CN113303114A - Method for reducing total tannin content of cyclobalanopsis glauca cutting slips - Google Patents
Method for reducing total tannin content of cyclobalanopsis glauca cutting slips Download PDFInfo
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- CN113303114A CN113303114A CN202110541548.7A CN202110541548A CN113303114A CN 113303114 A CN113303114 A CN 113303114A CN 202110541548 A CN202110541548 A CN 202110541548A CN 113303114 A CN113303114 A CN 113303114A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
Abstract
The invention relates to the technical field of cyclobalanopsis glauca cutting slips, in particular to a method for reducing the total tannin content of cyclobalanopsis glauca cutting slips, which comprises the following steps: and (3) cutting treatment: collecting current-year branches of cyclobalanopsis glauca with full buds, cutting the branches into small sections of 10-15 cm to obtain cutting slips, obliquely cutting the lower ends of the cutting slips, horizontally cutting the upper ends of the cutting slips, and cleaning; preparing a soaking solution: dissolving polyvinylpyrrolidone powder in 70% acetone aqueous solution, and fully dissolving and uniformly mixing to obtain a soaking solution; and (3) cutting and soaking: and (3) putting the lower end of the treated cutting into a soaking solution, soaking for 24 hours under the ultrasonic condition, and washing to be clean. The PVP is combined with the tannin in cyclobalanopsis glauca to form a complex, the complex is insoluble in water and other organic solvents, and the complex is easy to remove after centrifugation, so that the tannin content is reduced; soaking the cutting slips by using acetone to remove part of tannin in the cutting slips and reduce tannin browning in the cutting slips; the tannin is reduced by ultrasonic assistance, so that the reduction effect is better.
Description
Technical Field
The invention relates to the technical field of cyclobalanopsis glauca cutting slips, in particular to a method for reducing the total tannin content of cyclobalanopsis glauca cutting slips.
Background
Tannins are widely present in the plant kingdom and distributed in various tissues and organs of plants, and are classified into hydrolyzed tannins, condensed tannins, and complex tannins according to their structure and properties. Tannin can affect the rooting of cuttings of plants. At present, the study on the survival rate of the cyclobalanopsis glauca cutting slips mainly comprises the steps of adjusting factors such as a cutting matrix and a rooting agent, and directly judging whether the cutting propagation method is good or bad by taking the survival rate of the cyclobalanopsis glauca cutting slips as a standard, for example, the invention patent with the publication number of CN103329711B, namely the cyclobalanopsis glauca twig cutting propagation method, improves the survival rate of cutting only by using the rooting agent, and does not study substances influencing cutting rooting from endogenous substances. At present, no method for reducing the total amount of tannin in cyclobalanopsis glauca cutting slips exists, and particularly, no method for reducing the total amount of tannin in cyclobalanopsis glauca cutting slips exists.
Disclosure of Invention
The invention provides a method for reducing the total tannin content of cyclobalanopsis glauca cutting slips, aiming at solving the problem that the total tannin content of the cyclobalanopsis glauca cutting slips cannot be reduced in the prior art.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for reducing the total content of tannin in cyclobalanopsis glauca cutting slips comprises the following steps:
step 1: cutting treatment
Collecting current-year branches of cyclobalanopsis glauca with full buds, cutting the branches into small sections of 10-15 cm to obtain cutting slips, obliquely cutting the lower ends of the cutting slips, horizontally cutting the upper ends of the cutting slips, and cleaning the cutting slips for later use;
step 2: preparation of the soak solution
Dissolving polyvinylpyrrolidone (PVP) powder in 70% acetone aqueous solution by mass, wherein the mass-volume ratio of the polyvinylpyrrolidone powder to the acetone aqueous solution is 1: 20-1: 30, and obtaining a soaking solution after fully dissolving and uniformly mixing;
and step 3: cutting and soaking
And (3) putting the lower end of the cutting obtained in the step (1) into a soaking solution, soaking for 24 hours under the ultrasonic condition, and then washing to be clean.
Further, the lower end of the cutting is obliquely cut at an angle of 45 degrees.
Furthermore, a cross with the depth of 0.2-0.3 cm is cut on the oblique shearing surface of the cutting shoot, so that the contact area between the bottom of the cutting shoot and the soaking solution can be increased, and the tannin removing effect is better.
Further, the ultrasonic power in the cutting soaking step is 200W, and the ultrasonic temperature is 30 ℃.
Furthermore, the liquid level of the soaking solution is located between 1/4 and 1/3 of the height of the cuttings. If the liquid level is higher than 1/3 of the height of the cutting, the soaked cutting is too long and is easy to damage the whole cutting; if the liquid level is lower than 1/4 of the height of the cutting, the soaked cutting is too short, and the extraction effect on tannin in the branches is not good.
Further, taking part of the cuttings before soaking and part of the cuttings after soaking to respectively determine the total amount of tannin, and the specific operation steps are as follows: cutting the cutting into 0.5cm small sections, deactivating enzymes, drying, crushing, sieving with a 80-mesh sieve, taking 0.5-1 g of sieved materials, adding the materials into acetone according to a material-liquid ratio of 1:20, carrying out ultrasonic extraction for 20min, centrifuging, taking supernate, repeatedly extracting for 3 times, combining the supernate and fixing the volume, measuring by using a Folin phenol reagent method, and obtaining the total content of tannin in the cutting by using a standard curve made by using gallic acid as a standard substance.
Further, the drying step is as follows: cutting into small segments, deactivating enzyme at 103 deg.C for 15min, and oven drying at 70 deg.C to constant weight.
Further, the ultrasonic power in the step of measuring the total amount of tannin is 200W, and the water bath temperature is 50 ℃.
Through the technical scheme, the invention has the beneficial effects that:
1. the PVP is combined with the tannin in cyclobalanopsis glauca to form a complex, and the complex is insoluble in water and other organic solvents and is easy to remove, so that the tannin content is reduced;
2. the method comprises the steps of soaking the cutting slips in acetone to remove part of tannin in the cutting slips and reduce tannin browning in the cutting slips;
3. the invention reduces the content of tannin by using ultrasonic wave assistance, and has better effect;
4. the test instrument is a conventional instrument, the operation method is simple, and the cost is low.
Detailed Description
The invention will be further illustrated with reference to specific embodiments:
example 1
A method for reducing the total content of tannin in cyclobalanopsis glauca cutting slips comprises the following steps:
1. cutting treatment
Collecting cyclobalanopsis glauca annual branches with plump buds, cutting into 10cm long branches, obliquely cutting the branches at 45 degrees at the lower ends, horizontally cutting the branches at the upper ends, and cleaning. The cuttings were divided into at least 8 groups, group 1 for initial total tannin amount measurement, group 2 for soaking of the cuttings of this example, and groups 3-8 for comparative examples 1-6.
2. Initial total tannin determination
Cutting the group 1 cutting into 0.5cm long segments, deactivating enzyme at 103 deg.C for 15min, oven drying at 70 deg.C to constant weight, pulverizing, sieving with 80 mesh sieve, weighing 0.5g of sieved material, adding 10mL of 70% acetone aqueous solution, ultrasonic extracting for 20min (ultrasonic power 200W, water bath temperature 50 deg.C), centrifuging to obtain supernatant, repeatedly extracting for 3 times, mixing the supernatants, and diluting to 50 mL. And (3) measuring by using a Folin phenol reagent method, and obtaining the initial content of the tannin in the cutting slips by using a standard curve made by taking gallic acid as a standard substance, wherein the initial content of the tannin in the cyclobalanopsis glauca is 6.44 mg/g.
3. Preparation of the soak solution
Dissolving 5g of PVP powder in 100ml of acetone aqueous solution with the mass fraction of 70%, and fully dissolving and uniformly mixing to obtain a soaking solution.
4. Cutting and soaking
And (3) putting the lower ends of the cuttings in the group 2 into a soaking solution, wherein the liquid level is 3.3cm away from the lower ends of the cuttings, and soaking for 24 hours under the ultrasonic condition, wherein the ultrasonic power is 200W, and the ultrasonic temperature is 30 ℃. And (5) washing after soaking.
5. Final total tannin determination: and (3) taking part of the soaked cutting slips, and measuring the reduced tannin content according to the measuring method in the step 2. The tannin content of cyclobalanopsis glauca after reduction is 2.03 mg/g.
Comparative example 1: taking the 3 rd group of cutting slips, the method is basically the same as the method of the embodiment 1, except that: the soaking solution does not contain PVP powder, and the soaking solution is 100ml of 70% acetone aqueous solution by mass fraction. The content of cyclobalanopetannin after soaking is 4.22 mg/g.
Comparative example 2: taking the 4 th group of cutting slips, the same as the embodiment 1, except that: the soaking solution does not contain acetone, and the soaking solution is 5g of PVP powder dissolved in 100ml of water. The content of cyclobalanopetannin after soaking is 5.18 mg/g.
Comparative example 3: taking the 5 th group of cutting slips, the same as the embodiment 1, except that: the cutting is soaked without ultrasonic. The content of cyclobalanopetannin after soaking is 5.86 mg/g.
Comparative example 4 the 6 th group of cuttings was taken, which was substantially the same as in example 1 except that the liquid level of the soaking solution was located 2.5cm below the cuttings. The content of cyclobalanopetannin after soaking is 2.53 mg/g.
Comparative example 5: the group 7 cutting slips were taken, and the same as in example 1 was used, except that the immersion liquid was located 2.5cm below the cutting slips, 5g of PVP powder was dissolved in 150ml of 70% acetone aqueous solution. The content of cyclobalanopetannin after soaking is 3.12 mg/g.
Comparative example 6 the 8 th group of cuttings was prepared, which was substantially the same as example 1 except that: the liquid level of the soaking solution is positioned at the position of 2.5cm below the lower end of the cutting, 4g of PVP powder is dissolved in 100ml of acetone aqueous solution with the mass fraction of 70%. The content of cyclobalanopetannin after soaking is 2.98 mg/g.
Therefore, the total amount of tannin in the cyclobalanopsis glauca cutting is reduced, the acetone effect is the best, and the PVP is the next place, and the ultrasonic effect is the minimum. After the combination of 3 factors, there is an interaction promoting effect.
Example 2
1. Cutting treatment
Collecting cyclobalanopsis glauca annual branches with plump buds, cutting into 15cm long branches with 45-degree lower ends, cutting 0.3cm deep crosses on the oblique cutting surfaces of the cuttings, and cutting the upper ends of the branches and cleaning. The cuttings were divided into 2 groups, the 1 st group was used for initial total tannin amount measurement, and the 2 nd group was used for soaking of the cuttings of this example.
2. Initial total tannin determination
The measurement method was the same as in example 1, and the initial content of tannin in cyclobalanopsis glauca was 6.38 mg/g.
3. Preparation of the soak solution
Dissolving 5g of PVP powder in 100ml of acetone aqueous solution with the mass fraction of 70%, and fully dissolving and uniformly mixing to obtain a soaking solution.
4. Cutting and soaking
And (3) putting the lower ends of the cuttings in the group 2 into a soaking solution, wherein the liquid level is 5cm away from the lower ends of the cuttings, and soaking for 24 hours under the ultrasonic condition, wherein the ultrasonic power is 200W, and the ultrasonic temperature is 30 ℃. And (5) washing after soaking.
5. Final total tannin determination: and (3) taking part of the soaked cutting slips, and measuring the reduced tannin content according to the measuring method in the step 2. The tannin content of cyclobalanopsis glauca after reduction is 2.11 mg/g.
Example 3
1. Cutting treatment
Collecting cyclobalanopsis glauca annual branches with plump buds, cutting into 12cm long branches with 45-degree lower ends, cutting 0.2cm deep crosses on the oblique cutting surfaces of the cuttings, and cutting the upper ends of the branches and cleaning. The cuttings were divided into 2 groups, the 1 st group was used for initial total tannin amount measurement, and the 2 nd group was used for soaking of the cuttings of this example.
2. Initial total tannin determination
The measurement method was the same as in example 1, and the initial content of tannin in cyclobalanopsis glauca was 6.52 mg/g.
3. Preparation of the soak solution
Dissolving 5g of PVP powder in 100ml of acetone aqueous solution with the mass fraction of 70%, and fully dissolving and uniformly mixing to obtain a soaking solution.
4. Cutting and soaking
And (3) putting the lower ends of the cuttings in the group 2 into a soaking solution, wherein the liquid level is 4cm away from the lower ends of the cuttings, and soaking for 24 hours under the ultrasonic condition, wherein the ultrasonic power is 200W, and the ultrasonic temperature is 30 ℃. And (5) washing after soaking. .
5. Final total tannin determination: and (3) taking part of the soaked cutting slips, and measuring the reduced tannin content according to the measuring method in the step 2. The tannin content of cyclobalanopsis glauca after reduction is 2.11 mg/g.
The above-mentioned embodiments are merely preferred embodiments of the present invention, which are merely illustrative and not restrictive, and it should be understood that other embodiments may be easily implemented by those skilled in the art by means of replacement or modification according to the technical contents disclosed in the specification, and therefore, all changes and modifications that come within the spirit and technical conditions of the present invention should be included in the claims of the present invention.
Claims (8)
1. A method for reducing the total content of tannin in cyclobalanopsis glauca cutting slips is characterized by comprising the following steps:
step 1: cutting treatment
Collecting current-year branches of cyclobalanopsis glauca with full buds, cutting the branches into small sections of 10-15 cm to obtain cutting slips, obliquely cutting the lower ends of the cutting slips, horizontally cutting the upper ends of the cutting slips, and cleaning the cutting slips for later use;
step 2: preparation of the soak solution
Dissolving polyvinylpyrrolidone powder in 70% acetone aqueous solution by mass, wherein the mass volume ratio of the polyvinylpyrrolidone powder to the acetone aqueous solution is 1: 20-1: 30, and obtaining a soaking solution after fully dissolving and uniformly mixing;
and step 3: cutting and soaking
And (3) putting the lower end of the cutting obtained in the step (1) into a soaking solution, soaking for 24 hours under the ultrasonic condition, and then washing to be clean.
2. The method for reducing the total tannin content in the cyclobalanopsis tenuipes cutting shoot as claimed in claim 1, wherein the lower end of the cutting shoot is cut at an angle of 45 °.
3. The method for reducing the total tannin content in the cyclobalanopsis glauca cutting shoot as claimed in claim 1, wherein a cross with a depth of 0.2-0.3 cm is cut on the oblique shearing surface of the cutting shoot.
4. The method for reducing the total tannin content in the cyclobalanopsis tenuipes cutting shoot as claimed in claim 1, wherein the ultrasonic power in the cutting shoot soaking step is 200W, and the ultrasonic temperature is 30 ℃.
5. The method for reducing the total tannin content in the cyclobalanopsis tenuipes cutting shoot as claimed in claim 1, wherein the liquid level of the soaking solution is between 1/4-1/3 of the cutting shoot height.
6. The method for reducing the total tannin content in the cyclobalanopsis glauca cutting slips of claim 1, wherein the total tannin content is measured in a part of the cutting slips before and after soaking, and the specific operation steps are as follows: cutting the cutting into 0.5cm small sections, deactivating enzymes, drying, crushing, sieving with a 80-mesh sieve, taking 0.5-1 g of sieved materials, adding the materials into acetone according to a material-liquid ratio of 1:20, carrying out ultrasonic extraction for 20min, centrifuging, taking supernate, repeatedly extracting for 3 times, combining the supernate and fixing the volume, measuring by using a Folin phenol reagent method, and obtaining the total content of tannin in the cutting by using a standard curve made by using gallic acid as a standard substance.
7. The method for reducing the total tannin content in the cyclobalanopsis tenuipes cutting shoot as claimed in claim 6, wherein the drying step comprises: cutting into small segments, deactivating enzyme at 103 deg.C for 15min, and oven drying at 70 deg.C to constant weight.
8. The method for reducing tannin content in said cyclobalanopsis tenuipes cutting shoot as claimed in claim 6, wherein said step of determining tannin content comprises ultrasonic power of 200W and water bath temperature of 50 ℃.
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| RU2190327C2 (en) * | 2000-07-10 | 2002-10-10 | Кубанский государственный аграрный университет | Method for accelerating the rooting of rose cuttings |
| JP2011244751A (en) * | 2010-05-27 | 2011-12-08 | Nippon Paper Industries Co Ltd | Method for producing clone seedling |
| CN103329711A (en) * | 2013-07-01 | 2013-10-02 | 江苏省林业科学研究院 | Method for cutting propagation of cyclobalanopsis glauca twigs |
| CN103999706A (en) * | 2014-06-24 | 2014-08-27 | 南京林业大学 | Method for increasing cutting rooting rate of pecans |
| CN107897470A (en) * | 2017-11-10 | 2018-04-13 | 青岛胶平食品有限公司 | A kind of kiwifruit piece treatment process and its inorganic agent |
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2021
- 2021-05-18 CN CN202110541548.7A patent/CN113303114A/en active Pending
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|---|---|---|---|---|
| RU2190327C2 (en) * | 2000-07-10 | 2002-10-10 | Кубанский государственный аграрный университет | Method for accelerating the rooting of rose cuttings |
| JP2011244751A (en) * | 2010-05-27 | 2011-12-08 | Nippon Paper Industries Co Ltd | Method for producing clone seedling |
| CN103329711A (en) * | 2013-07-01 | 2013-10-02 | 江苏省林业科学研究院 | Method for cutting propagation of cyclobalanopsis glauca twigs |
| CN103999706A (en) * | 2014-06-24 | 2014-08-27 | 南京林业大学 | Method for increasing cutting rooting rate of pecans |
| CN107897470A (en) * | 2017-11-10 | 2018-04-13 | 青岛胶平食品有限公司 | A kind of kiwifruit piece treatment process and its inorganic agent |
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