阅读说明：本技术 一种组织培养快速繁育非洲双色野鸢尾的方法 (method for rapid breeding of African dual-color wild iris through tissue culture ) 是由 申瑞雪 李秋静 林丹 卿霞 周天宇 唐忠炳 张文昊 庞玉华 秦祥 于 2019-09-20 设计创作，主要内容包括：本发明涉及一种组织培养快速繁育非洲双色野鸢尾的方法,包括：(1)无菌材料的获得；(2)芽的分化和增殖；(3)生根培养；(4)炼苗与移栽。与现有技术相比,本发明采用组培快速繁育的方法,可以在短时间内获得大量株型整齐,性状优良的植株,以满足市场的大量需求。(The invention relates to a method for tissue culture and rapid propagation of African dual-color wild iris, which comprises the following steps: (1) obtaining a sterile material; (2) differentiation and proliferation of shoots; (3) rooting culture; (4) hardening and transplanting the seedlings. Compared with the prior art, the invention adopts a tissue culture rapid breeding method, can obtain a large number of plants with regular plant types and excellent properties in a short time, so as to meet a large number of demands of the market.)

1. A method for tissue culture and rapid propagation of African iris tectorum is characterized by comprising the following steps:

(1) obtaining of sterile Material

Collecting young stem segments with stem tips of the female parent of the African double-color wild iris in 3 months in spring, washing, sequentially soaking in 75% alcohol and 1 ‰ mercuric chloride, washing with sterile water, cutting the stem tips into small segments, and inoculating on stem tip induction culture medium;

(2) Differentiation and proliferation of shoots

After the buds are inoculated on a stem tip induction culture medium for 15 days, the base parts of the buds begin to expand, obvious callus can be seen in 25 days, the callus is cultured for 30 days, and the callus with the cut buds is placed in an adventitious bud multiplication culture medium for culture;

(3) Rooting culture

Taking seedlings in an adventitious bud multiplication culture medium, cutting robust plantlets, transferring the plantlets into a rooting culture medium to induce rooting, differentiating a plurality of white root primordia at the seedling base part after 25 days, growing to 2-3.5cm after 35 days, and ensuring that the rooting rate is 95%;

(4) hardening and transplanting seedlings

when the root system grows to 2-3cm, selecting aseptic seedlings with developed and strong root systems, placing the aseptic seedlings indoors at the temperature of 14-16 ℃ and the natural illumination and humidity for a week, taking out the seedlings, cleaning root agar, transplanting the seedlings onto a seedbed, watering the planted seedlings thoroughly, opening a greenhouse every day, keeping enough ventilation, watering when the soil is dry, and ensuring that the transplanting survival rate can reach 95%.

2. The method for tissue culture of rapidly propagating Iris africana according to claim 1, wherein in step (1), the good Iris africana plants with pure variety, robust plants, good growth vigor and no plant diseases and insect pests are selected as the parent.

3. The method for tissue culture of rapidly propagating Iris floribunda according to claim 1, wherein the washing method in step (1) comprises: flushing the collected explants under the running water to clean dust on the surfaces of the explants; then, washing with soapy water, and shaking the bottle during washing to ensure that the explants fully contact with the soapy water; and washing with running water after washing.

4. the method for tissue culture of rapidly propagating Iris floribunda, according to claim 1, wherein in step (1), the shoot tip induction medium contains macro-elements, iron salt, inositol, organic substances, trace elements, 6-BA, NAA,

The macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, and the 6-BA content is 2.0mg/L, NAA and 0.1 mg/L;

the macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

the pH value of the stem tip induction culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the stem tip induction culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 1800-2200 Lx.

5. the method of claim 1, wherein in step (2), the adventitious bud propagation medium contains macro-elements, iron salt, inositol, organic substances, trace elements, 6-BA, NAA,

The macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, the 6-BA content is 1.0mg/L, NAA and 0.1mg/L,

The macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the adventitious bud propagation culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the adventitious bud propagation culture medium, the culture temperature of the adventitious bud propagation culture medium is 21-23 ℃, and the illumination is 1800-2200 Lx.

6. the method for tissue culture of rapidly propagating Iris floribunda according to claim 1, wherein in step (3), the rooting medium contains macroelements, iron salt, inositol, organic substances, trace elements, IBA, NAA,

The macroelement content is 0.2212g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, and the trace element content is 0.2708g/1000L, IBA, the trace element content is 3.0mg/L, NAA, and the trace element content is 0.3 mg/L;

the macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the rooting culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the rooting culture medium, the culture temperature of the rooting culture medium is 21-23 ℃, and the illumination is 1800-.

7. The method for tissue culture of rapidly propagating Iris floribunda according to claim 1, wherein in step (4), the substrate on the seedbed is a mixture of turf and perlite in a volume ratio of 1:1,

Uniformly stirring the matrix, spreading the matrix on a seedbed, wherein the spreading thickness is 20cm, and spraying 500 times of sodium diurethane liquid medicine on the spread matrix for disinfection.

8. the method according to claim 1, wherein in step (4), the seedlings are transplanted after the roots of the African iris sanguinea are spread by using a tool jack and then the seedlings are planted by using forceps, and the planting depth is that the matrix covers the base of the seedlings and no root is exposed.

9. the method of claim 1, wherein in step (4), the application of liquid medicine is performed once a week, and carbendazim and Yinong-Dasheng Dou M45 are used alternately to prevent drug resistance and reduce the number of times in sunny days.

Technical Field

the invention relates to a plant tissue culture breeding method, in particular to a method for tissue culture rapid breeding of African double-color wild iris.

Background

iris floribunda (with the scientific name: Dietes bicolor (Steud.) Sweet ex Klatt) is a perennial root herb of Iris in Iridaceae. The plant height is about 1-1.5m, and the plant grows in clusters and has rootstocks; the leaf is basal, sword-shaped, leathery, light green, has obvious middle ribs and is 100-150cm long. In spring and summer, the scape is extracted from the leaf cluster, is slender and has branches. Fleur-de-lis, light yellow; the quilt has 6 quilts and 3 wide quilts, brown patches are embedded in the base parts of the quilts, each patch is provided with an orange halo edge, and orange spots are scattered at the bottom of each patch and serve as honey marks of insects so as to attract the insects to pollinate. The flower blooms for only one day, but the inflorescence continuously sprouts buds, so that the flower blossoms and falls off constantly, and the ornamental period is longer.

chinese patent CN102239805A discloses a tissue culture rapid propagation method of oriole tail, comprising: the method comprises the following steps: preparing a culture medium, including a basic culture medium; a bud induction medium; subculture multiplication medium; strong seedling culture medium; rooting culture medium; step two: culturing the virus-free tissue culture plantlets of the warburg tail, including selection and sterilization of explants; carrying out bud induction culture; bud multiplication culture; strong seedling culture; rooting culture; step three: hardening and transplanting the tissue culture seedlings, including hardening the seedlings; and (5) transplanting and culturing. The method remarkably reduces the contamination rate of oriole explants by adjusting the types and the content of culture medium hormones and adding antibiotics, so that the inoculation survival rate reaches more than 90 percent, the induced differentiation rate of the explants is also obviously improved, the induced differentiation rate is increased from 70-80 percent to 90 percent in the prior art, and the strong seedling rate of oriole tissue culture is improved.

chinese patent CN109588318A relates to a method for tissue culture and rapid breeding of plantain, which comprises (1) obtaining sterile materials; (2) differentiation and proliferation of shoots; (3) rooting culture; (4) hardening and transplanting the seedlings. The method can obtain a large number of plants with regular plant types and excellent properties in a short time so as to meet a large number of demands of the market. The plant stem tip is selected according to the material category, and the culture conditions are as follows: shoot tip induction medium: macroelements, iron salt, inositol, organic, trace elements, 6-BA2.0 and NAA 0.2; adventitious bud propagation medium: ② macroelement, iron salt, inositol, organic, microelement, 6-BA0.2 and NAA 0.1; rooting culture medium: ③ macroelement, ferric salt, inositol, organic, microelement and NAA 0.1.

The two-color wild iris in Africa originally produces south Africa, grows under the condition of half shading to sunny, has strong drought resistance, can be applied to greening in parks, commercial buildings and roadside, and has large market demand. The African wild iris adopts propagation modes such as division and the like, and has low propagation coefficient and survival rate. By adopting the tissue culture rapid breeding method, a large number of plants with regular plant types and excellent properties can be obtained in a short time, so as to meet a large number of demands of the market.

the "fast propagation technique research for tissue culture of iris" (author congratulatory peak, duffin, yunming, wanglizhen, from Hubei institute of ecological engineering and occupational technology) of "Jianxi agricultural journal" at stage 27 of 2015 introduces: the research of tissue culture and rapid propagation technology is carried out by taking the stem tip of the newly germinated bud of the German Iris (Iris germanica L.) variety of the golden doll as an explant. The results show that: the best results were obtained with 0.1% HgCl2 for explant sterilization for 7 min; the most suitable culture medium for primary culture, secondary culture and rooting culture is MS +6-BA1.5mg/L + IAA0.2mg/L, MS +6-BA2.0mg/L + IAA 0.5mg/L, 1/2MS +6-BA0.2 mg/L + IAA 1.5mg/L respectively; the multiplication coefficient of subculture by adopting a bud cluster containing 4-5 buds is highest; the optimal matrix for transplanting the test-tube plantlets is perlite, turf and garden soil (1:1:1), and the transplanting survival rate reaches 91.6%.

disclosure of Invention

the invention aims to overcome the defects in the prior art and provide a method for tissue culture to rapidly propagate the African double-color wild iris.

the purpose of the invention can be realized by the following technical scheme:

A method for tissue culture and rapid propagation of African iris biflora comprises the following steps:

(1) Obtaining of sterile Material

collecting young stem segments with stem tips of the female parent of the African double-color wild iris in 3 months in spring, washing, sequentially soaking in 75% alcohol and 1 ‰ mercuric chloride, washing with sterile water, cutting the stem tips into small segments, and inoculating on stem tip induction culture medium;

(2) differentiation and proliferation of shoots

after the buds are inoculated on a stem tip induction culture medium for 15 days, the base parts of the buds begin to expand, obvious callus can be seen in 25 days, the callus is cultured for 30 days, and the callus with the cut buds is placed in an adventitious bud multiplication culture medium for culture;

(3) Rooting culture

taking seedlings in an adventitious bud multiplication culture medium, cutting robust plantlets, transferring the plantlets into a rooting culture medium to induce rooting, differentiating a plurality of white root primordia at the seedling base part after 25 days, growing to 2-3.5cm after 35 days, and ensuring that the rooting rate is 95%;

(4) Hardening and transplanting seedlings

When the root system grows to 2-3cm, selecting aseptic seedlings with developed and strong root systems, placing the aseptic seedlings in a room with the temperature of 15 ℃, natural illumination and humidity for a week, taking out the seedlings, cleaning root agar, transplanting the seedlings onto a seedbed, watering the planted seedlings thoroughly, opening a greenhouse every day, keeping enough ventilation, properly watering when the soil is dry, and ensuring that the transplanting survival rate can reach 95%.

in the step (1), a good African double-color wild iris plant which is pure in variety, thick and strong in plant, good in growth vigor and free of diseases and insect pests is selected as a matrix.

In the step (1), the washing method comprises the following steps: washing the collected explants under fluid water, and cleaning dust on the surfaces of the explants for 10 min; washing with soapy water for 15min, and shaking the bottle during washing to make the explant fully contact with the soapy water; and washing with running water after washing.

In the step (1), the stem tip induction culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, 6-BA and NAA,

The macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, and the 6-BA content is 2.0mg/L, NAA and 0.1 mg/L.

the macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the stem tip induction culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the stem tip induction culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 1800-2200, preferably 2000 Lx.

In the step (2), the adventitious bud propagation culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, 6-BA and NAA,

The macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, the 6-BA content is 1.0mg/L, NAA and 0.1mg/L,

The macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the adventitious bud propagation culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the adventitious bud propagation culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 1800-2200, preferably 2000 Lx.

In the step (3), the rooting culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, IBA and NAA,

the macroelement content is 0.2212g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, and the trace element content is 0.2708g/1000L, IBA, the trace element content is 3.0mg/L, NAA, and the trace element content is 0.3 mg/L;

the macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

the pH value of the rooting culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the rooting culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, the illumination is 1800-.

In the step (4), the substrate on the seedbed is a mixture of turf and perlite in a volume ratio of 1:1,

Uniformly stirring the substrate, spreading the substrate on a seedbed, wherein the spreading thickness is about 20cm, and spraying 500 times of sodium diurethane liquid medicine on the spread substrate for disinfection.

In the step (4), a tool is used for inserting holes during transplanting, then the seedlings are planted after the roots of the seedlings are spread by forceps, and the planting depth is that the base parts of the seedlings are covered by the matrix to prevent the roots from exposing.

In the step (4), because the soil is moist in rainy days and the seedlings are easy to mildew and die, the liquid medicine needs to be applied once every week, and the carbendazim and the Dow Yinong-Dasheng M45 are alternately used to prevent the occurrence of drug resistance and reduce the times in sunny days.

The method for fast breeding iris through tissue culture is particularly suitable for breeding the African double-color wild iris variety.

Because the bred plants are different, the tissue culture conditions of different plant cultures are different, and the requirements for culture media are different, the method for quickly breeding the plantain by tissue culture, which is related to Chinese patent CN109588318A, is not suitable for the African double-color wild iris in the application, and the application changes the culture conditions, especially researches the element proportion in the culture media, and finds out the culture medium composition which is most suitable for the culture of the African double-color wild iris.

Compared with the tissue culture and rapid propagation method of iris in the background technology, the propagation culture medium and the rooting culture medium in the method are different, the environmental conditions are different, 30g/L of sucrose and 3g/L of agar are added into the rooting culture medium, the temperature range is reduced to 21-23 ℃, and the illumination condition is about 2000 Lx. The cultivated African double-color wild iris seedlings are robust, and the transplanting survival rate is 95%. The invention adopts a tissue culture rapid breeding method, can obtain a large number of plants with regular plant types and excellent properties in a short time, so as to meet a large number of demands of the market.

Detailed Description

The present invention will be described in detail with reference to specific examples.

a method for tissue culture and rapid propagation of African iris biflora comprises the following steps:

(1) obtaining of sterile Material

Collecting young stem segments with stem tips of the female parent of the African double-color wild iris in 3 months in spring, washing, sequentially soaking in 75% alcohol and 1 ‰ mercuric chloride, washing with sterile water, cutting the stem tips into small segments, and inoculating on stem tip induction culture medium;

(2) Differentiation and proliferation of shoots

after the buds are inoculated on a stem tip induction culture medium for 15 days, the base parts of the buds begin to expand, obvious callus can be seen in 25 days, the callus is cultured for 30 days, and the callus with the cut buds is placed in an adventitious bud multiplication culture medium for culture;

(3) Rooting culture

taking seedlings in an adventitious bud multiplication culture medium, cutting robust plantlets, transferring the plantlets into a rooting culture medium to induce rooting, differentiating a plurality of white root primordia at the seedling base part after 25 days, growing to 2-3.5cm after 35 days, and ensuring that the rooting rate is 95%;

(4) Hardening and transplanting seedlings

When the root system grows to 2-3cm, selecting aseptic seedlings with developed and strong root systems, placing the aseptic seedlings in a room with the temperature of 15 ℃, natural illumination and humidity for a week, taking out the seedlings, cleaning root agar, transplanting the seedlings onto a seedbed, watering the planted seedlings thoroughly, opening a greenhouse every day, keeping enough ventilation, properly watering when the soil is dry, and ensuring that the transplanting survival rate can reach 95%.

in the step (1), a good African double-color wild iris plant which is pure in variety, thick and strong in plant, good in growth vigor and free of diseases and insect pests is selected as a matrix.

In the step (1), the washing method comprises the following steps: washing the collected explants under fluid water, and cleaning dust on the surfaces of the explants for 10 min; washing with soapy water for 15min, and shaking the bottle during washing to make the explant fully contact with the soapy water; and washing with running water after washing.

in the step (1), the stem tip induction culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, 6-BA and NAA,

The macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, and the 6-BA content is 2.0mg/L, NAA and 0.1 mg/L.

The macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The stem tip induction culture medium has the pH value of 5.8, 30g/L of cane sugar and 3g/L of agar are added into the stem tip induction culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 2000 Lx.

In the step (2), the adventitious bud propagation culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, 6-BA and NAA,

the macroelement content is 0.4423g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, the trace element content is 0.2708g/1000L, the 6-BA content is 1.0mg/L, NAA and 0.1mg/L,

the macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the adventitious bud multiplication culture medium is 5.8, 30g/L of cane sugar and 3g/L of agar are added into the adventitious bud multiplication culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 2000 Lx.

In the step (3), the rooting culture medium contains macroelements, ferric salt, inositol, organic substances, trace elements, IBA and NAA,

The macroelement content is 0.2212g/L, the iron salt content is 0.0651g/L, the inositol content is 0.01g/L, the organic matter content is 0.031g/L, and the trace element content is 0.2708g/1000L, IBA, the trace element content is 3.0mg/L, NAA, and the trace element content is 0.3 mg/L;

The macroelements comprise one or more of the following substances: KNO3, NH4NO3, MgSO4, CaCl2, KH2PO 4; the iron salt is FeSO4 & 7H 2O; the organic substance comprises one or more of the following substances: pyridoxine hydrochloride, thiamine hydrochloride, glycine, nicotinic acid; the trace elements comprise one or more of the following substances: MnSO4 & 4H2O, ZnSO4 & 7H2O, H3BO3, KI, Na2MoO4 & 2H2O, CuSO4 & 5H2O, CoCl2 & 5H 2O;

The pH value of the rooting culture medium is 5.8, 30g/L of sucrose and 3g/L of agar are added into the rooting culture medium, the culture temperature of the stem tip induction culture medium is 21-23 ℃, and the illumination is 2000 Lx.

In the step (4), the substrate on the seedbed is a mixture of turf and perlite in a volume ratio of 1:1,

uniformly stirring the substrate, spreading the substrate on a seedbed, wherein the spreading thickness is about 20cm, and spraying 500 times of sodium diurethane liquid medicine on the spread substrate for disinfection.

In the step (4), a tool is used for inserting holes during transplanting, then the seedlings are planted after the roots of the seedlings are spread by forceps, and the planting depth is that the base parts of the seedlings are covered by the matrix to prevent the roots from exposing.

In the step (4), because the soil is moist in rainy days and the seedlings are easy to mildew and die, the liquid medicine needs to be applied once every week, and the carbendazim and the Dow Yinong-Dasheng M45 are alternately used to prevent the occurrence of drug resistance and reduce the times in sunny days.

the method for rapid propagation of plant tissue culture provided by the invention is particularly suitable for propagating the African double-color wild iris variety.

the previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.