A method for cultivating salt-tolerant sorghum varieties

Abstract

The present invention belongs to the technical field of sorghum, and specifically relates to a method for cultivating salt-tolerant sorghum varieties, comprising the following steps: S1, washing sorghum seeds with sterile water, disinfecting, and then soaking in a composite salt-tolerant inducer aqueous solution; S2, selecting a culture medium containing sodium chloride, evenly sowing the soaked seeds on the culture medium for cultivation, regularly checking the humidity of the culture medium, replenishing water, and timely removing sick and weak seedlings; S3, transplanting to a test field, conducting field management, and applying topdressing within one month after transplanting; S4, multi-generation selection and breeding. The present invention activates the defense mechanism and growth regulation system of seed cells by pretreating seeds with a composite salt-tolerant inducer. Modified chitosan enhances the stability of the cell wall and improves the survival ability of seeds under salt stress, while indoleacetic acid optimizes the root structure and enhances the adaptability of plants to adversity.

Description

Technical Field

[0001] The invention belongs to the technical field of sorghum, and in particular relates to a method for cultivating a salt-tolerant sorghum variety. Background Art

[0002] With the intensification of global climate change and land salinization, the cultivation and research of salt-tolerant crops are particularly important. Among many crops, sorghum has attracted widespread attention for its multiple resistance, especially salt tolerance. At present, research on sorghum salt tolerance has made certain progress. Researchers have used different methods, such as physiological and biochemical analysis, molecular biology technology and genetic research, to explore the physiological response and adaptation mechanism of sorghum under salt stress. These studies not only involve the morphological structure and growth status of sorghum, but also its physiological and biochemical characteristics and the expression regulation of related genes. In the identification of sorghum salt tolerance, the strength of its salt tolerance is mainly evaluated by observing the growth performance, changes in physiological indicators, and the effects on yield and quality of sorghum under salt stress. There are significant differences in salt tolerance among different varieties of sorghum, which are not only reflected in different growth stages, but also affected by environmental conditions and genetic factors. The regulation of protective enzymes is one of the important mechanisms for sorghum to cope with salt stress. Under salt stress, enzymes such as peroxidase and ascorbate peroxidase can eliminate toxic substances produced by the stress, thereby ensuring the normal growth and development of sorghum under a certain salt-alkali concentration. Osmotic regulation is also the main physiological mechanism for sorghum to adapt to salt stress. Sorghum regulates cell osmotic potential by accumulating substances that are harmless to protoplasts in cells to cope with salt stress. Solutes mainly include proline, betaine and soluble sugars, which play an important role in sorghum's salt tolerance.

[0003] In the existing technology, the traditional breeding methods lack precise directionality and rely on a large number of population screening and trial and error processes, so the efficiency is relatively low. In addition, the traditional breeding methods mainly select based on the performance under the current environmental conditions, so the varieties cultivated are less adaptable to future environmental changes, especially in the face of climate change and soil salinization. Summary of the invention

[0004] The purpose of the present invention is to provide a method for cultivating salt-tolerant sorghum varieties in view of the deficiencies of the prior art.

[0005] The purpose of the present invention can be achieved through the following technical solutions:

[0006] A method for cultivating a salt-tolerant sorghum variety comprises the following steps:

[0007] S1. Collect sorghum seeds without pests and damage, wash the seeds with sterile water for 3-5 times, each washing time is 5-10 minutes, soak the seeds in 1-2% sodium hypochlorite solution for 10-15 minutes, then rinse with sterile water, and then soak in the composite salt tolerance inducer aqueous solution for 30-40 minutes;

[0008] S2. Select a culture medium containing 0.5-1% sodium chloride to simulate salinized soil conditions, sow the soaked seeds evenly on the culture medium for cultivation, regularly check the humidity of the culture medium, replenish water, and remove sick and weak seedlings in time;

[0009] S3. When the seedlings grow to the 3-4 leaf stage, they are transplanted to the experimental field. Different salt concentration areas should be set in the experimental field. The salt concentration should be set to 0.4% and 0.8%. Transplanting should be carried out on cloudy days or in the evening. Keep the soil moist during transplanting to ensure that the root system of the seedlings is intact. Carry out field management and apply topdressing within one month after transplanting.

[0010] S4. Measure the yield of each plant at harvest, select plants whose yield at a salt concentration of 2% can still reach more than 70% of that at a salt concentration of 0.5%, breed the selected salt-tolerant plants, and cultivate a stable salt-tolerant sorghum variety through multiple generations of selection and breeding.

[0011] Furthermore, in step S1, the concentration of the composite salt tolerance inducer aqueous solution is 0.5-1.5% (w / v), the soaking time is 24-48 hours, and the solution is stirred every 6-8 hours.

[0012] The composite salt-tolerant inducer aqueous solution in step S1 is prepared by the following steps:

[0013] A1. Slowly add chitosan powder into 1% acetic acid solution and stir until completely dissolved. At a stirring speed of 300-400 rpm, add 2-acrylamido-2-methylpropanesulfonic acid, control the reaction temperature to 60-70°C, and the reaction time to 4-6h. After the reaction is completed, precipitate with excess ethanol, filter, wash, and vacuum dry at 60°C for 12-24h to obtain modified chitosan;

[0014] A2. Add modified chitosan to dimethyl sulfoxide, add indoleacetic acid at a stirring speed of 300-400 rpm, control the reaction temperature to 40-50°C, add N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, react for 24-30 hours, and after the reaction, precipitate with excess deionized water, filter, wash, and prepare a 5-10% composite salt-resistant inducer aqueous solution with deionized water.

[0015] The culture medium in step S2 comprises the following raw materials by mass fraction: 1.5-3% agar, 0.5-1% sodium chloride, 0.02-0.03% calcium chloride, 0.01-0.02% magnesium sulfate, 0.2-0.5% ammonium nitrate, 0.15-0.35% potassium dihydrogen phosphate, 0.001% thiamine and 0.001% riboflavin, and the balance is water, and the pH value is adjusted to 5.5-6.5.

[0016] The conditions for sowing and culturing on the culture medium in step S2 are: humidity controlled at 60-70%, temperature controlled at 25-30° C., light intensity at 2000-3000 l x, and light duration at 16 h / day.

[0017] The strategy of topdressing in step S3 is as follows: on the 10th day after transplanting, 1.5-2.5 kg of urea, 4-6 kg of diammonium phosphate and 5-7 kg of potassium chloride are applied per mu; on the 20th day after transplanting, 7-10 kg of urea are applied per mu; and on the 25th day after transplanting, 5-7 kg of potassium chloride are applied per mu.

[0018] In the step A1, the usage ratio of chitosan, acetic acid and 2-acrylamido-2-methylpropanesulfonic acid is (10-15) g: (100-120) mL: (3.5-4.5) g.

[0019] In the step A2, the usage ratio of modified chitosan, dimethyl sulfoxide, indoleacetic acid, N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is (10-12) g: (100-110) mL: (0.2-0.3) g: (0.05-0.07) g: (0.01-0.02) g.

[0020] Beneficial effects of the present invention:

[0021] 1. In the technical solution of the present invention, after the chitosan is modified, the active groups on its molecular chain can interact with the negative charges on the cell wall, thereby enhancing the stability of the cell wall and helping to maintain the normal function of the cell under salt stress. In addition, the modified chitosan also acts as a signal molecule to activate the defense mechanism in the plant body and improve the plant's resistance to salt stress. Indoleacetic acid is a plant growth hormone that plays a role in regulating cell elongation and division in the plant body. Under salt stress, indoleacetic acid can help plants adjust their root structure and enhance their ability to absorb water and nutrients, thereby improving the plant's salt tolerance.

[0022] 2. In the technical solution of the present invention, the seeds are pretreated with a composite salt-tolerance inducer to activate the defense mechanism and growth regulation system of the seed cells. Modified chitosan enhances the stability of the cell wall and improves the survival ability of seeds in a salt stress environment, while indoleacetic acid optimizes the root structure and enhances the adaptability of plants to adversity. By setting a gradually increasing salt concentration in the culture medium and the experimental field, a gradual adaptation process is provided for the seedlings, which helps to screen out plants with naturally high salt tolerance and ensure that these plants can still maintain high yields in a high-salt environment. Through comprehensive seed treatment, gradual adaptation to the salt stress environment, systematic field management, and selective breeding, an efficient and comprehensive salt-tolerant sorghum variety breeding system has been formed, which is more targeted and efficient than traditional methods. DETAILED DESCRIPTION

[0023] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] Example 1

[0025] The composite salt-tolerant inducer aqueous solution comprises the following steps to prepare:

[0026] A1. Slowly add 100 g chitosan powder into 100 mL of 1% acetic acid solution and stir until completely dissolved. At a stirring speed of 300 rpm, add 35 g 2-acrylamide-2-methylpropane sulfonic acid. Control the reaction temperature to 60 ° C and the reaction time to 4 h. After the reaction is completed, precipitate with excess ethanol, filter, wash, and vacuum dry at 60 ° C for 12 h to obtain modified chitosan.

[0027] A2. Add 100 g of modified chitosan to 1000 mL of dimethyl sulfoxide, add 2 g of indoleacetic acid at a stirring speed of 300 rpm, control the reaction temperature to 40°C, add 0.5 g of N-hydroxysuccinimide and 0.1 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, react for 24 h, and after the reaction is completed, precipitate with excess deionized water, filter, wash, and use deionized water to prepare a 5% composite salt-resistant inducer aqueous solution.

[0028] Example 2

[0029] The composite salt-tolerant inducer aqueous solution comprises the following steps to prepare:

[0030] A1. Slowly add 125 g chitosan powder into 110 mL of 1% acetic acid solution and stir until completely dissolved. At a stirring speed of 350 rpm, add 40 g 2-acrylamide-2-methylpropane sulfonic acid. Control the reaction temperature to 65 ° C and the reaction time to 5 h. After the reaction is completed, precipitate with excess ethanol, filter, wash, and vacuum dry at 60 ° C for 18 h to obtain modified chitosan.

[0031] A2. Add 110 g of modified chitosan to 1050 mL of dimethyl sulfoxide, add 2.5 g of indoleacetic acid at a stirring speed of 350 rpm, control the reaction temperature to 45 ° C, add 0.6 g of N-hydroxysuccinimide and 0.15 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, and react for 26 hours. After the reaction is completed, precipitate with excess deionized water, filter, wash, and use deionized water to prepare a composite salt-resistant inducer aqueous solution with a concentration of 7.5%.

[0032] Example 3

[0033] The composite salt-tolerant inducer aqueous solution comprises the following steps to prepare:

[0034] A1. Slowly add 150 g chitosan powder into 120 mL of 1% acetic acid solution and stir until completely dissolved. At a stirring speed of 400 rpm, add 45 g 2-acrylamide-2-methylpropane sulfonic acid. Control the reaction temperature to 70 ° C. The reaction time is 6 h. After the reaction is completed, precipitate with excess ethanol, filter, wash, and vacuum dry at 60 ° C for 24 h to obtain modified chitosan.

[0035] A2. Add 120g of modified chitosan to 1100mL of dimethyl sulfoxide, add 2-3g of indoleacetic acid at a stirring speed of 400rpm, control the reaction temperature to 50°C, add 0.7g of N-hydroxysuccinimide and 0.2g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and react for 30h. After the reaction, precipitate with excess deionized water, filter, wash, and use deionized water to prepare a 10% composite salt-tolerant inducer aqueous solution.

[0036] Example 4

[0037] A method for cultivating a salt-tolerant sorghum variety comprises the following steps:

[0038] S1. Collect sorghum seeds without pests and diseases and damage, wash the seeds with sterile water for 3 times, each washing time is 5 minutes, soak the seeds in 1% sodium hypochlorite solution for 10 minutes after washing, then rinse with sterile water, and then soak them in the composite salt tolerance inducer aqueous solution prepared in Example 1 for 30 minutes;

[0039] S2. Select a culture medium containing 0.5% sodium chloride to simulate salinized soil conditions, sow the soaked seeds evenly on the culture medium, keep the humidity of the culture medium at 60%, control the temperature at 25°C, the light intensity at 2000l x, and the light time at 16h / day. Check the humidity of the culture medium regularly, replenish water, and remove sick and weak seedlings in time;

[0040] S3. When the seedlings grow to the 3-leaf stage, they are transplanted to the experimental field. Different salt concentration areas should be set in the experimental field. The salt concentration is set to 0.4% and 0.8%. Transplanting is carried out on cloudy days or in the evening. The soil is kept moist during transplanting to ensure that the root system of the seedlings is intact. Field management is carried out. On the 10th day after transplanting, 1.5 kg of urea, 4 kg of diammonium phosphate and 5 kg of potassium chloride are applied per mu. On the 20th day after transplanting, 7 kg of urea is applied per mu. On the 25th day after transplanting, 5 kg of potassium chloride is applied per mu.

[0041] S4. Measure the yield of each plant at harvest, select plants whose yield at a salt concentration of 2% can still reach more than 70% of that at a salt concentration of 0.5%, breed the selected salt-tolerant plants, and cultivate a stable salt-tolerant sorghum variety through multiple generations of selection and breeding.

[0042] Example 5

[0043] A method for cultivating a salt-tolerant sorghum variety comprises the following steps:

[0044] S1. Collect sorghum seeds without pests and diseases and damage, wash the seeds with sterile water for 4 times, each washing time is 7.5 minutes, soak the seeds in 1.5% sodium hypochlorite solution for 12.5 minutes, then rinse with sterile water, and then soak them in the composite salt tolerance inducer aqueous solution prepared in Example 2 for 35 minutes;

[0045] S2. Select a culture medium containing 0.75% sodium chloride to simulate salinized soil conditions, sow the soaked seeds evenly on the culture medium, keep the humidity of the culture medium at 65%, control the temperature at 27°C, the light intensity at 2500l x, and the light time at 16h / day. Check the humidity of the culture medium regularly, replenish water, and remove sick and weak seedlings in time;

[0046] S3. When the seedlings grow to the 3-leaf stage, they are transplanted to the experimental field. Different salt concentration areas should be set in the experimental field. The salt concentration is set to 0.4% and 0.8%. Transplanting is carried out on cloudy days or in the evening. The soil is kept moist during transplanting to ensure that the root system of the seedlings is intact. Field management is carried out. On the 10th day after transplanting, 2kg of urea, 5kg of diammonium phosphate and 6kg of potassium chloride are applied per mu. On the 20th day after transplanting, 8.5kg of urea is applied per mu. On the 25th day after transplanting, 6kg of potassium chloride is applied per mu.

[0047] S4. Measure the yield of each plant at harvest, select plants whose yield at a salt concentration of 2% can still reach more than 70% of that at a salt concentration of 0.5%, breed the selected salt-tolerant plants, and cultivate a stable salt-tolerant sorghum variety through multiple generations of selection and breeding.

[0048] Example 6

[0049] A method for cultivating a salt-tolerant sorghum variety comprises the following steps:

[0050] S1. Collect sorghum seeds without pests and diseases and damage, wash the seeds with sterile water for 5 times, each washing time is 10 minutes, soak the seeds in 2% sodium hypochlorite solution for 15 minutes, then rinse with sterile water, and then soak them in the composite salt tolerance inducer aqueous solution prepared in Example 3 for 40 minutes;

[0051] S2. Select a culture medium containing 1% sodium chloride to simulate salinized soil conditions, sow the soaked seeds evenly on the culture medium, keep the humidity of the culture medium at 70%, control the temperature at 30°C, the light intensity at 3000l x, and the light time at 16h / day. Check the humidity of the culture medium regularly, replenish water, and remove sick and weak seedlings in time;

[0052] S3. When the seedlings grow to the 4-leaf stage, they are transplanted to the experimental field. Different salt concentration areas should be set in the experimental field. The salt concentration is set to 0.4% and 0.8%. Transplanting should be carried out on cloudy days or in the evening. Keep the soil moist during transplanting to ensure that the root system of the seedlings is intact. Field management should be carried out. On the 10th day after transplanting, 2.5kg of urea, 6kg of diammonium phosphate and 7kg of potassium chloride should be applied per mu. On the 20th day after transplanting, 10kg of urea should be applied per mu. On the 25th day after transplanting, 7kg of potassium chloride should be applied per mu.

[0053] S4. Measure the yield of each plant at harvest, select plants whose yield at a salt concentration of 2% can still reach more than 70% of that at a salt concentration of 0.5%, breed the selected salt-tolerant plants, and cultivate a stable salt-tolerant sorghum variety through multiple generations of selection and breeding.

[0054] Comparative Example 1

[0055] The difference between this comparative example and Example 4 is that a 5% chitosan aqueous solution is used instead of the composite salt-tolerance inducer aqueous solution, and the remaining steps are the same as those of Example 4.

[0056] Comparative Example 2

[0057] The difference between this comparative example and Example 5 is that 7.5% modified chitosan aqueous solution is used instead of the composite salt tolerance inducer aqueous solution, and the remaining steps are the same as those in Example 5.

[0058] According to the methods of Examples 4-6 and Comparative Examples 1-2, salt tolerance cultivation tests were conducted, including seed treatment, sowing and cultivation, planting and management, harvesting and data recording, measuring the yield of each plant, and calculating the yield ratio under each salt concentration. The results are shown in Table 1:

[0059] Table 1. Test results of Examples 4-6 and Comparative Examples 1-2

[0060]

[0061] The salt tolerance index (STI) of Example 4-6 was calculated with reference to DB21 / T 3222-2020 "Technical Regulations for Identification of Salt and Alkali Tolerance of Sorghum". The salt-tolerant sorghum varieties cultivated in Example 4-6 were identified according to the method of DB21 / T 3222-2020, and a control group without stress treatment was set. The results are shown in Table 2:

[0062] Table 2. Identification results of Examples 4-6

[0063] project Example 4 Example 5 Example 6 Relative germination rate 0.85 0.82 0.87 Relative germination index 0.91 0.86 0.92 Relative root length 0.75 0.70 0.78 Relative leaf fresh weight 0.65 0.63 0.68 Relative leaf dry weight 0.71 0.65 0.73 STI 0.82 0.79 0.83

[0064] As can be seen from Table 1 and Table 2, the average yield of Examples 4-6 is significantly higher than that of Comparative Examples 1-2, indicating that the yield of sorghum under salt stress is significantly improved after the seeds are treated with the composite salt tolerance inducer aqueous solution. At a salt concentration of 2%, the yield of the examples can still reach more than 70% of that at a salt concentration of 0.5%, showing good salt tolerance.

[0065] The salt tolerance index (STI) calculated by DB21 / T 3222-2020 "Technical Regulations for Identification of Salt and Alkali Tolerance of Sorghum" shows that the sorghum varieties of Examples 4-6 all have high salt tolerance. Indicators such as relative germination rate, relative germination index, relative root length, relative leaf fresh weight and relative leaf dry weight all performed well, indicating that the composite salt tolerance inducer has a positive effect on improving the growth of sorghum under salt stress, and may improve the resistance of seeds to salt stress by improving the physiological and biochemical environment inside the seeds. Specifically, ingredients such as chitosan and modified chitosan may enhance the stability of cell membranes and reduce the damage of salt to cells. The composite salt tolerance inducer may have stimulated the stress resistance mechanism inside sorghum seeds, enabling them to better regulate growth under salt stress and maintain a high germination rate and growth rate.

[0066] In the description of the specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0067] The above contents are merely examples and explanations of the present invention. Those skilled in the art may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the invention or exceed the scope defined by the claims, they shall all fall within the protection scope of the present invention.

Claims

1. A method for breeding salt-tolerant sorghum varieties, characterized in that: The following steps are involved: S1. Collect sorghum seeds without pests and damage, wash the seeds with sterile water for 3-5 times, each washing time is 5-10 minutes, soak the seeds in 1-2% sodium hypochlorite solution for 10-15 minutes, then rinse with sterile water, and then soak in the composite salt tolerance inducer aqueous solution for 30-40 minutes; S2. Select a culture medium containing 0.5-1% sodium chloride to simulate salinized soil conditions, sow the soaked seeds evenly on the culture medium for cultivation, regularly check the humidity of the culture medium, replenish water, and remove sick and weak seedlings in time; S3. When the seedlings grow to the 3-4 leaf stage, they are transplanted to the experimental field. Different salt concentration areas should be set in the experimental field. The salt concentration should be set to 0.4% and 0.8%. Transplanting should be carried out on cloudy days or in the evening. Keep the soil moist during transplanting to ensure that the root system of the seedlings is intact. Carry out field management and apply topdressing within one month after transplanting. S4. Measure the yield of each plant at harvest, select plants whose yield at a salt concentration of 0.8% can still reach more than 70% of that at a salt concentration of 0.4%, breed the selected salt-tolerant plants, and cultivate a stable salt-tolerant sorghum variety through multi-generation selection and breeding; The composite salt-tolerant inducer aqueous solution in step S1 is prepared by the following steps: A1. Slowly add chitosan powder into 1% acetic acid solution and stir until completely dissolved. At a stirring speed of 300-400 rpm, add 2-acrylamido-2-methylpropanesulfonic acid, control the reaction temperature to 60-70°C, and the reaction time to 4-6h. After the reaction is completed, precipitate with excess ethanol, filter, wash, and vacuum dry at 60°C for 12-24h to obtain modified chitosan; A2. Add modified chitosan to dimethyl sulfoxide, add indoleacetic acid at a stirring speed of 300-400 rpm, control the reaction temperature to 40-50°C, add N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, react for 24-30 hours, and after the reaction, precipitate with excess deionized water, filter, wash, and prepare a 5-10% composite salt-resistant inducer aqueous solution with deionized water.

2. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: The concentration of the composite salt tolerance inducer aqueous solution in step S1 is 0.5-1.5% (w / v), and the soaking time is 24-48 hours, during which the solution is stirred every 6-8 hours.

3. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: The culture medium in step S2 includes the following raw materials by mass fraction: Agar 1.5-3%, sodium chloride 0.5-1%, calcium chloride 0.02-0.03%, magnesium sulfate 0.01-0.02%, ammonium nitrate 0.2-0.5%, potassium dihydrogen phosphate 0.15-0.35%, thiamine 0.001% and riboflavin 0.001%, the balance is water, and the pH value is adjusted to 5.5-6.

5.

4. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: The conditions for sowing and culturing on the culture medium in step S2 are: humidity controlled at 60-70%, temperature controlled at 25-30° C., light intensity at 2000-3000 lx, and light duration at 16 h / day.

5. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: The strategy of topdressing in step S3 is as follows: on the 10th day after transplanting, 1.5-2.5 kg of urea, 4-6 kg of diammonium phosphate and 5-7 kg of potassium chloride are applied per mu; on the 20th day after transplanting, 7-10 kg of urea are applied per mu; and on the 25th day after transplanting, 5-7 kg of potassium chloride are applied per mu.

6. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: In step A1, the usage ratio of chitosan, acetic acid and 2-acrylamido-2-methylpropanesulfonic acid is (10-15) g: (100-120) mL: (3.5-4.5) g.

7. The method for breeding salt-tolerant sorghum varieties according to claim 1, characterized in that: In step A2, the usage ratio of modified chitosan, dimethyl sulfoxide, indoleacetic acid, N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is (10-12) g: (100-110) mL: (0.2-0.3) g: (0.05-0.07) g: (0.01-0.02) g.

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