Method for increasing total alkaloid content of pinellia tuber and / or yield of pinellia tuber

Abstract

The invention discloses a method for increasing the total alkaloid content of pinellia tuber and / or the yield of pinellia tuber, and belongs to the technical field of planting of traditional Chinese medicinal materials. The method comprises the step of spraying a betaine solution to pinellia ternate plant leaves in the pinellia ternate planting process. By applying 20 mmol / L of betaine solution, the total alkaloid content of the pinellia tuber can be increased by 82.85%, the fresh weight of the tuber can be increased by 31.29%, the dry weight of the tuber can be increased by 33.30%, and the diameter of the tuber can be increased by 12.70%. The method is easy to operate, low in input cost and environmentally friendly, the quality and yield of pinellia ternate tubers can be remarkably improved, and economic benefits of pinellia ternate planting production are increased.

Description

Technical Field

[0001] The invention belongs to the technical field of Chinese medicinal material planting, and particularly relates to application of betaine in improving the total alkaloid content of pinellia tubers and / or the yield of pinellia tubers, and a method for improving the total alkaloid content of pinellia tubers and / or the yield of pinellia tubers. Background Art

[0002] Pinellia ternata is an important member of the treasure house of traditional Chinese medicine. Its source plant is Pinellia ternata (Thunb.) Breit. of the Araceae family. Pinellia ternata is a major Chinese medicinal material with important economic value. Its tubers are used as medicine. It contains a variety of active ingredients including alkaloids, flavonoids, organic acids, etc. It has antitussive and expectorant, anti-ulcer, anti-tumor, anti-arrhythmic, sedative and hypnotic effects. The level of alkaloid content in Pinellia ternata is an important indicator for evaluating the quality of Pinellia ternata medicinal materials. With the widespread application of Pinellia ternata in the field of traditional Chinese medicine and the pressure of long-term export abroad, its market demand has increased year by year. Pinellia ternata is widely distributed in my country and has abundant resources, but due to changes in the ecological environment, excessive excavation and the use of herbicides, its wild resource reserves have sharply decreased, and even disappeared in some areas. Therefore, wild Pinellia ternata has long been unable to meet market demand, and artificially cultivated products have become the main source of commercial Pinellia ternata.

[0003] Betaine (C5H 11 NO2) is a quaternary ammonium water-soluble alkaloid, which is commonly found in animals and plants and is an important intermediate product of animal and plant metabolism. In plants, the current research on betaine mainly focuses on stress physiology. A large number of studies have shown that betaine can improve the tolerance of plants to various adversities such as drought, high temperature, low temperature, high salt, heavy metals, etc. Its mechanism of action includes: participating in osmotic regulation, protecting photosynthetic machinery, removing active oxygen, enhancing the stability of biofilm, maintaining enzyme activity and biomacromolecule function, etc. However, under non-stress conditions, there are few reports on the effect of betaine on the content of the main active ingredients of medicinal plants and their yield. Summary of the invention

[0004] The present invention finds that spraying a betaine solution can significantly increase the total alkaloid content of the tubers of Pinellia ternata and increase the tuber yield of Pinellia ternata. Based on this, the present invention aims to provide an application of betaine in increasing the total alkaloid content of the tubers of Pinellia ternata and / or the tuber yield of Pinellia ternata, and to provide a method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the tuber yield of Pinellia ternata.

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

[0006] Application of betaine in increasing the total alkaloid content of tubers of pinellia and / or the yield of pinellia. The application can be achieved by spraying a betaine solution on the pinellia plants. The concentration of the betaine solution is preferably 5-80 mmol / L, more preferably 5-40 mmol / L, further preferably 5-20 mmol / L, and more preferably 20 mmol / L.

[0007] A method for increasing the total alkaloid content of pinellia tubers and / or the yield of pinellia, comprising the following steps: during the planting process of pinellia, spraying a betaine solution on the leaves of the pinellia plants. The concentration of the betaine solution is preferably 5-80 mmol / L, more preferably 5-40 mmol / L, further preferably 5-20 mmol / L, and more preferably 20 mmol / L.

[0008] Furthermore, the method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the yield of Pinellia ternata comprises the following steps: spraying the betaine solution when the three leaves of the Pinellia ternata plant are fully unfolded, spraying once every 7 days, and spraying 6-7 times in total. The time for spraying the betaine solution is preferably between 9 and 10 a.m.; during spraying, the leaves of the Pinellia ternata plant are evenly covered with the betaine solution.

[0009] The invention has the following advantages and beneficial effects: the invention increases the total alkaloid content of the tubers of pinellia by 82.85%, the fresh weight of the tubers by 31.29%, the dry weight of the tubers by 33.30%, and the diameter of the tubers by 12.70% by applying a 20 mmol / L betaine solution. The invention uses betaine in the planting of pinellia, and can simultaneously increase the total alkaloid content and yield of the tubers of pinellia, and the operation is simple, the input cost is low, and the invention is green and environmentally friendly. The invention provides a new method and idea for improving the quality and increasing the yield of pinellia, and increases the economic benefits of the planting and production of pinellia. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The results show the effects of spraying different concentrations of betaine solution on the total alkaloid content of Pinellia tubers. T5, spraying 5mmol / L betaine solution; T20, spraying 20mmol / L betaine solution; T80, spraying 80mmol / L betaine solution. The numbers in the figure represent the mean ± standard error, and different letters indicate significant differences (P<0.05).

[0011] Figure 2 The effects of spraying different concentrations of betaine solution on the fresh weight (a), dry weight (b) and diameter (c) of tubers of Pinellia ternata. T5, spraying 5mmol / L betaine solution; T20, spraying 20mmol / L betaine solution; T80, spraying 80mmol / L betaine solution. The numbers in the figure represent the mean ± standard error, and different letters indicate significant differences (P<0.05). DETAILED DESCRIPTION

[0012] The present invention is further described below in conjunction with specific examples, but the present invention is not limited to the following examples. The methods described are all conventional methods unless otherwise specified.

[0013] Example 1

[0014] Select dry pinellia tubers of uniform size and sow them in pots, and water them according to actual conditions.

[0015] Four treatment groups were set up: pure water as control (marked as: CK); betaine solution with a concentration of 5mmol / L prepared with pure water (marked as: T5); betaine solution with a concentration of 20mmol / L prepared with pure water (marked as: T20); betaine solution with a concentration of 80mmol / L prepared with pure water (marked as: T80). When the three leaves were fully expanded, plants with basically the same growth were selected, and the four prepared solutions were evenly sprayed on the leaves of Pinellia ternata at 9:00-10:00 in the morning, once every 7 days, for a total of 6 times. After the last application, the plants grew for 7 days before measuring various indicators. The total alkaloid content of the tubers of Pinellia ternata was repeated 5 times in each group, and the related indicators of the tuber yield of Pinellia ternata were repeated 20 times in each group.

[0016] The detection methods of various indicators are as follows:

[0017] 1. Determination of total alkaloid content

[0018] (1) Solution preparation

[0019] Citric acid-sodium citrate buffer solution: 32 mL 0.1 M citric acid solution and 68 mL 0.1 M sodium citrate solution, adjust the pH to 5.40.

[0020] 0.05% bromothymol blue: Weigh 0.025 g of bromothymol blue, dissolve it in 0.8 mL of 0.05 M NaOH solution, and dilute it to 50 mL with water to obtain a 0.05% bromothymol blue aqueous solution, which is ready for use.

[0021] (2) Preparation of reference solution

[0022] Accurately weigh 0.0022g of ephedrine hydrochloride standard, place it in a 100mL volumetric flask, add water to the scale, shake well, and obtain a concentration of 22μg / mL ephedrine hydrochloride standard solution, accurately measure 1, 2, 3, 4, 5mL of the above solution, each add water to 5mL, add 5mL of citric acid-sodium citrate buffer solution of pH=5.40 in sequence, then accurately add 0.05% bromothymol blue solution 1mL, chloroform 10mL in sequence, fully shake for 1min, place stratification, let stand for 1h, and take the chloroform layer for standby use. Measure the absorbance of the chloroform layer at a wavelength of 414nm. Take the concentration of ephedrine hydrochloride as the horizontal coordinate and the absorbance as the vertical coordinate, and regress to obtain the regression equation of the standard curve.

[0023] (3) Preparation and determination of test solution

[0024] Take the dried Pinellia tuber to constant weight, grind it into powder with a medicine roller, pass it through an 80-mesh sieve, weigh about 0.1000g of the Pinellia tuber powder, put it in a 50mL centrifuge tube, add 0.5mL 25%-28% concentrated ammonia water and 10mL chloroform, soak it overnight, and then extract it in an ice bath with ultrasonic for 1h. Centrifuge at 12000rpm for 10min, take the supernatant, and accurately add 5mL of citric acid-sodium citrate buffer solution with pH=5.40, 1mL of 0.05% bromothymol blue solution, and 5mL of water in sequence, shake it thoroughly for 1min, let it stand for 1h, take the chloroform layer as the test solution, and measure the absorbance value at a wavelength of 414nm.

[0025] (4) Calculation of results

[0026] The sample concentration was calculated using the obtained standard curve: C = (ABS-0.0589) / 0.06, R 2 =0.9952.

[0027] Where: C refers to sample concentration, ABS refers to sample absorbance.

[0028] The total alkaloid content is calculated by the formula: X = (C × 10 -6 ) / (W×F)×100%.

[0029] Where: X is the total alkaloid content; W refers to the sample weight, and F refers to the dilution factor (0.1).

[0030] 2. Method for determining the yield of Pinellia tuber

[0031] After the pinellia tubers are harvested, they are washed and dried, and their fresh weight is measured with an electronic balance, and their diameter is measured with a vernier caliper. Finally, the tubers are dried in an oven to a constant weight, and their dry weight is measured with an electronic balance.

[0032] The results of total alkaloid content determination in Pinellia tubers are as follows Figure 1Compared with CK, the total alkaloid content of T5 treatment group increased by 58.74%, the total alkaloid content of T20 treatment group increased by 82.85%, and the total alkaloid content of T80 treatment group increased by 24.10%.

[0033] The results of the determination of the yield of the tubers of Pinellia ternata are as follows Figure 2 Compared with CK, the fresh weight of the tubers of Pinellia ternata in the T5 treatment group increased by 17.89%, the fresh weight of the tubers of Pinellia ternata in the T20 treatment group increased by 31.29%, and the fresh weight of the tubers of Pinellia ternata in the T80 treatment group decreased by 15.03%. Compared with CK, the dry weight of the tubers of Pinellia ternata in the T5 treatment group did not change significantly, the dry weight of the tubers of Pinellia ternata in the T20 treatment group increased by 33.30%, and the dry weight of the tubers of Pinellia ternata in the T80 treatment group decreased by 24.18%. Compared with CK, the diameter of the tubers of Pinellia ternata in the T5 treatment group increased by 6.80%, the diameter of the tubers of Pinellia ternata in the T20 treatment group increased by 12.70%, and the diameter of the tubers of Pinellia ternata in the T80 treatment group did not change significantly.

[0034] The above results show that spraying a certain concentration of betaine solution can significantly increase the total alkaloid content of tubers and the yield of Pinellia tubers, and the concentration of the betaine solution is preferably 20 mmol / L.

[0035] The invention has low investment cost, obvious effect, is green and environmentally friendly, and can significantly increase the total alkaloid content and yield of the pinellia tubers, thereby improving the quality and supply of the pinellia medicinal materials.

[0036] The above specific implementation modes are not limitations of the present invention, and the present invention is not limited to the above implementation modes. Any changes, modifications, additions, substitutions, combinations and simplifications made within the essence and principles of the present invention are within the protection scope of the present invention.

Claims

1. Application of betaine in increasing the total alkaloid content of Pinellia tubers and / or the yield of Pinellia.

2. The use according to claim 1, characterized in that: This is achieved by spraying betaine solution on the pinellia plants.

3. The use according to claim 2, characterized in that: The concentration of the betaine solution is 5-80 mmol / L.

4. The use according to claim 2, characterized in that: The concentration of the betaine solution is 20mmol / L.

5. A method for increasing the total alkaloid content of pinellia tubers and / or the yield of pinellia, characterized in that: The following steps are involved: During the planting process of Pinellia ternata, spray betaine solution on the leaves of the Pinellia ternata plants.

6. The method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the yield of Pinellia ternata according to claim 5, characterized in that: The following steps are involved: When the three leaves of the Pinellia plant are fully expanded, start spraying the betaine solution, spray once every 7 days, for a total of 6-7 times.

7. The method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the yield of Pinellia ternata according to claim 5 or 6, characterized in that: The time for spraying the betaine solution is from 9:00 to 10:00 in the morning.

8. The method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the yield of Pinellia ternata according to claim 5 or 6, characterized in that: The concentration of the betaine solution is 5-80 mmol / L.

9. The method for increasing the total alkaloid content of the tubers of Pinellia ternata and / or the yield of Pinellia ternata according to claim 5 or 6, characterized in that: The concentration of the betaine solution is 20mmol / L.

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