A planting method for Morina nepalensis D. Don var. delavayi (Franch.) W. W. Smith

Abstract

A planting method of *Dipsacus inermis* Wall. var. *macranthus* W. C. Chen provided by the present invention realizes effective reflowering of *Dipsacus inermis* Wall. var. *macranthus* W. C. Chen by controlling seed harvesting, embedding and seed production, seedling raising, overwintering management and flower induction. It plays a positive role in the conservation of this plant with both medicinal and ornamental values and the development of exploration and utilization work. It has good development, utilization and popularization prospects in the utilization of similar plants. Since there is no report on any cultivation of *Dipsacus inermis* Wall. var. *macranthus* W. C. Chen at the present stage, it is the first in China.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and more specifically, to a cultivation method of Acanthocalyx nepalensis subsp. delavayi. Background Art

[0002] Acanthocalyx nepalensis subsp. delavayi, also known as Acanthocalyx delavayi, is a perennial herbaceous plant of the family Dipsacaceae and the genus Acanthocalyx. Compared with Acanthocalyx nepalensis distributed in the same area of this genus, it has an approximate appearance but larger flowers, with a diameter of up to 1.2 - 1.5 cm; the corolla lobes are oblong, 5 - 6 mm long, with a slightly concave apex, and the corolla tube is relatively wide, 4 - 5 mm wide. Acanthocalyx nepalensis subsp. delavayi is mainly distributed in southeastern Tibet, western Sichuan, and northwestern Yunnan in the mountainous areas of southwestern China, and often grows in alpine meadows at an altitude of 3000 - 4200 m, and is occasionally distributed in forest margins.

[0003] The flower shape of Acanthocalyx nepalensis subsp. delavayi is beautiful and can be developed for ornamental purposes. The whole plant can be used medicinally. In traditional Tibetan medicine, its whole herb can treat indigestion, and has the functions of strengthening the stomach, inducing vomiting, and detumescence. The aerial part can be used for joint pain, urinary incontinence, low back pain, and dizziness; externally, it can treat sores and suppurative wounds, etc. In the southwestern region, it is also a traditional tonic medicinal material of ethnic minorities such as the Naxi, Lisu, and Yi ethnic groups. There is a tradition among the Naxi people of stewing the roots with meat or chicken for daily physical conditioning, and it is one of the traditional "ginseng-like" medicinal materials of the Naxi ethnic group.

[0004] At the present stage, due to the limited vegetation survey coverage, the low level of understanding and utilization of resource plants, and the backward development of ethnic medicinal plants, there are no reports and technical solutions related to the research and development of any conservation technologies for Acanthocalyx nepalensis subsp. delavayi. The breeding, cultivation, and planting of Acanthocalyx nepalensis subsp. delavayi are still blank. It is urgent to conduct research on this species to protect its wild resources while meeting the development needs as a medicinal plant and an ornamental flower. Summary of the Invention

[0005] To solve or partially solve the problems existing in the related technologies, the present invention provides a cultivation method of Acanthocalyx nepalensis subsp. delavayi.

[0006] The present application provides a cultivation method of Acanthocalyx nepalensis subsp. delavayi, and the method includes the following steps:

[0007] S1: Seed collection and storage: Collect the slightly split fruit pods at the top of Acanthocalyx nepalensis subsp. delavayi, place them in a drying cabinet with an air humidity of 15% and a temperature of 5°C for 20 days, then take them out, shake off the seeds and collect them;

[0008] S2: Seed germination acceleration: In November or December, mix the seeds collected in S1 with leaf mold substrate at a volume ratio of 1:1 to obtain a mixture. Place the mixture in a 400-mesh screen bag, soak it until water-saturated, then place it in a breathable box and conduct light-avoiding germination acceleration at a room temperature of 18°C. The particle size of the leaf mold substrate is 0.5 - 2 mm.

[0009] S3: Sowing and seedling hardening: Regularly turn the screen bag in S2. After observing the growth of radicles in the screen bag, immediately take out all the radicles and the substrate, and broadcast them on the cultivation bed surface. Then, cover the whole with a layer of leaf mold with a thickness of 1 cm and a particle size of 5 - 20 mm, and maintain the soil volume water content of the growth layer substrate at 50% - 60% until the seedlings grow true leaves.

[0010] S4: Seedling overwintering management: Keep the daily air temperature in the planting area above 5°C, and at the same time maintain the soil volume water content in the planting area not less than 40%. Cover the soil surface with crushed withered leaves, and the covering thickness is 2 cm. The particle size of the crushed withered leaves is 2 - 4 cm.

[0011] S5: Flowering management: After the daily effective accumulated temperature is greater than 0°C and before the seedlings' leaves grow, spray a 100 μmol / L ABA solution once every 10 days until the new leaves of the seedlings grow. After 3 - 4 true leaves, independently spray boron foliar fertilizer until the leaves are completely wet. Spray once every 5 days, and stop spraying after the inflorescence axis elongates.

[0012] Furthermore, all sides of the breathable box in S2 are perforated for ventilation, and the area ratio of the single-sided ventilation pore diameter is 20% - 30%. The screen bag is fully covered with vermiculite. The particle size of the vermiculite is 3 - 5 mm.

[0013] Furthermore, the surface of the cultivation bed in S3 is composed of gravel, perlite, and Kanuma soil mixed at a volume ratio of 1:1:1; the lower part of the cultivation bed is composed of garden soil and river sand mixed at a volume ratio of 1:1. The thickness of the gravel is 2 cm, and the diameter grade is 1 - 1.5 cm; the diameter of the river sand does not exceed 5 cm.

[0014] Furthermore, the cultivation bed in S3 is placed in an outdoor environment with strong inter-annual variations; the 5-year average minimum temperature of the outdoor environment is between -10 and -5°C; the 5-year average maximum temperature of the outdoor environment is 20 - 25°C.

[0015] Furthermore, the boron foliar fertilizer in S5 is a 2500-fold solution of sodium octaborate tetrahydrate.

[0016] Furthermore, during the spraying period in S5, the ground drip irrigation method is used to keep the soil volume water content in the planting area not less than 40%.

[0017] Furthermore, the broadcasting thickness in S3 does not exceed 0.5 cm; the covering degree of the broadcasting is 50%.

[0018] Advantageous technical effects of the present invention:

[0019] The method provided by the present invention effectively elaborates on the key technologies in the process of plant propagation and flowering through processes such as seed collection and storage, seed germination, sowing and seedling hardening, overwintering management of seedlings, and flowering management. It has important value for the protection and expansion of the wild populations of this species, the development of the flower industry, the exploration of medicinal plant resources, the improvement of new technologies, as well as the corresponding new varieties, new production material products, and high-quality technical service capabilities. This method has the advantages of wide applicability and simple operation, and is the first of its kind in China, effectively filling the technical gap. Specific embodiments

[0020] The optional embodiments of the present invention application will be described in more detail below. Although the optional embodiments of the present invention application are presented, it should be understood that the present invention application can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention application more thorough and complete, and to be able to fully convey the scope of the present invention application to those skilled in the art.

[0021] For clarity, the following is a detailed description through Example 1 below. Unless otherwise specified, the test materials and reagents used in the following Example 1 are all purchased from the market.

[0022] Example 1

[0023] A method for planting Dipsacus spinosus var. megalanthus, the method comprising the following steps:

[0024] (1) In September 2020, at the periphery of Yulong Snow Mountain in Lijiang City, Yunnan Province, collect the slightly split fruit pods at the top of Dipsacus spinosus var. megalanthus, place them in a drying cabinet with an air humidity of 15% and a temperature of 5°C for 20 days, and then take them out, shake off the seeds and collect them.

[0025] (2) In November - December of the same year, mix the seeds of Dipsacus spinosus var. megalanthus with a leaf mold substrate (0.5 - 2 mm) that has passed through an 8 - mesh sieve and is not filtered by a 40 - mesh sieve in a volume ratio of 1:1, place them in a 400 - mesh plastic screen bag, fully soak them until water - saturated, then place them in a breathable plastic box filled with vermiculite with a particle size of 3 - 5 mm. The screen bag should be fully covered by vermiculite so that it is not visible to the naked eye; the plastic box should ensure that all sides are perforated for ventilation, and the area ratio of the single - side ventilation hole diameter is 20% - 30%. All plastic boxes are subjected to dark germination at a room temperature of 18°C.

[0026] (3) Regularly turn the gauze bags in the box. After observing the growth of radicles, immediately take out all the mixtures (substrate + seeds) and evenly spread them on the cultivation bed surface at a thickness not exceeding 0.5 cm and a coverage of 50%. The surface of the cultivation bed is composed of gravel, perlite, and Kanuma soil with a thickness of 2 cm and a particle size of 1 - 1.5 cm, mixed in a volume ratio of 1:1:1; except for the surface layer, the lower part is a mixture of garden soil and river sand (removing large particles with a diameter exceeding 5 cm, no sieving required) in a volume ratio of 1:1. After sowing, cover the entire surface of the bed with a layer of leaf mold soil with a thickness of 1 cm that cannot pass through a 4-mesh sieve, and water to keep the volumetric water content of the growth layer substrate soil at 50% - 60% until the seedlings grow true leaves. The bed should be placed in an outdoor area with strong interannual variations in air temperature. The average minimum temperature over 5 years is between -10 and -5 °C, and the average maximum temperature over 5 years is 20 - 25 °C to complete the growth of the seedlings in the current year.

[0027] (4) After the temperature drops in winter, use a ground-inserted tunnel shed to keep the planting area warm, maintaining the daytime air temperature in the planting area above 5 °C, and at the same time keeping the volumetric water content of the soil in the planting area not less than 40%. Cover the surface of the bed with withered leaves for heat preservation. The withered leaf covering layer is made into coarse particles (4 cm ≥ particle size ≥ 2 cm) by a material crusher, with a covering thickness of 2 cm. Observe in parallel groups of three with 100 plants in each group, and synchronously count the lodging rate.

[0028] (5) When the temperature warms up in the second year and the effective accumulated temperature with a daily temperature ≥ 10 °C is greater than 0 °C, before the seedlings' leaves grow, spray a 100 μmol / L ABA solution once every 10 days until the new leaves of the seedlings grow. After 3 - 4 true leaves, independently spray with a boron foliar fertilizer (2500-fold solution of sodium octaborate tetrahydrate) until the leaves are completely wet. Keep the leaves completely wet during each spraying, and spray once every 5 days until the inflorescence axis elongates and then stop. During the spraying period, use drip irrigation on the ground to keep the volumetric water content of the soil in the planting area not less than 40%. Observe in parallel groups of three with 100 plants in each group and count the flowering rate.

[0029] Control Example 1

[0030] According to the planting method of Morina nepalensis D. Don var. delavayi (Franch.) W. W. Smith provided in Example 1, after implementing steps (1) to (3) of Example 1, do not perform heat preservation treatment, expose the seedlings outdoors to overwinter by lodging, and then implement step (5) of Example 1, and sample and count the underground biomass after the seedlings turn green.

[0031] Control Example 2

[0032] According to a method for planting Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang provided in Example 1, after implementing steps (1) to (4) of Example 1, ABA solutions with different concentrations (10 μmol / L, 50 μmol / L, 200 μmol / L, 500 μmol / L) were sprayed, and no ABA spraying treatment (only spraying sterile water) was done. Only boron foliar fertilizer was sprayed after the seedlings turned green. Taking every 100 plants as a group, three parallel groups were used to statistically analyze the flowering rate.

[0033] Table 1 Effects of different overwintering methods on the underground biomass of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang seedlings

[0034]

[0035] As can be seen from Table 1, the heat preservation treatment has a significant enhancing effect on the accumulation of the underground biomass of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang. On the one hand, keeping the temperature in the cultivation area can significantly reduce the lodging rate of seedlings in winter and maintain the assimilation of some seedlings in winter. On the other hand, the underground part of the plant will also continue to grow in winter.

[0036] Table 2 Effects of different hormone treatment methods on the underground biomass of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang seedlings

[0037]

[0038] As can be seen from Table 2, spraying with a 100 μmol / L ABA solution can significantly increase the flowering rate of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang. At too low a concentration, the plant responds insensitively, while at too high a concentration, an obvious inhibitory effect will occur. However, even under the condition of suitable ABA solution treatment, the overall flowering rate of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang is still not high. This is mainly because for two-year-old seedlings, even though they have the conditions for flower bud differentiation, due to the limitation of physiological age, they cannot achieve full re-flowering.

[0039] To sum up, a method for planting Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang provided by the present invention realizes effective re-flowering of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang by controlling seed harvesting, embedding and seed production, seedling raising, overwintering management and flower induction. It plays a positive role in the conservation of this plant with both medicinal and ornamental values, as well as the development and utilization work. It has good development, utilization and popularization prospects in the utilization of similar plants. And since there is no report on any cultivation of Dipsacus inermis Wall. var. macranthus (Diels) C. Y. Wu & W. T. Wang at present, the present invention is the first in China.

[0040] The various embodiments 1 of the present invention application have been described above. The above description is exemplary and not exhaustive, and is also not limited to the disclosed embodiments 1. Many modifications and variations will be obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The choice of terms used herein is intended to best explain the principles of the embodiments, the practical application, or the improvement of the technology in the market, or to enable other ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A method for planting Dipsacus asper, characterized in that: The method comprises the following steps: S1: Seed collection and storage: Collect the slightly cracked fruit pods at the top of Dipsacus asper, place them in a drying cabinet with an air humidity of 15% and 5°C for 20 days, take out the seeds, shake them off and collect them; S2: Seed germination: In November and December, the seeds collected in S1 were mixed with leaf humus matrix at a volume ratio of 1:1 to obtain a mixture; the mixture was placed in a 400-mesh gauze bag, soaked until saturated with water, and then placed in a ventilation box for light-proof germination at room temperature of 18°C; the particle size of the leaf humus matrix was 0.5-2mm; S3: sowing and hardening seedlings: regularly turn over the mesh bag in S2, and when the radicle in the mesh bag is observed to grow, immediately take out all the radicle and matrix, and spread them on the cultivation bed; then cover the whole with a layer of leaf mold with a thickness of 1 cm and a particle size of 5-20 mm, and maintain the volume moisture content of the growth layer matrix soil at 50%-60% until the seedlings grow true leaves; The surface of the cultivation bed is composed of gravel, perlite, and Kanuma soil mixed in a volume ratio of 1:1:1; the lower part of the cultivation bed is composed of garden soil and river sand mixed in a volume ratio of 1:1; the thickness of the gravel is 2 cm and the diameter is 1-1.5 cm; the diameter of the river sand does not exceed 5 cm; S4: Winter management of seedlings: keep the daytime air temperature in the planting area above 5°C, and maintain the soil volume moisture content in the planting area not less than 40%; and cover the soil surface with crushed dead leaves with a covering thickness of 2 cm; the particle size of the crushed dead leaves is 2 to 4 cm; S5: Flowering management: After the daily effective accumulated temperature is greater than 0℃, before the leaves of the seedlings grow out, spray 100μmol / L ABA solution once every 10 days until the new leaves of the seedlings grow out; after 3-4 true leaves, use boron foliar fertilizer to spray independently until the leaves are completely moistened; spray once every 5 days until the inflorescence axis is elongated.

2. The method according to claim 1, characterized in that In S2, all sides of the air-permeable box are perforated for ventilation, and the area of ​​the ventilation holes on a single side accounts for 20%-30%; the mesh bag is fully covered with vermiculite; the particle size of the vermiculite is 3-5 mm.

3. The method according to claim 1, characterized in that The cultivation bed surface in S3 is placed in an outdoor environment with strong interannual variation; the 5-year average minimum temperature of the outdoor environment is between -10 and -5°C; the 5-year average maximum temperature of the outdoor environment is 20-25°C.

4. The method according to claim 1, characterized in that: The boron foliar fertilizer in S5 is 2500 times diluted sodium octaborate tetrahydrate.

5. The method according to claim 1, characterized in that During the spraying in S5, the surface drip irrigation method is used to keep the soil volume moisture content in the planting area not less than 40%.

6. The method according to claim 1, characterized in that The sowing thickness in S3 does not exceed 0.5 cm; the sowing coverage is 50%.

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