humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 8001 x; the formula of the culture medium for subculture proliferation is as follows: adding 6-BA and IAA on the basis of the improved MS culture medium, wherein the concentration of the 6-BA is 2mg/L, and the concentration of the IAA is 0.2 mg/L;

(5) sterile rooting:

and (4) when the zanthoxylum nitidum obtained in the step (4) grows to 2-3cm high from the seedling, carrying out induced rooting. Cutting off the single plant, inoculating the single plant into a rooting culture medium, and performing rooting culture for 40 days to obtain the Zanthoxylum nitidum seedlings; the rooting culture conditions were as follows: the culture temperature is 28 +/-2 ℃, the humidity is 85 +/-10 percent, the illumination time is 13 hours/day, and the illumination intensity is 7001 x; the formula of the culture medium for sterile rooting is as follows: adding NAA and IBA on the basis of the improved MS culture medium, wherein the concentrations are 1mg/L and 0.5mg/L respectively;

after the Zanthoxylum nitidum seedlings grow to the root length of 2-4cm and the height of 5-8cm in the step (5), opening the bottle caps of the culture bottles shown in figure 1, hardening and culturing indoors for 6 days, then cleaning the root culture medium, planting the seedlings in seedling bags with the diameter of 5cm and filled with peat soil, placing the seedlings in a plastic greenhouse, keeping the substrate wet, and transplanting the seedlings to a field after the plants survive.

Example 2

A method for culturing and rapidly propagating zanthoxylum nitidum tissues comprises the following steps:

(1) selecting and disinfecting explants:

selecting young stems of healthy zanthoxylum nitidum as explants;

removing redundant leaves, washing dirt on the surfaces of the branches with tap water, and then sucking the branches with paper towels;

thirdly, soaking the workpiece on a clean bench for 25s by using 70 percent alcohol, and then washing the workpiece for 1 time and 3min by using sterile water;

fourthly, sterilizing for 15min by using 0.3 mass percent mercuric chloride solution containing 0.01 mass percent of Tween-20, and finally washing for 5 times by using sterile water, wherein each time lasts for 3min to obtain a sterilized explant;

(2) preparation of improved MS culture medium:

the formula of the improved MS culture medium is as follows: the dosage of ammonium nitrate is reduced to 825mg/L, and other components are unchanged.

(3) Obtaining a sterile explant and inducing adventitious buds:

cutting the disinfected explant obtained in the step (1) on a superclean workbench by using a sterile scalpel blade to cut the part of the two ends of the disinfected explant, wherein the part of the two ends of the disinfected explant is contacted with a mercuric chloride solution, inserting the lower biological end of the axillary bud of the stem into a culture medium for inducing the adventitious bud according to the direction that the top biological end is upward, placing the axillary bud in an artificial climate box for culturing for 4 weeks, and inducing to obtain adventitious buds of the Zanthoxylum nitidum; the induction culture conditions were as follows: the temperature is 28 +/-2 ℃, the relative humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 8001 x; the formula of the culture medium for adventitious bud induction is as follows: adding 6-BA and IAA on the modified MS culture medium; wherein the concentration of 6-BA is 1mg/L, and the concentration of IAA is 0.1 mg/L;

(4) subculture proliferation:

cutting the adventitious buds of the Zanthoxylum nitidum obtained in the step (3) from the explant, cutting off leaves, inoculating a subculture medium, and performing subculture for 4 weeks to obtain cluster buds of the Zanthoxylum nitidum; the subculture conditions were as follows: the temperature is 28 +/-2 ℃, the humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 9001 x; the formula of the culture medium for subculture proliferation is as follows: adding 6-BA and IAA on the basis of the improved MS culture medium, wherein the concentration of the 6-BA is 1mg/L, and the concentration of the IAA is 0.1 mg/L;

(5) sterile rooting:

and (4) when the zanthoxylum nitidum obtained in the step (4) grows to 2-3cm high from the seedling, carrying out induced rooting. Cutting off the single plant, inoculating the single plant into a rooting culture medium, and performing rooting culture for 42 days to obtain the Zanthoxylum nitidum seedlings; the rooting culture conditions were as follows: the culture temperature is 28 +/-2 ℃, the humidity is 85 +/-10 percent, the illumination time is 14 hours/day, and the illumination intensity is 7001 x; the formula of the culture medium for sterile rooting is as follows: adding NAA and IBA on the basis of the improved MS culture medium, wherein the concentrations are 0.5mg/L and 0.5mg/L respectively;

and (5) after the Zanthoxylum nitidum seedlings grow to the root length of 1-2cm and the height of the seedlings is 5-8cm, opening the bottle caps of culture bottles, hardening and culturing indoors for 7 days, then cleaning a root culture medium, planting the seedlings in seedling culture bags with the diameter of 5cm and filled with peat soil, placing the seedlings in a plastic greenhouse, keeping the substrate moist, and transplanting the seedlings to a field after the plants survive.

Example 3

A method for culturing and rapidly propagating zanthoxylum nitidum tissues comprises the following steps:

(1) selecting and disinfecting explants:

selecting young stems of healthy zanthoxylum nitidum as explants;

removing redundant leaves, washing dirt on the surfaces of the branches with tap water, and then sucking the branches with paper towels;

thirdly, soaking the workpiece on a clean bench for 20s by using 70 percent alcohol, and then washing the workpiece for 1 time and 3min by using sterile water;

fourthly, sterilizing for 15min by using 0.1 mass percent mercuric chloride solution containing 0.01 mass percent of Tween-20, and finally washing for 5 times by using sterile water, wherein each time lasts for 3min, so as to obtain a sterilized explant;

(2) preparation of improved MS culture medium:

the formula of the improved MS culture medium is as follows: the dosage of ammonium nitrate is reduced to 825mg/L, and other components are unchanged.

(3) Obtaining a sterile explant and inducing adventitious buds:

cutting the disinfected explant obtained in the step (1) on a superclean workbench by using a sterile scalpel blade to cut the part of the two ends of the disinfected explant, wherein the part of the two ends of the disinfected explant is contacted with a mercuric chloride solution, inserting the lower biological end of the axillary bud of the stem into a culture medium for inducing the adventitious bud according to the direction that the top biological end is upward, placing the axillary bud in an artificial climate box for culturing for 6 weeks, and inducing to obtain adventitious buds of the Zanthoxylum nitidum; the induction culture conditions were as follows: the temperature is 28 +/-2 ℃, the relative humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 6501 x; the formula of the culture medium for adventitious bud induction is as follows: adding 6-BA and IAA on the modified MS culture medium; wherein the concentration of 6-BA is 3mg/L, and the concentration of IAA is 0.3 mg/L;

(4) subculture proliferation:

cutting the adventitious buds of the Zanthoxylum nitidum obtained in the step (3) from the explant, cutting off leaves, inoculating a subculture medium, and performing subculture for 4 weeks to obtain cluster buds of the Zanthoxylum nitidum; the subculture conditions were as follows: the temperature is 28 +/-2 ℃, the humidity is 85 +/-10%, the illumination time is 12 hours/day, and the illumination intensity is 9001 x; the formula of the culture medium for subculture proliferation is as follows: adding 6-BA and IAA on the basis of the improved MS culture medium, wherein the concentration of the 6-BA is 3mg/L, and the concentration of the IAA is 0.2 mg/L;

(5) sterile rooting:

and (4) when the zanthoxylum nitidum obtained in the step (4) grows to 2-3cm high from the seedling, carrying out induced rooting. Cutting off the single plant, inoculating the single plant into a rooting culture medium, and performing rooting culture for 36 days to obtain the Zanthoxylum nitidum seedlings; the rooting culture conditions were as follows: the culture temperature is 28 +/-2 ℃, the humidity is 85 +/-10 percent, the illumination time is 14 hours/day, and the illumination intensity is 8001 x; the formula of the culture medium for sterile rooting is as follows: adding NAA and IBA on the basis of the improved MS culture medium, wherein the concentrations are 0.5mg/L and 0.5mg/L respectively;

and (5) after the Zanthoxylum nitidum seedlings grow to the root length of 1-2cm and the height of the seedlings is 5-8cm, opening the bottle caps of culture bottles, hardening and culturing indoors for 7 days, then cleaning a root culture medium, planting the seedlings in seedling culture bags with the diameter of 5cm and filled with peat soil, placing the seedlings in a plastic greenhouse, keeping the substrate moist, and transplanting the seedlings to a field after the plants survive.

Experiment on influence of 6-BA and culture medium on growth of Zanthoxylum nitidum

1. Influence of different concentrations of 6-BA on multiplication coefficient of radix Zanthoxyli

Based on the process of example 1, the culture was carried out while changing the concentration of 6-BA only and adding 6-BA concentrations of 1mg/L and 3mg/L, respectively, and the culture results are shown in FIG. 2, in which the concentrations of 6-BA from left to right were 1, 2 and 3mg/L, respectively, and the propagation coefficients were 2.32, 3.84 and 4.56.

2. Effect of modified MS Medium on plant growth

The control group supplemented with the MS medium was cultured based on the process of example 1, and the results of the culture are shown in FIG. 3, which shows modified MS medium and MS medium from left to right.

The above embodiments illustrate various embodiments of the present invention in detail, but the embodiments of the present invention are not limited thereto, and those skilled in the art can achieve the objectives of the present invention based on the disclosure of the present invention, and any modifications and variations based on the concept of the present invention fall within the scope of the present invention, which is defined by the claims.