Method for promoting premna microphylla seed germination

Abstract

The invention relates to the technical field of plant cultivation, and discloses a method for promoting premna microphylla seed germination, which comprises the following steps: S1, material preparation: picking fruits of plants with robust growth vigor and no disease, taking out large and full seeds in the fruits, and placing the seeds in a ventilated place for airing; s2, gibberellin soaking: soaking the seeds aired in S1 in a gibberellin solution; the concentration of the gibberellin solution is 100 ppm to 500 ppm; the soaking time is 1-6 hours; s3, auxin soaking: soaking the seeds soaked in the gibberellin solution in auxin; the concentration of the auxin is 100 ppm to 500 ppm; the soaking time is 1-6 hours; s4, hole tray sowing: after the seeds soaked in the gibberellin solution are soaked and disinfected with carbendazim, hole tray sowing is carried out in the substrate. The gibberellin in the method can effectively break dormancy of premna microphylla seeds, promote early germination of the seeds and remarkably improve the germination rate of the seeds; the auxin can effectively improve the germination rate.

Description

Technical Field

[0001] The invention relates to the technical field of plant cultivation, and in particular to a method for promoting germination of tofu shiitake seeds. Background Art

[0002] Tofu Chai, also known as Guanyin Chai, stinking yellow broom, and rotten maiden, is a perennial deciduous shrub of the Verbenaceae family. The plant is distributed in South China, Central South, East China, Sichuan, Guizhou and other places in my country. It grows under forests, forest edges, roadsides and ditches below 700 meters above sea level, and is scattered under natural conditions. Tofu Chai has the characteristics of both medicine and food, and its whole body can be used. Pectin can be extracted from leaves and young branches, and can be processed into fairy tofu or refreshing drinks as green food raw materials; roots and stems have the effects of clearing heat and detoxification, anti-inflammatory and swelling, hemostasis, as well as enhancing immunity and anti-oxidation. It has high medicinal value and has formed an industrial chain.

[0003] However, there are certain difficulties in sexual reproduction of Tofu Chai, and seed propagation has not been successful so far. This seriously restricts the renewal of wild populations and the development of the industry. Therefore, how to break through seed propagation has become a major problem in current research. According to literature, asexual propagation methods are currently mostly used for cultivation, such as cuttings, rapid proliferation of tissue culture, and in vitro solution culture, while reports on Tofu Chai seed propagation are very rare. Previously, some scholars used low temperature treatment and warm water soaking to treat Tofu Chai seeds but failed to make them germinate. The author inferred that it may be dormant, and exogenous hormones are an important method to relieve seed dormancy and promote germination. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides a method for promoting the germination of tofu shiitake seeds, which solves the problem of difficulty in sexual reproduction of tofu shiitake in the prior art.

[0005] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention is: a method for promoting the germination of tofu shiitake seeds, comprising the following steps:

[0006] S1, material preparation: pick fruits from healthy and disease-free plants, take out the large and plump seeds in the fruits and place them in a ventilated place to dry;

[0007] S2, soaking in gibberellin: soak the dried seeds in S1 in a gibberellin solution; the concentration of the gibberellin solution is 100ppm to 500ppm; the soaking time is 1h to 6h;

[0008] S3, auxin soaking: the seeds soaked in the gibberellin solution are placed in the auxin solution for soaking; the auxin concentration is 100ppm to 500ppm; the soaking time is 1h to 6h;

[0009] S4, plug tray sowing: After the seeds soaked in gibberellin solution are disinfected with carbendazim, plug tray sowing is carried out in the substrate.

[0010] The beneficial effects of the present invention are as follows: by picking fruits of healthy and disease-free plants and selecting large and full seeds as materials, the genetic excellence and physiological activity of the seeds are fundamentally guaranteed. Subsequently, soaking in a gibberellin solution can break the dormancy of the seeds; auxin in the plant body mainly promotes cell elongation and participates in regulating the formation of plant morphology. Auxin treatment with an appropriate concentration can enhance the root development of seedlings, improve the stress resistance (such as drought resistance and disease resistance) and growth potential of seedlings; soaking and disinfecting the seeds with carbendazim before sowing in plug trays is a key measure to prevent seed bacteria and reduce the occurrence of seedling diseases.

[0011] Furthermore, the above method for promoting the germination of tofu shiitake seeds, S1 specifically comprises: picking fruits from healthy and disease-free plants, soaking the fruits in clean water, rubbing the peel after the pulp softens, crushing the pulp, separating the peel from the seeds, and selecting large and full seeds and placing them in a ventilated place to dry.

[0012] The beneficial effects of adopting the above further scheme are as follows: by soaking the fruit in clean water, rubbing the peel and crushing the flesh after the flesh softens, the peel, flesh and other impurities can be effectively separated from the seeds. The purity of the seeds is greatly improved, the pulp residues and possible microbial contamination attached to the surface of the seeds are removed, and the risk of disease caused by impurities during the germination of the seeds is reduced. At the same time, large and full seeds are selected for drying to ensure the high quality of the seeds.

[0013] Furthermore, in the above method for promoting the germination of tofu shiitake seeds, the matrix in S4 includes peat and perlite, and the weight ratio of peat to perlite is 2:1.

[0014] The beneficial effects of adopting the above further scheme are: peat, as a high-quality organic medium, has extremely high porosity and water holding capacity, and is rich in organic matter and microorganisms. When it is mixed with perlite in a ratio of 2:1, it can form a loose, porous, and well-ventilated matrix environment. This matrix is ​​conducive to the respiration of seed roots, reduces the root rot caused by lack of oxygen, and provides favorable conditions for the healthy germination of seeds.

[0015] Furthermore, in the above method for promoting germination of tofu shiitake seeds, the substrate in S4 is kept moist.

[0016] The invention has the following beneficial effects: the tofu shiitake seeds have the characteristic of long-term dormancy and the phenomenon of germination every other year. In the natural state, the seeds are sown in July of the current year and start to germinate in March of the next year, and the germination rate is about 20%. Gibberellic acid can effectively break the dormancy of the seeds, promote early germination of the seeds, and significantly improve the germination rate of the seeds. The seeds can germinate in the first year, but the germination is less, and the germination is mainly concentrated in the second year. The optimal treatment is 500ppm gibberellin soaking for 1 hour, the seeds start to germinate on the 15th day, and the germination rate can reach 61.11% in the second year; auxin cannot break the dormancy of the seeds, the seeds do not germinate in the current year, but can effectively improve the germination rate. The optimal treatment is 200ppm auxin soaking for 1 hour, the seeds start to germinate in March of the second year, and the germination rate reaches 55.56%. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Flowchart of this method. DETAILED DESCRIPTION

[0018] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0019] The fruit of the tofu taro is a berry-like drupe, which is obovate to nearly spherical, green when young and purple-black when mature. Each drupe contains one seed, which is ovoid, with a hard seed coat, irregular stripes and tumor-like protrusions on the surface, and a beak-like notch at the tip. It was observed to be naturally formed, not artificially caused, and this special structure is significantly different from other plants in the same family. During the experiment, it was found that when the seeds germinate, the radicle grows downward from there, and then the hypocotyl continues to elongate, pushing the seed coat out of the ground, and then the seed coat falls off and cotyledons grow. The average length of tofu taro seeds is 4.20mm, width is 2.91mm, thickness is 2.95mm, and the thousand-grain weight is 10.152g.

[0020] Example 1

[0021] like Figure 1 As shown, this embodiment provides a method for promoting the germination of tofu shiitake seeds. This embodiment adopts sowing as soon as the fruits are harvested. The fruits are harvested from the tofu shiitake nursery in Xiashanmen Village, Yongkang City, Zhejiang Province in June 2022. Plants with strong growth and no diseases are selected. The harvested fruits are brought back to the laboratory and soaked in clean water. After the pulp is softened, the peel is rubbed and the pulp is crushed. The peel is separated from the seeds, and large and plump seeds are selected and placed in a ventilated place to dry for later use.

[0022] The dried seeds were placed in a gibberellin solution and soaked. The concentration of the gibberellin solution was set to 100ppm, 200ppm, 300ppm, 400ppm, and 500ppm, with a total of 5 gradients. The soaking time was set to 1h, 2h, 4h, and 6h, with a total of 4 gradients. A total of 20 groups were treated, with 30 seeds in each group. The 20 groups of seeds soaked in the gibberellin solution were placed in the same concentration of auxin and soaked for the same time. After the treated seeds were soaked and disinfected with carbendazim, they were sown in a hole tray in the substrate. Each group was labeled. The substrate was peat and perlite, and the weight ratio of peat to perlite was 2:1. During the experiment, the substrate was kept moist, the seed germination was observed every week, and the seed germination rate was counted.

[0023] Example 2

[0024] In this embodiment, the fruit is sown as it is harvested. The fruits are harvested from the Doufucha nursery in Xiashanmen Village, Yongkang City, Zhejiang Province in June 2022. Plants with strong growth and no diseases are selected. The harvested fruits are brought back to the laboratory and soaked in clean water. After the pulp is softened, the peel is rubbed and the pulp is crushed to separate the peel from the seeds. Large and plump seeds are selected and placed in a ventilated place to dry for later use.

[0025] Set up 20 groups, each with 30 seeds, and soak the 20 groups of seeds in the same concentration of gibberellin solution for the same time. Soak the seeds soaked in the gibberellin solution in auxin. The concentration of auxin is set to 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, and a total of 5 gradients. The soaking time is set to 1h, 2h, 4h, and 6h, and a total of 4 gradients. After the treated seeds are soaked and disinfected with carbendazim, they are sown in plug trays in the substrate. Each group is labeled. The substrate is peat and perlite, and the weight ratio of peat to perlite is 2:1. Keep the substrate moist during the experiment, observe the germination of seeds every week, and count the seed germination rate.

[0026] Comparative Example

[0027] This comparative example adopts the method of sowing immediately after harvesting. The fruits were harvested from the Doufuchai Nursery in Xiashanmen Village, Yongkang City, Zhejiang Province in June 2022. Healthy and disease-free plants were selected. The harvested fruits were brought back to the laboratory and soaked in clean water. After the pulp softened, the peel was rubbed and the pulp was crushed to separate the peel from the seeds. Large and plump seeds were selected and placed in a ventilated place to dry for later use.

[0028] Twenty groups were set up, each with 30 seeds. These 20 groups were not treated in any way, and were sown in plug trays in the substrate. Each group was labeled. The substrate was peat and perlite, and the weight ratio of peat to perlite was 2:1. During the experiment, the substrate was kept moist, and the germination of seeds was observed every week, and the seed germination rate was calculated.

[0029] According to the above two embodiments and one comparative example, different gibberellin seed soaking treatment results and different auxin seed soaking treatment results were obtained, wherein the gibberellin seed soaking treatment results are shown in Table 1, in which GA3 100 is a gibberellin solution with a concentration of 100 ppm, GA3 200 is a gibberellin solution with a concentration of 200 ppm, GA3 300 is a gibberellin solution with a concentration of 300 ppm, GA3 400 is a gibberellin solution with a concentration of 400 ppm, and GA3 500 is a gibberellin solution with a concentration of 500 ppm; the auxin seed soaking treatment results are shown in Table 2, in which IAA100 is an auxin with a concentration of 100 ppm, IAA200 is an auxin with a concentration of 200 ppm, IAA300 is an auxin with a concentration of 300 ppm, IAA400 is an auxin with a concentration of 400 ppm, and IAA500 is an auxin with a concentration of 500 ppm. Seed germination rate (%) in the table = number of seeds germinated / total number of tested seeds × 100%; experimental data were analyzed using Microsoft Excel software and SPSS software.

[0030] Table 1:

[0031]

[0032] Table 2:

[0033]

[0034]

[0035] As shown in Table 1, under natural conditions, the seeds of the tofu shiitake mushroom do not germinate until April of the following year after being sown at the end of June of the current year, which takes about 9 months. This shows that the seeds of the tofu shiitake mushroom have a long-term dormancy and have the characteristic of germinating every other year. This is similar to some plants of the genus Ilex in the Aquifoliaceae family. Studies have shown that the reason for the dormancy of plants of this genus is that the embryo is not yet mature when the seeds mature and that the seed endosperm and embryo contain germination inhibitors. However, the reason for the dormancy of tofu shiitake mushroom seeds has not yet been found.

[0036] GA3 200+2h and GA3 300+2h had the fastest germination, and the seeds began to germinate on the 7th day, 263 days earlier than the control group. High concentration (400-500ppm) gibberellin concentration was more stable than low concentration (100-300ppm) in relieving the dormancy of tofu shiitake seeds, and the seeds could germinate in the same year. The germination rate was best in GA3 500+1h treatment, and the seeds began to germinate on the 14th day. The germination rate could reach 61.33% in the second year, which was more than 3 times that of the control group. The next treatments were GA3 100+1h, GA3 200+1h, and GA3 200+2h, with germination rates above 50%. The germination rate showed a trend of gradually decreasing with the increase of soaking time to a certain extent, and soaking seeds with gibberellin for 6h had an inhibitory effect on seed germination.

[0037] Gibberellic acid, as one of the most important plant hormones, can accelerate cell division and elongation, relieve the inhibition caused by abscisic acid in seeds, increase the activity of hydrolases in seeds, mainly α-amylase, decompose most of the storage materials, provide non-structural carbohydrates and other nutrients for seed germination, and then transport them to new organs for growth, thus promoting seed germination. The experimental results show that the treatment of seed soaking with gibberellins has a significant effect on relieving the dormancy of tofu shiitake seeds and improving the germination rate, and the seeds can germinate in the same year.

[0038] As shown in Table 2, auxin seed soaking has no significant effect on breaking the dormancy of tofu shiitake seeds, and the seeds began to germinate after 270 days. Auxin can effectively improve the germination rate. The variance analysis data showed that there were extremely significant differences between different treatment groups (p = 0.000). The optimal treatment was IAA200 + 1h, with the highest germination rate of 55.33%, followed by IAA300 + 2h, IAA400 + 4h, IAA400 + 1h, etc., with germination rates exceeding 50%, which is more than twice that of the control group. Auxin soaking for 6h inhibited seed germination.

Claims

1. A method for promoting the germination of tofu shiitake seeds, characterized in that: The following steps are involved: S1, material preparation: pick fruits from healthy and disease-free plants, take out the large and plump seeds in the fruits and place them in a ventilated place to dry; S2, soaking in gibberellin: soaking the seeds dried in S1 in a gibberellin solution; the concentration of the gibberellin solution is 100ppm to 500ppm; the soaking time is 1h to 6h; S3, auxin soaking: placing the seeds soaked in the gibberellin solution in the auxin solution; the auxin concentration is 100ppm to 500ppm; the soaking time is 1h to 6h; S4, plug tray sowing: After the seeds soaked in gibberellin solution are disinfected with carbendazim, plug tray sowing is carried out in the substrate.

2. The method for promoting germination of tofu shiitake seeds according to claim 1, characterized in that: The S1 specifically comprises: picking fruits from healthy and disease-free plants, soaking the fruits in clean water, rubbing the peels after the pulp is softened, crushing the pulp, separating the peels from the seeds, and selecting large and full seeds and placing them in a ventilated place to dry.

3. The method for promoting germination of tofu shiitake seeds according to claim 1, characterized in that: The matrix in S4 includes peat and perlite, and the weight ratio of the peat to perlite is 2:

1.

4. The method for promoting germination of tofu shiitake seeds according to claim 1, characterized in that: The substrate in S4 was kept moist.

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