Method for rapid propagation of altai hawthorn

Abstract

The application provides a method for rapidly propagating Altai hawthorn: the Altai hawthorn is rapidly propagated by sequentially performing primary culture, propagation subculture, rooting culture, seedling raising and field transplanting on the Altai hawthorn. The method for rapidly propagating the Altai hawthorn has the advantages of high increment coefficient, short rooting time and high transplanting survival rate, and helps to realize the factory cultivation of the Altai hawthorn tissue culture.

Description

Technical Field

[0001] The invention belongs to the field of plant tissue culture, and particularly relates to a method for rapidly propagating Altai hawthorn. Background Art

[0002] Altai hawthorn (Crataegusaltaica (Loudon) Lange) is produced in the central and northern parts of Xinjiang, China. It is a wild fruit tree resource in the Tianshan Mountain area. It is cold-resistant, drought-resistant, and resistant to barrenness. It has many and dense flowers, white flowers, and a diameter of 12-15mm. The fruit is golden yellow and spherical, with a diameter of 8-10mm. The flowering period is from May to June and the fruiting period is from August to September. Because of its beautiful appearance and wide adaptability, it is an important garden ornamental and barren mountain afforestation tree species in my country. At the same time, Altai hawthorn can be used as the rootstock of edible hawthorn and other Rosaceae ornamental tree species to improve the cold resistance, drought resistance and adaptability of the scion. At present, the seedling breeding of Altai hawthorn in production mostly adopts seed propagation, but the outer layer of Altai hawthorn seed is hard and lignified, resulting in a low seed germination rate, a long time consumption, and different seed sources. The seedlings produced are uneven, which has seriously hindered the promotion and application of "Altai" hawthorn.

[0003] Therefore, developing a rapid propagation method for Altai hawthorn becomes a problem that demands urgent solution. Summary of the Invention

[0004] In order to solve the above problems, the purpose of the present invention is to provide a method for rapid propagation of Altai hawthorn, so as to solve the current problems of low efficiency, long time consumption and uneven seedlings of Altai hawthorn seed propagation.

[0005] In order to achieve the above object, the present invention provides a method for rapidly propagating Altai hawthorn, comprising the following steps:

[0006] 1) Selection and disinfection of explants;

[0007] 2) inoculating the explants treated in step 1) into a primary culture medium for primary culture; wherein the primary culture medium comprises: MS + 0.5 mg / L 6-BA + 30 g / L sucrose + 6 g / L agar, and adjusting the pH to 5.8;

[0008] 3) Using the well-grown explants obtained in step 2) as test materials, cutting 2 cm new stem segments from the test materials and inoculating them on a subculture medium for propagation and subculture to obtain tissue culture plantlets; wherein the subculture medium consists of MS + 0.5 mg / L 6-BA + 0.2 mg / L IAA + 30 g / L sucrose + 6 g / L agar, adjusted to a pH of 5.8;

[0009] 4) cut 5cm of the well-proliferated and well-grown tissue culture seedlings in the subculture of step 3), cut off the leaves at 1cm from the cut end, and then inoculate into a rooting culture medium for rooting culture to obtain the rooted tissue culture seedlings; wherein the rooting culture is divided into two stages, the first stage is dark culture for 6-10 days, the medium used in the dark culture is 1 / 2MS+0.5mg / L IBA+0.5mg / L NAA+20g / L sucrose+6g / L agar, and the pH value is adjusted to 5.8; the second stage is light culture for 8-10 days, the medium used in the light culture is 1 / 2MS+0.001mg / L IBA+20g / L sucrose+6g / L agar, and the pH value is adjusted to 5.8;

[0010] 5) when the length of the roots of the rooted tissue culture seedlings reaches 2-3cm, move to a greenhouse for closed bottle hardening for 3 days, open the bottle for hardening for 3 days, then take out the rooted tissue culture seedlings in the bottle, wash off the culture medium, perform root disinfection treatment with 0.2% carbendazim, transplant into a nutrient pot, spray 5-8mg / L gibberellin, and the substrate is fine coconut coir with a particle size of 1-3mm: coarse coconut coir with a size of 1cm block: peat soil: perlite = 1:1:2:1, and transplant into a field after the root system is fully developed.

[0011] Further, the steps of selecting and disinfecting the explant of step 1) are as follows: select several one-year-old V. tangutorium branches, remove the leaves, trim the branches into stem segments of about 5cm, and only keep 1 axillary bud per segment; place the trimmed stem segments with buds into a tissue culture bottle, wrap the bottle opening with a single layer of gauze, tighten with an elastic band, add 1 drop of washing-up liquid the size of a soybean on the gauze, and place under running water for 1h; then immerse the washed stem segments with buds in 75% ethanol for disinfection treatment for 30s, constantly shake the bottle during the treatment, then wash with sterile water for 4 times for about 60s each time; immerse in 1% sodium hypochlorite containing 5‰ Tween 20 for 4min, constantly shake the bottle during the treatment, wash with sterile water for 4-5 times for 60s each time until there is no foam in the liquid; place the stem segments with buds after disinfection treatment on sterile filter paper to absorb the surface moisture, and then inoculate into the primary culture medium for induction.

[0012] Further, the culture conditions of the primary culture in step 2) are as follows: temperature 25℃, humidity 55-75%, light intensity 1500lx, and light time 14h / d; the culture conditions of the subculture in step 3) are as follows: temperature 25℃, humidity 55-75%, light intensity 1500lx, and light time 14h / d.

[0013] Further, the temperature of the dark culture in step 4) is 18-26℃, and the humidity is 55-75%; the temperature of the light culture is 18-26℃, the humidity is 55-75%, the light intensity is 1800-2200lx, and the light time is 16-20h / d.

[0014] The application also provides application of the rapid propagation method of the above-mentioned Altai hawthorn in hawthorn breeding.

[0015] The application has the following characteristics:

[0016] In the application, the explants are washed with running water for 1 hour, treated with 75% ethanol for 30 seconds, and treated with 1% sodium hypochlorite for 4 minutes, and under the treatment, the explant contamination rate is 0%, the mortality rate is 0%, and the survival rate is 100%.

[0017] In the application, the primary induction medium used is MS+0.5 mg / L 6-BA+30 g / L sucrose+6 g / L agar, pH 5.8, and under the formula, the initiation rate of the tissue culture seedlings reaches 100%, and the growth state is good.

[0018] In the application, the subculture medium used is MS+0.5 mg / L 6-BA+0.2 mg / L IAA+30 g / L sucrose+6 g / L agar, pH 5.8, the proliferation coefficient is 5.067, the plant height is 4.107 cm, the tissue culture seedlings grow vigorously in the medium, have strong branching ability, are mostly bush seedlings, have strong stems, and have dark green leaves.

[0019] In the application, the rooting culture is divided into two stages: dark culture for 6-10 days, the culture medium is 1 / 2MS+0.5 mg / L IBA+0.5 mg / L NAA+20 g / L sucrose+6 g / L agar, the pH value is 5.8, the culture condition is temperature 18-26 DEG C and humidity 55-75%, light culture for 8-10 days, the culture medium is 1 / 2MS+0.001 mg / L ABA+20 g / L sucrose+6 g / L agar, the pH value is 5.8, the culture condition is temperature 18-26 DEG C, humidity 55-75%, light time 16-20 h / d, and light intensity 1800-2200 lx. The rooting time is about 15 days, the rate is 96.67%, the average number of roots is 6.067, and the average root length is 2.295 cm. The rooting is stable, the root system is much and strong, and the seedlings grow well. In the application, ABA is added to the culture medium in the light culture stage to promote rooting, because a small amount of ABA is found to be helpful to rooting.

[0020] In the application, fine coconut husk (particle size 1-3 mm) : coarse coconut husk (1 cm block) : peat soil : perlite = 1:1:2:1 is used as the transplanting substrate, and 5-8 mg / L gibberellin is sprayed, so that the seedling growth is significantly improved, and the transplanting survival rate is improved.

[0021] The application has the following beneficial effects:

[0022] The application provides a method for rapidly propagating Altai hawthorn, wherein the subculture period is 25 days, the increment coefficient is 5.067, the rooting time is not more than 15 days, the rooting rate is more than 95%, the survival rate of field transplanting after seedling acclimatization is greater than 90%, and the method provides a fast and efficient technical method for Altai hawthorn tissue culture factory seedling. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 It is a photo of the axillary bud germination condition in the primary culture in the embodiment 1 of the application.

[0024] Figure 2 It is a photo of the tissue culture seedling growth condition when the plant hormones KT and IBA are in different proportions in the subculture in the embodiment 1 of the application.

[0025] Figure 3 It is a photo of the tissue culture seedling growth condition when the plant hormones 6-BA+IAA are in different proportions in the subculture in the embodiment 1 of the application.

[0026] Figure 4 It is the bud proliferation condition in the subculture in the embodiment 1 of the application.

[0027] Figure 5 It is the rooting condition of the tissue culture seedling when different plant hormone proportions are used in the rooting culture in the embodiment 1 of the application.

[0028] Figure 6 It is the growth condition of the rooting tissue culture seedling transplanted into the substrate in the embodiment 1 of the application. DETAILED DESCRIPTION

[0029] The embodiments of the application will be described in detail below, and the advantages and features of the application can be more easily understood by those skilled in the art, so that the protection scope of the application can be more clearly and explicitly defined.

[0030] If not particularly specified, the technical means used in the embodiments is the conventional means well known by those skilled in the art.

[0031] Materials:

[0032] 1. The formula of MS medium is: macroelements: KNO3 1900 mg / L; NH4NO3 1650 mg / L, MgSO4·7H2O 370 mg / L, KH2PO4 170 mg / L, CaCl2·2H2O 440 mg / L; microelements MnSO4·4H2O 22.3 mg / L, ZnSO4·7H2O 8.6 mg / L, H3BO3 6.2 mg / L, KI 0.83 mg / L, Na2MoO4·2H2O 0.25 mg / L, CuSO4·5H2O 0.025 mg / L, CoCl2·6H2O 0.025 mg / L, EDTA-Na2·2H2O 37.25 mg / L, FeSO4·7H2O 27.8 mg / L; organic substances: myo-inositol 100 mg / L, nicotinic acid VB3 0.5 mg / L, VB1 0.1 mg / L, VB6 0.5 mg / L, glycine 2 mg / L.

[0033] 2. The formula of 1 / 2MS medium is: macroelements: KNO3 950 mg / L; NH4NO3 825 mg / L, MgSO4·7H2O 185 mg / L, KH2PO4 85 mg / L, CaCl2·2H2O 220 mg / L; microelements MnSO4·4H2O 22.3 mg / L, ZnSO4·7H2O 8.6 mg / L, H3BO3 6.2 mg / L, KI 0.83 mg / L, Na2MoO4·2H2O 0.25 mg / L, CuSO4·5H2O 0.025 mg / L, CoCl2·6H2O 0.025 mg / L, EDTA-Na2·2H2O 37.25 mg / L, FeSO4·7H2O 27.8 mg / L; organic substances: myo-inositol 100 mg / L, nicotinic acid VB3 0.5 mg / L, VB1 0.1 mg / L, VB6 0.5 mg / L, glycine 2 mg / L.

[0034] 3. 6-BA (6-benzylaminopurine) was purchased from Sigma-Aldrich, product number B3408

[0035] 4. Sucrose was purchased from Sigma-Aldrich, product number: V900116.

[0036] 5. Agar was purchased from Sigma-Aldrich, product number: A1296.

[0037] 6. IAA (Indole-3-acetic acid) was purchased from Sigma-Aldrich, product number: I2886.

[0038] 7. IBA (indole butyric acid) was purchased from Sigma-Aldrich Company, product number: I5386.

[0039] 8. NAA (naphthalene acetic acid) was purchased from Sigma-Aldrich Company, product number: N0640.

[0040] 9. ABA (abscisic acid) was purchased from Sigma-Aldrich Company, product number: 90769, ABA was used to promote the growth of main roots and lateral roots.

[0041] 10. Gibberellin was purchased from Sigma-Aldrich Company, product number: G7645, gibberellin was used to promote the growth of stems.

[0042] 11. KT (kinetin) was purchased from Sigma-Aldrich Company, product number K3378.

[0043] The proliferation coefficient measured in the embodiment of the application refers to the number of effective seedlings differentiated from 1 seedling in one culture period, which is counted and averaged.

[0044] The survival rate referred to in the application refers to the germination and no death of explants after being put into bottles.

[0045] Example 1

[0046] A tissue culture and rapid propagation method of Altai hawthorn includes establishing an Altai hawthorn sterile system, conducting subculture and rooting culture, and conducting field transplanting of the rooted tissue culture seedlings after domestication, and the specific steps are as follows:

[0047] 1) Selection and disinfection treatment of Altai hawthorn explants

[0048] Collect several one-year-old Altai hawthorn branches, remove the leaves, and only keep the axillary buds; trim the branches into stem segments of about 5 cm, and only keep 1 axillary bud in each segment; place the trimmed bud-containing stem segments into tissue culture bottles, wrap the bottle opening with a single layer of mesh cloth, tighten it with an elastic band, add 1 drop of washing-up liquid the size of a soybean on the mesh cloth, and place it under running water for 0-1 h; soak the washed bud-containing stem segments in 75% ethanol for disinfection treatment for 20-30 s, constantly shake the bottle body during the treatment, then wash with sterile water for 4 times, each time for about 60 s; soak in 1% sodium hypochlorite (containing 5‰ Tween 20) for 4-14 min, constantly shake the bottle body during the treatment, then wash with sterile water for 4-5 times, each time for about 60 s, until there is no foam in the liquid. Dry the surface moisture of the disinfected bud-containing stem segments on sterile filter paper, and then inoculate them into the primary culture medium for induction.

[0049] 2) Primary culture of Altai hawthorn

[0050] The primary culture medium is MS + 0.5 mg / L 6-BA + 30 g / L sucrose + 6 g / L agar, pH 5.8, and the culture conditions are: temperature 25°C, humidity 55-75%, light 1500 lx, and light time 14 h / d.

[0051] The primary culture can test the contamination rate and the initiation rate (germination rate) of Altai hawthorn explants. As shown in Table 1, the effects of different disinfection treatments on the contamination rate and the initiation rate of Altai hawthorn explants, Table 1 Effects of different disinfection treatments on the contamination rate and the initiation rate of Altai hawthorn explants

[0052]

[0053] As can be seen from Table 1, T8, T10, T11 and T12 can all achieve ideal explant contamination rate and initiation rate, but considering the disinfection work efficiency and contamination rate, the disinfection effect of rinsing with running water for 1 h, 75% alcohol treatment for 30 s, and 1% sodium hypochlorite treatment for 4 min is the best (T10), as shown in Figure 1 the photos of Altai hawthorn axillary bud germination under primary culture using T10.

[0054] 3) Propagation and subculture of Altai hawthorn

[0055] The well-grown explants obtained by primary culture were used as test materials, and new stem segments of 2 cm or more were cut and inoculated on the subculture medium for propagation and subculture.

[0056] KT and 6-BA belong to cytokinins, and IAA and IBA belong to auxins. Cytokinins and auxins generally need to be added in the subculture propagation medium, but the specific addition of which cytokinin and which auxin and the proportion of addition need to be experimentally screened

[0057] To screen the plant hormones and concentrations in the subculture medium, different plant hormone ratios and concentrations were set. Hormone formula one: KT + IBA, with 0.5, 1, 1.5 mg / L of KT and 0.2, 0.5, 1 mg / L of IBA gradient treatment; hormone formula two: 6-BA + IAA, with 0.5, 1 mg / L of 6-BA and 0.2, 0.5, 1 mg / L of IAA gradient treatment, as shown in Tables 2 and 3, and Tables 2 and 3 are the effects of different plant hormone ratios on Altai hawthorn bud proliferation, Figure 2 are photos of Altai hawthorn bud proliferation using different group treatments of Table 2, Figure 3 are photos of Altai hawthorn bud proliferation using different group treatments of Table 3.

[0058] Table 2 Effects of different plant hormone KT and IBA ratios on bud proliferation

[0059]

[0060] Note: The data in the table are mean ± standard error; different lower case letters in the same column indicate significant difference at P<0.05 level

[0061] Table 3 Effects of different combinations of plant hormones 6-BA and IAA on bud proliferation

[0062]

[0063]

[0064] Note: The data in the table are mean ± standard error; different lower case letters in the same column indicate significant difference at P<0.05 level.

[0065] From Table 2 and Table 3, it can be seen that the combination of 6-BA and IAA is more conducive to the bud proliferation of Altai hawthorn tissue culture seedlings. Considering the proliferation coefficient and the growth state of the plants, the most suitable subculture medium for Altai hawthorn tissue culture seedlings is MS+0.5mg / L 6-BA+0.2mg / L IAA.

[0066] Therefore, the medium for subculture of Altai hawthorn is MS+0.5mg / L 6-BA+0.2mg / L IAA+30g / L sucrose+6g / L agar, pH 5.8, and the culture conditions are: temperature 25℃, humidity 55-75%, light intensity 1500lx, and light time 14h / d. MS+0.5mg / L 6-BA+0.2mg / L IAA. Figure 4 For the bud proliferation in subculture, from Figure 4 It can be seen that the proliferation coefficient is high, the seedlings are clump-shaped, the stems are strong, and the leaves are dark green.

[0067] 4) Rooting culture of Altai hawthorn

[0068] About 5cm of tissue culture seedlings with good proliferation effect and growth state in subculture were cut, the leaves at the cut end of 1cm were removed, and then inoculated into the rooting medium for rooting.

[0069] IAA, IBA and NAA all belong to auxin plant hormones, and rooting culture generally needs to add auxin, but the specific auxin to be added and the proportion need to be selected by experiment.

[0070] To screen the plant hormones and their concentrations in rooting medium in dark culture stage, two groups of hormone formula were set. Group A was added with IBA and NAA for experiment, the concentration gradient of IBA was set as 0.5 mg / L, and the concentration gradient of NAA was set as 0.2, 0.5 and 1 mg / L. Group B was added with IAA and IBA for experiment, the concentration gradient of IBA was set as 0.1 and 0.3 mg / L, and the concentration gradient of IAA was set as 0.4 and 1 mg / L. The specific grouping is shown in Table 4. Table 4 is the effect of different plant hormone ratios on the rooting of Altai hawthorn tissue culture seedlings in dark culture stage. Figure 5 The photos of the rooting of Altai hawthorn tissue culture seedlings in Table 4 groups are shown in the following.

[0071] Table 4 is the effect of different plant hormone ratios on the rooting of tissue culture seedlings in dark culture stage.

[0072]

[0073]

[0074] Note: The data in the table are mean ± standard error; different lowercase letters in the same column indicate significant difference at the level of P < 0.05.

[0075] As can be seen from Table 4, considering the rooting rate, rooting number, root length and growth condition of the plant, 1 / 2MS + 0.5 mg / L IBA + 0.5 mg / L NAA was selected as the best rooting medium for Altai hawthorn tissue culture seedlings in dark culture stage. Figure 5

[0076] Therefore, the specific steps of rooting culture of Altai hawthorn are as follows: first, dark culture for 6-10 days, the culture medium is 1 / 2MS + 0.5 mg / L IBA + 0.5 mg / L NAA + 20 g / L sucrose + 6 g / L agar, the pH value is 5.8, the culture condition is temperature 18-26℃ and humidity 55-75%; then, light culture for 8-10 days, the culture medium is 1 / 2MS + 0.001 mg / L IBA + 20 g / L sucrose + 6 g / L agar, the pH value is 5.8, the culture condition is temperature 18-26℃, humidity 55-75%, light 16-20 h / d and light intensity 1800-2200 lx; here, it was found through experiment that a small amount of ABA can change from inhibition to promotion of rooting.

[0077] 4) Hardening and field transplanting of Altai hawthorn rooting seedlings

[0078] ​When the root length of the rooted culture of Altai hawthorn reaches 2-3 cm, the seedlings are moved to a greenhouse for 3 days of closed bottle hardening, 3 days of open bottle hardening, then the rooted seedlings in the bottle are taken out, the culture medium is washed off, the roots are disinfected with 0.2% carbendazim, transplanted into a nutrient pot, the substrate is fine coconut husk (particle size 1-3 mm) : coarse coconut husk (1 cm block) : peat soil : perlite = 1 : 1 : 2 : 1, 5-8 mg / L gibberellin is sprayed, and after the root system develops completely, the seedlings are transplanted into a field, as shown in Fig. 2. Figure 6 As can be seen from Fig. 2, the rooted seedlings of Altai hawthorn grow well and reach the transplanting standard. Figure 6 As can be seen from Fig. 2, the rooted seedlings of Altai hawthorn grow well and reach the transplanting standard.

[0079] As can be seen from the above examples, the method for rapidly propagating Altai hawthorn provided by the present application has simple operation steps and low cost, and is conducive to the popularization and application of Altai hawthorn, and provides an effective way for factory seedling raising.

[0080] The above examples only express several embodiments of the present application, and the description is relatively specific and detailed, but it should not be understood as a limitation on the scope of the patent of the present application. It should be pointed out that for ordinary skilled persons in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. A method for rapid propagation of Hippophae rhamnoides L. var. sinensis K. Koch, characterized by, The method comprises the following steps: 1) selection and disinfection of explants; 2) inoculating the explants treated in step 1) into a primary culture medium for primary culture; wherein the primary culture medium is MS+0.5 mg / L 6-BA+30 g / L sucrose+6 g / L agar, and the pH value is adjusted to 5.8; 3) taking the well-grown explants obtained in step 2) as test materials, cutting 2 cm of the new stem segments on the test materials, inoculating the stem segments into a subculture medium for propagation and subculture, and obtaining tissue culture seedlings; wherein the subculture medium is MS+0.5 mg / L 6-BA+0.2 mg / L IAA+30 g / L sucrose+6 g / L agar, and the pH value is adjusted to 5.8; 4) cutting 5 cm of the tissue culture seedlings with good proliferation effect and growth state in the subculture in step 3), cutting off the leaves at a distance of 1 cm from the cut, and then inoculating the tissue culture seedlings into a rooting culture medium for rooting culture, to obtain rooted tissue culture seedlings; wherein the rooting culture is divided into two stages, the first stage is dark culture for 6-10 days, the medium used in the dark culture is 1 / 2 MS+0.5 mg / L IBA+0.5 mg / L NAA+20 g / L sucrose+6 g / L agar, and the pH value is adjusted to 5.8; the second stage is light culture for 8-10 days, the medium used in the light culture is 1 / 2 MS+0.001 mg / L ABA+20 g / L sucrose+6 g / L agar, and the pH value is adjusted to 5.8; 5) when the length of the roots of the rooted tissue culture seedlings reaches 2-3 cm, the rooted tissue culture seedlings are moved to a greenhouse for bottle hardening for 3 days, and then open bottle hardening for 3 days, then the rooted tissue culture seedlings in the bottle are taken out, the medium is washed off, the roots are disinfected with 0.2% carbendazim, and the rooted tissue culture seedlings are transplanted into a nutrient pot and sprayed with 5-8 mg / L gibberellin, the substrate is fine coconut coir with a particle size of 1-3 mm: coarse coconut coir with a block size of 1 cm: peat soil: perlite = 1:1:2:1, and the rooted tissue culture seedlings are transplanted into a field after the root system is fully developed; wherein, the step 1) is to select and disinfect the explants: a plurality of one-year-old Aralia decaisneana branches are selected, the leaves are removed, the branches are trimmed into 5 cm stem segments, and only one axillary bud is retained on each stem segment; the trimmed stem segments with buds are placed in a culture bottle, the bottle opening is wrapped with a single layer of gauze, an elastic band is tightened, 1 drop of washing-up liquid as big as a soybean is added on the gauze, and the culture bottle is placed under a tap for water flushing for 1 h; then the washed stem segments with buds are soaked in 75% ethanol for disinfection treatment for 30 s, the bottle body is shaken constantly during the treatment, and then the stem segments with buds are washed with sterile water for 4 times, 60 s each time; the stem segments with buds are soaked in 1% sodium hypochlorite containing 5% Tween 20 for 4 min, the bottle body is shaken constantly during the treatment, and the stem segments with buds are washed with sterile water for 4-5 times, 60 s each time, until there is no foam in the liquid; the stem segments with buds after disinfection treatment are placed on sterile filter paper to absorb surface moisture, and then inoculated into the primary culture medium for induction.

2. The method of claim 1, wherein, The culture condition of the primary culture in the step 2) is: temperature 25 ℃, humidity 55-75%, illumination 1500 lx, and illumination time 14 h / d; the culture condition of the subculture in the step 3) is: temperature 25 ℃, humidity 55-75%, illumination 1500 lx, and illumination time 14 h / d.

3. The method of claim 1, wherein, The temperature of the dark culture in the step 4) is 18-26 ℃, and the humidity is 55-75%; the temperature of the light culture is 18-26 ℃, the humidity is 55-75%, the illumination intensity is 1800-2200 lx, and the illumination time is 16-20 h / d.

4. Application of the method for rapid propagation of Altai hawthorn according to any one of claims 1-3 in hawthorn breeding.

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