Method for improving budding ability of cotton top soil in saline-alkali soil by exogenous application of 6-benayl aminopurine

Abstract

The invention relates to the field of saline-alkali soil treatment, aims to solve the technical problem of how to promote the growth of cotton in saline-alkali soil, and provides application of 6-benayl aminopurine in improving the salt tolerance of cotton in a high-salt environment. The method for enhancing the salt tolerance of the cotton is preliminarily established, reference is provided for promoting growth of the cotton in the saline-alkali soil, and application of the cotton in saline-alkali soil remediation is further promoted.

Description

Technical Field

[0001] This invention relates to the field of saline-alkali land management, and specifically discloses a method for improving the ability of cotton to sprout from the soil in saline-alkali land by applying exogenous 6-benzylaminopurine (6-BA, a synthetic cytokinin). Background Technology

[0002] Salt stress is a major global environmental factor limiting plant growth and crop productivity, affecting more than 6% of the world's land area (approximately 800 million hectares). Poor irrigation practices, improper fertilizer application, and industrial pollution increase soil salinity. Seed germination is considered the most critical stage in the life cycle because it is highly vulnerable to damage, disease, and environmental stress. Sodium ions (Na+) in saline-alkali soil solutions... + High salt concentrations inhibit seed germination primarily through osmotic stress, oxidative stress, and ion toxicity, manifesting as reduced germination rate and prolonged germination time. Salt stress leads to excessive reactive oxygen species, damaging proteins, lipids, nucleic acids, or cell structures, generating oxidative stress, and altering plant hormone balance.

[0003] Cotton plays a vital role in agricultural economic development. As a pioneer crop for saline-alkali land, cotton is also an extremely important fiber crop and a significant oilseed crop (accounting for 35% of global fiber consumption). Given its wide range of applications and considerable economic value, studying cotton development under salt stress is of great significance.

[0004] Seed germination and emergence require hypocotyl elongation. Hypocotyl elongation is crucial for the early morphological development of seedlings. The hypocotyl is one of the main tissues formed during plant embryonic development; it connects the seed embryo to the root system and eventually develops into the plant's stem. The hypocotyl plays a vital role in supporting and transporting water and nutrients during plant growth. It also transports substances beneficial to seedling growth, development, and adaptation to environmental conditions. For example, the hypocotyl contains hormones that can enhance seedling resistance and adaptability to environmental conditions such as water and temperature. Therefore, the ability of the hypocotyl to elongate directly affects seedling growth and development, ultimately influencing cotton yield and quality. However, currently, there are very few measures to promote cotton seed germination and emergence in saline-alkali soils. Summary of the Invention

[0005] To improve the salt tolerance of cotton, this application promotes the growth of the hypocotyl after cotton seed germination by exogenously adding 6-benzylaminopurine, thereby promoting the emergence of cotton from the soil in a high-salt environment and thus promoting the application of cotton in the remediation of saline-alkali land.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A first aspect of the present invention is the application of 6-benzylaminopurine. The application of 6-benzylaminopurine is any one or more of the following:

[0007] Y1. Application in improving the salt tolerance of cotton in saline-alkali land;

[0008] Y2. Application in promoting cotton seed germination in saline-alkali land;

[0009] Y3. Application in relieving the inhibition of cotton hypocotyl growth by saline-alkali soil;

[0010] Y4. Application in improving the ability of cotton to sprout from the soil in saline-alkali land;

[0011] Y5. Application of cotton in the remediation of saline-alkali land.

[0012] In the above applications, the 6-benzylaminopurine is prepared as a solution, and the content of the 6-benzylaminopurine in the solution can be 0.2 μM.

[0013] In the above applications, the solvent of the solution can be water.

[0014] In the above applications, 6-benzylaminopurine is used during cotton seed germination.

[0015] The present invention also provides a method for improving the salt tolerance of cotton, comprising the step of adding 6-benzylaminopurine during cotton seed germination.

[0016] In the above method, the 6-benzylaminopurine is prepared as a solution when used, and the content of the 6-benzylaminopurine in the solution can be 0.2 μM.

[0017] In the above method, the solvent of the solution can be water.

[0018] This invention has initially established a method for enhancing the salt tolerance of cotton, providing a reference for promoting cotton growth in saline-alkali land and further promoting the application of cotton in the remediation of saline-alkali land. Attached Figure Description

[0019] Figure 1 These are photographs of cotton seeds from eight materials in Example 1 of this invention germinating for three days under three different treatments (ddH2O, NaCl, and NaCl+6-BA). Detailed Implementation

[0020] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0021] In the quantitative experiments described below, three replicate experiments were conducted, and the average value of the results was taken.

[0022] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0023] The cotton material Xinqiu No. 1 used in the following examples has been approved by the state, with the approval number being Guoshenmian 2006002, and is available to the public for purchase.

[0024] The cotton material Xinyan 96-48 used in the following examples has been approved by the state, with the approval number being Guoshen Mianma 2002005, and is available to the public for purchase.

[0025] The cotton material GK50 used in the following examples was approved by Hebei Province in 2007 and is available to the public for purchase.

[0026] The cotton material Ji Mian 228 used in the following examples is described in the non-patent literature "Meng Jianchao, Ge Chaohong. High-yield and disease-resistant new cotton variety Ji 228.2008", which is available to the public from the applicant to repeat this experiment.

[0027] The cotton material in the following examples is described in non-patent literature "Functional study of cotton cell wall remodeling related genes GhEXLB1 and GhGRP1 under salt stress", which can be obtained from the applicant to replicate this experiment.

[0028] The cotton material in the following examples is described in non-patent literature "Functional study of cotton cell wall remodeling related genes GhEXLB1 and GhGRP1 under salt stress", which can be obtained from the applicant to replicate this experiment.

[0029] The cotton material TM-1 in the following examples was obtained from the National Cotton Germplasm Bank (Anyang), with the intermediate bank number Genebank M140053. It is available to the public from the National Cotton Germplasm Bank (Cotton Research Institute, Chinese Academy of Agricultural Sciences).

[0030] The cotton material S9612 in the following examples is described in the non-patent literature "Functional study of cotton cell wall remodeling related genes GhEXLB1 and GhGRP1 under salt stress", which can be obtained by the public from the applicant to replicate this experiment.

[0031] Example 1

[0032] Plant hormones are important for seed germination. During the exogenous application of plant hormones, many genes are activated, thereby generating tolerance in plants.

[0033] This invention provides a method for alleviating cotton salt stress and promoting hypocotyl elongation by applying exogenous 6-BA (synthetically synthesized 6-benzylaminopurine).

[0034] 1. Material preparation

[0035] Eight samples of upland cotton were randomly selected. The names and sources of the materials are shown in Table 1.

[0036] Table 1. Names and Sources of Cotton Materials

[0037]

[0038] Cotton seeds were delinted with concentrated sulfuric acid, dried, and then selected seeds of uniform size and plumpness. They were sterilized with a 75% ethanol solution for 5-10 minutes, then rinsed repeatedly with distilled water 5 times (1 minute each time), and soaked in distilled water for 24 hours to germinate, resulting in white-tinged seeds.

[0039] Vertical double-layer filter paper method: Lay the filter paper flat on a plastic tray and moisten it with liquid. Place the seeds (30 seeds of one variety) on the filter paper, with the growing points facing the same direction and spaced 5-7 cm apart. Roll up the filter paper horizontally along the seed position, ensuring the seeds are rolled in the middle of the paper, forming a straight cylinder. Place the cylindrical filter paper vertically in a beaker with the seed growing points facing down. After 24 hours of natural growth, add the liquid to the bottom of the beaker to keep the filter paper moist throughout the germination period. Place the beaker in an incubator at 28°C in darkness for 24 hours.

[0040] 2. Screening of salt stress conditions

[0041] Seeds of the 'Zhong 9807' variety with white tips were used as materials for screening under salt stress conditions.

[0042] Dissolve NaCl in distilled water to prepare five salt solutions of different concentrations, as follows (based on NaCl concentration): 0 mM, 50 mM, 100 mM, 150 mM, and 200 mM.

[0043] Seeds of 9807 that have developed white color were cultured using the vertical double-layer filter paper method described above. The liquids used were the five different concentrations of NaCl solution described above, with 30 seeds treated with each concentration of NaCl solution, in 30 replicates.

[0044] Seed germination was observed after 3 and 5 days of cultivation. It was observed that the growth of hypocotyls in cotton seeds treated with 100mM, 150mM, and 200mM NaCl solutions was significantly inhibited after 5 days. However, considering that the germination rate of cotton seeds treated with 150mM and 200mM NaCl solutions was relatively low, 100mM NaCl solution was selected for subsequent treatment.

[0045] 3. Screening of treatment conditions for 6-benzylaminopurine

[0046] Seeds of the Zhong 9807 variety with white tips were used as materials for screening of 6-benzylaminopurine treatment concentrations.

[0047] NaCl and 6-BA (Solepro, product number A8170) were dissolved in distilled water. The concentration of NaCl was set to 100 mM, and seven concentrations of 6-BA were prepared (0 μM, 0.1 μM, 0.2 μM, 0.5 μM, 1.0 μM, 5.0 μM, and 10.0 μM), resulting in the following seven solutions:

[0048] A1. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 0μM 6-BA.

[0049] A2. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 0.1μM 6-BA.

[0050] A3. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 0.2μM 6-BA.

[0051] A4. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 0.5μM 6-BA.

[0052] A5. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 1.0μM 6-BA.

[0053] A6. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 5.0μM 6-BA.

[0054] A7. The solvent is water, and the types and concentrations of solutes are: 100mM NaCl and 10.0μM 6-BA.

[0055] Seeds of 9807 that showed white color were cultured using the vertical double-layer filter paper method described above. The liquids added to the bottom of the beaker were solutions of A1, A2, A3, A4, A5, A6 and A7, respectively. Each concentration of NaCl solution was used to treat 30 seeds, i.e., 30 replicates.

[0056] Seed germination was observed after 3 and 5 days of cultivation. Treatment with A3 solution (containing 0.2 μM 6-BA) for 3 days effectively alleviated salt stress and promoted hypocotyl elongation. Therefore, 0.2 μM 6-BA was selected as the treatment condition for 6-benzylaminopurine.

[0057] 4. Effects of 6-Benzylaminopurine on cotton seed germination in high-salt environments

[0058] Using the budding seeds of the eight cotton materials in Table 1 as materials for the experiment, the seeds were germinated using the vertical double-layer filter paper method described above. Three treatment groups were set up for each cotton variety as follows:

[0059] Treatment 1 (ddH2O): The liquid used was ddH2O, and the culture was carried out for 3 days.

[0060] Treatment 2 (NaCl): The liquid used was a 100mM NaCl solution (solvent was water, and the type and concentration of solute were 100mM NaCl), and the solution was cultured for 3 days.

[0061] Treatment 3 (NaCl + 6-BA): The liquid used was 100mM NaCl + 0.2μM 6-BA (solvent was water, and the solutes and their concentrations were 100mM NaCl and 0.2μM 6-BA), and the mixture was incubated for 3 days.

[0062] Each cotton material was treated with 30 seeds, i.e., 30 replicates.

[0063] Phenotypic results of each cotton material after 3 days of cultivation are shown in the figure. Figure 1 ImageJ software and GraphPad Prism were used to statistically analyze the hypocotyl lengths of each material and perform a significance analysis. The results are shown in Table 2.

[0064] Table 2. Statistical table of hypocotyl length for different upland cotton varieties under three treatments.

[0065]

[0066] Note: Data in the table are mean ± standard deviation, with 30 replicates. Different lowercase letters after the data in the same row indicate significant differences between treatments (P<0.05).

[0067] The results showed that hypocotyl elongation was significantly inhibited in all eight cotton samples under 100 mM NaCl treatment, while the addition of 6-benzylaminopurine significantly alleviated the inhibition of hypocotyl elongation in all eight samples. This indicates that exogenous addition of 6-benzylaminopurine can alleviate the inhibition of hypocotyl elongation in cotton under NaCl stress, enhance the salt tolerance of cotton, and improve the sprouting ability of cotton in saline-alkali soil.

[0068] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

Application of 1,6-benzylaminopurine in improving the salt tolerance of cotton in saline-alkali soil. Application of 2,6-benzylaminopurine in promoting cotton seed germination in saline-alkali soil. Application of 3,6-benzylaminopurine in relieving the inhibition of cotton hypocotyl growth by saline-alkali soil. Application of 4,6-benzylaminopurine in improving the sprouting ability of cotton in saline-alkali soil. Application of 5,6-benzylaminopurine in the remediation of saline-alkali land using cotton.

6. The application according to any one of claims 1-5, characterized in that: The 6-benzylaminopurine is used during cotton seed germination.

7. A method for improving the salt tolerance of cotton, characterized in that: This includes the step of adding 6-benzylaminopurine during cotton seed germination.

8. The method according to claim 7, characterized in that: The 6-benzylaminopurine is added in a solution.

9. The method according to claim 8, characterized in that: The concentration of 6-benzylaminopurine in the solution is 0.2 μM.

10. The method according to claim 9, characterized in that: The solvent for the solution is water.

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