Method for improving germination rate of Alisma orientale Juzepczuk seed

Abstract

The present invention relates to a method for improving the germination rate of seeds of *Alisma orientale* (Juzepczuk). Since the method according to the present invention provides optimal conditions for effectively improving the germination rate of seeds, it is expected that the effect of reducing labor and increasing productivity can be anticipated by efficiently inducing the germination of *Alisma orientale*.

Description

Technology Field

[0001] The present invention relates to the plantain ( Alisma orientale This relates to a method for improving the germination rate of seeds. Background Technology

[0002] Plantain Alisma orientale Alisma orientale (*Jūzepczuk*) is a semi-aquatic perennial plant belonging to the Alismataceae family that grows in mud and shallow water. As a traditional medicinal herb in North and East Asia, Alisma orientale has long been used to treat edema, diuresis, lipid dysregulation, inflammatory diseases, diabetes, and tumors. The main components of Alisma orientale are known to include starch, protein, furfural, resin, inorganic salts, and the triterpenoid alisol. It is rich in potassium (K) and possesses the efficacy of significantly increasing potassium excretion. Furthermore, Alisma orientale has been reported to reduce blood cholesterol, lower blood pressure and blood sugar, exhibit hepatoprotective activity against carbon tetrachloride-induced hepatotoxicity, and demonstrate excellent antibacterial, antioxidant, and immune-enhancing activities. Although it is recognized as a useful plant, its very low seed germination rate poses significant challenges for mass production.

[0003] Methods to increase the germination rate of plant seeds include chemical, physical, biological, light, and priming treatments; however, it is reported that the optimal conditions vary significantly depending on the type of plant, and no research has yet been reported on the optimal conditions for improving the seed germination rate of Alisma orientale.

[0004] Meanwhile, Korean Registered Patent No. 2255802 discloses a "method for improving the germination rate of seeds of halophyte plants" and Korean Registered Patent No. 2374478 discloses a "seed treatment method for improving the germination rate of seeds of Salicornia europaea," but there is no description of a method for improving the germination rate of seeds of Alisma orientale according to the present invention. The problem to be solved

[0005] The present invention was derived from the above requirements, and the inventor completed the invention by confirming that when seeds of Alisma orientale were disinfected and washed, then subjected to a first pretreatment with 50% (v / v) sulfuric acid and a second pretreatment with 0.3% (v / v) potassium nitrate, and then immersed in distilled water to induce germination under light conditions for 24 hours, the germination rate increased significantly compared to the untreated control group. means of solving the problem

[0006] To solve the above problem, the present invention ( Alisma orientale A method for improving the germination rate of seeds of *Plantago asiatica* is provided, comprising the step of sequentially pre-treating seeds of *Plantago asiatica* with sulfuric acid and potassium nitrate, followed by immersion treatment.

[0007] In addition, the present invention

[0008] (1) Step of disinfecting and washing the seeds of the plantain;

[0009] (2) A step of first pre-treating the above-mentioned disinfected seeds with 40~60% (v / v) sulfuric acid for 4~6 minutes;

[0010] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.2~0.4% (v / v) potassium nitrate for 20~28 hours; and

[0011] (4) A step of immersing the seeds that have undergone the second pretreatment in sterile water to induce germination; thereby providing a method for pretreating seeds of Alisma plantain to improve germination rate.

[0012] In addition, the present invention provides seeds of Alisma orientale with an improved germination rate, which are pretreated by the above method.

[0013] In addition, the present invention provides a plantain plant derived from the above plantain seeds. Effects of the invention

[0014] Since the method according to the present invention provides optimal conditions for effectively improving the germination rate of seeds, it is expected that the effects of reduced labor and increased productivity can be anticipated by efficiently inducing the germination of Alisma orientale. Brief explanation of the drawing

[0015] Figure 1 shows the germination rate of Alisma plantain seeds according to photoperiod (light treatment conditions). L: light condition, D: dark condition. Figure 2 shows the germination rate of Alisma orientale seeds according to sulfuric acid pretreatment. Control: Untreated group. Figure 3 shows the germination rate of Alisma plantain seeds according to potassium nitrate pretreatment. Control: Untreated group. Figure 4 shows the germination rate of Alisma orientale seeds according to immersion treatment. Control: Untreated group. Figure 5 shows the results of a photograph (A) and germination rate (B) of Alisma orientale seeds pretreated with 50% (v / v) sulfuric acid, 0.3% (v / v) potassium nitrate, and then immersed in distilled water. Control: Untreated group. Specific details for implementing the invention

[0016] To achieve the objective of the present invention, the present invention (Plantago asiatica) Alisma orientale A method for improving the germination rate of seeds of *Plantago asiatica* is provided, comprising the step of sequentially pre-treating seeds of *Plantago asiatica* with sulfuric acid and potassium nitrate, followed by immersion treatment.

[0017] The method for improving the germination rate of seeds of Alisma orientale according to the present invention preferably,

[0018] (1) Step of disinfecting and washing the seeds of the plantain;

[0019] (2) A step of first pre-treating the above-mentioned disinfected seeds with 40~60% (v / v) sulfuric acid for 4~6 minutes;

[0020] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.2~0.4% (v / v) potassium nitrate for 20~28 hours; and

[0021] (4) A step of inducing germination by immersing the above-mentioned secondary pretreated seeds in sterile water; may be included,

[0022] More preferably,

[0023] (1) Step of disinfecting and washing the seeds of the plantain;

[0024] (2) A step of first pre-treating the above-mentioned disinfected seeds with 50% (v / v) sulfuric acid for 5 minutes;

[0025] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.3% (v / v) potassium nitrate for 24 hours; and

[0026] (4) A step of inducing germination by immersing the above-mentioned secondary pretreated seeds in sterile water; may be included, but is not limited thereto.

[0027] In a method for improving the germination rate of seeds of Alisma orientale according to one embodiment of the present invention, the germination induction of step (4) may be performed under light conditions of 16 to 24 hours, preferably under light conditions of 24 hours, but is not limited thereto.

[0028] The present invention also,

[0029] (1) Plantain ( Alisma orientale Step of disinfecting and washing the seeds of Juzepczuk;

[0030] (2) A step of first pre-treating the above-mentioned disinfected seeds with 40~60% (v / v) sulfuric acid for 4~6 minutes;

[0031] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.2~0.4% (v / v) potassium nitrate for 20~28 hours; and

[0032] (4) A step of immersing the seeds that have undergone the second pretreatment in sterile water to induce germination; thereby providing a method for pretreating seeds of Alisma plantain to improve germination rate.

[0033] The pretreatment method for Alisma plantain seeds to improve germination rate according to the present invention is preferably,

[0034] (1) Step of disinfecting and washing the seeds of the plantain;

[0035] (2) A step of first pre-treating the above-mentioned disinfected seeds with 40~60% (v / v) sulfuric acid for 4~6 minutes;

[0036] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.2~0.4% (v / v) potassium nitrate for 20~28 hours; and

[0037] (4) A step of inducing germination by immersing the above-mentioned secondary pretreated seeds in sterile water; may be included,

[0038] More preferably,

[0039] (1) Step of disinfecting and washing the seeds of the plantain;

[0040] (2) A step of first pre-treating the above-mentioned disinfected seeds with 50% (v / v) sulfuric acid for 5 minutes;

[0041] (3) A step of secondarily pretreating the seeds that have undergone the first pretreatment with 0.3% (v / v) potassium nitrate for 24 hours; and

[0042] (4) A step of inducing germination by immersing the above-mentioned secondary pretreated seeds in sterile water; may be included, but is not limited thereto.

[0043] The present invention also provides seeds of Alisma plantain that have been pretreated by the above method and have an improved germination rate.

[0044] The seeds of the plantain pretreated by another method of the present invention are characterized by a germination rate that is increased by about 10 times compared to seeds that have not been treated at all.

[0045] The present invention also provides a plantain plant derived from the above plantain seeds.

[0047] The present invention will be explained in detail below through examples. However, the following examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following examples.

[0049] Example 1. Analysis of germination rate of Alisma plantain seeds according to photoperiod (light treatment)

[0050] Alisma plantain seeds were disinfected with 70% (v / v) ethanol for 1 minute, twice with 2% (v / v) sodium hypochlorite for 15 minutes, and then washed five times with sterile water. 100 disinfected seeds were randomly selected and placed in Petri dishes containing filter paper. 3 ml of sterile water was added, and germination was induced for 14 days under light treatment conditions of 0, 8, 16, or 24 hours. The seed germination rate was then measured according to ISTA (International Seed Testing Association) standards. The experiment was performed in four replicates, and the average value was calculated.

[0051] As a result, it was confirmed that germination rates of 2.4% to 2.9% were observed under 0 or 8 hours of light treatment, 7.1% under 16 hours of light treatment, and 7.2% under 24 hours of light treatment (Fig. 1).

[0052] Based on the above results, it was found that the seeds of Alisma plantain are photophilic seeds whose germination is promoted by light, and to establish optimal conditions for the germination of Alisma plantain seeds, all subsequent experiments were conducted under 24-hour light treatment conditions.

[0054] Example 2. Analysis of germination rate of Alisma plantain seeds according to sulfuric acid pretreatment

[0055] Alisma plantain seeds were disinfected with 70% (v / v) ethanol for 1 minute, followed by two rounds of disinfection with 2% (v / v) sodium hypochlorite for 15 minutes each, and then washed five times with sterile water. One hundred disinfected seeds were randomly selected, pretreated with 0% (v / v), 25% (v / v), or 50% (v / v) sulfuric acid solutions for 5 minutes, and then washed three times with sterile water. The seeds were then placed in Petri dishes containing filter paper, 3 ml of sterile water was added, and germination was induced for 14 days under 24-hour light exposure conditions. Finally, the seed germination rate was measured according to ISTA standards. The experiment was performed in four replicates, and the average values ​​were calculated.

[0056] As a result, it was confirmed that the untreated group showed a germination rate of 8.6%, the 25% (v / v) sulfuric acid treated group showed a germination rate of 29.9%, and the 50% (v / v) sulfuric acid treated group showed a germination rate of 47.6% (Fig. 2).

[0057] Based on the above results, it was determined that the seed coat of Alisma plantain inhibits embryonic growth of the seeds, and it was found that the germination rate of Alisma plantain can be improved by removing the seed coat through sulfuric acid pretreatment, and that 50% (v / v) sulfuric acid pretreatment is the optimal condition.

[0059] Example 3. Analysis of germination rate of Alisma plantain seeds according to potassium nitrate pretreatment

[0060] Alisma plantain seeds were disinfected with 70% (v / v) ethanol for 1 minute, twice with 2% (v / v) sodium hypochlorite for 15 minutes, and then washed five times with sterile water. 100 disinfected seeds were randomly selected and pretreated in 0.1% (v / v), 0.2% (v / v), or 0.3% (v / v) potassium nitrate solution for 24 hours, followed by washing three times with sterile water. The seeds were then placed in Petri dishes containing filter paper, 3 ml of sterile water was added, and germination was induced for 14 days under 24-hour light treatment conditions. Finally, the seed germination rate was measured according to ISTA standards. The experiment was performed in four replicates, and the average value was calculated.

[0061] As a result, it was confirmed that the untreated group showed a germination rate of 7.6%, the 0.1% (v / v) potassium nitrate treatment group showed a germination rate of 48.2%, the 0.2% (v / v) potassium nitrate treatment group showed a germination rate of 48.6%, and the 0.3% (v / v) potassium nitrate treatment group showed a germination rate of 50% (Fig. 3).

[0062] Based on the above results, it was found that the germination rate of Alisma plantain can be improved by promoting seed germination through potassium nitrate pretreatment to break seed dormancy, and that 0.3% (v / v) potassium nitrate pretreatment is the optimal condition.

[0064] Example 4. Analysis of germination rate of Alisma plantain seeds according to immersion treatment

[0065] Alisma plantain seeds were disinfected with 70% (v / v) ethanol for 1 minute, followed by two rounds of disinfection with 2% (v / v) sodium hypochlorite for 15 minutes each, and then washed five times with sterile water. One hundred disinfected seeds were randomly selected and placed in Petri dishes containing filter paper. 20 ml of sterile water was added, and germination was induced for 14 days under 24-hour light treatment conditions. The seed germination rate was then measured according to ISTA standards. The experiment was performed in four replicates, and the average value was calculated.

[0066] As a result, it was confirmed that the untreated group showed a germination rate of 13.8%, and the soaked treated group showed a germination rate of 41.5% (Fig. 4).

[0067] Based on the above results, it was found that increasing the moisture content of the seeds through soaking treatment promotes seed germination and improves the germination rate of Alisma orientale.

[0069] Example 5. Analysis of Seed Germination Rate of Alisma plantain according to combined treatment

[0070] Seeds of *Plantago asiatica* were disinfected with 70% (v / v) ethanol for 1 minute, twice with 2% (v / v) sodium hypochlorite for 15 minutes, and then washed five times with sterile water. One hundred disinfected seeds were randomly selected, pretreated in 50% (v / v) sulfuric acid solution for 5 minutes, and then pretreated in 0.3% (v / v) potassium nitrate solution for 24 hours. Afterward, the seeds were placed in Petri dishes containing filter paper, 20 ml of sterile water was added, and germination was induced for 14 days under 24-hour light treatment conditions. The seed germination rate was then measured according to ISTA standards. The experiment was performed in four replicates, and the average value was calculated.

[0071] As a result, the untreated group showed a germination rate of 8.6%, and the combined treatment group, which was pretreated with 50% (v / v) sulfuric acid and 0.3% (v / v) potassium nitrate followed by immersion treatment, showed a germination rate of 80.3%, confirming that the germination rate increased by approximately 10 times compared to the untreated group (Fig. 5, Table 1).

[0072] Analysis of Seed Germination Rate of Alisma plantain according to combined treatment division GP(%) MGT(day) GSI T50(day) Untreated group 8.63±1.05 6.12±0.23 0.67±0.09 19.57±1.96 Complex treatment area 80.30±3.68 1.91±0.21 8.05±0.73 0.92±0.13

[0073] GP (germination percentage): germination rate, MGT (mean germination time): mean germination time, GSI (germination speed index): germination speed index, T50: time required to reach a germination rate of 50%.

[0075] Based on the above results, it was found that the optimal condition for improving the germination rate of Alisma plantain seeds is to first pretreat with 50% (v / v) sulfuric acid, second pretreat with 0.3% (v / v) potassium nitrate, and then immerse in distilled water to induce germination under light conditions for 24 hours.

Claims

Claim 1 delete Claim 2 (1) Plantain ( Alisma orientale A method for improving the germination rate of seeds of *Plantago asiatica*, characterized by comprising: (1) a step of disinfecting and washing the seeds of *Plantago asiatica*; (2) a step of first pre-treating the disinfected seeds with 40-60% (v / v) sulfuric acid for 4-6 minutes; (3) a step of second pre-treating the first pre-treated seeds with 0.2-0.4% (v / v) potassium nitrate for 20-28 hours; and (4) a step of immersing the second pre-treated seeds in sterile water and culturing them under light conditions for 16-24 hours to induce germination. Claim 3 delete Claim 4 (1) Plantain ( Alisma orientale A method for pre-treating seeds of Alisma plantain to improve germination rate, comprising: (1) a step of disinfecting and washing the seeds of Alisma plantain; (2) a step of first pre-treating the disinfected seeds with 40-60% (v / v) sulfuric acid for 4-6 minutes; (3) a step of second pre-treating the first pre-treated seeds with 0.2-0.4% (v / v) potassium nitrate for 20-28 hours; and (4) a step of immersing the second pre-treated seeds in sterile water and culturing them under light conditions for 16-24 hours to induce germination. Claim 5 delete Claim 6 delete

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