Germination culture method of artemisia selengensis seeds

Abstract

The invention relates to the technical field of plant culture, in particular to a germination culture method of artemisia selengensis seeds. The method specifically comprises the following steps: soaking seeds of Artemisia worshiana in distilled water, and then placing the seeds in a disinfectant for disinfection treatment; the disinfectant is a potassium permanganate solution with the mass fraction of 0.5%-1%, a sodium hypochlorite solution with the mass fraction of 5%-10% or a hydrogen peroxide solution with the mass fraction of 10%-30%; the disinfection treatment conditions are as follows: the soaking time is 10 to 30 minutes; cleaning the disinfected artemisia selengensis seeds with distilled water, absorbing moisture, placing the cleaned artemisia selengensis seeds on the surface of filter paper, and performing germination culture at the temperature of 20-30 DEG C. According to the method, the germination rate of the seeds can be effectively increased, the mildew rate is remarkably reduced, and the problem that the germination rate is relatively low when the method in the prior art is used for treating the artemisia anomala seeds is solved.

Description

Technical Field

[0001] The present invention relates to the technical field of plant cultivation, and particularly relates to a method for germinating and culturing Artemisia halodendron seeds. Background Art

[0002] Artemisia halodendron belongs to the genus Artemisia of the Compositae family and is a perennial herbaceous plant. It has attracted much attention due to its role in wind prevention and sand fixation. Artemisia halodendron is mainly distributed in the Horqin Sandy Land and the Hulunbuir Sandy Land and is one of the main sand-fixing plants. The above-ground part of Artemisia halodendron plays an important role in traditional Mongolian medicine. It is bitter in taste and cool in nature, and has multiple pharmacological effects such as insecticidal, analgesic, drying "xie ri wu su", antispasmodic and detumescence, and is widely used in the treatment of diseases such as chronic bronchitis.

[0003] As the starting stage of the plant life cycle, seed germination is not only a crucial link in growth and development but also the weakest period for the plant body to resist the external environment and is extremely vulnerable to external environmental interference. Currently, the main known factors affecting seed germination are temperature and humidity. However, in the study of Artemisia halodendron seed germination, based on the special growth environment of Artemisia halodendron seeds, drought stress is also one of the key factors affecting germination.

[0004] For example, Document 1: Man Da, Study on the Germination and Drought Resistance Physiological Characteristics of Artemisia halodendron Seeds [D]. Inner Mongolia Agricultural University, 2017. During the study of Artemisia halodendron seed germination in Document 1, mainly petri dish germination was used, and PEG-6000 was used to conduct solution drought stress on Artemisia halodendron seeds. The results showed that drought stress also had a certain impact on the germination rate. However, after the seeds of Artemisia halodendron were treated by the method in Document 1, the germination rate was still relatively low, only reaching 87% - 88%. Summary of the Invention

[0005] In order to solve the problem that the existing technology method for treating Artemisia halodendron seeds has a relatively low germination rate, the purpose of the present invention is to provide a method for germinating and culturing Artemisia halodendron seeds. The present invention can improve the germination rate of Artemisia halodendron seeds, thereby enhancing the application effect of Artemisia halodendron seeds in ecological restoration.

[0006] To achieve the above purpose, the technical solution of the present invention is as follows.

[0007] The present invention provides a method for germinating and culturing Artemisia halodendron seeds, comprising the following steps:

[0008] Soak the Artemisia halodendron seeds in distilled water and then place them in a disinfectant for disinfection; the disinfectant is a potassium permanganate solution with a mass fraction of 0.5% - 1%, a sodium hypochlorite solution with a mass fraction of 5% - 10% or a hydrogen peroxide solution with a mass fraction of 10% - 30%; the conditions for the disinfection treatment are: soaking time 10 min - 30 min.

[0009] Wash the disinfected Artemisia halodendron seeds with distilled water and blot them dry. Then place the washed Artemisia halodendron seeds on the surface of filter paper and conduct germination cultivation at a temperature of 20°C - 30°C.

[0010] This invention mainly conducts disinfection treatment on the Artemisia halodendron seeds after soaking. By selecting appropriate disinfectants and disinfection treatment conditions, and determining appropriate germination conditions, it can not only effectively solve the problem of endophyte contamination, but also effectively improve the germination rate of the seeds, significantly reduce the mildew rate, and solve the problem of relatively low germination rate in the existing technology for treating Artemisia halodendron seeds.

[0011] Preferably, the disinfectant is a potassium permanganate solution with a mass fraction of 1% or a sodium hypochlorite solution with a mass fraction of 10%.

[0012] This invention selects a potassium permanganate solution with a mass fraction of 1% for disinfection treatment and germination research. The results show that the germination rate of Artemisia halodendron seeds is 96%, the germination index is 42.45, and the vigor index is 206.98. This invention selects a sodium hypochlorite solution with a mass fraction of 10% for disinfection treatment and germination research. The results show that the germination rate of Artemisia halodendron seeds is 97.33%, the germination index is 40.31, and the vigor index is 149.97. The test results show that the method of this invention for disinfection treatment and germination research on Artemisia halodendron seeds can effectively increase the germination rate of Artemisia halodendron seeds to over 96%, and the germination index reaches over 40, and the vigor index reaches over 150.

[0013] Preferably, the conditions for the disinfection treatment are: soaking time 20 min.

[0014] Preferably, the conditions for the germination cultivation are: cultivation temperature 20°C, humidity 80%, light 12 h, darkness 12 h. The light intensity refers to natural light.

[0015] Preferably, 2 layers of filter paper are laid.

[0016] Preferably, during the germination cultivation, it also includes: keeping the filter paper moist; the degree of moisture of the filter paper should be such that no water flows out when pressed with a finger.

[0017] Preferably, the germination cultivation is based on the standard that the radicle grows 1 mm.

[0018] Advantages of the present invention:

[0019] 1. The present invention mainly disinfects the seeds of Artemisia halodendron after soaking. By selecting appropriate disinfectants and disinfection conditions, and determining appropriate germination conditions, it can not only effectively solve the problem of endophyte contamination, but also effectively improve the germination rate of seeds, significantly reduce the mildew rate, and solve the problem that the existing technology has a relatively low germination rate when treating the seeds of Artemisia halodendron.

[0020] 2. The method of the present invention disinfects and conducts germination research on the seeds of Artemisia halodendron. The results show that the method of the present invention can effectively increase the germination rate of the seeds of Artemisia halodendron to over 96%, and the germination index reaches over 40, and the vigor index reaches over 150. Description of the drawings

[0021] Figure 1 It is a comparison diagram of the effects of the treatment group and the control group in the disinfection soaking test of Artemisia halodendron seeds. Among them, A is a diagram of the difference ratio of 5 indicators between treatment groups 1 to 27 and the control group respectively; B is a Venn analysis diagram of the influence of treatment groups 1 to 27 and the control group on 5 indicators. UP indicates that a single indicator is significantly greater than the control group; MID indicates that there is no significant difference between a single indicator and the control group; DOWN indicates that a single indicator is significantly less than the control group.

[0022] Figure 2 It is a line graph showing the daily germination number of Artemisia halodendron seeds in the treatment groups treated with different disinfectants changing with the germination time. Among them, A is a line graph showing the daily germination number of Artemisia halodendron seeds in the treatment group treated with sodium hypochlorite changing with the germination time; B is a line graph showing the daily germination number of Artemisia halodendron seeds in the treatment group treated with potassium permanganate changing with the germination time; C is a line graph showing the daily germination number of Artemisia halodendron seeds in the treatment group treated with hydrogen peroxide changing with the germination time. T1 to T27 respectively represent treatment groups 1 to 27; CK represents the control group.

[0023] Figure 3 It is a correlation analysis diagram between indicators. Among them, * represents a significant level at 0.05, p ≤ 0.05; ** represents a significant level at 0.01, p ≤ 0.01; *** represents a significant level at 0.001, p ≤ 0.001.

[0024] Figure 4 It is a picture of the germination of Artemisia halodendron seeds in two control groups. Among them, (a) is a picture of the germination of Artemisia halodendron seeds in control group one; (b) is a picture of the germination of Artemisia halodendron seeds in control group two.

[0025] Figure 5It is a picture of the germination of Artemisia halodendron seeds in the theoretically optimal treatment group of KMnO4 and the theoretically optimal treatment group of H2O2. Among them, (a) is a picture of the germination of Artemisia halodendron seeds in the theoretically optimal treatment group of KMnO4; (b) is a picture of the germination of Artemisia halodendron seeds in the theoretically optimal treatment group of H2O2.

[0026] Figure 6 It is a picture of the germination of Artemisia halodendron seeds in the theoretically optimal treatment group of NaClO and the practically optimal treatment group of NaClO. Among them, (a) is a picture of the germination of Artemisia halodendron seeds in the theoretically optimal treatment group of NaClO; (b) is a picture of the germination of Artemisia halodendron seeds in the practically optimal treatment group of NaClO. Detailed implementation manners

[0027] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0028] Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the scope of protection of the present invention.

[0029] The present invention mainly disinfects the Artemisia halodendron seeds after soaking. By selecting appropriate disinfectants and disinfection conditions, it can not only effectively solve the problem of endophyte contamination, but also effectively improve the germination rate of seeds and significantly reduce the mildew rate.

[0030] The technical solution of the present invention will be further described below through specific embodiments.

[0031] In the following embodiments, unless otherwise specified, the methods are all conventional methods; the reagents and materials, unless otherwise specified, can all be purchased in the market.

[0032] Example 1

[0033] A method for germinating and culturing Artemisia halodendron seeds includes the following steps:

[0034] S1. Seed soaking: Select Artemisia halodendron seeds that are uniform in size, plump in appearance and undamaged. Place the Artemisia halodendron seeds in a petri dish with a diameter of 90 mm and soak them in distilled water for 2 h to make them fully absorb water.

[0035] S2. Disinfection treatment: Use a potassium permanganate solution with a mass fraction of 1% as the disinfectant, and place the soaked Artemisia halodendron seeds in the disinfectant for disinfection treatment. The conditions for disinfection treatment are: soaking time is 20 min.

[0036] S3. Germination culture: The disinfected Artemisia halodendron seeds are washed 3 times repeatedly with distilled water, and the water is blotted dry with absorbent paper. Then, the washed Artemisia halodendron seeds are neatly arranged on the surface of a petri dish lined with two layers of filter paper and transferred to an incubator for germination culture. The conditions for germination culture are a culture temperature of 20°C, a humidity of 80%, 12 hours of light, and 12 hours of darkness. During the germination culture process, keep the filter paper moist; the degree of moistness of the filter paper should be such that no water flows out when pressed with a finger.

[0037] The Artemisia halodendron seeds are observed daily, and the number of germinated seeds is recorded. Using the emergence of a 1-mm radicle as the germination standard, continuous observation is carried out for 10 days, and the germination rate, germination potential, germination index, vigor index, and mildew rate of the Artemisia halodendron seeds are calculated. The results show that after treatment with the method of Example 1, the germination rate of the Artemisia halodendron seeds is 96.00%, the germination potential is 62.33%, the germination index is 42.45, the vigor index is 206.72, and the mildew rate is 0.33%.

[0038] Example 2

[0039] A method for the germination culture of Artemisia halodendron seeds, comprising the following steps:

[0040] S1. Seed soaking: Select Artemisia halodendron seeds that are of the same size, plump in appearance, and undamaged. Place the Artemisia halodendron seeds in a petri dish with a diameter of 90 mm and soak them in distilled water for 2 hours to allow them to fully absorb water.

[0041] S2. Disinfection treatment: Using a 10% sodium hypochlorite solution by mass fraction as the disinfectant, place the soaked Artemisia halodendron seeds in the disinfectant for disinfection treatment. The conditions for disinfection treatment are: soaking time of 20 minutes.

[0042] S3. Germination culture: The disinfected Artemisia halodendron seeds are washed 3 times repeatedly with distilled water, and the water is blotted dry with absorbent paper. Then, the washed Artemisia halodendron seeds are neatly arranged on the surface of a petri dish lined with two layers of filter paper and transferred to an incubator for germination culture. The conditions for germination culture are a culture temperature of 20°C, a humidity of 80%, 12 hours of light, and 12 hours of darkness. During the germination culture process, keep the filter paper moist; the degree of moistness of the filter paper should be such that no water flows out when pressed with a finger.

[0043] The Artemisia halodendron seeds are observed daily, and the number of germinated seeds is recorded. Using the emergence of a 1-mm radicle as the germination standard, continuous observation is carried out for 10 days, and the germination rate, germination potential, germination index, vigor index, and mildew rate of the Artemisia halodendron seeds are calculated. The results show that after treatment with the method of Example 1, the germination rate of the Artemisia halodendron seeds is 97.33%, the germination potential is 56.00%, the germination index is 40.31, the vigor index is 149.97, and the mildew rate is 0%.

[0044] In summary, the above analysis shows that by using potassium permanganate with a mass fraction of 1% and sodium hypochlorite solution with a mass fraction of 10% as disinfectants, and taking the culture temperature of 20°C and the soaking time of 20 minutes as the conditions for disinfection treatment, the present invention can not only effectively solve the problem of endophyte contamination, but also effectively improve the germination rate of seeds and significantly reduce the mildew rate. Especially when using potassium permanganate solution with a mass fraction of 1% as the disinfectant, and taking the culture temperature of 20°C and the soaking time of 20 minutes as the conditions for disinfection treatment, it can ensure that the germination index and vigor index of Artemisia halodendron seeds reach the highest level.

[0045] Example 3

[0046] In the embodiment of the present invention, through the design of an orthogonal experiment, the disinfection effects and germination indexes of three disinfectants, namely NaClO, KMnO4, and H2O2, on Artemisia halodendron seeds under different treatment conditions such as different disinfectant concentrations, different culture temperatures, and different soaking times are studied, and the optimal treatment combination for Artemisia halodendron seeds is screened, in order to provide technical support for the disinfection and germination of Artemisia halodendron seeds and provide materials for the further development and utilization of Artemisia halodendron. Given the significant ecological benefits and medicinal values of Artemisia halodendron, the development prospect of Artemisia halodendron is very broad. Therefore, exploring the technology to improve the germination of Artemisia halodendron seeds is of great significance for its artificial breeding.

[0047] A method for germinating and culturing Artemisia halodendron seeds includes the following steps:

[0048] S1. Seed soaking: Select Artemisia halodendron seeds that are of the same size, plump in appearance, and without damage. Place the Artemisia halodendron seeds in a petri dish with a diameter of 90 mm and soak them in distilled water for 2 hours to allow them to fully absorb water.

[0049] S2. Disinfection treatment: Select a suitable disinfectant, and then place the soaked Artemisia halodendron seeds in the disinfectant for disinfection treatment. The disinfectant is potassium permanganate solution with a mass fraction of 0.5% - 1%, sodium hypochlorite solution with a mass fraction of 5% - 10%, or hydrogen peroxide solution with a mass fraction of 10% - 30%; the conditions for the disinfection treatment are: the soaking time is 10 minutes - 30 minutes.

[0050] S3. Germination and culture: Wash the disinfected Artemisia halodendron seeds with distilled water repeatedly for 3 times, blot the water with absorbent paper, and then neatly arrange the washed Artemisia halodendron seeds on the surface of a petri dish lined with 2 layers of filter paper, and transfer them to an incubator for germination and culture; the conditions for germination and culture are a culture temperature of 20°C - 30°C, a humidity of 80%, a light of 12 hours, and darkness of 12 hours. During the germination and culture process, keep the filter paper moist; the degree of moisture of the filter paper should be such that no water flows out when pressed with a finger.

[0051] The following is the germination test of Artemisia halodendron seeds according to the above method to explore the effects of different disinfection conditions on the germination of Artemisia halodendron seeds.

[0052] 1. Materials and methods.

[0053] Artemisia halodendron seeds were collected from Naiman Banner, Tongliao City, Inner Mongolia Autonomous Region in January 2023.

[0054] An orthogonal experiment design was adopted, with the types of disinfectants, the concentrations of disinfectants, the soaking time, and the culture temperature as influencing factors. An orthogonal experiment with 27 treatment groups of 3 factors and 3 levels was designed for 3 disinfectants using an orthogonal table, as shown in Table 1. Together with the control group, there were a total of 28 treatment groups, as shown in Table 2, and each treatment group was repeated 3 times. In the experiment, 50 seeds were placed in each petri dish, each treatment group had 3 replicates, and each replicate had 100 seeds.

[0055] Table 1 Factor levels of orthogonal experiment design

[0056]

[0057]

[0058] Note: The concentration of the disinfectant is factor A; the soaking time is factor B; the culture temperature is factor C.

[0059] Table 2 Treatment combinations of Artemisia halodendron seed disinfection test

[0060]

[0061]

[0062] Note: Treatment group 28 is the control group; " / " indicates none.

[0063] 2. Determine the thousand-seed weight and morphological indexes of seeds.

[0064] Determination of thousand-seed weight: Randomly take out 1000 seeds from the purified seeds, weigh them with an electronic balance with a precision of one ten-thousandth, repeat 5 times, and calculate the average value and standard deviation.

[0065] Determination of seed morphological indexes: Randomly select 50 relatively plump seeds, and measure the length, width, and thickness of each seed under an optical microscope, and calculate the average value and standard deviation.

[0066] It was observed that Artemisia halodendron seeds are small, oblong in shape, and have parallel stripes on the surface.

[0067] The measurement results showed that the 1000-seed weight of Artemisia halodendron seeds was 364.26 mg ± 7.90 mg, the length was 2.19 mm ± 0.23 mm, the width was 0.67 mm ± 0.07 mm, and the thickness was 0.48 mm ± 0.06 mm.

[0068] 3. Determination of related indicators.

[0069] The Artemisia halodendron seeds were observed daily, and the number of germinated seeds was recorded. With the radicle growing 1 mm as the germination standard, continuous observation was carried out for 10 days, and the average fresh weight of the seedlings was measured.

[0070] Germination rate, the full English name is Germination Rate, abbreviated as GR. Germination potential, the full English name is Germination Energy, abbreviated as GE. Germination index, the full English name is Germination Index, abbreviated as GI. Vitality index, the full English name is Vitality Index, abbreviated as VI. Mildew rate, the full English name is Midew Rate, abbreviated as MR. Calculation formulas for germination rate, germination potential, germination index, vitality index, and mildew rate: Germination rate = (cumulative number of germinated seeds / total number of seeds) × 100%;

[0071] Germination potential = (number of normally germinated seeds at the initial stage of germination (the 3rd day) / total number of seeds) × 100%;

[0072] Germination index = Σ(Gt / Dt); Vitality index = Σ(Gt / Dt) × S;

[0073] Mildew rate = (number of mildewed seeds / total number of seeds) × 100%;

[0074] Among them, the initial stage of germination refers to the 3rd day of germination; Gt represents the number of germinated seeds within Dt time; Dt represents time, in days; S represents the average fresh weight of the seedlings.

[0075] 4. Data statistical analysis.

[0076] Data statistics and analysis were carried out using Excel software and SPSS 25.0 software for data analysis, and one-way analysis of variance was used to compare the index differences between different treatment groups; GraphPad Prism 10.0 and Origin 2021 were used for graph drawing.

[0077] 5. Analysis of variance of the orthogonal experiment on soaking and disinfection of Artemisia halodendron seeds, and the experimental results are shown in Table 3.

[0078] Table 3 Results of the orthogonal experiment on soaking and disinfection of Artemisia halodendron seeds

[0079]

[0080]

[0081] Note: Different lowercase letters in the same column indicate significant differences between treatment groups, P < 0.05.

[0082] As can be seen from Table 3, there are significant differences in germination rate, germination potential, germination index, vigor index, and mildew rate among the 28 treatment groups. The treatment group with the highest germination rate is treatment group 6; the treatment group with the highest germination potential is treatment group 6; the treatment group with the highest germination index is treatment group 16; the treatment group with the highest vigor index is treatment group 17; the treatment groups with the lowest mildew rate are treatment groups 3, 7, 8, 12, and 24, all of which are 0%.

[0083] Combined with Figure A of Figure 1 Table 3 shows that the number of treatment groups with a single index significantly better than the control group is as follows: there are 15 treatment groups for germination rate, 21 treatment groups for germination potential and germination index, 17 treatment groups for vigor index, and 5 treatment groups for mildew rate. The number of treatment groups with no significant difference from the control group in a single index is as follows: there are 10 treatment groups for germination rate, 2 treatment groups for germination potential, germination index, and vigor index, and 17 treatment groups for mildew rate. The number of treatment groups with a single index significantly worse than the control group is as follows: there are 2 treatment groups for germination rate, 4 treatment groups for germination potential and germination index, 8 treatment groups for vigor index, and 5 treatment groups for mildew rate.

[0084] Combined with Figure 1 Figure B of Table 3 shows that by taking the intersection of the 5 treatment groups with a single index better than the control group, the treatment groups with a single index significantly better than the control group are as follows: the germination rate of 15 treatment groups, namely treatment groups 1, 2, 4, 6, 8, 9, 10, 11, 13, 15, 17, 18, 19, 24, 26; the germination potential of 21 treatment groups, namely treatment groups 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 24, 26; the germination index of 21 treatment groups, namely treatment groups 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 24, 26; the vigor index of 17 treatment groups, namely treatment groups 2, 3, 4, 5, 6, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 24, 26; the mildew rate of 5 treatment groups, namely treatment groups 3, 7, 8, 12, 24.

[0085] 6. Influence of different disinfection soaking combinations on the germination potential of Artemisia halodendron seeds.

[0086] As Figure 2 can be seen, different disinfectant soaking treatments will have a certain impact on the germination time and daily germination number of Artemisia halodendron seeds. Due to the large temperature difference during germination, the uniformity of the germination potential of the three disinfectants will also vary, indicating that temperature has a certain impact on seed germination.

[0087] When disinfecting and soaking with three kinds of disinfectants, most of them reached the germination peak on the 2nd to 3rd day. There were a total of 18 treatment groups that reached the germination peak on the 2nd day: among them, there were 9 in the NaClO treatment group, namely T1, T2, T3, T4, T5, T6, T7, T8, and T9; there were 7 in the KMnO4 treatment group, namely T11, T12, T13, T14, T16, T17, and T18; there were 2 in the H2O2 treatment group, namely T24 and T25.

[0088] There were a total of 5 treatment groups that reached the germination peak on the 3rd day: among them, there were 2 in the KMnO4 treatment group, namely T10 and T15; there were 3 in the H2O2 treatment group, namely T19, T23, and T26.

[0089] There were a total of 3 treatment groups that reached the germination peak on the 4th day: among them, there were 2 in the H2O2 treatment group, namely T22 and T27; there was 1 in the control group, namely T28.

[0090] There was a total of 1 treatment group that reached the germination peak on the 5th day: there was 1 in the H2O2 treatment group, namely T20. There was a total of 1 treatment group that reached the germination peak on the 6th day: there was 1 in the H2O2 treatment group, namely T21. The daily germination number of seeds treated with NaClO and KMnO4 was higher than that of the control group on the 2nd day and lower than that of the control group on the 4th day; the germination number of seeds treated with H2O2 was partially higher than that of the control group on the 2nd to 3rd day and lower than that of the control group on the 4th day.

[0091] From Figure 2 Figure A, it can be seen that on the 3rd day, the daily germination number of seeds in 6 treatment groups was higher than that of the control group, namely T1, T2, T4, T6, T8, and T9. From Figure 2 Figure B, it can be seen that on the 3rd day, the daily germination number of seeds in 5 treatment groups was higher than that of the control group, namely T10, T11, T13, T15, and T17. From Figure 2 Figure C, it can be seen that the germination number of 3 treatment groups was significantly higher than that of the control group and other treatment groups from the 2nd to 3rd day, namely T19, T24, and T26. This shows that the disinfection treatment with 3 kinds of disinfectants has a certain promoting effect on the germination of Artemisia halodendron seeds.

[0092] 7. Correlation analysis among various statistical indicators.

[0093] In order to screen out representative indicators to reduce the evaluation difficulty, Pearson correlation analysis was carried out on 5 single indicators. From Figure 3It can be seen that the germination rate is extremely significantly positively correlated with the germination potential, germination index, and vigor index, and the correlation coefficients are 0.78, 0.73, and 0.59 in sequence; the germination rate is extremely significantly negatively correlated with the mildew rate, and the correlation coefficient is -0.51. The germination potential is extremely significantly positively correlated with the germination index and vigor index, and the correlation coefficients are 0.90 and 0.79 respectively. The germination potential is extremely significantly negatively correlated with the mildew rate, and the correlation coefficient is -0.76. The germination index is extremely significantly positively correlated with the vigor index, and the correlation coefficient is 0.77. The germination index is extremely significantly negatively correlated with the mildew rate, and the correlation coefficient is -0.70. The vigor index is significantly negatively correlated with the mildew rate, and the correlation coefficient is -0.64. This indicates that the relationship between these three disinfectants and the germination of Artemisia halodendron seeds is very close. Since the germination index and vigor index additionally cover the time-axis information compared with the other two groups of indicators, and the result value of the germination rate is larger than that of the germination potential, it is more conducive to evaluation. The mildew rate has an extremely significant negative correlation with the other four indicators, indicating that the mildew rate covers contradictory information. Therefore, it is finally decided to use the germination rate, germination index, and vigor index as evaluation indicators for subsequent evaluation.

[0094] 8. Range analysis of the orthogonal test for the disinfection and soaking of Artemisia halodendron seeds.

[0095] 8.1. Disinfection effect of seeds treated by soaking with NaClO.

[0096] Table 4 Orthogonal test results of the disinfection and soaking treatment with sodium hypochlorite

[0097] Treatment group Factor A Factor B Factor C Germination rate / % Germination index Vigor index 1 5.0％ 10 min 20℃ 95.00 41.46 78.06 2 5.0％ 20 min 25℃ 94.00 38.00 126.91 3 5.0％ 30 min 30℃ 71.00 36.12 123.31 4 7.5％ 10 min 25℃ 90.33 35.92 145.85 5 7.5％ 20 min 30℃ 72.33 39.87 124.00 6 7.5％ 30 min 20℃ 97.67 42.44 167.62 7 10.0％ 10 min 30℃ 70.00 37.39 91.61 8 10.0％ 20 min 20℃ 97.33 40.31 149.97 9 10.0％ 30 min 25℃ 92.33 37.69 151.89

[0098] Table 5 Analysis of the germination rate of the orthogonal test results of the disinfection and soaking treatment with sodium hypochlorite / %

[0099]

[0100]

[0101] Table 6 Analysis of the germination index of the orthogonal test results of the disinfection and soaking treatment with sodium hypochlorite

[0102] Average value of germination index Factor A Factor B Factor C k1 38.53 38.26 41.40 k2 39.41 39.39 37.20 k3 38.46 39.75 37.79 R 0.95 1.49 4.20

[0103] Table 7 Analysis of the vigor index of the orthogonal test results of the disinfection and soaking treatment with sodium hypochlorite

[0104] Average value of vigor index Factor A Factor B Factor C k1 109.43 105.17 131.88 k2 145.82 133.63 141.55 k3 131.16 147.61 112.97 R 36.39 42.44 28.58

[0105] Note: k1, k2, and k3 respectively represent the average values of the sum of the indicators at each level of each factor; R represents the range.

[0106] As can be seen from the results in Table 5, the order of the influence degrees of the three factors in the sodium hypochlorite disinfection and soaking treatment on the germination rate of Artemisia halodendron seeds is: Factor C > Factor B > Factor A; from the k values, the best treatment combination for the influence of the sodium hypochlorite disinfection and soaking treatment on the germination rate of Artemisia halodendron seeds is A2B2C1.

[0107] As can be seen from the results in Table 6, the order of the influence degrees of the three factors in the sodium hypochlorite disinfection and soaking treatment on the germination index of Artemisia halodendron seeds is: Factor C > Factor A > Factor B; from the k values, the best treatment combination for the influence of the sodium hypochlorite disinfection and soaking treatment on the germination index of Artemisia halodendron seeds is A1B2C1.

[0108] As can be seen from the results in Table 7, the order of the influence degrees of the three factors in the sodium hypochlorite disinfection and soaking treatment on the vigor index of Artemisia halodendron seeds is: Factor B > Factor A > Factor C; from the k values, the best treatment combination for the influence of the sodium hypochlorite disinfection and soaking treatment on the vigor index of Artemisia halodendron seeds is A2B3C2.

[0109] Overall, the best combination for the influence of the sodium hypochlorite disinfection and soaking treatment on the germination of Artemisia halodendron seeds is A2B2C1, that is, the disinfectant concentration is 7.5% NaClO solution, the soaking time is 20 min, and the cultivation temperature is 20 °C. Analysis shows that a disinfectant concentration of 7.5% NaClO solution can ensure the highest germination rate and vigor index of Artemisia halodendron seeds, and a soaking time of 20 min and a cultivation temperature of 20 °C can ensure the highest germination rate and germination index of Artemisia halodendron seeds.

[0110] 8.2, Disinfection effect of seeds treated by soaking with KMnO4.

[0111] Table 8 Orthogonal test results of potassium permanganate disinfection and soaking treatment

[0112] Treatment group Factor A Factor B Factor C Germination rate / % Germination index Vigor index 10 0.5％ 10 min 20℃ 96.33 37.93 84.69 11 0.5％ 20 min 25℃ 83.00 31.38 132.13 12 0.5％ 30 min 30℃ 64.33 31.07 125.98 13 0.75％ 10 min 25℃ 86.00 34.89 155.62 14 0.75％ 20 min 30℃ 74.00 39.62 131.14 15 0.75％ 30 min 20℃ 97.00 38.30 126.40 16 1.0％ 10 min 30℃ 70.00 42.61 197.73 17 1.0％ 20 min 20℃ 96.00 42.45 206.72 18 1.0％ 30 min 25℃ 87.00 36.67 147.41

[0113] Table 9 Germination rate analysis of orthogonal test results of potassium permanganate disinfection and soaking treatment

[0114]

[0115]

[0116] Table 10 Germination index analysis of orthogonal test results of potassium permanganate disinfection and soaking treatment

[0117] Average value of germination index Factor A Factor B Factor C k1 33.46 38.48 39.56 k2 37.60 37.82 34.31 k3 40.58 35.35 37.77 R 7.12 3.13 5.25

[0118] Table 11 Vigor index analysis of orthogonal test results of potassium permanganate disinfection and soaking treatment

[0119] Average value of vigor index Factor A Factor B Factor C k1 114.27 146.01 139.27 k2 137.72 156.66 145.05 k3 183.95 132.26 151.62 R 69.68 24.40 12.35

[0120] Note: k1, k2, and k3 respectively represent the average values of the index sums at each level of each factor; R represents the range.

[0121] As can be seen from the results in Table 9, the order of the influence degrees of the three factors in the potassium permanganate disinfection and soaking treatment on the germination rate of Artemisia halodendron seeds is: Factor C > Factor A > Factor B; from the k values, the best treatment combination for the influence of the potassium permanganate disinfection and soaking treatment on the germination rate of Artemisia halodendron seeds is A2B2C1.

[0122] As can be seen from the results in Table 10, the order of the influence degrees of the three factors in the potassium permanganate disinfection and soaking treatment on the germination index of Artemisia halodendron seeds is: Factor A > Factor C > Factor B; from the k values, the best treatment combination for the influence of the potassium permanganate disinfection and soaking treatment on the germination index of Artemisia halodendron seeds is A3B1C1.

[0123] As can be seen from the results in Table 11, the order of the influence degrees of the three factors in the potassium permanganate disinfection and soaking treatment on the vigor index of Artemisia halodendron seeds is: Factor A > Factor B > Factor C; from the k values, the best treatment combination for the influence of the potassium permanganate disinfection and soaking treatment on the vigor index of Artemisia halodendron seeds is A3B2C3.

[0124] Comprehensively, the best combination for the influence of the potassium permanganate disinfection and soaking treatment on the germination of Artemisia halodendron seeds is A3B2C1, that is, a KMnO4 solution with a disinfectant concentration of 1.0%, an immersion time of 20 min, and a cultivation temperature of 20°C. Analysis shows that a KMnO4 solution with a disinfectant concentration of 1.0% can ensure the highest germination index and vigor index of Artemisia halodendron seeds, an immersion time of 20 min can ensure the highest germination rate and vigor index of Artemisia halodendron seeds, and a cultivation temperature of 20°C can ensure the highest germination rate and germination index of Artemisia halodendron.

[0125] 8.3. Disinfection effect of seeds treated by H2O2 immersion disinfection.

[0126] Table 12 Orthogonal test results of hydrogen peroxide disinfection and soaking treatment

[0127]

[0128]

[0129] Table 13 Germination rate analysis of orthogonal test results of hydrogen peroxide disinfection and soaking treatment

[0130] Average value of germination rate Factor A Factor B Factor C k1 63.00 74.78 95.67 k2 76.00 72.33 62.67 k3 77.45 69.33 58.11 R 14.45 5.45 37.56

[0131] Table 14 Germination index analysis of orthogonal test results of hydrogen peroxide disinfection and soaking treatment

[0132] Average value of germination index Factor A Factor B Factor C k1 20.52 24.94 39.73 k2 24.40 21.77 12.38 k3 23.05 21.26 15.85 R 3.88 3.68 27.35

[0133] Analysis of the vitality index of the orthogonal test results of hydrogen peroxide disinfection and immersion treatment in Table 15

[0134] Average value of vigor index Factor A Factor B Factor C k1 45.54 58.70 116.47 k2 85.74 68.13 40.56 k3 74.13 78.57 48.38 R 40.20 19.87 75.91

[0135] Note: k1, k2, and k3 represent the average values of the sum of the indicators at each level of each factor; R represents the range.

[0136] As can be seen from the results in Table 13, the order of the influence degree of the three factors of hydrogen peroxide disinfection and immersion treatment on the germination rate of Artemisia halodendron seeds is: Factor C > Factor A > Factor B; from the k value, the best treatment combination for the influence of hydrogen peroxide disinfection and immersion treatment on the germination rate of Artemisia halodendron seeds is A3B1C1.

[0137] As can be seen from the results in Table 14, the order of the influence degree of the three factors of hydrogen peroxide disinfection and immersion treatment on the germination index of Artemisia halodendron seeds is: Factor C > Factor A > Factor B; from the k value, the best treatment combination for the influence of hydrogen peroxide disinfection and immersion treatment on the germination index of Artemisia halodendron seeds is A2B1C1.

[0138] As can be seen from the results in Table 15, the order of the influence degree of the three factors of hydrogen peroxide disinfection and immersion treatment on the vitality index of Artemisia halodendron seeds is: Factor C > Factor A > Factor B; from the k value, the best treatment combination for the influence of hydrogen peroxide disinfection and immersion treatment on the vitality index of Artemisia halodendron seeds is A2B3C1.

[0139] Comprehensively, the best combination for the influence of hydrogen peroxide disinfection and immersion treatment on the germination of Artemisia halodendron seeds is A2B1C1, that is, the disinfectant concentration is 20% H2O2 solution, the soaking time is 10 min, and the culture temperature is 20°C. A disinfectant concentration of 20% H2O2 solution can ensure the highest germination index and vitality index of Artemisia halodendron seeds. A soaking time of 10 min can ensure the highest germination rate and germination index of Artemisia halodendron seeds. A culture temperature of 20°C can ensure the highest germination rate, germination index, and vitality index of Artemisia halodendron seeds.

[0140] 9. Comprehensive comparison of the optimal combinations.

[0141] Since the temperature of the three theoretical best combinations is 20°C, for this reason, soaking in distilled water for 30 min and a culture temperature of 25°C are used as control group one; soaking in distilled water for 30 min and a culture temperature of 20°C are used as control group two. Subsequently, the two control groups were compared, and it was found that there was no significant difference between the two control groups in the three indicators. Based on the above analysis, the comparison between the best combinations is shown in Table 16 and Figures 4 to 6 .

[0142] Table 16 Comparison between the best combinations

[0143]

[0144]

[0145] Note: Different lowercase letters in the same column indicate significant differences between treatment groups, P < 0.05.

[0146] As can be seen from Table 16, by comparing the theoretical optimal treatment groups of KMnO4, H2O2, NaClO, the actual optimal treatment group, and Control Group 1 and Control Group 2, it was found that among the three theoretical optimal treatment groups, the NaClO group and the KMnO4 group were significantly superior to the two control groups in terms of germination rate, germination index, and vigor index. The H2O2 group had no significant difference in germination rate compared with the two control groups, but was significantly inferior to the two control groups in terms of germination index and vigor index.

[0147] When comparing the theoretical optimal treatment group of NaClO with the actual optimal treatment group of NaClO, the actual optimal treatment group was superior to the theoretical optimal treatment group in terms of germination rate and germination index. The mildew of the seeds in the theoretical optimal treatment group of H2O2 might be relatively large, and the errors caused by human factors and environmental factors could not be used as the evaluation criteria.

[0148] Among the theoretical optimal treatment groups, the KMnO4 theoretical optimal treatment group was the best in terms of the three indicators. The specific conditions for disinfection treatment were a KMnO4 solution with a disinfectant concentration of 1.0%, an immersion time of 20 min, and a cultivation temperature of 20°C. By further comparing with the actual optimal treatment group of NaClO, it was found that the theoretical optimal KMnO4 in the two combinations was significantly superior to the actual optimal NaClO in terms of vigor index. Finally, it was decided to select the KMnO4 theoretical optimal treatment group as the optimal combination for soaking and disinfecting Artemisia halodendron, and its disinfection treatment conditions were: a KMnO4 solution with a disinfectant concentration of 1.0%, an immersion time of 20 min, and a cultivation temperature of 20°C.

[0149] Combined with the above analysis, explore the disinfection treatment mechanism of three disinfectants on Artemisia halodendron seeds:

[0150] Sodium hypochlorite, with the chemical formula NaClO, mainly sterilizes by the chlorine gas decomposed by itself. Due to its characteristics of low toxicity, high efficiency, and easy volatilization of the residual liquid, it can be applied to the disinfection of Artemisia halodendron seeds. However, because sodium hypochlorite has certain corrosiveness, therefore, when using NaClO for the disinfection treatment of Artemisia halodendron seeds, NaClO with a mass fraction ≤ 10.0% is used.

[0151] Potassium permanganate, with the chemical formula KMnO4, is a strong oxidant. Potassium permanganate solution has a strong bactericidal and disinfecting effect, which can effectively solve the problem of endophyte contamination. Moreover, the potassium and manganese ions in the potassium permanganate solution can activate the endoenzyme system of Artemisia serrata seeds, effectively shortening the germination period of Artemisia serrata seeds, thereby achieving the purpose of rapid germination. The above test proves that potassium permanganate with a mass fraction of 1% as a disinfectant can have a good disinfection effect on Artemisia serrata seeds, effectively solve the problem of endophyte contamination, and no inhibitory effect on seedling growth is found.

[0152] Hydrogen peroxide, with the chemical formula H2O2, also known as hydrogen peroxide, can break seed dormancy at an appropriate concentration and promote germination. However, in the study of the embodiments of the present invention, the effect of hydrogen peroxide in promoting germination seems to be less stable than that of the other two disinfectants. Figure 2 Figure C. Due to the large differences in the results of hydrogen peroxide disinfection treatments and the high rate of mildew, it is not considered as a suitable disinfectant for Artemisia argyi seeds.

[0153] Since temperature can directly affect the enzyme activity in seeds, it is an important condition that affects seed germination. Different plant seeds have different optimum temperatures for germination. Even for the same species, the optimum temperatures may vary due to the different growing regions. For example, the Masson pine seeds grown in high-cold and warmer areas have different requirements for culture temperature. Temperature variation can promote seed enzyme activity, thereby increasing the germination rate, but too high a temperature can cause damage to the enzyme structure, and too low a temperature can cause insufficient enzyme activity. From the above test results of the embodiments of the present invention, it can be seen that when the temperature is 20°C, the germination rate and related indicators of Artemisia selengensis seeds in the best combination of three disinfectants are optimal, as shown in Table 7.

[0154] In summary, the embodiment of the present invention selects germination rate, germination index and germination index to comprehensively evaluate the disinfection effect of Artemisia argyi seeds treated with different disinfectants. It can be seen that disinfectant concentration, immersion time and ambient temperature are the main factors affecting seed germination.

[0155] The results showed that the germination index and vitality index of Artemisia serrata seeds could be improved by using KMnO4 solution with a disinfectant concentration of 1.0%, immersion time of 20 minutes and culture temperature of 20℃; the germination rate of Artemisia serrata seeds could be improved by using NaClO solution with a disinfectant concentration of 10%, immersion time of 20 minutes and culture temperature of 20℃. After comprehensive comparison, it was determined that the optimal combination of disinfectant concentration of 1.0% KMnO4 solution, immersion time of 20 minutes and culture temperature of 20℃ was used as the disinfection treatment for Artemisia serrata seeds, which could effectively solve the problem of endophyte contamination of Artemisia serrata seeds and improve the germination rate of Artemisia serrata seeds, thereby improving its application effect in ecological restoration.

[0156] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for germination and cultivation of Artemisia argyi seeds, characterized in that: The following steps are involved: The seeds of Artemisia chaenomelis are soaked in distilled water, and then placed in a disinfectant for disinfection; the disinfectant is a potassium permanganate solution with a mass fraction of 0.5% to 1%, a sodium hypochlorite solution with a mass fraction of 5% to 10%, or a hydrogen peroxide solution with a mass fraction of 10% to 30%; the conditions of the disinfection treatment are: the soaking time is 10 minutes to 30 minutes; The sterilized Artemisia rapa seeds were washed with distilled water and the water was dried. The washed Artemisia rapa seeds were then placed on the surface of filter paper and cultured for germination at a temperature of 20°C to 30°C.

2. The germination and cultivation method of Artemisia chaenomelis seeds according to claim 1, characterized in that: The disinfectant is a 1% by mass potassium permanganate solution or a 10% by mass sodium hypochlorite solution.

3. The germination and cultivation method of Artemisia chaenomelis seeds according to claim 1, characterized in that: The conditions for the disinfection treatment are: soaking time 20 minutes.

4. The germination and cultivation method of Artemisia chaenomelis seeds according to claim 1, characterized in that: The conditions of the germination culture are: culture temperature 20° C., humidity 80%, light 12 h, and darkness 12 h.

5. The germination and cultivation method of Artemisia chaenomelis seeds according to claim 1, characterized in that: The filter paper is laid in 2 layers.

6. The germination and cultivation method of Artemisia chaenomelis seeds according to claim 1, characterized in that: The germination culture process also includes: keeping the filter paper moist; the filter paper should be moist enough that no water flows out when pressed with a finger.

7. The method for germination and cultivation of Artemisia chaenomelis seeds according to claim 1, characterized in that: The germination standard for germination culture is when the radicle grows to 1mm.

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