A method for improving the vigor of sweet and sticky corn seeds under hypoxia stress

Abstract

The invention discloses a method for improving the vitality of sweet and sticky corn seeds under hypoxia stress, the method comprising: preparing a compound agent; the compound agent uses water as a solvent and contains 60-1000 mg / L reduced glutathione. ‑1 , Vitamin B2 12~200mg·L ‑1 ; Place the sweet / glutinous corn seeds between the germination papers soaked with the compound agent, and wash off the surface agent after swelling for 22-26 hours at 23-25°C, and then slowly dry in a fume hood for 24 hours before sowing to obtain sweet and glutinous corn seeds with high vitality. The method of the present invention delivers the compound agent containing reduced glutathione and vitamin B2 into the sweet and glutinous corn seeds in a slow swelling manner, which can effectively improve the physiological metabolic activities of the seeds during germination and improve the vitality of the seeds when encountering hypoxia stress.

Description

Technical Field

[0001] The invention relates to the technical field of seed treatment, in particular to a method for improving the vitality of sweet and sticky corn seeds under hypoxia stress. Background Art

[0002] Corn (Zea mays L.) is an important cash crop, food crop and feed crop in my country, and plays a vital role in the supply and demand of grain and the development of animal husbandry in my country (Zhao Min, Zhou Shuxin, Cui Yanhong. Research on the application of plant growth regulators in corn production in my country [J]. Corn Science, 2006(01):127-131.). Corn is highly sensitive to water. my country has a monsoon climate, and water often accumulates in the fields during the rainy season, which causes the roots of corn plants to be unable to breathe normally, and toxic substances such as ethanol, lactic acid, and MDA (malondialdehyde) accumulate; the stomata of the plants are closed, photosynthesis and transpiration are significantly reduced, energy supply is insufficient, material assimilation slows down, the growth rate of aboveground dry matter decreases, seedlings develop slowly or even stop growing, and yields decrease (Fageria NK, Carvalho GD, Santos A B. Chemistry of lowland rice soils and nutrient availability [J]. Communications in Soil Science and Plant Analysis, 2011, 42: 1913-1933.). During the entire growth process of corn, different growth stages have different sensitivities to external stress. The critical period of germination and morphological establishment is more sensitive to stress than the subsequent development period, and the seed germination process is more susceptible to external environmental influences (Pérez-Ramos IM, Maraňón. Effects of waterlogging on seedgermination of three Mediterraneanoak species: ecological implications [J]. Acta Oecologica, 2009, 35: 422-428.). Among them, oxygen is an important condition for seed germination. Flooding and waterlogging will reduce the oxygen content in the soil air, thereby affecting seed germination and reducing seed vitality.

[0003] At present, people mainly use gene regulation and chemical reagents to alleviate crops from hypoxia stress; studies have shown that soybean gene GmALAAT1 overexpression plants can effectively resist hypoxia stress (Fu Jiayu, Wang Chunsheng, Wang Xinyu, Zhang Ye, Zhang Hanzhu, Dong Baozhu, Yang Zhuoyu, Wang Piwu. Cloning and identification of hypoxia tolerance function of soybean alanine aminotransferase gene GmALAAT1 [J]. Journal of Jilin Agricultural University: 1-8.). The wheat TaERF2 gene can quickly respond to ethylene transcription factors after stress, and its expression increases significantly, improving plant resistance (Song Guicheng, Zhou Miaoping, Yu Guihong, Yao Jinbao, Ma Hongxiang. Expression analysis of wheat ethylene transcription factor TaER F2 in response to moisture stress [J]. Journal of Nuclear Agricultural Sciences, 2022, 36(05): 876-884.). Current research on exogenous agents for plants is mostly focused on the seedling stage. For example, spraying 200 μM melatonin on the leaves and watering 600 μM melatonin on the roots can significantly alleviate the stress of apple seedlings in adversity (Nicolas E. Castro-Duque, Cristhian C. Chávez-Arias and Hermann Restrepo-Díaz. Foliar Glycine Betaine or Hydrogen Peroxide Spra ysnAmeliorate Waterlogging Stress in Cape Gooseberry[J]. Plants, 2020, 9: 644-662.). Applying 25 ppm of ordinary silicon particles and nano-silicon in the form of aeration to the nutrient solution of grape seedlings can increase the biomass of grape seedlings under hypoxia stress (Zheng Xiaodong, Zhou Jingzhe, Tan Dun-Xian, et al. Melatonin Improves Waterlogging Tolerance of Malus baccata (Linn.) Borkh. Seedlings by Maintaining Aerobic Respiration, Photosynthesis and ROS Imitation [J]. Frontiers in Plant Science, 2017, 5 (8): 483-493.). However, scholars have paid less attention to hypoxia stress during seed germination, and most of them use a single agent to treat seeds to alleviate hypoxia stress during germination, and the treatment effects vary greatly. Summary of the invention

[0004] The invention provides a method for improving the vigor of sweet and sticky corn seeds under hypoxia stress. The method can effectively alleviate the stress damage of germination under hypoxia conditions to sweet corn seeds and sticky corn seeds, promote seed germination and seedling growth, and the method of use is simple.

[0005] The specific technical solutions are as follows:

[0006] The present invention provides a method for improving the vigor of sweet and sticky corn seeds under hypoxia stress, comprising the following steps:

[0007] (1) Preparing a compound medicine; the compound medicine uses water as a solvent and contains 60 to 1000 mg / L of reduced glutathione. -1 , Vitamin B2 12~200mg·L -1 ;

[0008] (2) The sweet / glutinous corn seeds are placed between germination papers soaked with the compound agent, and the surface agent is cleaned after being imbibed at 23-25° C. for 22-26 hours. The seeds are then placed in a fume hood and slowly dried for 24 hours before being sown to obtain sweet / glutinous corn seeds with high vitality.

[0009] Furthermore, in step (1), the compound medicine contains 125-500 mg / L reduced glutathione in water as solvent. -1 , Vitamin B2 25~100mg·L -1 .

[0010] Furthermore, in step (1), the compound medicine contains 250 mg·L reduced glutathione in water as solvent. -1 , Vitamin B2 50mg·L -1 .

[0011] Furthermore, the time for the seeds to swell between germination papers soaked with the compound agent is 24 hours.

[0012] Furthermore, the slow drying time is 24 hours.

[0013] Furthermore, in step (1), the preparation method of the compound medicine is to directly dissolve reduced glutathione and vitamin B2 in water, stir them evenly and set them aside.

[0014] Furthermore, in step (2), the germination paper is fully soaked in the compound agent, and no water drops fall after being lifted, and two layers of the soaked germination paper are placed in a covered germination box; the corn seeds are then disinfected with 1% sodium hypochlorite solution, washed with clean water, placed on the soaked germination paper, and then covered with a layer of germination paper soaked in the agent, and the germination box is covered with a lid, and swelled at 25° C. for 24 hours; after the swelling is completed, the seeds are taken out, the surface agent is cleaned, and then placed in a fume hood to slowly dry for 24 hours.

[0015] The seeds are placed between germination papers soaked with a compound agent to swell. The seeds can absorb water slowly and evenly. Experiments have found that compared with the seed soaking treatment, the slow swelling between papers will not inhibit the normal respiration of the seeds and can effectively avoid the damage caused by the swelling of the seeds.

[0016] The specific method includes the following steps:

[0017] (A) fully soaking the germination paper in the compound agent until no water drops after being lifted; placing two layers of the soaked germination paper in a germination box with a cover;

[0018] (B) The corn seeds were disinfected with 1% sodium hypochlorite solution by mass, washed with clean water, placed on germination paper soaked with the agent, covered with another layer of germination paper soaked with the agent, and covered with a germination box lid, and imbibed at 25°C for 24 hours;

[0019] (C) After the imbibition is completed, the corn seeds are taken out, the surface agents are cleaned, and then they are placed in a fume hood to slowly dry for 24 hours to obtain sweet and sticky corn seeds with high vitality.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) The present invention uses reduced glutathione and vitamin B2 as the basic agents, which are dissolved in a certain amount of water. After treatment, the reduced glutathione and vitamin B2 can effectively improve the resistance of sweet corn and waxy corn seeds to low oxygen environments, promote seed germination and seedling growth and development, and improve economic benefits without polluting the environment.

[0022] (2) The seed treatment method provided by the present invention can be used to pre-treat sweet corn and glutinous corn seeds before field sowing. When sudden rainfall causes field waterlogging or continuous rainy weather occurs, it helps to improve seed resistance, increase field emergence rate and seedling uniformity, and reduce economic losses.

[0023] (3) The preparation method of the compound medicine of the present invention is simple, and water is used as the matrix, which is highly environmentally friendly and safe, requires a small amount of medicine, and has low cost and is suitable for promotion. DETAILED DESCRIPTION

[0024] The present invention will be further described below in conjunction with specific embodiments. The following are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto.

[0025] In the following examples, the seeds of sweet corn varieties "S40-4-3-1-2-3" and "S60-4-1-3-2-2-1" and the seeds of waxy corn varieties "N32-3-2-3-2" and "N52-1-2-3-2" were all provided by Zhejiang University. Reduced glutathione and vitamin B2 were purchased from Shanghai Aladdin Reagent Co., Ltd.

[0026] Example 1

[0027] 1. Test materials: "S40-4-3-1-2-3" seeds

[0028] 2. Test reagents: 1% sodium hypochlorite solution, vitamin B2, reduced glutathione, NBT, ascorbic acid, 30% H2O2 solution, thiobarbituric acid, trichloroacetic acid, PBS buffer. All the above reagents were purchased from Shanghai Aladdin Reagent Company.

[0029] 3. Test methods

[0030] Soak the corn seeds in 1% sodium hypochlorite solution for 10 minutes, rinse with clean water, drain and set aside.

[0031] (1) Seed treatment method

[0032] Using water as solvent, reduced glutathione and vitamin B2 were dissolved in water according to the types and dosages of agents required for different treatments to obtain a compound agent combination that can be used directly (Table 1, treatments 1 to 5). At the same time, single agent and clean water treatments were set for comparison (treatments 6 to 8). Treatments 1 to 8 were all dried after swelling, and hypoxia stress treatment was performed during seed germination; dry seeds were used as controls (CK1, CK2), of which CK1 seeds were directly germinated after disinfection, and CK2 seeds were disinfected and hypoxia stress treatment was performed during seed germination. The specific treatment methods are shown in Table 1.

[0033] Table 1 Treatment number, drug combination and treatment method

[0034]

[0035]

[0036] Soak the germination paper fully in the solution of the agent in treatments 1 to 8 or in clean water, and place two layers of soaked germination paper in a covered germination box until no water drops down after lifting it. Then disinfect the corn seeds with 1% sodium hypochlorite solution, wash them with clean water, and evenly place them on the soaked germination paper. Then cover them with a layer of germination paper soaked in the agent, cover them with a lid, and imbibe at 25°C for 24 hours.

[0037] After the imbibition is completed, take out the seeds, clean the surface agent, and place them in a fume hood to slowly dry for 24 hours before using them for sowing.

[0038] (2) Hypoxia stress

[0039] The test seeds that need hypoxia stress were placed in different beakers according to different treatments (treatments 1 to 8, CK2), deionized water was added to cover the seeds, and then placed in a dark environment at 25°C for 48 hours to simulate hypoxia stress. After the stress ended, the seeds could be used for germination test.

[0040] (3) Germination test

[0041] The germination test was carried out in accordance with the provisions of GB / T3534.1~3543.7-1995 "Inspection Procedures for Crop Seeds", using the paper roll method. 50 seeds were divided into two rows and evenly placed on two layers of germination paper moistened with clean water, and then a piece of germination paper moistened with clean water of the same size was covered on the seeds, the bottom was folded 2 cm, rolled into a paper roll, and the upper end was fixed with a rubber band, and placed vertically in the germination box, and cultured at 25°C for 7 days with 18 hours of light and 6 hours of darkness. 100 seeds were used as a replicate (2 paper rolls), and each treatment was repeated three times. In addition, in order to ensure the consistency of germination water absorption between CK1 and treatments 1~8 and CK2, and to synchronize the germination process as much as possible, CK1 was placed in bed for germination one day in advance, and this day was used as pretreatment time and was not included in the total number of culture days.

[0042] Determination of seed vitality and vigor index: The day of seed placement is regarded as the first day, the germination potential is calculated on the fourth day, and the germination rate is calculated on the seventh day. The calculation method is as follows:

[0043] Germination potential (%) = (number of seeds that germinated normally after 4 days of culture / number of seeds tested) × 100

[0044] Germination rate (%) = (number of seeds that germinated normally at the end of the 7th day of culture / number of seeds tested) × 100

[0045] After the germination test, 10 normal seedlings were randomly selected from each replicate to measure the root length and seedling height. After the seed residues were stripped from the seedlings, the surface moisture was absorbed with absorbent paper, and the fresh weight was weighed. Then the whole seedling was placed in a clean aluminum box and pre-dried at 85°C for 30 minutes, and then dried at 105°C for 8 hours, and the dry weight was weighed.

[0046] (4) Enzyme activity assay

[0047] According to the results of the vitality data analysis of the germination test, the best combination of agents, a single agent and two controls were selected to conduct hypoxia stress and paper roll germination tests again to detect the relevant indicators of the antioxidant enzyme system. Each replicate had 50 seeds, which were placed in a paper roll; each treatment was repeated three times.

[0048] Preparation of crude enzyme extract: After germination for 24 hours, the seeds of the paper roll used for enzyme activity determination were taken out, rinsed in clean water, and the surface moisture was wiped dry. Ten seeds were randomly selected from each paper roll (repeated) and fully ground into powder in liquid nitrogen; 0.2 g of fresh powdered sample was weighed and placed in a 10 ml centrifuge tube, 5 ml of PBS buffer was added, and after sufficient shaking, the tube was centrifuged at 25,000 r / min for 10 min, and 4 ml of supernatant was taken, which was the crude extract required for enzyme determination.

[0049] SOD activity determination: Nitroblue tetrazolium (NBT) photoreduction method was used. 100 μL of crude enzyme extract was added to 3 mL of NBT reaction solution and illuminated for 20 min. 3 mL of NBT reaction solution plus 100 μL of extraction buffer was used as control. Colorimetry was performed at a wavelength of 560 nm.

[0050] POD activity determination: Add 2700μL PBS, 100μL ascorbic acid, and 100μL H2O2 to 100μL enzyme solution, shake well, and quickly transfer to a cuvette. Use a spectrophotometer to measure the change in absorbance at 340nm within 1 minute. A decrease of 0.1 in absorbance per minute is considered one unit of enzyme activity.

[0051] CAT activity determination: using hydrogen peroxide colorimetry, add 2800μL PBS and 100μL H2O2 to 100μL enzyme solution, shake well, and quickly transfer to a colorimetric dish, and determine its activity using a kinetic method at a wavelength of 240nm. A decrease in absorbance of 0.01 per minute is considered one unit of enzyme activity.

[0052] Determination of MDA content: Take 1.5 mL of enzyme solution and mix it with 2.5 mL of 20% trichloroacetic acid (TCA) solution containing 0.5% thiobarbituric acid (TBA), boil it in a boiling water bath for 10 min, cool it quickly, centrifuge it for 10 min, and measure the absorbance of the supernatant at 532, 600, and 450 nm, respectively.

[0053] Statistical analysis: SPSS 26.0 software was used to analyze the experimental data with Duncan method (P﹤0.05).

[0054] 4. Test results

[0055] The results of this example are shown in Table 2. Comparing the data of CK1 and CK2, under hypoxia stress, the germination potential and germination rate of the seeds were significantly reduced, which indicates that hypoxia stress reduces the vigor of corn seeds and inhibits the germination of corn seeds; after treatment with the agent of this example, the seed vigor was improved to varying degrees, among which the germination potential and germination rate of treatment 3 were 36.67% and 43.33%, respectively, which were 10 and 14.44 percentage points higher than those of CK2, respectively, and both reached significant differences; the germination potential and germination rate of treatment 4 were 38.89% and 40.00%, respectively, which were 12.22 and 11.11 percentage points higher than those of CK2, respectively, and both reached significant differences.

[0056] Compared with CK1, from the perspective of seedling growth, the root length, seedling height, fresh weight and dry weight of CK2 under hypoxia stress all decreased to varying degrees, but there was no significant difference in seedling height and fresh weight, while the root length and dry weight were significantly reduced by 33.97% and 26.32%, indicating that the hypoxic environment has a greater impact on the root growth and dry matter accumulation of seedlings. Compared with CK2, after treatment with the agent of this embodiment, the root length, seedling height, fresh weight and dry weight of the seedlings were improved to varying degrees, among which treatment 3 performed best, and the improvement of the four indicators was better than other treatments (including single agent and clear water treatment), and reached or exceeded CK1, and the root length was significantly better than other treatments and CK2. It shows that treatment 3 can promote the root growth and dry matter accumulation of corn seeds under hypoxia stress, and resist the adverse effects of stress.

[0057] Based on the above germination and seedling growth conditions, the present embodiment treated with 3 compound agents (250 mg·L -1 Reduced glutathione +50mg·L -1 Vitamin B2) has the best effect on improving seed vitality under hypoxia stress.

[0058] Table 2 Effects of treatment with chemicals on seed vigor of S40-4-3-1-2-3 under hypoxia stress

[0059]

[0060] Note: The lowercase letters after the data in the table indicate the significant difference between the data of different treatments in the same column (P﹤0.05), the same below.

[0061] To further study the mechanism of improving seed vitality under hypoxic stress, we measured and analyzed the indicators related to the antioxidant enzyme system. SOD, POD, and CAT are enzymes closely related to plant stress resistance. These enzymes can remove substances that are destructive to cells, such as hydrogen peroxide and reactive oxygen free radicals, and enhance the resistance of plants to stress; MDA is one of the commonly used indicators to measure the degree of oxidative stress, which can reflect the degree of plant membrane lipid peroxidation. The MDA content in plants will increase in a hypoxic environment, and excessive accumulation will affect the normal life activities of plants and even cause plant death. From the data listed in Table 3, it can be seen that compared with CK2, the SOD, POD and CAT activities of treatment 3 were significantly improved, among which SOD and POD were significantly increased by 51.31% and 19.10%, respectively, while the MDA content was significantly reduced by 56.28%; at the same time, compared with the single-agent treatment, the compound agent treatment 3 was more effective in improving the activities of POD, SOD and CAT, among which the SOD activity was significantly higher than the two single agents, resisting the adverse effects of stress and ensuring the normal germination and growth of corn seeds.

[0062] Table 3 Effects of treatment with chemicals on the antioxidant enzyme system of S40-4-3-1-2-3 seeds under hypoxia stress

[0063]

[0064] Example 2

[0065] In Example 1, it was confirmed that the method of the present invention can improve the vitality of corn seeds under hypoxia stress, and the effect is better than that of a single agent. In this example, the effect of the method of the present invention is further verified.

[0066] 1. Test materials: "S60-4-1-3-2-2-1" seeds

[0067] 2. Test reagent: same as in Example 1.

[0068] 3. Test method: Same as Example 1.

[0069] 4. Test results

[0070] In this example, under hypoxia stress, the germination potential and germination rate of the seeds also decreased significantly, further indicating that hypoxia stress reduced the vigor of corn seeds and inhibited seed germination. After treatment with the agent of this example, the seed vigor was improved to varying degrees, among which the germination potential and germination rate of the compound agent treatment 3 were significantly higher than those of CK2, single agent treatments 6 and 7, and water, reaching 57.78% and 63.33%, respectively, which was 22.22 and 27.77 percentage points higher than that of CK2; and the germination potential was no significantly different from that of the unstressed CK1 (Table 4).

[0071] From the perspective of seedling growth, hypoxia stress inhibited the normal growth of the root system of corn seeds, and the root length of CK2 was 24.88% lower than that of CK1. After treatment with the agent of this embodiment, the root length, seedling height, fresh weight and dry weight of the seedlings were improved to varying degrees, among which treatment 3 performed the best, and there was no significant difference in the four indicators compared with CK1, and the improvement was significantly better than other treatments (including single agent and clear water treatment). The root length and fresh weight were significantly increased by 31.81% and 57.14% respectively compared with CK2 (Table 4).

[0072] Based on the above germination and seedling growth, this example further shows that the treatment 3 compound (250mg·L -1 Reduced glutathione +50mg·L -1 Vitamin B2) can effectively improve the vitality of corn seeds under hypoxia stress.

[0073] Table 4 Effects of treatment with chemicals on seed vigor of S60-4-1-3-2-2-1 under hypoxia stress

[0074]

[0075] In order to further verify the mechanism of treatment 3 in improving seed vitality, the antioxidant enzyme system during seed germination was compared and analyzed, and the results are shown in Table 5. Compared with CK2, the activities of SOD, POD, and CAT in treatment 3 were significantly increased by 96.65%, 9.80%, and 27.78%, respectively, and the MDA content was significantly reduced by 32.19%. There were no significant differences between the activities of SOD and POD in treatment 6 and CK2, but the CAT activity increased significantly by 11.11%, and the MDA content decreased significantly by 22.60%. There were no significant differences between the activities of SOD and POD in treatment 7 and CK2, but the CAT activity increased significantly by 16.27%, and the MDA content decreased significantly by 25.34%. Therefore, the promoting effect of treatment 3 on the activities of the three enzymes was better than that of treatments 6 and 7, and the effect of reducing the MDA content was the best. Compared with single agents, compound agents can better promote the normal physiological activities of corn seeds under hypoxia stress.

[0076] Table 5 Effects of treatment with chemicals on the antioxidant enzyme system of S60-4-1-3-2-2-1 seeds under hypoxia stress

[0077]

[0078]

[0079] Example 3

[0080] In Examples 1 and 2, it was confirmed that the method of the present invention can improve the vitality of corn seeds under hypoxia stress, and the effect is better than that of a single agent. The effect of the method is further verified in this example.

[0081] 1. Test materials: "N32-3-2-3-2" seeds

[0082] 2. Test reagent: same as in Examples 1 and 2.

[0083] 3. Test method: Same as Example 1 and Example 2.

[0084] 4. Test results

[0085] In this example, under hypoxia stress, the germination potential and germination rate of the seeds also decreased significantly, further indicating that hypoxia stress reduced the vigor of corn seeds and inhibited seed germination. After treatment with the agent of this example, the seed vigor was improved to varying degrees, among which the germination potential and germination rate of the compound agent treatment 3 were significantly higher than those of CK2, single agent treatments 6 and 7, and clear water, reaching 47.78% and 53.33% respectively, which was significantly higher than CK2 by 12.22 and 11.11 percentage points; and the germination potential was no significantly different from that of the unstressed CK1 (Table 6).

[0086] From the perspective of seedling growth, hypoxia stress has an inhibitory effect on the root length and seedling height of the seedlings to a certain extent, but it has not reached a significant level; after treatment with the agent of this example, the root length and seedling height of the seedlings were improved to varying degrees compared with CK2, but there were no significant differences in the four indicators including fresh weight and dry weight, which may be related to the enhanced resistance of this glutinous corn variety to the hypoxic environment after emergence (Table 6).

[0087] Based on the above germination and seedling growth, this example further shows that the treatment 3 compound (250mg·L -1 Reduced glutathione +50mg·L -1 Vitamin B2) can effectively improve the vitality of corn seeds under hypoxia stress.

[0088] Table 6 Effects of treatment with chemicals on seed vigor of N32-3-2-3-2 under hypoxia stress

[0089]

[0090] In order to further verify the mechanism of treatment 3 in improving seed vitality, the seed antioxidant enzyme system was compared and analyzed, and the results are shown in Table 7. Compared with CK2, the SOD activity of treatment 3 increased by 60.70%, the POD activity increased by 10.41%, and the CAT activity and MDA content did not change significantly; the SOD activity of treatments 6 and 7 increased by 17.21% and 12.60%, respectively, the CAT activity decreased, and the remaining two indicators did not change significantly. This may be due to the fact that CAT and the other two enzymes play a similar role in resisting stress, resulting in feedback regulation in the plant body. Therefore, based on the four indicators of the antioxidant enzyme system, treatment 3 is better than a single agent in improving enzyme activity, and can better promote the normal physiological activities of corn seeds under hypoxia stress.

[0091] Table 7 Effects of treatment with chemicals on the antioxidant enzyme system of N32-3-2-3-2 seeds under hypoxia stress

[0092]

[0093] Example 4

[0094] In Examples 1, 2 and 3, it was confirmed that the method of the present invention can improve the vitality of corn seeds under hypoxia stress, and the effect is better than that of a single agent treatment. The effectiveness of the method is further verified in this example.

[0095] 1. Test materials: "N52-1-2-3-2" seeds

[0096] 2. Test reagent: Same as Example 1, Example 2 and Example 3.

[0097] 3. Test method: Same as Example 1, Example 2 and Example 3.

[0098] 4. Test results

[0099] As can be seen from Table 8, hypoxia stress has a significant inhibitory effect on the germination of the seeds of this corn variety, and the germination potential and germination rate of CK2 decreased by 12.22 and 18.89 percentage points respectively compared with CK1. After being treated with the agent of this embodiment, the seed vigor was improved to varying degrees, among which the germination potential and germination rate of the compound agent treatment 3 were significantly higher than those of CK2, single agent treatments 6 and 7, and water, reaching 63.33% and 67.78% respectively, which was significantly higher than CK2 by 15.55 and 14.45 percentage points; and there was no significant difference in germination potential and germination rate compared with the unstressed CK1.

[0100] From the perspective of seedling growth, hypoxia stress has an inhibitory effect on the root length and seedling height of the seedlings to a certain extent, but it has not reached a significant level; after treatment with the agent of this example, the root length and seedling height of the seedlings were improved to varying degrees compared with CK2, but there were no significant differences in the four indicators including fresh weight and dry weight, which may be related to the enhanced resistance of glutinous corn varieties to low oxygen environments after emergence (Table 8).

[0101] Based on the above germination and seedling growth, this example further shows that the treatment 3 compound (250mg·L -1 Reduced glutathione +50mg·L -1 Vitamin B2) can effectively improve the vitality of corn seeds under hypoxia stress.

[0102] Table 8 Effects of treatment with chemicals on seed vigor of N52-1-2-3-2 under hypoxia stress

[0103]

[0104]

[0105] To further verify the mechanism of treatment 3 in improving seed vitality, the seed antioxidant enzyme system was compared and analyzed, and the results are shown in Table 9. Compared with CK2, the SOD and POD activities of seeds in treatment 3 were significantly increased by 42.17% and 4.9%, and the MDA content was significantly reduced by 31.51%; the SOD activities of treatments 6 and 7 were increased by 17.60% and 16.93%, respectively, and the POD activity and MDA content did not change significantly. Therefore, based on the four indicators of the antioxidant enzyme system, treatment 3 is better than a single agent in improving enzyme activity and can better promote the normal physiological activities of corn seeds under hypoxia stress.

[0106] Table 9 Effects of treatment with chemicals on the antioxidant enzyme system of N52-1-2-3-2 seeds under hypoxia stress

[0107]

[0108] Example 5

[0109] Examples 1, 2, 3 and 4 all verify that the method of the present invention can effectively improve the vigor of corn seeds under hypoxia stress. This example verifies the effect of the method on seed germination under non-stress conditions (normal conditions).

[0110] 1. Test materials: seeds of "S40-4-3-1-2-3", "S60-4-1-3-2-2-1", "N32-3-2-3-2" and "N52-1-2-3-2"

[0111] 3. Test reagent: Same as Example 1, Example 2, Example 3 and Example 4.

[0112] 3. Test method: The combination of agents, active ingredients and concentrations of treatment 3 are the same as those of Examples 1, 2, 3 and 4, but no hypoxia stress treatment is performed; the seed germination test and related index determination are the same as those of Examples 1, 2, 3 and 4.

[0113] 4. Test results

[0114] In this embodiment, the seeds of the four varieties treated by the method of the present invention without hypoxia stress had no obvious effect on seed vitality and vigor when they germinated under normal circumstances, and significantly improved the germination potential and germination rate of N32-3-2-3-2, and also had a good promoting effect on the seedling growth of S40-4-3-1-2-3 (Table 10). Therefore, the treatment of the method of the present invention has no negative impact on the germination and seedling growth of sweet and waxy corn seeds, and when encountering hypoxia stress, it can effectively improve the physiological metabolic activities of corn seeds and improve the vitality of seeds.

[0115] Table 10 Effect of the method of the present invention on corn seed germination under non-hypoxia stress

[0116]

[0117]

Claims

1. A method for improving the vigor of sweet and sticky corn seeds under hypoxia stress, characterized in that: The following steps are involved: (1) Preparing a compound medicine; the compound medicine uses water as a solvent and contains 125 to 500 mg / L of reduced glutathione -1 , Vitamin B2 25~100 mg·L -1 (2) placing the sweet / glutinous corn seeds between germination papers soaked with the compound agent, allowing them to swell at 23-25°C for 22-26 hours, then cleaning the surface agent, and then slowly drying them in a fume hood for 24 hours before sowing to obtain sweet / glutinous corn seeds with high vitality.

2. The method for improving the vigor of sweet and sticky corn seeds under hypoxia stress according to claim 1, characterized in that: In step (1), the compound medicine uses water as solvent and contains 250 mg·L reduced glutathione. -1 , Vitamin B250 mg·L -1 .

3. The method for improving the vigor of sweet and sticky corn seeds under hypoxia stress according to claim 1, characterized in that: The steps include: (A) Soaking the germination paper in the compound agent until no water drops after lifting it; placing two layers of soaked germination paper in a germination box with a cover; (B) Disinfect corn seeds with 1% sodium hypochlorite solution, wash with clean water, place on germination paper soaked in the agent, cover with another layer of germination paper soaked in the agent, cover with the lid of the germination box, and imbibe at 25°C for 24 h; (C) After the imbibition is complete, the corn seeds are taken out, the surface agents are cleaned, and then they are placed in a fume hood to slowly dry for 24 hours to obtain sweet and sticky corn seeds with high vitality.

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