Method for promoting germination of viola tianshanica seeds

Abstract

The invention belongs to the technical field of medicinal plant cultivation, and particularly relates to a method for promoting viola tianshanica seed germination. The invention provides a method for promoting germination of viola tianshanica seeds, which comprises the following steps: soaking disinfected viola tianshanica seeds by using a gibberellin solution to obtain soaked seeds, and performing germination culture; the concentration of the gibberellin solution is 100 to 750 mg / L; the soaking treatment time is 4 to 24 hours. The viola tianshanensis seeds are soaked with gibberellin, dormancy of the viola tianshanensis seeds can be broken, germination is promoted, and the germination rate is increased.

Description

Technical Field

[0001] The invention belongs to the technical field of medicinal plant cultivation, and particularly relates to a method for promoting germination of Viola tianshanica seeds. Background Art

[0002] Tianshan violet (Viola tianschanica Maxim.) is a plant of the genus Viola of the Violaceae family, also known as Tibetan violet. Its scientific name was revised to Viola kunawarensis Royle in the Flora of China. It is a perennial dwarf herb with no aboveground stems and a height of 2.5 to 6 cm. The rhizome is shortened and relatively sturdy, with short internodes and dense nodes; the root is conical, brown or pale, slender, and usually unbranched. The leaves are all basal and rosette-shaped; the leaves are thick and papery, ovate, round or oblong, 0.5 to 2 cm long, 2 to 5 mm wide, obtuse at the apex, cuneate or broad cuneate at the base, slightly decurrent, the margin is entire or sparsely shallowly crenate, glabrous on both sides, and the main vein is obviously raised; the petiole is slightly longer or nearly equal to the leaf blade; the stipules are membranous, whitish, 1 / 2 to 2 / 3 connate to the petiole, the free part is lanceolate, the apex is gradually pointed, and the margin is sparsely glandular tassels. Flowers are small, dark blue-purple; pedicels are thin and straight, slightly longer than or nearly as long as leaves, with 2 bracts slightly above the middle; bracts are nearly opposite, linear or narrowly lanceolate, acuminate at the apex, with sparse glandular tassels at the lower part of the margin; sepals are oblong or ovate-lanceolate, 3-4mm long, 1-1.5mm wide, obtuse at the apex, with very short appendages at the base, obtuse at the end with 3 veins, and narrow membranous margins; petals are oblong-obovate, 7-10mm long, obtuse at the apex, slightly Narrow, lateral petals without hair, lower petals slightly shorter, with white veins; spur very short, sac-shaped, slightly longer or not longer than the appendages of the calyx; anthers about 1.5mm long, appendages at the top of the connective about 1mm long, spurs on the back of the two stamens below very short, only 0.4mm long; ovary ovoid, about 1.5mm long, smooth and hairless; style club-shaped, base obviously geniculate, top blunt and without edges, extending a very short beak forward; beak end with thinner stigma hole. Capsule oval, 5-7mm long. Flowering period June-July, fruiting period July-August.

[0003] Tianshan Viola is a high-cold, high-altitude plant distributed in Gansu, Qinghai, Xinjiang, northwest Sichuan, and Tibet. It grows in high mountains and subalpine meadows at an altitude of 2900 to 4500 meters, or in subalpine shrubs, and is often found in rock crevices or shaded and wet places beside gravel piles. In recent years, due to the influence of natural environment and human factors, the wild resources of Tianshan Viola have been severely damaged, the distribution range has gradually shrunk, the population has gradually decreased, and it is facing the danger of depletion. In addition, the germination rate of Tianshan Viola is low and the dormancy time is long in the natural state. Therefore, improving the germination rate of Tianshan Viola seeds and developing large-scale artificial planting are the inevitable choices for resource development, utilization and protection. However, the current domestic research on Tianshan Viola mainly focuses on medicinal value and chemical composition, and there are no reports on the germination mechanism of Tianshan Viola seeds and the effect of improving seed germination rate. Summary of the invention

[0004] The invention aims to make up for the deficiencies of the prior art, break the dormancy of Viola tianshanica seeds, promote the germination of Viola tianshanica seeds, and improve the germination rate of Viola tianshanica seeds.

[0005] The present invention provides a method for promoting germination of Viola tianshanica seeds, comprising the following steps:

[0006] The sterilized Viola tianshanica seeds are soaked in a gibberellin solution to obtain soaked seeds; and the soaked seeds are subjected to germination culture;

[0007] The concentration of the gibberellin solution is 100-750 mg / L; the soaking treatment time is 4-24 hours.

[0008] Preferably, the concentration of the gibberellin solution is 200-500 mg / L.

[0009] Preferably, the concentration of the gibberellin solution is 300 mg / L.

[0010] Preferably, the soaking treatment time is 8 hours.

[0011] Preferably, the seeds of Viola tianshanica are first disinfected using a sodium hypochlorite solution to obtain first disinfected seeds;

[0012] The first disinfected seeds are disinfected for the second time using an ethanol solution with a volume concentration of 65% to 75% to obtain disinfected Viola tianshanica seeds.

[0013] Preferably, the first disinfection method includes a first immersion, and the first immersion time is 5 to 15 minutes.

[0014] Preferably, the second disinfection method includes a second immersion; the second immersion is performed 2 to 3 times, and the duration of each immersion is 5 to 60 seconds.

[0015] Preferably, the germination culture method includes: after dark treatment for 4 days, culturing at 25° C. and a light-dark ratio of 12h / 12h.

[0016] Preferably, the humidity is maintained at 80-90% during the germination culture process.

[0017] Preferably, the germination culture time is 14 to 30 days.

[0018] Beneficial effects:

[0019] The present invention provides a method for promoting the germination of Viola tianshanica seeds, comprising the following steps: soaking the sterilized Viola tianshanica seeds with a gibberellin solution, obtaining the soaked seeds and then culturing them for germination; the concentration of the gibberellin solution is 100-750 mg / L; and the soaking time is 4-24 hours. The present invention soaks the Viola tianshanica seeds with gibberellin, which can increase the activity of multiple hydrolytic molds in the Viola tianshanica seeds to promote the decomposition of storage substances, and at the same time, can hinder the degradation of intercellular dormancy release signal transduction plasmodesmata callose, thereby transporting the signal substance to the apical meristem, breaking the dormancy of the Viola tianshanica seeds, promoting the germination of the Viola tianshanica seeds, and improving the germination rate. DETAILED DESCRIPTION

[0020] The present invention provides a method for promoting germination of Viola tianshanica seeds, comprising the following steps:

[0021] The sterilized Viola tianshanica seeds are soaked in a gibberellin solution to obtain soaked seeds; and the soaked seeds are subjected to germination culture;

[0022] The concentration of the gibberellin solution is 100-750 mg / L; the soaking treatment time is 4-24 hours.

[0023] As an embodiment, the present invention uses sodium hypochlorite solution to perform a first disinfection on Viola tianshanica seeds to obtain first disinfected seeds. As an embodiment, the first disinfection method includes a first soaking, and the first soaking time is 5 to 15 minutes; as another embodiment, the first soaking time of the present invention is 10 minutes. As an embodiment, the mass concentration of the sodium hypochlorite solution of the present invention is 1%.

[0024] As an embodiment, after obtaining the first disinfected seeds, the present invention uses an ethanol solution with a volume concentration of 65% to 75% to perform a second disinfection on the first disinfected seeds to obtain disinfected Tianshan Viola seeds; as another embodiment, after obtaining the first disinfected seeds, the present invention uses an ethanol solution with a volume concentration of 75% to perform a second disinfection on the first disinfected seeds to obtain disinfected Tianshan Viola seeds. As an embodiment, the second disinfection method of the present invention includes a second soaking. As an embodiment, the number of times of the second soaking of the present invention is 2 to 3 times, and the time of each soaking is 5 to 60 seconds; as another embodiment, the time of each soaking of the present invention is 10 to 50 seconds; as another embodiment, the time of each soaking of the present invention is 20 to 40 seconds; as another embodiment, the time of each soaking of the present invention is 30 seconds.

[0025] As an embodiment, the Viola tianshanica seeds of the present invention include Viola tianshanica seeds with full grains after natural maturity and indoor shade drying.

[0026] The invention sequentially utilizes sodium hypochlorite solution and 65%-75% volume concentration ethanol solution to disinfect the tianshanica violet seeds, which has low toxicity and is easy to remove, can accelerate the germination of the tianshanica violet seeds, and improve the germination rate of the tianshanica violet seeds.

[0027] After obtaining the sterilized Tianshan Viola seeds, the present invention soaks the sterilized Tianshan Viola seeds with a gibberellin solution to obtain soaked seeds. The concentration of the gibberellin solution of the present invention is 100-750 mg / L; and the soaking time is 4-24 hours.

[0028] As an embodiment, the concentration of the gibberellin solution of the present invention is 200-500 mg / L; as another embodiment, the concentration of the gibberellin solution of the present invention is 300 mg / L. As an embodiment, the soaking treatment time of the present invention is 8 hours.

[0029] After obtaining the soaked seeds, the present invention performs germination culture on the soaked seeds. As an embodiment, the present invention places the soaked seeds in a culture dish containing a double layer of virgin wood pulp paper for germination culture. The present invention utilizes a double layer of virgin wood pulp paper, which has good moisture retention, air permeability, heat preservation and water absorption compared to other materials, such as filter paper, and can further reduce seed rot.

[0030] As an embodiment, the germination culture method of the present invention includes: after 4 days of light-proof treatment, culturing at 25°C and a light-dark ratio of 12h / 12h. As an embodiment, the germination culture time of the present invention is 14 to 30 days; as another embodiment, the germination culture time of the present invention is 15 to 25 days; as another embodiment, the germination culture time of the present invention is 16 to 20 days; as another embodiment, the germination culture time of the present invention is 18 days. As an embodiment, the humidity is maintained at 80 to 90% during the germination culture process of the present invention.

[0031] Due to its own characteristics, the germination rate of Viola tianshanica is low under natural conditions. And conventional seed germination methods are difficult to be applied to Viola tianshanica. For example, the inventor tried to use the seed coat peeling experiment, and found that the seeds of Viola tianshanica were easy to rot during cultivation, and the cumulative 30-day germination rate was only 4%; the inventor tried to use the seed coat peeling experiment, and found that the cumulative 30-day germination rate of Viola tianshanica seeds was only 6%; the inventor tried to use 4°C dry storage for 2 weeks, 1 month, 2 months, 4 months and 6 months before sowing, and found that the cumulative 30-day germination rates of Viola tianshanica seeds were 0, 0.67%, 0, 0 and 0 respectively; the inventor tried to use 4°C sand storage for 2 weeks, 1 month, 2 months, 4 months and 6 months The inventor tried to sow the seeds after 2 weeks, 1 month, 2 months, 4 months and 6 months of dry storage at -20°C, and found that the cumulative 30d germination rates of the seeds of Viola tianshanica were 0, 1.33%, 0, 0 and 0, respectively; the inventor tried to sow the seeds after 2 weeks, 1 month, 2 months, 4 months and 6 months of dry storage at -20°C, and found that the cumulative 30d germination rates of the seeds of Viola tianshanica were 0, 0.67%, 0, 0 and 0, respectively; the inventor tried to sow the seeds after 2 weeks, 1 month, 2 months, 4 months and 6 months of sand storage at -20°C, and found that the cumulative 30d germination rates of the seeds of Viola tianshanica were 0, 0.67%, 0, 0 and 0, respectively. The present invention soaks the seeds of Viola tianshanica with gibberellins, and can accelerate the germination of the seeds of Viola tianshanica in a short time and improve the germination rate.

[0032] In order to further illustrate the present invention, a method for promoting germination of Viola tianshanica seeds provided by the present invention is described in detail below in conjunction with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0033] Example 1

[0034] 1. Test materials

[0035] Test seeds: In May 2024, the seeds of Tianshan violets that were artificially planted and naturally matured (transplanted and grown as seedlings) in the Chinese medicinal materials greenhouse in Taxkorgan County, Xinshi District, Urumqi, Xinjiang were harvested. After the harvested seeds were dried indoors, the seeds with full grains and not attacked by insects and pathogens were selected as test materials;

[0036] Experimental reagents: Gibberellic acid (GA3, purity ≥ 95%), purchased from BASF Biotechnology Co., Ltd. in Hefei, item number C0200, dissolved in alcohol at room temperature, generally 50 mL of 75% alcohol or 60% liquor is used to dissolve each gram for more than 1 hour;

[0037] Sodium hypochlorite solution (Antichloride, NaClO) was purchased from Shanghai Test (Sinopharm Chemical Reagent Co., Ltd.) with batch number PN: 20230116.

[0038] 2. Preprocessing

[0039] The experiment was conducted in August 2024. The experimental seeds were placed in a small gauze bag, put into a small beaker, and soaked and disinfected in 1% sodium hypochlorite for 5 minutes. After rinsing with sterile water three times, they were soaked and disinfected in 75% volume concentration alcohol twice, rinsed clean, and obtained pretreated seeds.

[0040] 3. Seed soaking

[0041] The pretreated seeds were soaked in 100, 200, 300, 500 and 750 mg / L gibberellin solutions for 8 h, with clear water as the control. Three replicates were set for each group, with 50 seeds in each replicate.

[0042] 4. Germination Culture

[0043] The soaked seeds of each treatment group were evenly arranged in a 100 mm culture dish covered with a double layer of virgin wood pulp paper, 150 seeds were placed in each 100 mm culture dish, water was added to cover the dish, and the dish was placed in an incubator for germination culture; wherein, during the germination culture process, an appropriate amount of water was added to the dish to keep the humidity of the dish at 80-90%; the light and temperature conditions of the incubator were set to avoid light for 4 days, and then light for 12 hours and darkness for 12 hours under a constant temperature of 25°C.

[0044] The number of germinated seeds was observed and recorded every day, and the germination rate of Viola truncatula seeds after germination and culture for 30 days was counted. The results are shown in Tables 1 and 2.

[0045] Table 1 Germination number and germination rate of Viola tianshanica seeds in different soaking treatments

[0046]

[0047]

[0048] Table 2 Germination number and germination rate of Viola tianshanica seeds in different soaking treatments

[0049]

[0050]

[0051] It can be seen from Table 1 and Table 2 that soaking the seeds of Viola tianshanica in gibberellin can improve the germination rate of the seeds. The germination rate of Viola tianshanica seeds is the highest when soaked in 300 mg / L gibberellin for 8 hours.

[0052] According to the above content, it can be seen that gibberellins can promote the germination of Viola tianshanica seeds and increase the seed germination rate.

[0053] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for promoting germination of Viola tianshanica seeds, characterized in that: The steps include: The sterilized Viola tianshanica seeds are soaked in a gibberellin solution to obtain soaked seeds; and the soaked seeds are subjected to germination culture; The concentration of the gibberellin solution is 100-750 mg / L; the soaking treatment time is 4-24 hours.

2. The method according to claim 1, characterized in that The concentration of the gibberellin solution is 200-500 mg / L.

3. The method according to claim 2, characterized in that The concentration of the gibberellin solution is 300 mg / L.

4. The method according to claim 1, characterized in that: The soaking time is 8 hours.

5. The method according to claim 1, characterized in that The method for preparing the sterilized Viola tianshanica seeds comprises: Using a sodium hypochlorite solution to first disinfect the Viola tianshanica seeds to obtain first disinfected seeds; The first disinfected seeds are disinfected for the second time using an ethanol solution with a volume concentration of 65% to 75% to obtain disinfected Viola tianshanica seeds.

6. The method according to claim 5, characterized in that The first disinfection method includes a first immersion, and the first immersion time is 5 to 15 minutes.

7. The method according to claim 5, characterized in that The second disinfection method includes a second immersion; the second immersion is performed 2 to 3 times, and the duration of each immersion is 5 to 60 seconds.

8. The method according to any one of claims 1 to 7, characterized in that: The germination culture method includes: after dark treatment for 4 days, culturing at 25° C. and a light-dark ratio of 12h / 12h.

9. The method according to claim 8, characterized in that During the germination culture, the humidity is maintained at 80-90%.

10. The method according to claim 8, characterized in that The germination culture time is 14 to 30 days.

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