CN114507271B - Small molecular protein for improving tobacco leaf quality - Google Patents
Small molecular protein for improving tobacco leaf quality Download PDFInfo
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- CN114507271B CN114507271B CN202210176701.5A CN202210176701A CN114507271B CN 114507271 B CN114507271 B CN 114507271B CN 202210176701 A CN202210176701 A CN 202210176701A CN 114507271 B CN114507271 B CN 114507271B
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Abstract
Aiming at the problems of' increasing planting cost, reducing fertilizer utilization rate, aggravating environmental pollution, improving potassium content and the like existing in the prior art by increasing the potassium fertilizer to improve the potassium content of tobacco leaves, the invention provides a small molecular protein for improving the quality of tobacco leaves, which is any one or a combination of more than two of 25 small molecular proteins (EPUT-1 to EPUT-25), and is prepared into an EPUT aqueous solution containing Silwet series organosilicon surfactant during use. The invention is proved by experiments: the EPUT solution can be sprayed on the leaf surfaces to obviously improve the potassium ion content in tobacco leaves, but the nitrogen content is not increased, and the effect is obvious in improving the quality of the tobacco leaves. Compared with the conventional potassium fertilizer increasing and applying mode, the mode of improving the potassium ion content in tobacco leaves by spraying the EPUT solution on leaf surfaces is simpler, more environment-friendly and more efficient.
Description
Technical Field
The invention belongs to the technical field of agricultural biology, and particularly relates to a small molecular protein for improving tobacco leaf quality.
Background
Tobacco is a potassium-like economic crop, and the application amount of the potassium fertilizer in field production is generally 2.5-3 times of that of the nitrogenous fertilizer. The potassium content of the tobacco leaves is one of the most direct key indexes for evaluating the quality of the tobacco, and increasing the potassium content of the tobacco leaves can obviously improve the yield and quality of the tobacco leaves, improve the combustibility of the tobacco leaves, improve the identity color of the tobacco leaves and reduce the tar content of the tobacco leaves. The nitrogen content of the tobacco leaves is inversely related to the quality of the tobacco leaves, the higher the nitrogen content of the tobacco leaves is, the worse the quality of the tobacco leaves is, and the nitrogen content of the tobacco leaves is reduced, so that the quality of the tobacco leaves can be improved. The potassium content of the high-quality tobacco leaves is generally not lower than 3%, and the potassium content of the tobacco leaves in the high-quality tobacco leaf production countries such as Brazil and the United states can reach 4% -6%; the average value of the potassium content of the tobacco leaves in China is less than 2 percent, and the average value of the potassium content of the tobacco leaves in Shandong province is only about 1.5 percent. The low content of potassium ions in tobacco leaves has become a key for limiting the improvement of tobacco leaf quality in Shandong even China.
The potassium absorption capacity of the tobacco root system and the potassium content in the soil are key factors influencing the potassium content of the tobacco leaf. The reasonable fertilization for increasing the effective potassium ion supply in the tobacco root zone is an important way for increasing the potassium ion content of tobacco leaves. However, the application of the potash fertilizer increases the planting cost, reduces the fertilizer utilization rate, aggravates the environmental pollution, and has limited effect on improving the potassium ion content of tobacco leaves by simply applying the potash fertilizer. Therefore, research and development of a technology for improving the utilization rate of the tobacco potash fertilizer has important significance for improving the potassium content of tobacco and promoting the green sustainable development of tobacco production.
Small molecular substances such as plant hormone, polypeptide and the like play a very important role in plant growth and development and nutrient absorption. The foliar spray of IAA, GA and other hormones can improve the potassium absorption of plants, the potassium content of plants and the fertilizer utilization rate. Recently, some small molecule proteins have been found to also function like hormones. For example, tobacco CLE can regulate plant senescence, and arabidopsis CLE25 can regulate plant drought resistance. Therefore, the novel micromolecular protein for regulating and controlling potassium ion absorption is screened and developed, and a proper in-vitro application method is established, so that the method is an emerging strategy for improving the utilization rate of tobacco potassium fertilizer and improving the content and quality of tobacco potassium ions.
The applicant has previously developed a small molecular substance (patent application number 2021110799970) for improving cotton yield and fiber quality, the effective component of which is any one or a combination of more than two of small molecular proteins ZLC1 to ZLC20, and the last 6 amino acid of the small molecular proteins is SPGMGH. The cotton leaves are sprayed in the full bloom stage by adopting the aqueous solution containing the small molecular substance ZLC, so that the nitrogen, phosphorus and potassium absorption capacity of cotton can be enhanced, the cotton growth can be promoted, and the cotton yield and the fiber quality (fiber length and fiber strength) can be improved. However, tobacco is a plant of the family Solanaceae of the order Melaleuca, whereas cotton is a plant of the family Malvaceae of the order Malvaceae, both belonging to different orders and having a far reaching relationship. The effect of increasing the potassium content of tobacco leaves is not obvious when the small molecular substance ZLC which is effective for cotton is applied to tobacco, but the nitrogen content of the tobacco leaves can be slightly increased (figure 4), and the effect of increasing the quality of the tobacco leaves is not obvious. Unlike the strategy of searching cotton to enhance the absorption of nitrogen, phosphorus and potassium, the method is to search small molecular substances in tobacco, which only increase the potassium content of tobacco leaves but not the nitrogen content. Plants typically absorb nitrogen from the root system in the form of nitrate, while plants typically absorb both potassium and nitrate ions in order to maintain the anion-cation balance. Therefore, in order to find the small molecular proteins for improving the potassium content of the tobacco leaves, the thought completely different from cotton is utilized, and finally, a large number of experiments prove that 25 small molecular proteins for improving the quality of the tobacco leaves are found, and the small molecular proteins are more excellent in improving the potassium ion content of the tobacco leaves.
Disclosure of Invention
Aiming at the problems of 'increasing planting cost, reducing fertilizer utilization rate, aggravating environmental pollution, improving potassium content and limited effect' and the like existing in the prior art that the potassium content of tobacco leaves is improved by additionally applying a potassium fertilizer, the invention provides a small molecular protein (Enhance potassium uptake in tobacco, EPUT) for improving the quality of tobacco leaves. The invention is proved by experiments: the EPUT solution can be sprayed on the leaf surfaces to obviously improve the potassium ion content in tobacco leaves, but the nitrogen content is not increased, and the effect is obvious in improving the quality of the tobacco leaves.
The first object of the invention is to provide a small molecular protein for improving tobacco leaf quality, which is characterized in that the small molecular protein is any one or more than two of 25 small molecular proteins, and the amino acid sequence is as follows:
EPUT-1：SIEVEKSGHSPGAGH；
EPUT-2：SIEVEKSGHSPGEGH；
EPUT-3：SIEVEKSGHSPGIGH；
EPUT-4：SIEVEKSGHSPGMGH；
EPUT-5：SIEVEKSGHSPGRGH；
EPUT-6：SIEVEKSGHSPGVGH；
EPUT-7：SIEVEKSGNSPGAGH；
EPUT-8：SIEVEKSGNSPGEGH；
EPUT-9：SIEVEKSGNSPGIGH；
EPUT-10：SIEVEKSGNSPGMGH；
EPUT-11：SIEVEKSGNSPGRGH；
EPUT-12：SIEVEKSGNSPGVGH；
EPUT-13：SIEVEKSGPSPGAGH；
EPUT-14：SIEVEKSGPSPGEGH；
EPUT-15：SIEVEKSGPSPGIGH；
EPUT-16：SIEVEKSGPSPGMGH；
EPUT-17：SIEVEKSGPSPGRGH；
EPUT-18：SIEVEKSGPSPGVGH；
EPUT-19：SIEVEKSGSSPGAGH；
EPUT-20：SIEVEKSGSSPGEGH；
EPUT-21：SIEVEKSGSSPGIGH；
EPUT-22：SIEVEKSGSSPGMGH；
EPUT-23：SIEVEKSGSSPGRGH；
EPUT-24：SIEVEKSGSSPGVGH；
EPUT-25：SIEVQKSGPSPGEGH。
the second purpose of the invention is to provide the application of the small molecular protein in improving the quality of tobacco leaves. The improvement of the quality of the tobacco leaves is realized mainly by improving the content of potassium ions in the tobacco leaves without increasing the nitrogen content.
The third object of the invention is to provide a method for improving the quality of tobacco leaves, which is characterized in that EPUT water solution is sprayed on leaf surfaces 5-10 days before the bud period of tobacco leaves, so that the absorption of potassium ions by the tobacco leaves is promoted, the potassium ion content in the tobacco leaves is improved, the nitrogen content is not increased, and the quality of the tobacco leaves and the proportion of the tobacco leaves with the same grade are improved.
Further, the EPUT solution is an aqueous solution of EPUT containing Silwet series organosilicon surfactant. A Silwet L-77 surfactant is preferably used.
Preferably, the working concentration of the EPUT solution is: the final concentration of the single EPUT or the mixture of EPUTs is 0.05-0.2g/L (preferably 0.1 g/L), and the concentration of silwet surfactant is 0.01-0.02%. The tobacco leaf spraying agent is prepared according to the proportion, and the tobacco leaf spraying agent is sprayed on the tobacco leaf by using a sprayer after being uniformly mixed.
Aiming at the field application, the method for improving the quality of tobacco leaves provided by the invention comprises the following specific steps: spraying the solution 5-10 days before the bud period of the tobacco, and spraying 15-20L of EPUT solution with the concentration of 0.05-0.2g/L per mu; the method is characterized in that the method is used for spraying before 10 am or after 4 pm, and EPUT solution can be uniformly sprayed on the front and back sides of tobacco leaves as much as possible during spraying.
The invention has the following beneficial effects:
1. at present, the potassium fertilizer is mainly added to improve the potassium ion content of tobacco leaves and the quality of tobacco leaves. Along with the large-scale application of the potash fertilizer, the utilization rate of the tobacco potash fertilizer is gradually reduced, environmental pollution is easy to cause, and the excessive application of the potash fertilizer has adverse effects on the growth of tobacco leaves and has limited increase range of the potassium content. The method breaks the limitation of the prior art, improves the potassium ion content of tobacco leaves and the tobacco quality and the proportion of the superior tobacco leaves by regulating and controlling the potassium ion absorption capacity of the tobacco, so that the product can improve the potassium ion content of the tobacco leaves and the tobacco quality, can also improve the potassium fertilizer utilization rate, reduces the environmental pollution and has wide market application prospect.
2. Experiments prove that after the EPUT solution is sprayed on the large tobacco Tian Shemian, the potassium content in tobacco leaves can be obviously improved, the nitrogen content cannot be increased, and the effect on improving the quality of tobacco leaves is obvious. Therefore, the invention has the functions of improving the utilization rate of the potash fertilizer, improving the quality of tobacco leaves and improving the proportion of the tobacco leaves, and is simpler, more environment-friendly and efficient compared with the conventional mode of applying the potash fertilizer, thereby having wide application prospect in the aspects of extracting potassium from tobacco and enhancing the flavor (quality).
Drawings
FIG. 1 is a graph showing the results of test examples of the effect of treatment of tobacco seedlings with EPUT (EPUT-1 to EPUT-25) solutions on the potassium ion content of tobacco leaves;
FIG. 2 is a graph showing the effect of treatment of field tobacco leaves (1 week before the bud phase) with EPUT (EPUT 2, EPUT 10) solution for test example two on the potassium ion content of tobacco leaves; wherein, A is the potassium ion content of the lower leaf, middle leaf and upper leaf of the tobacco sampled on day 7 and 23; panel B shows the potassium content of the middle and upper leaves of tobacco sampled on day 8 and 17;
FIG. 3 is a graph showing the effect of test example three-purpose EPUT solutions (EPUT 2, EPUT 10) on the proportion of higher smoke when treating tobacco leaves in the field (1 week before the bud phase);
FIG. 4 is a graph showing the results of test example four treatments of tobacco seedlings with ZLC mixed solution and EPUT mixed solution, on the effect of potassium ion content and nitrogen content of tobacco leaves; wherein, the A graph shows the potassium ion content result, and the B graph shows the total nitrogen content result.
Note that: CK is clean water sprayed with silwet L-77 surfactant with the same concentration, EPUT (EPUT 2, EPUT 10) is water solution containing silwet L-77 surfactant sprayed with EPUT (EPUT 2, EPUT 10); the ZLC mixed solution is an aqueous solution containing silwet L-77 surfactant mixed by spraying ZLC.
Detailed Description
The technical scheme of the present invention and the technical effects thereof are further described below with reference to specific embodiments and drawings, the following description is only for explaining the present invention, but not limiting the present invention in any way, and any changes or substitutions based on the present invention are all within the scope of the present invention. The method of the invention is a conventional method in the art unless specifically stated otherwise. The reagents of the invention are commercially available unless otherwise specified.
Example 1: determination and synthesis of amino acid sequence of small molecular protein
(1) Determination of amino acid sequence of small molecule protein
The inventor searches the protein sequence in Solanaceae plants (tobacco, tomato, eggplant and the like) by utilizing a bioinformatics means, combines the expression abundance and the sequence specificity of the protein gene in tobacco, selects 56 polypeptide sequences to carry out tobacco leaf spraying experiments, and discovers 7 polypeptides for improving the potassium ion content of the tobacco leaves and arranging the potassium ion content at the front 7 positions, wherein the front 8 amino acids of 6 polypeptides are SIEVQKSG, and the front 8 amino SIEVEKSG of another polypeptide; on the basis that the first 8 amino acids are SIEVEKSG, the 9 amino acids and the 13 amino acids are respectively replaced, 72 different polypeptide sequences are designed, then the 72 polypeptides are subjected to experimental screening, and finally 24 polypeptides with better potassium ion content improving capability are obtained, and the other polypeptide with the first 8 amino acids being SIEVQKSG is added, so that 25 small molecule proteins with the sequences are found, and the potassium ion content of tobacco leaves can be obviously improved.
Thus, 25 small molecule protein sequences were ultimately determined. The first 8 amino acids of these small molecule proteins are conserved, as follows: SIEV E (Q) KSG. The 25 small molecule protein sequences were respectively named: EPUT-1 through EPUT-25. Remarks: EPUT is an acronym for Enhance potassium uptake in tobacco, meaning increasing tobacco potassium ion absorption.
(2) Synthesis and preservation of small molecule proteins
And synthesizing small molecular protein by Fmoc solid-phase polypeptide synthesis method. At least 100mg of crude product is expected to be synthesized, calculated as 10mg of the target small molecule polypeptide. The synthesis steps are as follows:
(1) 0.3g of Fmoc-C terminal first amino acid-king resin was taken, added to a 1.5ml centrifuge tube, swollen with DCM for 20 min, and then the DCM was filtered off with suction.
(2) Adding 20% piperidine/DMF solution with the volume of 3 times of resin, blowing nitrogen for 30 minutes, and pumping out; then, the resin of the previous step was washed 5 times with 2 times the resin volume of DMF.
(3) Taking 0.27mmol of Fmoc-C terminal second amino acid-OH, 0.54mmol DIPEA,0.26mmol HBTU, adding a proper amount of solvent DMF, and reacting for 30 minutes to obtain Fmoc-C terminal second amino acid-C terminal first amino acid-king resin. Amino acid: DIPEA: HBTU: resin=3: 6:2.85:1 (molar ratio); the resin of the previous step was then washed 3 times with 2 volumes of DMF;then adding 20% pip/DMF solution with 3 times of resin volume, blowing nitrogen for 30 minutes, pumping out, removing Fmoc group to obtain H 2 N-second amino acid-first amino acid-king resin; finally, the resin of the previous step was washed 5 times with 2 volumes of DMF.
(4) And (3) repeating the step (3), wherein 1 amino acid can be added for each repetition of the step (3), and repeating the step according to the sequence requirement of the small molecule protein to obtain the final target small molecule polypeptide.
(5) Washing the resin 3 times with methanol and draining the resin; then adding 6 times of resin volume of cutting fluid (volume ratio: trifluoroacetic acid: anisole: 1, 2-ethanedithiol: phenol: water=87.5%: 2.5%) into the mixture, shaking the mixture for 2 hours, filtering the resin, precipitating filtrate with ice anhydrous diethyl ether, washing the precipitate with ice anhydrous diethyl ether for 3 times, and finally placing the precipitate into a vacuum drying kettle, and drying the precipitate at normal temperature for 24 hours to obtain a small molecular protein crude product.
(6) Purifying by HPLC instrument to obtain small molecule protein with purity of more than 95%:
first, the crude small molecule protein was analyzed by a C18 analytical column with a rapid gradient. The mobile phase is: aqueous phase a:0.1% tfa/water; organic phase B:0.1% tfa/acetonitrile; gradient: 5% -70%,20 min, and the main peak of the crude product is about 7 min.
Next, small molecule protein separation was performed using a C18 preparative column, and the target peak was collected. And (3) measuring mass spectrum, wherein the molecular weight of the collected target peak is consistent with the molecular weight of the target small molecular protein, and measuring the HPLC purity to be more than 95% is preliminary qualified.
Then, the collected target peak solution is frozen by liquid nitrogen, and then is placed on a freeze dryer for vacuum drying to be in powder form, 1mg of the freeze-dried powder small molecular protein is taken and dissolved by water, HPLC and MS retest are carried out, the HPLC purity is more than 95%, and the MS molecular weight is not changed to be finally qualified.
The small molecular proteins can be stored at-80deg.C for about 2 years and at-20deg.C for about half a year.
Example 2: preparation and application of foliar fertilizer EPUT
10g of EPUT (single substance or mixture) is taken and dissolved by 1L of deionized water to prepare 100x mother liquor, and the mother liquor can be placed below-20 ℃ for 1-2 weeks, and is prepared at room temperature for use.
When the EPUT mother liquor is applied in fields, 100ml of the EPUT mother liquor is taken, the EPUT mother liquor is diluted by tap water, the final dilution volume is kept to 10L, then 1ml of silwet L-77 surfactant is added, and the mixture is uniformly mixed, so that the final concentration of the EPUT mother liquor (single EPUT or EPUT mixture) reaches 0.10g/L, and the EPUT working liquor contains 1/10000 of silwet L-77 surfactant.
Leaf surface spraying is carried out 1 week before the bud period of the tobacco, and the spraying amount is 15-20L/mu; spraying before 10 am or after 4 pm is selected. When in spraying, the front and back sides of the tobacco leaves can be uniformly sprayed with EPUT solution as much as possible.
Test example one, effect of foliar spray of EPUT solution on field cotton growth and yield
The full and full special fragrant 301 tobacco seeds are sown in a plastic box (with the diameter of 10cm and the height of 10 cm), then placed in a climatic chamber, the temperature of the greenhouse is 28 ℃, 1 true leaf is grown after germination, and 1 robust tobacco seedling is remained in each pot. After the 6 th true leaves of the tobacco grow out, selecting tobacco seedlings with consistent growth vigor for treatment, setting 25 treatments of spraying EPUT (EPUT-1 to EPUT-25 total 25 types) solutions, taking clear water sprayed with silwet L-77 surfactant with the same concentration as a control, setting 3 repetitions for each treatment, and setting 1 tobacco seedling for each repetition. After 10 days of treatment, each treated tobacco leaf was collected, dried, weighed, and then the potassium ion content was determined. In the test, the EPUT solution is 25 small molecular protein solutions respectively containing EPUT-1 to EPUT-25 (the preparation method is shown in example 2), the final concentration is 0.10g/L, and when the EPUT solution is sprayed, the front and back sides of tobacco leaves can be uniformly sprayed with the EPUT solution as much as possible, and the spraying amount of each plant is 10ml.
Test results: after 10 days of tobacco treatment, samples were taken to determine leaf potassium ion content, and it was found that spraying EPUT significantly increased tobacco leaf potassium ion content compared to control CK (FIG. 1). It is proved that the foliar spray EPUT can promote the tobacco root system to absorb potassium ions and increase the leaf potassium content, wherein the foliar spray EPUT-2 and EPUT-10 solution has the best effect of increasing the leaf potassium content.
Test example II, influence of foliar spray of EPUT solution on Potassium ion content of field tobacco leaves
And (3) sowing the filled and full special-flavor 301 tobacco seeds in a seedling tray for 20 months in 2021, growing the seedling tray in a tobacco seedling greenhouse of tobacco company in Fuxiang, and transplanting the tobacco seedlings to a field for 25 days in 4 months. The tobacco is sprayed for 6 months and 30 days just before bud emergence for one week, and after 4 pm, 0.10g/L EPUT solution 15L is sprayed on each mu of leaf surfaces. 2 treatments of spraying EPUT (EPUT-2 and EPUT-10) were set, and the spraying of clear water containing the same concentration of silwet L-77 surfactant was used as a control, and the treatments were repeated 3 times in a random block arrangement. Cell area 60m 2 4 rows of areas are adopted, the row spacing is 120cm, and the plant spacing is 50cm. And daily management is carried out on tobacco according to the local tobacco planting habit. And respectively taking the lower leaf, the middle leaf and the upper leaf of the tobacco at the day of 7 months and 23 days, and respectively taking the middle leaf and the upper leaf of the tobacco at the day of 8 months and 17 days for potassium ion measurement. The preparation methods of EPUT-2 and EPUT-10 solutions in the test are shown in example 2, and the EPUT solutions can be uniformly sprayed on the front and back surfaces of the tobacco leaves as much as possible during spraying.
Test results: as compared with control CK, EPUT-2 and EPUT-10 solutions had both increased potassium ion content of lower, middle and upper leaves of the sampled tobacco for 7 months and 23 days (FIG. 2A), and also increased potassium ion content of middle and upper leaves of the sampled tobacco for 8 months and 17 days (FIG. 2B), and EPUT-2 and EPUT-10 were not significantly different in effect in increasing potassium ion content, and EPUT-2 was slightly better in increasing potassium ion content of the middle leaf of the tobacco, but slightly worse in increasing potassium ion content of the lower and upper leaves than EPUT-10.
Test example Effect of three-leaf surface spraying EPUT solution on field tobacco leaf quality
And (3) sowing the filled and full special-flavor 301 tobacco seeds in a seedling tray for 20 months in 2021, growing the seedling tray in a tobacco seedling greenhouse of tobacco company in Fuxiang, and transplanting the tobacco seedlings to a field for 25 days in 4 months. The tobacco is sprayed for 6 months and 30 days just before bud emergence for one week, and after 4 pm, 0.10g/L EPUT solution 15L is sprayed on each mu of leaf surfaces. 2 treatments of spraying EPUT (EPUT-2 and EPUT-10) were set, and the spraying of clear water containing the same concentration of silwet L-77 surfactant was used as a control, and the treatments were repeated 3 times in a random block arrangement. Cell area60m 2 4 rows of areas are adopted, the row spacing is 120cm, and the plant spacing is 50cm. And daily management is carried out on tobacco according to the local tobacco planting habit. And harvesting tobacco leaves according to a local planting mode, baking, and finally counting the proportion of the tobacco leaves with equal proportion. The preparation methods of EPUT-2 and EPUT-10 solutions in the test are shown in example 2, and the EPUT solutions can be uniformly sprayed on the front and back surfaces of the tobacco leaves as much as possible during spraying.
Test results: both EPUT-2 and EPUT-10 solution treatments increased the proportion of superior tobacco leaves compared to control CK (FIG. 3).
Remarks: the evaluation criteria for the top tobacco leaves were: tobacco leaves are classified into 42 grades according to growing parts (upper part, middle part and lower part) and leaf colors (lemon yellow, orange and red brown); chinese orange one C1F, chinese orange two C2F, chinese orange three C3F, chinese orange one C1L, chinese orange two C2L, chinese orange one B1F, chinese orange two B2F, chinese orange one B1L, chinese orange one B1R, and finished one H1F and lower orange one X1F are the upper tobacco leaves.
Test example IV, effect of foliar spray ZLC and EPUT solutions on tobacco leaf Potassium ion content and Nitrogen content
The full and full special fragrant 301 tobacco seeds are sown in a plastic box (with the diameter of 10cm and the height of 10 cm), then placed in a climatic chamber at the temperature of 28 ℃ for example, and after 1 true leaf grows out after germination, thinning, and keeping 1 robust tobacco seedling in each pot. After the 6 th true leaves of the tobacco grow out, selecting tobacco seedlings with consistent growth vigor for treatment, setting treatment of spraying ZLC (ZLC-1 to ZLC-20 total 20 types of equal weight mixture) solution and treatment of spraying EPUT (EPUT-1 to EPUT-25 total 25 types of equal weight mixture) solution, taking clear water sprayed with silwet L-77 surfactant with the same concentration as a control, and setting 3 replicates for each treatment, and 1 tobacco seedling for each replicate. After 10 days of treatment, each treated tobacco leaf was collected, dried, weighed, and then the potassium ion content and nitrogen content were determined. The preparation method of the EPUT solution in the test is shown in the example 2, the preparation method of the ZLC solution is the same as that of the patent with the application number of 2021110799970, the final concentration of the EPUT solution and the ZLC solution is 0.10g/L, and the front and the back of tobacco leaves can be uniformly sprayed with the liquid medicine as much as possible during spraying, and the spraying amount of each plant is 10ml.
Test results: after 10 days of tobacco treatment, sampling is carried out to determine the potassium ion content of leaves, and the potassium ion content of the leaves of the tobacco is obviously improved by spraying EPUT solution compared with control CK, but the potassium increasing effect of spraying ZLC solution is not obvious (figure 4A); spraying the EPUT solution slightly reduced the tobacco nitrogen content, but spraying the ZLC solution slightly increased the tobacco nitrogen content (FIG. 4B). Because the potassium content and the nitrogen content in the tobacco leaf respectively show positive correlation and negative correlation with the quality of the tobacco leaf. The small molecular protein EPUT can obviously improve the potassium ion content in tobacco leaves, and slightly reduce the nitrogen content, so that the effect is obvious in improving the quality of the tobacco leaves. Meanwhile, compared with the conventional potassium fertilizer increasing mode, the method for improving the quality of tobacco leaves by spraying the EPUT solution on the tobacco leaves is simpler, more environment-friendly and more efficient, so that the method has a wide application prospect in improving the quality of tobacco leaves.
Claims (9)
1. The small molecular protein for improving the quality of tobacco leaves is characterized by being any one or a combination of more than two of 25 small molecular proteins, and the amino acid sequence of the small molecular protein is as follows:
EPUT-1：SIEVEKSGHSPGAGH；
EPUT-2：SIEVEKSGHSPGEGH；
EPUT-3：SIEVEKSGHSPGIGH；
EPUT-4：SIEVEKSGHSPGMGH；
EPUT-5：SIEVEKSGHSPGRGH；
EPUT-6：SIEVEKSGHSPGVGH；
EPUT-7：SIEVEKSGNSPGAGH；
EPUT-8：SIEVEKSGNSPGEGH；
EPUT-9：SIEVEKSGNSPGIGH；
EPUT-10：SIEVEKSGNSPGMGH；
EPUT-11：SIEVEKSGNSPGRGH；
EPUT-12：SIEVEKSGNSPGVGH；
EPUT-13：SIEVEKSGPSPGAGH；
EPUT-14：SIEVEKSGPSPGEGH；
EPUT-15：SIEVEKSGPSPGIGH；
EPUT-16：SIEVEKSGPSPGMGH；
EPUT-17：SIEVEKSGPSPGRGH；
EPUT-18：SIEVEKSGPSPGVGH；
EPUT-19：SIEVEKSGSSPGAGH；
EPUT-20：SIEVEKSGSSPGEGH；
EPUT-21：SIEVEKSGSSPGIGH；
EPUT-22：SIEVEKSGSSPGMGH；
EPUT-23：SIEVEKSGSSPGRGH；
EPUT-24：SIEVEKSGSSPGVGH；
EPUT-25：SIEVQKSGPSPGEGH。
2. use of the small molecule protein of claim 1 for improving tobacco leaf quality.
3. The use according to claim 2, wherein the improvement in tobacco quality is achieved mainly by increasing the potassium ion content of the tobacco without increasing the nitrogen content.
4. A method for improving the quality of tobacco leaves is characterized in that the aqueous solution of the small molecular protein EPUT as defined in claim 1 is sprayed on leaf surfaces 5-10 days before the bud period of the tobacco leaves to promote the absorption of potassium ions by the tobacco leaves, so that the content of potassium ions in the tobacco leaves is improved, the nitrogen content is not increased, and the quality of the tobacco leaves and the proportion of the tobacco leaves with higher quality are improved.
5. The method of claim 4, wherein the aqueous solution of small molecule protein EPUT is an aqueous EPUT solution containing Silwet series silicone surfactant.
6. A method for improving tobacco leaf quality as claimed in claim 5 wherein the Silwet series of silicone surfactant is Silwet L-77 surfactant.
7. The method for improving tobacco leaf quality according to claim 5, wherein the working concentration of the aqueous solution of the small molecule protein EPUT is as follows: the final concentration of the single EPUT or the mixture of EPUTs is 0.05-0.2g/L, and the concentration of the Silwet series of organosilicon surfactant is 0.01-0.02%.
8. A method of improving tobacco leaf quality as claimed in claim 7 wherein the final concentration of the single EPUT or EPUT blend is 0.1g/L.
9. The method for improving the quality of tobacco leaves according to claim 7, which is specifically used in a field and comprises the following steps: spraying the solution 5-10 days before the bud period of the tobacco, and spraying 15-20L of EPUT solution with the concentration of 0.05-0.2g/L per mu; the method is characterized in that the method is used for spraying before 10 am or after 4 pm, and EPUT solution can be uniformly sprayed on the front and back sides of tobacco leaves as much as possible during spraying.
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| WO2001035770A1 (en) * | 1999-11-19 | 2001-05-25 | Philip Morris Products Inc. | A method for reduction of tobacco specific nitrosamines |
| CN113754732A (en) * | 2021-09-15 | 2021-12-07 | 山东省农业科学院 | Small molecular substance for improving cotton yield and fiber quality |
| CN113980988A (en) * | 2021-11-19 | 2022-01-28 | 云南省烟草农业科学研究院 | NtYpi1 gene capable of regulating and controlling potassium content of tobacco leaves and application thereof |
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| US8227573B2 (en) * | 2005-05-20 | 2012-07-24 | University Of Kentucky Research Foundation | Utility of phylloplanins as antibiotics, selective fungicides and for enhancing microbial resistance in plants |
| CN101548673B (en) * | 2009-05-12 | 2012-07-18 | 湖北省烟草科研所 | Tobacco alkalinity reducing agent used in field tobacco cultivation |
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| WO2001035770A1 (en) * | 1999-11-19 | 2001-05-25 | Philip Morris Products Inc. | A method for reduction of tobacco specific nitrosamines |
| CN113754732A (en) * | 2021-09-15 | 2021-12-07 | 山东省农业科学院 | Small molecular substance for improving cotton yield and fiber quality |
| CN113980988A (en) * | 2021-11-19 | 2022-01-28 | 云南省烟草农业科学研究院 | NtYpi1 gene capable of regulating and controlling potassium content of tobacco leaves and application thereof |
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