Method for improving transplanting survival rate of polyphylla prataes in vitro seedling

Abstract

The present application belongs to the field of plant biotechnology, and discloses a method for improving the survival rate of transplanting of tissue culture seedlings of Polygonatum cyathopetalum, comprising the following steps: taking elongated buds obtained from tissue culture of Polygonatum cyathopetalum as explants, separating the explants from adventitious bud clusters, treating the bud bases with a rooting promoting liquid, inoculating in a culture medium for synchronous induction culture of adventitious roots and microtubers, transferring the tissue culture bottles containing the regenerated plants of Polygonatum cyathopetalum with induced microtubers and adventitious roots to a plastic greenhouse under natural light for domestication, transplanting the treated tissue culture seedlings of Polygonatum cyathopetalum to a substrate for seedling culture under a forest to obtain regenerated plants of Polygonatum cyathopetalum. The present application has the beneficial effect of providing a method for effectively improving the survival rate of transplanting of tissue culture seedlings of Polygonatum cyathopetalum, and providing certain technical support for industrialized production and transplanting of tissue culture seedlings of Polygonatum cyathopetalum.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of plant biotechnology, and in particular to a method for improving the survival rate of tissue culture seedlings of Polygonatum cyrtonema. BACKGROUND

[0002] Polygonatum cyrtonema Hua, also known as "Xianren Yuliang", is a perennial herbaceous plant of the Liliaceae Polygonatum genus, and its rhizome is used as medicine, which has the effects of tonifying qi and yin, invigorating the spleen, moistening the lungs, and nourishing the kidney. As one of the "ten major Anhui medicines", Polygonatum cyrtonema has been widely cultivated in Anhui Province, and is a traditional bulk medicinal material with the values of medicine, food, ornamental and health care. In recent years, with the development of traditional Chinese medicine and the health industry, the medicinal and health care values of Polygonatum cyrtonema have been increasingly recognized by people, and its market prospect is very broad.

[0003] The lack of excellent varieties and effective variety propagation techniques is a bottleneck problem restricting the development of the Polygonatum cyrtonema industry. Traditional sowing and tuber propagation methods have the problems of low propagation efficiency, long propagation cycle, and propagation being limited by materials and seasons, etc., and it is difficult to meet the increasing demand of the Polygonatum cyrtonema seedling market. Plant tissue culture technology has important significance in the conservation and factory propagation of medicinal plant resources germplasm due to its high propagation efficiency, short propagation cycle, and the advantages of not being limited by seasons and time. At present, there have been many reports on the use of plant tissue culture technology in the rapid propagation of Polygonatum cyrtonema seedlings, but in the process of tissue culture production and planting of Polygonatum cyrtonema seedlings, the low survival rate of tissue culture seedlings in the field after transplanting has become a key factor restricting the industrialized production of Polygonatum cyrtonema tissue culture seedlings.

[0004] Therefore, in view of the above problems, on the basis of the tissue culture propagation of excellent strain seedlings of Polygonatum cyrtonema, there is an urgent need for a method for improving the survival rate of tissue culture seedlings of Polygonatum cyrtonema to meet the production needs of large-scale planting of Polygonatum cyrtonema tissue culture seedlings. SUMMARY

[0005] The technical problem to be solved by the present application is how to improve the survival rate of tissue culture seedlings of Polygonatum cyrtonema.

[0006] The present application solves the above technical problems by the following technical means:

[0007] A method for improving the survival rate of tissue culture seedlings of Polygonatum cyrtonema, comprising the following steps:

[0008] (1) Synchronous induction culture of adventitious roots and microtubers: taking the elongated buds obtained by tissue culture of Polygonatum cyrtonema as explants, separating them from the adventitious bud clusters, treating the bud bases with a rooting promoting liquid, and then inoculating them into a culture medium for synchronous induction culture of adventitious roots and microtubers;

[0009] (2) The pre-transplanting treatment of the tissue culture seedlings of Polygonatum cyathopetalum: the tissue culture bottles containing the regenerated plants of Polygonatum cyathopetalum with induced microtubers and adventitious roots are transferred to a plastic greenhouse under natural light for acclimatization;

[0010] (3) The transplanting and seedling management of the tissue culture seedlings of Polygonatum cyathopetalum: the treated tissue culture seedlings of Polygonatum cyathopetalum are transplanted into a substrate and cultivated under a forest to obtain the regenerated plants of Polygonatum cyathopetalum.

[0011] Preferably, in (1), the Polygonatum cyathopetalum is Polygonatum cyathopetalum Qiyuan.

[0012] Preferably, in (1), the rooting promoting solution is an aqueous solution added with NAA at a final concentration of 500-1000 mg / L, IBA at a final concentration of 250-1000 mg / L and ABA at a final concentration of 10-100 mg / L.

[0013] Preferably, in (1), the rooting promoting solution treatment lasts for 60-180 s.

[0014] Preferably, in (1), the medium is 1 / 4-2MS medium added with 10-60 g / L sucrose and 7.0 g / L agar.

[0015] Preferably, in (1), the synchronous induction culture of adventitious roots and microtubers is performed in a greenhouse with a temperature of 25±2℃, a light intensity of 2000-2500 lx, a light cycle of 14 / 10 h (light / dark) and one or more of blue light, red light and white light.

[0016] Preferably, in (1), the synchronous induction culture lasts for 6 weeks.

[0017] Preferably, in (2), the greenhouse acclimatization lasts for 7-10 d.

[0018] Preferably, in (2), the treated tissue culture seedlings are further soaked in a 0.01%-0.05% potassium permanganate solution added with 10-60 μM melatonin for 5-10 min.

[0019] Preferably, in (3), the method further comprises the following steps: the treated tissue culture seedlings of Polygonatum cyathopetalum are transplanted into a seedling raising bag containing garden soil: perlite: nutrient soil at a volume ratio of 4-8:1-2:1-4 and cultivated under a forest at an altitude of 300-600 m, 1 / 8-1 / 2MS macroelements are sprayed on the leaves after the transplanting, a sunshade net is covered, the sunshade net is removed after 4 weeks, and 1 / 8-1 / 2MS macroelements are sprayed on the leaves and water is poured every 2 weeks during the period to obtain the regenerated plants of Polygonatum cyathopetalum.

[0020] The present application has the following advantages:

[0021] The method for improving the transplant survival rate of the tissue culture seedlings of Polygonatum cyrtonema has the following outstanding advantages: the developed induction synchronization technology of adventitious roots and microtubers of the tissue culture elongated buds of Polygonatum cyrtonema and the cultivation and management technology realize the synchronous induction of the adventitious roots and microtubers of the tissue culture seedlings of Polygonatum cyrtonema and the direct cultivation under the forest, greatly improve the transplant survival rate of the tissue culture seedlings of Polygonatum cyrtonema, simplify the transplanting steps of the tissue culture seedlings, and achieve a high survival rate of 96.8% under the forest, thereby providing important technical support for the industrialized production and planting of the tissue culture seedlings of Polygonatum cyrtonema. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 The regenerated plants of Polygonatum cyrtonema obtained by adopting the conventional tissue culture technology for 6 weeks in the embodiment 1 of the present application only have the formation of adventitious roots without the formation of microtubers;

[0023] Figure 2 The regenerated plants of Polygonatum cyrtonema obtained by adopting the synchronous induction technology of adventitious roots and microtubers for 4 weeks in the embodiment 1 of the present application;

[0024] Figure 3 The regenerated plants of Polygonatum cyrtonema obtained by adopting the synchronous induction technology of adventitious roots and microtubers for 6 weeks in the embodiment 1 of the present application;

[0025] Figure 4 The cleaning and air-drying of the regenerated plants of Polygonatum cyrtonema in the embodiment of the present application;

[0026] Figure 5 The regenerated plants of Polygonatum cyrtonema transplanted for 1 month in the embodiment 1 of the present application;

[0027] Figure 6 The complete regenerated plants of Polygonatum cyrtonema transplanted for 2 months in the embodiment 1 of the present application. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme in the embodiments of the present application will be described clearly and completely in combination with the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0029] The test materials and reagents used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0030] The specific technologies or conditions not mentioned in the embodiments can be carried out according to the technologies or conditions described in the literature in the art or according to the product instructions.

[0031] Embodiment 1

[0032] A method for improving the survival rate of polyphylla tuber propagation seedlings after transplanting, the specific operation is as follows:

[0033] (1) The uniform and healthy polyphylla tuber propagation elongated buds with a height of about 2.5-3.0 cm (obtained from the Chinese patent application document with application number 202211199919.9, i.e. the polyphylla tuber adventitious buds obtained after step (4) of the example 1 of the application document) are separated from the adventitious bud cluster. A part is directly inoculated into the adventitious root induction medium and culture conditions involved in the example 2(6) of the Chinese patent application document with application number 202211199916.5 to induce adventitious roots as the control group; another part is treated by immersing the base in an aqueous solution containing NAA with a final concentration of 1000 mg / L, IBA with a final concentration of 750 mg / L and ABA with a final concentration of 25 mg / L for 90 s, and then inoculating the treated elongated buds into 1 / 2MS medium containing 40 g / L sucrose and 7.0 g / L agar to induce adventitious roots and microtubers synchronously in a greenhouse with a temperature of 25±2℃, a light intensity of 2000 lx, a light cycle of 14 / 10 h (light / dark), and blue light quality. After 6 weeks of light culture, the control group polyphylla tuber elongated buds induced adventitious roots at the base, but no microtubers formed, with an average of 6.8 adventitious roots per explant, and the average length of adventitious roots was 2.1 cm( Figure 1 )。 The polyphylla tuber propagation seedlings are taken out and washed under running water, and then soaked in a 0.05% potassium permanganate solution containing 40 μM melatonin for 7.5 min. The treated polyphylla tuber propagation seedlings are transplanted into a seedling raising bag containing garden soil, perlite and nutrient soil in a volume ratio of 6:1:2 and cultivated under the forest at an altitude of 300-600 m. After transplanting, 1 / 4MS macroelements are sprayed on the leaves. Cover the shading net, and remove the shading net after 4 weeks. During this period, water and 1 / 4MS macroelements are sprayed every 2 weeks, and the average survival rate after 2 months of transplanting is only 75.6%. For the experimental group of polyphylla tuber propagation elongated buds, adventitious roots are formed at the base of the stems accompanied by the formation of microtubers after 4 weeks of light culture( Figure 2 ), and the synchronous induction rate of adventitious roots and microtubers reaches 96.5% after 2 weeks of continuous light culture, with an average of 12.3 adventitious roots per explant, and the average length of adventitious roots is 5.4 cm( Figure 3 ).

[0034] (2) The test tube containing the regenerated plant of Qiyuan Rhizoma Polygonati with induced microtuber and strong adventitious root was moved to a plastic greenhouse under natural light for 7-10 days. Then the test tube containing the regenerated plant of Huangjing Rhizoma Polygonati was taken out and washed under running water to clean the root, and then soaked in 40 μM melatonin added with 0.05% KMnO4 solution for 7.5 min. The surface water was dried, and the test tube was ready for use. Figure 4

[0035] (3) The treated test tube containing the regenerated plant of Huangjing Rhizoma Polygonati was transplanted into a seedling bag containing garden soil: perlite: nutrient soil with a volume ratio of 6:1:2, and then cultured under a forest with an altitude of 300-600 m. After transplantation, the leaves were sprayed with 1 / 4 MS macroelements. The seedling bag was covered with a shading net, and the shading net was removed after 4 weeks. During the period, water was poured and 1 / 4 MS macroelements were sprayed every 2 weeks. After 1 month of transplantation, the original test tube leaves of the regenerated plant of Huangjing Rhizoma Polygonati were shed, and new leaves grew out Figure 5 ) After 2 months of transplantation, the plant was strong and green, with an average plant height of 7.3 cm, and an average survival rate of 96.8% after transplantation Figure 6 ).

[0036] Example 2

[0037] This example tests the effect of exogenous plant growth regulators on the synchronous induction of adventitious root and microtuber of elongated bud of Huangjing Rhizoma Polygonati. The specific steps are as follows:

[0038] ​(1) After separating the uniformly growing, robust tissue culture elongation shoots of Polygonatum multiflorum with a height of about 2.5-3.0 cm (obtained by the method described in Example 1) from the adventitious shoot clusters, the bases were immersed in aqueous solutions containing different concentrations of NAA (0, 250, 500, 750 and 1000 mg / L), IBA (0, 250, 500, 750 and 1000 mg / L) and ABA (0, 10, 25, 50, 75 and 100 mg / L) for 60 s. Then, the treated elongation shoots were inoculated into MS medium containing 30 g / L sucrose and 7.0 g / L agar and cultured under LED blue light conditions at a temperature of 25±2℃, a light intensity of 2000 lx, and a photoperiod of 14 / 10 h (light / dark) for the simultaneous induction culture of adventitious roots and micro-tubers. During the cultivation period, the induction of adventitious roots and microtubers was observed continuously. After 6 weeks of light cultivation, the induction rate of adventitious roots and microtubers, the average number of adventitious roots produced per explant, and the length of adventitious roots were counted. The results are shown in Table 1. The results indicate that during the synchronous induction of adventitious roots and microtubers in Polygonatum cyrtonema, exogenous plant growth regulators NAA, IBA, and ABA played a synergistic promoting role within a certain concentration range. High concentrations of ABA inhibited the formation of adventitious roots and microtubers. Among them, when the base of the elongated shoot of Polygonatum cyrtonema was placed in an aqueous solution with a final concentration of 1000 mg / L NAA, 750 mg / L IBA, and 25 mg / L ABA for 60 s, the induction effect of adventitious roots was the best, with a synchronous induction efficiency of 77.9% for adventitious roots and microtubers. On average, 5.2 adventitious roots were produced per explant, and the average length of the adventitious roots was 4.4 cm.

[0039] Table 1. Effects of different exogenous NAA, IBA, and ABA concentrations on the synchronous induction of adventitious roots and micro-tuberization in tissue culture of Polygonatum odoratum.

[0040]

[0041] Note: Data are averages. Each treatment contained 120 explants and was repeated three times.

[0042] Example 3

[0043] This embodiment tested the effects of exogenous NAA, IBA, and ABA treatment time and sucrose concentration on the synchronous induction of adventitious roots and micro-tubers from tissue-cultured Polygonatum cyrtonema. The specific steps are as follows:

[0044] (1) The uniform and robust multiple-flowered Polygonum cy-clopetalum plantlets with the height of 2.5-3.0 cm were separated from the adventitious bud clusters (the method of obtaining was described in Example 1) and the base of the elongated buds was treated in the water solution with the concentration of 1000 mg / L NAA, 750 mg / L IBA and 25 mg / L ABA for different time (30, 60, 90, 120 and 180 s), then the treated elongated buds were inoculated in the MS medium with different concentrations of sucrose (20, 30, 40 and 60 g / L) and 7.0 g / L agar at the temperature of 25±2℃, the light intensity of 2000 lx and the light cycle of 14 / 10 h (light / dark) under the LED blue light condition to induce the synchronous induction of adventitious roots and microtubers. The induction of adventitious roots and microtubers was observed during the culture, and the induction rate of adventitious roots and microtubers, the number of adventitious roots produced by each explant and the length of adventitious roots were counted after the light culture for 6 weeks. The research results were shown in Table 2, and the research results showed that the treatment time of exogenous plant growth regulators and the concentration of sucrose had important regulatory effects on the synchronous induction of adventitious roots and microtubers in the process of synchronous induction of adventitious roots and microtubers of multiple-flowered Polygonum cy-clopetalum. In a certain range, with the extension of the treatment time of plant growth regulators and the increase of the concentration of sucrose, the synchronous induction rate of adventitious roots and microtubers of multiple-flowered Polygonum cy-clopetalum gradually increased. When the base of the elongated buds of multiple-flowered Polygonum cy-clopetalum was placed in the water solution with the final concentration of 1000 mg / L NAA, 750 mg / L IBA and 25 mg / L ABA for 90 s and then inoculated in the MS medium with 40 g / L sucrose and 7.0 g / L agar, the induction effect of adventitious roots was the best, the synchronous induction efficiency of adventitious roots and microtubers was 89.4%, 8.3 adventitious roots were produced by each explant on average, and the average length of adventitious roots was 5.1 cm. Then, with the extension of the treatment time and the increase of the concentration of sucrose, the synchronous induction efficiency of adventitious roots and microtubers showed a downward trend.

[0045] Table 2 Effects of different treatment time of exogenous plant growth regulators and the concentration of sucrose in the medium on the synchronous induction of adventitious roots and microtubers of multiple-flowered Polygonum cy-clopetalum plantlets

[0046]

[0047] Note: The data were the average values, each treatment contained 120 explants, and each treatment was repeated three times.

[0048] Example 4

[0049] In this example, the effects of macronutrients in the medium and LED light quality on the synchronous induction of adventitious roots and microtubers of multiple-flowered Polygonum cy-clopetalum plantlets were tested. The specific steps were as follows:

[0050] (1) The uniform and robust P. polyphylla var. glabrescens elongated buds (obtained by the method described in Example 1) with a height of about 2.5-3.0 cm were separated from the adventitious bud clusters, and the base was treated by immersion in an aqueous solution containing 1000 mg / L NAA, 750 mg / L IBA and 25 mg / L ABA for 90 s, and then the treated elongated buds were inoculated into MS medium containing 40 g / L sucrose, 7.0 g / L agar and different concentrations of macroelements (1 / 4, 1 / 2, 1) and cultured under different LED light qualities (white light, red light, blue light and red and blue light quality ratio of 3:1) at a temperature of 25±2℃, light intensity of 2000 lx and light cycle of 14 / 10 h (light / dark). The induction of adventitious roots and microtubers was observed during the culture period, and the induction rate of adventitious roots and microtubers, the number of adventitious roots produced by each explant and the length of adventitious roots were counted after 6 weeks of light culture. The results are shown in Table 3, and the results show that the content of macroelements in the medium and the LED light quality have important regulatory effects on the synchronous induction of adventitious roots and microtubers of P. polyphylla var. glabrescens. Compared with white light, LED blue light can effectively promote the synchronous formation of adventitious roots and microtubers of P. polyphylla var. glabrescens. Further research found that within a certain range, with the decrease of the content of macroelements in the medium, the synchronous induction rate of adventitious roots and microtubers of P. polyphylla var. glabrescens gradually increased. When the base of the elongated buds of P. polyphylla var. glabrescens was placed in an aqueous solution containing 1000 mg / L NAA, 750 mg / L IBA and 25 mg / L ABA with a final concentration of 1000 mg / L NAA, 750 mg / L IBA and 25 mg / L ABA for 90 s and then inoculated into 1 / 2MS medium containing 40 g / L sucrose and 7.0 g / L agar, the induction effect of adventitious roots was the best, the synchronous induction efficiency of adventitious roots and microtubers was 96.5%, 12.3 adventitious roots were produced per explant on average, and the average length of adventitious roots was 5.4 cm.

[0051] Table 3 Effect of LED light quality and macroelement content in the medium on the synchronous induction of adventitious roots and microtubers of P. polyphylla var. glabrescens tissue culture elongated buds

[0052]

[0053] Note: The data are the average values, each treatment contains 120 explants, and each treatment is repeated three times.

[0054] Example 5

[0055] In this example, the effects of exogenous melatonin concentration and transplanting substrate ratio on the survival rate of P. polyphylla var. glabrescens tissue culture seedlings were tested. The specific steps are as follows:

[0056] The tissue culture bottles containing the regenerated plants of Qiyuan polygonatum with induced microtubers and robust adventitious roots obtained in Examples 1-4 were moved to a plastic greenhouse under natural light for 7 days; then the polygonatum tissue culture seedlings were taken out and the roots were washed under running water, and then were immersed in a 0.05% potassium permanganate solution with different concentrations (0, 10, 20, 40 and 60 uM) of melatonin for 7.5 min, and after the surface moisture was dried, the seedlings were transplanted into a seedling raising bag containing different proportions of nutrient medium (garden soil: perlite: nutrient soil = 4-8: 1-2: 1-4) and were cultured to grow into seedlings under the forest at an altitude of 300-600 m. After transplantation, the leaves were sprayed with 1 / 4MS macroelements. Cover the sunshade net, and remove the sunshade net after 4 weeks. During this period, water and 1 / 4MS macroelements were sprayed every 2 weeks. The survival rate of transplanted seedlings was counted after 2 months of transplantation.

[0057] Table 4 Effect of exogenous melatonin concentration and transplanting medium ratio on the survival rate of transplanted polygonatum cyrtonema tissue culture seedlings

[0058]

[0059] Note: The data are the average values, each treatment contains 120 explants, and each treatment is repeated three times.

[0060] The research results are shown in Table 4, and the research results show that during the transplanted process of polygonatum cyrtonema tissue culture seedlings, the exogenous melatonin concentration and the nutrient medium ratio have an important influence on the survival rate of transplanted polygonatum cyrtonema tissue culture seedlings. The research results show that a certain concentration of melatonin soaking treatment can effectively improve the survival rate of transplanted polygonatum cyrtonema tissue culture seedlings. It is speculated that the reason may be that an appropriate concentration of melatonin can effectively promote the growth of polygonatum cyrtonema tissue culture seedlings and improve the stress tolerance to the environment. The research results show that after the polygonatum cyrtonema tissue culture seedlings are treated with a 0.05% potassium permanganate solution with 40 uM melatonin and transplanted into a nutrient medium with Vgarden soil: Vperlite: Vnutrient soil = 6:1:2, the survival rate of transplanted seedlings is the highest after 2 months of culture, which is 96.8%.

[0061] The above examples are only used to illustrate the technical solutions of the present application, but not limit it; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that the technical solutions recorded in the foregoing examples can be modified, or some technical features can be replaced by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for improving the survival rate of tissue culture seedlings of Polygonatum multiflorum during transplantation, characterized in that: Includes the following steps: (1) Synchronous induction culture of adventitious roots and micro-tuber: The elongated shoots obtained from the tissue culture of Polygonatum multiflorum were used as explants. They were separated from the adventitious shoot clusters, and the base of the shoots was treated with rooting promotion solution for 60-180 s. Then, they were inoculated into the culture medium for synchronous induction culture of adventitious roots and micro-tuber. The rooting promotion solution was an aqueous solution with a concentration of 500-1000 mg / L NAA, 250-1000 mg / L IBA and 10-100 mg / L ABA. The culture medium was 1 / 4-2 MS medium with 20-60 g / L sucrose and 7.0 g / L agar. (2) Pre-transplanting treatment of Polygonatum tissue culture seedlings: transfer the tissue culture bottles containing Polygonatum multiflorum regenerated plants with induced micro-tuber and adventitious roots to a plastic greenhouse under natural light outdoors for acclimatization. (3) Transplanting and seedling management of Polygonatum tissue culture seedlings: The treated Polygonatum tissue culture seedlings were transplanted into the substrate and cultivated under the forest to obtain multi-flowered Polygonatum regenerated plants.

2. The method for improving the survival rate of transplanted Polygonatum multiflorum tissue culture seedlings according to claim 1, characterized in that: In (1), the Polygonatum multiflorum is Polygonatum sibiricum from Qiyuan.

3. The method for improving the survival rate of Polygonatum multiflorum tissue culture seedlings after transplanting according to claim 1, characterized in that: In (1), adventitious roots and micro-tubers were synchronously induced and cultured in a greenhouse with a temperature of 25±2℃, a light intensity of 2000~2500lx, a photoperiod of 14h light / 10h darkness, and a light quality of one or more of blue light, red light, and white light.

4. The method for improving the survival rate of Polygonatum multiflorum tissue culture seedlings after transplanting according to claim 1, characterized in that: In (1), the synchronous induction culture time is 6 weeks.

5. The method for improving the survival rate of Polygonatum multiflorum tissue culture seedlings after transplanting according to claim 1, characterized in that: In (2), the acclimatization time in the greenhouse is 7-10 days.

6. The method for improving the survival rate of Polygonatum multiflorum tissue culture seedlings after transplanting according to claim 1, characterized in that: In (2), it also includes soaking the tissue culture seedlings after greenhouse domestication in a potassium permanganate solution with a concentration of 10-60 μM melatonin and a mass fraction of 0.01%-0.05% for 5-10 minutes.

7. The method for improving the survival rate of Polygonatum multiflorum tissue culture seedlings after transplanting according to any one of claims 1-6, characterized in that: In (3), the specific steps include: transplanting the treated Polygonatum tissue culture seedlings into seedling bags containing garden soil: perlite: nutrient soil volume ratio = 4-8: 1-2: 1-4 under the forest at an altitude of 300-600m for seedling cultivation; after transplanting, spray the leaves with 1 / 8-1 / 2 MS macronutrients, cover with a shade net, remove the shade net after 4 weeks, and during this period, water once every 2 weeks and spray with 1 / 8-1 / 2 MS macronutrients once to obtain Polygonatum multiflorum regenerated plants.

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