A method for inducing flowering of morning glory in test tube

Abstract

The invention belongs to the field of morning glory cultivation, and particularly relates to a method for inducing morning glory test tube flowering. By adding a homogenate filtrate of geckos leaves into a conventional flower bud induction culture medium and a flower preservation culture medium, a flowering rate of 100% for the first morning glory flower, an average flowering rate of about 90% for the second flower, and an average flowering rate of 80% for the third flower of the morning glory is achieved, thereby greatly improving the flowering rate, greatly extending the flowering viewing time, improving the ornamental value of the test tube flower, and greatly improving the economic value of the test tube flower.

Description

Technical Field

[0001] The invention belongs to the field of morning glory cultivation, and particularly relates to a method for inducing morning glory test tube flowering. Background Art

[0002] The test tube flowering technology uses plant tissue culture technology to complete the flowering process of plants in a culture container under artificially controlled sterile environment conditions. There are still relatively few species of flowering plants, and only a few species of plants that can grow healthily in special test tubes have been developed, which is difficult to satisfy consumers' curiosity. At present, domestic test tube plants can be roughly divided into two categories: one is the flower-viewing type, such as Chinese orchids, cockscombs, green boxes, dianthus, dendrobium, etc.; the other is the leaf-viewing type, such as golden thread vine, aloe, cactus, etc. However, the varieties of flower-viewing test tube plants are relatively single and need to be developed urgently.

[0003] Iris Ipomoea quamoclit L.) is an annual weak twining herbaceous plant of the genus Ipomoea in the family Convolvulaceae. The leaves are ovate or oblong, deeply pinnate to the midrib, and often have pseudostipules at the base; the inflorescence is axillary, composed of a few flowers in a cyme, the flowers are upright, the pedicels are longer than the calyx, and thicken into a rod shape when fruiting. The name of the morning glory is because its leaves resemble bird feathers and it is a creeping herb. As morning glory has a high ornamental value, it has gradually become one of the important materials for in vitro flowering. Summary of the invention

[0004] The invention provides a method for inducing flowering of morning glory in a test tube, which greatly prolongs the viewing time of the flowering period and improves the ornamental value of the test tube flower.

[0005] The technical solution of the present invention is achieved in this way:

[0006] A culture medium combination for inducing flowering of morning glory in a test tube, characterized by comprising the following components:

[0007] Flower bud induction medium: modified MS + abscisic acid 0~1 mg / L + Gecko wood leaves 25~100 g / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8; in the modified MS, NH4NO3 825mg / L, KNO3 950mg / L, KH2PO4340mg / L, H3BO3 24.8mg / L, the other components and concentrations refer to MS medium;

[0008] Flowering preservation medium: MS + naphthaleneacetic acid 0.5 mg / L + paclobutrazol 0-0.05 mg / L + Gecko styracifolia leaves 50-100 g / L + Shannong No. 1 endophyte type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH=5.8; The filtrate prepared from the Gecko styracifolia leaves is added to the flower bud induction medium and the flowering preservation medium.

[0009] Furthermore, the method for preparing the filtrate of the leaves of the Gecko styraciflua is as follows: fresh leaves of the Gecko styracifluaflua without diseases and insect pests are selected, and the leaves are cleaned and removed of impurities; distilled water is added, the leaves are crushed, and the filtrate is obtained by filtering.

[0010] Furthermore, the composition of the flower bud induction culture medium is: modified MS + abscisic acid 0.5 mg / L + Gecko sphaerocephala leaves 75-100 g / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8.

[0011] Furthermore, the composition of the flowering preservation medium is: MS + naphthaleneacetic acid 0.5 mg / L + paclobutrazol 0.02-0.05 mg / L + gecko leaves 50-100 g / L + Shannong No. 1 endophyte type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH=5.8.

[0012] Furthermore, the composition of the flower bud induction culture medium is: modified MS + abscisic acid 0.5 mg / L + Gecko sphaerocephala leaves 75 g / L + Shannong No. 1 endophytic bacteria type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8; the composition of the flower preservation culture medium is: MS + naphthaleneacetic acid 0.5 mg / L + paclobutrazol 0.05 mg / L + Gecko sphaerocephala leaves 75 g / L + Shannong No. 1 endophytic bacteria type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH=5.8.

[0013] A method for inducing flowering of morning glory in a test tube comprises the following steps:

[0014] (1) Seed disinfection and inoculation

[0015] Take the morning glory seeds and sterilize them; on a sterile workbench, inoculate the sterilized morning glory seeds into a test tube and place them on the surface of the seed germination medium;

[0016] (2) Proliferation culture

[0017] After the seeds germinate but before true leaves grow, cut the buds and place them in proliferation medium for 15 days;

[0018] (3) Inducing flower bud differentiation

[0019] The seedlings were cut into pieces with 1-2 buds, inoculated into the above-mentioned flower bud induction medium, and cultured for 15 days, and then placed in a culture room for culture;

[0020] (4) Preservation of mature flowers

[0021] The plants cultured in the flower bud induction medium for 15 days were cut off from the callus and root system at the base, inoculated into the above-mentioned flower preservation medium for culture, and placed in a culture room for culture.

[0022] Furthermore, in the step (1), the disinfection method of morning glory seeds is as follows: take morning glory seeds, first rinse with tap water for 30 minutes, then rinse with 1% detergent for 3 times, then transfer to a sterile workbench, soak in 75% alcohol for 1 minute, then rinse with sterile water for 5 times, then soak in 15% sodium hypochlorite for 8 minutes, then rinse with sterile water for 5 times; the composition of morning glory seed germination culture medium is: MS + naphthaleneacetic acid 0.05-0.5 mg / L + Shannong No. 1 explant type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH = 5.8; culture until the seed bud height is 2-3 cm.

[0023] Furthermore, in the step (2), the composition of the proliferation medium is: proliferation medium: MS + naphthaleneacetic acid 0.05~0.5 mg / L + 6-BA 0.5~1 mg / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 30 g / L + carrageenan 7 g / L, pH = 5.8.

[0024] Furthermore, in the steps (1)-(4), the temperature in the culture room is controlled at 24°C, the light intensity is 2000Lx, and the light treatment period is 12h-dark treatment period is 12h.

[0025] Beneficial effects of the present invention:

[0026] The present invention uses mature seeds of morning glory as explants, the seeds are disinfected, aseptically inoculated into a culture medium, and after germination, they undergo the four steps of proliferation, flower bud induction, and flower preservation and cultivation. The flowering rate of morning glory can reach 60% after conventional flower bud induction and flower cultivation, and basically no second flower blooms, and the flowering period is short. The homogenate filtrate of the leaves of the gecko is added to the conventional flower bud induction culture medium and the flower preservation culture medium, so that the flowering rate of the first flower of morning glory is 100%, the flowering rate of the second flower is about 90% on average, and the flowering rate of the third flower is 80% on average, which greatly prolongs the flowering period, improves the ornamental value of the test tube flower, and greatly improves the economic value of the test tube flower. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0028] Figure 1The left picture shows the germination of morning glory seeds, the induction of flower buds, and the flowering and withering.

[0029] Figure 2 This is a picture of the flower buds induced by adding only ABA to morning glory.

[0030] Figure 3 This is a picture of the test tube flowering and the second flower bud.

[0031] Figure 4 The left picture shows the successful flowering, and the right picture shows the flower buds falling off and the failure to bloom. DETAILED DESCRIPTION

[0032] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] Example 1

[0034] (1) Preparation and packaging of culture medium (1L for 30 bottles)

[0035] Culture medium formula:

[0036] 1. Seed germination medium: MS + naphthaleneacetic acid (NAA) 0.05~0.5 mg / L + Shannong No. 1 explant type (broad-spectrum biocide, Shandong Zhike Biotechnology Co., Ltd.) 0.5 mL / L + sucrose 25 g / L + activated carbon 0.5 g / L + carrageenan 7 g / L, pH=5.8.

[0037] 2. Proliferation medium: MS + naphthaleneacetic acid (NAA) 0.05~0.5 mg / L + 6-BA 0.5~1 mg / L + Shannong No. 1 endophyte type (broad-spectrum biocide, Shandong Zhike Biotechnology Co., Ltd.) 0.5 mL / L + sucrose 30 g / L + carrageenan 7 g / L, pH=5.8.

[0038] 3. Flower bud induction medium: modified MS + abscisic acid (ABA) 0.5 mg / L + Gecko leaves 75 g / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH = 5.8. Among them, in the modified MS, NH4NO3 825 mg / L, KNO3 950 mg / L, KH2PO4 340 mg / L, H3BO3 24.8 mg / L, the other components and concentrations refer to MS medium.

[0039] 4. Flower preservation medium: MS + naphthaleneacetic acid (NAA) 0.5 mg / L + paclobutrazol 0.05 mg / L + Gecko sphaerocephala leaf homogenate filtrate 75 g / L + Shannong No. 1 endophytic bacteria type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH=5.8.

[0040] Shou Gong Wood ( Sauropus androgynus (L.) Merr.) leaves The filtrate prepared by homogenizing the leaves of the gecko was added to the flower bud induction medium and the flower preservation medium.

[0041] The method of making and adding the filtrate of the leaves of the Gecko Wood is as follows:

[0042] Take the addition of 100g / L of Gecko leaves as an example: select 100g of fresh Gecko leaves without diseases and insect pests, wash and remove impurities; add 100mL of distilled water and homogenize in a wall breaker for 10 minutes, and filter with filter paper to obtain the filtrate. In the preparation process of flower bud induction medium or flower preservation medium, take the obtained filtrate and add it to the corresponding culture medium, then add distilled water to make the volume to 1000ml, and divide it into 30 bottles for sterilization. If the addition amount of leaves is 25g / L, take 25g of leaves and make it in the same way as above.

[0043] Every 100g edible part (dry matter) of Gecko leaves contains 0.32g of mineral elements phosphorus, 3.22g of potassium, 0.02g of sodium, 0.72g of calcium, 0.49g of magnesium, and 0.08g of iron; contains 16.58μg / g of trace elements copper, 82.13μg / g of zinc, 82.13μg / g of manganese, 4.94μg / g of selenium, 24.97μg / g of iodine, and 19.94μg / g of molybdenum; and contains 24.16g of amino acids.

[0044] MS medium: NH4NO3 1650mg / L, KNO3 1900mg / L, CaCl2·2H2O 440mg / L, MgSO4 · 7H2O370mg / L, KH2PO4 170mg / L, KI 0.83mg / L, H3BO3 6.2mg / L, MnSO4·4H2O 22.3mg / L (MnSO4·H2O is 1.6900mg / L), ZnSO4 · 7H2O 8.6mg / L, Na2MoO4·2H2O 0.25mg / L, CuSO4·5H2O 0.025mg / L, CoCl2·6H2O 0.025mg / L, FeSO4·7H2O 27.8mg / L, Na2·EDTA 37.3mg / L, inositol 100mg / L, niacin 0.5mg / L, thiamine hydrochloride 0.1mg / L, pyridoxine hydrochloride 0.5mg / L, glycine 2.0mg / L, sucrose 30000mg / L, agar 10000mg / L, pH 5.8mg / L.

[0045] (2) Seed disinfection and inoculation

[0046] Rinse the morning glory seeds with tap water for 30 minutes, then rinse with 1% detergent 3 times, then transfer to a sterile workbench and soak in 75% alcohol for 1 minute, then rinse with sterile water 5 times, then soak in 15% sodium hypochlorite for 8 minutes, then rinse with sterile water 5 times. On a sterile workbench, inoculate the sterilized seeds into a test tube and place it on the surface of the seed germination medium. The culture room controls the temperature at 24°C, the light intensity at 2000Lx, and the light treatment is 12h-dark treatment for 12h. Cultivate until the height of the seeds is about 2-3cm after germination.

[0047] (3) Proliferation culture.

[0048] After the seeds germinate and before true leaves grow, they are about 2-3 cm tall, cut and placed in proliferation medium for 15 days. The culture room is controlled at 24°C, the light intensity is 2000Lx, and the light treatment is 12h-dark treatment 12h.

[0049] (4) Inducing flower bud differentiation

[0050] The seedlings were cut into pieces with 1-2 buds, inoculated into the flower bud induction medium and cultured for 15 days, and then cultured in a culture room with a controlled temperature of 24°C, a light intensity of 2000 Lx, and a light treatment of 12 hours and a dark treatment of 12 hours.

[0051] (5) Preservation of mature flowers

[0052] The plants cultured in the flower bud induction medium for 15 days had the callus and root system at the base cut off and were inoculated into the flower preservation medium for culture in a culture room (24°C, 2000Lx, 12h light-12h dark).

[0053] The whole cultivation process Figure 1 shown.

[0054] Example 2-13

[0055] The cultivation method is the same as that in Example 1, except that the addition amounts of ABA and Gecko sphaerocephala leaves (added after homogenization into filtrate) in the flower bud induction medium are different, and the addition amounts of paclobutrazol and Gecko sphaerocephala leaves (added after homogenization into filtrate) in the flowering medium are different. The specific composition is as follows:

[0056] Table 1 Composition of culture medium for Examples 2-13

[0057] Serial number Flower bud induction medium Flower preservation medium Example 2 ABA / 0mg / L+leaves / 0g / L Paclobutrazol / 0mg / L+leaves / 0g / L Example 3 ABA / 0.5mg / L+leaves / 25g / L Paclobutrazol / 0mg / L+leaves / 50g / L Example 4 ABA / 0.5mg / L+leaves / 25g / L Paclobutrazol / 0mg / L+leaves / 75g / L Example 5 ABA / 0.5mg / L+leaves / 50g / L Paclobutrazol / 0mg / L+leaves / 100g / L Example 6 ABA / 0.5mg / L+leaves / 75g / L Paclobutrazol / 0.02mg / L+leaves / 0g / L Example 7 ABA / 0.5mg / L+leaves / 100g / L Paclobutrazol / 0.02mg / L+leaves / 50g / L Example 8 ABA / 0.5mg / L+leaves / 75g / L Paclobutrazol / 0.02mg / L+leaves / 75g / L Example 9 ABA / 0.5mg / L+leaves / 75g / L Paclobutrazol / 0.02mg / L+leaves / 100g / L Example 10 ABA / 0.5mg / L+leaves / 0g / L Paclobutrazol / 0.05mg / L+leaves / 0g / L Embodiment 11 ABA / 0.5mg / L+leaves / 75g / L Paclobutrazol / 0.05mg / L+leaves / 50g / L Example 12 ABA / 0mg / L+leaves / 75g / L Paclobutrazol / 0.05mg / L+leaves / 75g / L Example 13 ABA / 0.5mg / L+leaves / 75g / L Paclobutrazol / 0.05mg / L+leaves / 100g / L

[0058] Note: The leaves in the table are from the Gecko tree.

[0059] 1. Induced flower bud differentiation test According to the culture medium preparation method and cultivation method of Example 1, the growth regulator ABA (abscisic acid) and Gecko sempervivum leaves (mixed and filtrated to add) were added to the flower bud induction culture medium to prepare the culture medium and carry out the test. The test treatments are shown in Table 2. The seedlings of the proliferation culture were cut into pieces with 1-2 buds in size, inoculated into the flower bud induction culture medium and cultured for 15 days, and placed in a culture room (24°C, 2000Lx, 12h light-12h dark treatment) for culture. The number of the first flower bud and the second flower bud of each treated plant was tracked and recorded. Each test was treated with 10 bottles, 2 plants in one bottle, and repeated 3 times. The data were averaged. The results are shown in Tables 2 and Figure 2 , Figure 3 .

[0060] Table 2 Effects of growth regulators on flower bud induction

[0061]

[0062] Note: The leaves in the table are from the Gecko tree.

[0063] Studies have shown that the probability of producing flower buds is extremely low without the addition of plant regulator ABA and filtrate of Gecko leaves; in the conventional flower bud induction medium with ABA added, the induction rate of the first flower bud is 66.7%, and the induction rate of the second flower bud is 8.3% ( Figure 2); After adding the filtrate of the homogenate of the leaves of the gecko, the flower bud induction rate can be significantly increased. Among them, ABA / 0.5mg / L+leaves / 75g / L has the best effect, with the first flower bud induction rate up to 100% and the second flower bud induction rate up to 86.7% ( Figure 3 ). This indicates that the addition of the homogenate filtrate of Gecko cycad leaves can significantly increase the flower bud induction rate of Cycas revoluta and induce Cycas revoluta to produce more flower buds.

[0064] 2. Flower preservation test

[0065] According to the medium preparation method of Example 1, paclobutrazol PP 333 The experiment was carried out by preparing the culture medium with two growth regulators, NAA and the filtrate of the homogenate of the leaves of the gecko. The experimental treatments are shown in Table 3. The plants cultured in the flower bud induction medium for 15 days were cut off the callus and root system at the base, inoculated into the flower preservation medium for culture, and cultured in the culture room (24℃, 2000Lx, 12h light-12h dark treatment). The number of the first flower, the second flower, and the third flower of each treated plant and the best viewing date of the plant were tracked and recorded. Each test treatment had 10 bottles, 2 plants in one bottle, and was repeated 3 times. The data were averaged. The results are shown in Tables 3 and Figure 4 The best viewing period starts from about 5-7 days after inoculation into the flower preservation medium, starting from the time when the inoculated plants take root and the leaves stretch, to the time when the flowers bloom and the first yellow leaves appear. Since the flowering conditions in each culture medium are different, some have one flower, and some have more than three, the best viewing period is calculated from the time when the flowers bloom and the first yellow leaves appear.

[0066] Flowering rate (%) = number of flowering trees / number of inoculated trees*100

[0067] Table 3 Effects of growth regulators on flower quantity and viewing period

[0068]

[0069] Note: The leaves in the table are from Gecko sempervivum; Treatment 1 is MS+NAA 0.5mg / L, and the other treatments are based on Treatment 1 with the addition of growth regulator PP 333 and leaves.

[0070] The study showed that in treatment 1, no growth regulator PP was added. 333 The third flower flowering rate is 0, and the ornamental period is short. The leaves will turn yellow and wilt after about 28 days of cultivation. Figure 4 , right).

[0071] From Table 3, it can be seen that the addition of PP 333It can significantly increase the number of days for viewing and the shelf life, which has a greater impact on the viewing period, but has little effect on the flowering rate. 333 Best preserved.

[0072] Treatments 6-8 and 10-12 added PP to the flower storage medium 333 The homogenate filtrate of the leaves of the gecko can effectively increase the flowering rate. 333 / 0.05mg / L+leaves / 75g / L culture medium has the best effect ( Figure 4 The left picture in the figure shows that the first and second flowers have a flowering rate of 100%, and the third flower has a flowering rate of 95%. The viewing period can reach 86 days. Gecko is cultivated in Hainan, Guangdong, Yunnan, Fujian and Sichuan provinces in my country. It is an edible woody vegetable with huge leaf yield. Adding it to test tube flower products not only has a good effect of inducing flowering, but is also economical and environmentally friendly. It can significantly increase the viewing period of test tube flowers and greatly improve economic benefits.

[0073] Based on the above experimental data, the addition of the homogenate filtrate of Gecko cycad leaves can not only increase the flower bud induction rate of Cardinal twig, but also significantly increase the flowering rate of the second and third flowers of Cardinal twig, significantly improving the best viewing period of Cardinal twig test tube flowers.

[0074] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A culture medium combination for inducing flowering of morning glory in a test tube, characterized in that: Includes the following components: Flower bud induction medium: modified MS + abscisic acid 0.5 mg / L + Gecko wood leaves 25-100 g / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8; in the modified MS, NH4NO3 825 mg / L, KNO3 950 mg / L, KH2PO4 340 mg / L, H3BO3 24.8 mg / L, the other components and concentrations refer to MS medium; Flower preservation medium: MS + naphthaleneacetic acid 0.5 mg / L + paclobutrazol 0-0.05 mg / L + Gecko leaves 50-100 g / L + Shannong No. 1 endophyte type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH = 5.8; The filtrate prepared by homogenizing the leaves of the gecko is added into a flower bud induction medium and a flower formation preservation medium.

2. The culture medium combination according to claim 1, characterized in that The method for preparing the filtrate of the leaves of the gecko shrub is as follows: fresh leaves of the gecko shrub without diseases and insect pests are selected, and the leaves are washed and impurities are removed; distilled water is added to homogenize the leaves, and the filtrate is obtained by filtering.

3. The culture medium combination according to claim 1, characterized in that The induction flower bud culture medium is composed of: modified MS + abscisic acid 0.5 mg / L + Gecko wood leaf 75-100 g / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.

8.

4. The culture medium combination according to claim 1, characterized in that The composition of the flowering preservation medium is: MS+naphthaleneacetic acid 0.5 mg / L+paclobutrazol 0.02-0.05 mg / L+Gecko sphaerocephala leaves 50-100 g / L+Shannong No. 1 endophytic bacteria type 1.0 mL / L+sucrose 25 g / L+carrageenan 9 g / L, pH=5.

8.

5. The culture medium combination according to claim 1, characterized in that The composition of the flower bud induction culture medium is: modified MS + abscisic acid 0.5 mg / L + Gecko Wood Leaves 75 g / L + Shannong No. 1 Endophytic Bacteria Type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8; the composition of the flower preservation culture medium is: MS + naphthaleneacetic acid 0.5 mg / L + paclobutrazol 0.05 mg / L + Gecko Wood Leaves 75 g / L + Shannong No. 1 Endophytic Bacteria Type 1.0 mL / L + sucrose 25 g / L + carrageenan 9 g / L, pH=5.

8.

6. A method for inducing flowering of morning glory in a test tube, characterized in that: The following steps are involved: (1) Seed disinfection and inoculation Take the morning glory seeds and sterilize them; on a sterile workbench, inoculate the sterilized morning glory seeds into a test tube and place them on the surface of the seed germination medium; (2) Proliferation culture After the seeds germinate but before true leaves grow, cut the buds and place them in a proliferation medium for 15 days; (3) Inducing flower bud differentiation Cutting the proliferated seedlings into pieces with 1-2 buds, inoculating them into the flower bud induction medium according to any one of claims 1-3 or 5, culturing them for 15 days, and culturing them in a culture room; (4) Preservation of mature flowers The plant cultured in the flower bud induction medium for 15 days has the callus and root system at the base cut off, and is inoculated into the flower preservation medium according to any one of claims 1 or 4-5 for culture, and is placed in a culture room for culture.

7. A method for inducing flowering of morning glory in a test tube as claimed in claim 6, characterized in that: In the step (1), the disinfection method of morning glory seeds is as follows: take morning glory seeds, rinse them with tap water for 30 minutes, then rinse them with 1% detergent for 3 times, then transfer them to a sterile workbench, soak them in 75% alcohol for 1 minute, then rinse them with sterile water for 5 times, then soak them in 15% sodium hypochlorite for 8 minutes, then rinse them with sterile water for 5 times; the composition of morning glory seed germination culture medium is: MS + naphthaleneacetic acid 0.05-0.5 mg / L + Shannong No. 1 explant type 0.5 mL / L + sucrose 25 g / L + carrageenan 7 g / L, pH=5.8; culture until the seed bud height is 2-3 cm.

8. The method for inducing flowering of morning glory in a test tube as claimed in claim 6, characterized in that: In the step (2), the composition of the proliferation medium is: proliferation medium: MS + naphthaleneacetic acid 0.05~0.5 mg / L + 6-BA 0.5~1 mg / L + Shannong No. 1 endophyte type 0.5 mL / L + sucrose 30 g / L + carrageenan 7 g / L, pH = 5.

8.

9. The method for inducing flowering of morning glory in a test tube as claimed in claim 6, characterized in that: In the steps (1) to (4), the temperature in the culture room is controlled at 24°C, the light intensity is 2000 Lx, and the light treatment is 12 hours and the dark treatment is 12 hours.

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