CN111758735A - Method for improving polysaccharide and total flavonoids of effective components of astragalus membranaceus - Google Patents

Method for improving polysaccharide and total flavonoids of effective components of astragalus membranaceus Download PDF

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CN111758735A
CN111758735A CN202010799928.6A CN202010799928A CN111758735A CN 111758735 A CN111758735 A CN 111758735A CN 202010799928 A CN202010799928 A CN 202010799928A CN 111758735 A CN111758735 A CN 111758735A
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paclobutrazol
astragalus
content
concentration
polysaccharide
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CN111758735B (en
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刘广娜
杨祥波
黄伟鹏
李海涛
党永康
郑家学
奚广生
王艳铃
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Jilin Agricultural Science and Technology College
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/64—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with three nitrogen atoms as the only ring hetero atoms
    • A01N43/647—Triazoles; Hydrogenated triazoles
    • A01N43/653—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
    • A01N43/36—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings
    • A01N43/38—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings condensed with carbocyclic rings

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Abstract

The invention discloses a method for improving astragalus active ingredient polysaccharide and total flavonoids, which is realized by applying indoleacetic acid and/or paclobutrazol, and comprises the following specific steps: the highest content of astragalus polysaccharide is 400mg/L of paclobutrazol, the content of astragalus root polysaccharide which is applied with indoleacetic acid is in an ascending trend between 100mg/L and 200mg/L, is in a descending trend between 200mg/L and 400mg/L, and is the highest content when 200 mg/L; paclobutrazol with the highest flavone content is applied to 400 mg/L; in the treatment with the growth regulator of indoleacetic acid, the content of flavone tends to increase between 100mg/L and 200mg/L, and tends to decrease between 200mg/L and 400mg/L, and reaches a maximum value at 200 mg/L. The invention provides a new method for improving the contents of polysaccharide and total flavone which are effective components of astragalus, and has a promoting effect on the yield of the astragalus.

Description

Method for improving polysaccharide and total flavonoids of effective components of astragalus membranaceus
Technical Field
The invention relates to the technical field of astragalus planting, in particular to a method for improving polysaccharides and total flavonoids as active ingredients of astragalus.
Background
Radix astragali (Astragalus root)Astragalus mongholicusBunge), a leguminous plant, one of the commonly used Chinese herbs, is the root of astragalus mongholicus or astragalus membranaceus of leguminous plants. Mainly produced in inner Mongolia, Shanxi and Heilongjiang provinces of China. Collected in spring and autumn, dried in the sun, sliced, unprocessed or stir-baked with honey. Sweet in nature and slightly warm in nature, entering lung, spleen, liver and kidney meridians. Has effects of promoting urination, expelling toxin, expelling pus, healing sore, and promoting granulation. The main components of radix astragali include total flavone and polysaccharide, and radix astragali polysaccharide is an immunopotentiator or regulator, and has antiviral, antitumor, antiaging, radioprotective, anti-stress, and antioxidant effects.
The indoleacetic acid as plant growth hormone can promote the growth of plant roots, promote the formation of adventitious roots, accelerate the speed of vegetative propagation and improve the fruit setting rate. Indoleacetic acid, as a plant growth hormone, has a variety of physiological effects, which are related to its concentration. At low concentrations, growth is promoted, and at high concentrations, growth is inhibited, even the plant dies. The experimental result shows that the application of indole acetic acid with proper concentration can promote the growth of roots and the accumulation of components and improve the yield. Zhanxihong et al confirmed that the application of indoleacetic acid with appropriate concentration has a promoting effect on the accumulation of polysaccharide content in Cordyceps sinensis.
Paclobutrazol is a plant growth regulator and has the effects of delaying plant growth, improving yield and the like. The results of applying paclobutrazol to sweet potatoes in different concentrations by dawn light and the like show that the paclobutrazol can increase the tuberous roots of the sweet potatoes and improve the yield of the sweet potatoes. The application state of the existing paclobutrazol in plant production also illustrates that the high-concentration paclobutrazol can promote the growth of plant roots.
At present, no relevant report research on the influence of indoleacetic acid and paclobutrazol on the contents of astragalus active ingredient polysaccharide and total flavone is available.
Disclosure of Invention
The inventor discovers through research that: the application of indoleacetic acid and/or paclobutrazol can improve the active ingredients of astragalus polysaccharide and total flavonoids, and specifically comprises the following steps: the highest content of astragalus polysaccharide is 400mg/L of paclobutrazol which is 1.7 times of blank control, and the lowest content of astragalus polysaccharide is 100mg/L of indoleacetic acid which is 1.238 times of blank control. The two growth regulators have promoting effect on the accumulation of polysaccharide content in radix astragali. In the comparison of the same growth regulator, the content of the astragalus root polysaccharide applied with the indoleacetic acid is in an ascending trend between 100mg/L and 200mg/L, is in a descending trend between 200mg/L and 400mg/L, has the maximum content at 200mg/L and is 1.535 times of that of a blank control, and the effect is best; the radix astragali polysaccharide applied with paclobutrazol increases along with the increase of the concentration, and the high concentration of paclobutrazol can promote the accumulation of the polysaccharide content. The effect of applying paclobutrazol is better than that of applying indoleacetic acid. In the treatment of applying the paclobutrazol growth regulator, the flavone content is increased along with the increase of the concentration, the maximum value is reached at 400mg/L, and the effect is the best; in the treatment of spraying the indoleacetic acid growth regulator, the flavone content is in an ascending trend between 100mg/L and 200mg/L, and in a descending trend between 200mg/L and 400mg/L, and reaches a maximum value at 200mg/L, which is 1.08 times of that of a blank control, so that the effect is best.
The invention provides a new method for improving the contents of polysaccharide and total flavone which are effective components of astragalus, and the method can promote the accumulation of the contents of the polysaccharide and the total flavone in the astragalus and has a promoting effect on the yield of the astragalus.
Drawings
FIG. 1 is a glucose standard curve.
Fig. 2 is a rutin standard curve.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.
Examples of the experiments
1 Material
Radix astragali Mongolici is grown for two years (transplanted to trial field of the left school district of Jilin agriculture science and technology institute in 2017).
Test method
2.1 field test of Astragalus membranaceus
This experiment was designed using random block. Selecting radix astragali with similar size and growth vigor and no insect pest, planting, and weighing. And transplanting the seedlings in the experimental field of the left school of Jilin agriculture science and technology institute in 2017 in 4 months. Measuring out 90 square meters of land, performing comprehensive soil preparation, and deeply ploughing the land to remove weeds. In the experiment, the concentration of indoleacetic acid and paclobutrazol are respectively set to be 100, 200 and 400mg/L, each concentration is repeated for three times, and the same amount of tap water is sprayed in contrast. Each ridge is 2 meters long, 1 meter wide, 20 centimeters in row spacing and 50 ridges. The astragalus bud heads are upward, soil is covered and watering is carried out after cultivation is finished, and field weeds are timely treated and disease and pest control is well carried out. Treating the leaves of the astragalus before blooming after the leaves of the astragalus are spread, and spraying the leaves once every ten days.
Collected in 2017 and 10 months. Sampling each ridge by adopting a five-point sampling method, and determining related indexes. Treating the components according to intervals, removing fibrous roots, cleaning, drying at 80 ℃, crushing, and sieving by a 60-mesh sieve for later use.
Determination of photosynthetic rate of astragalus membranaceus
Photosynthetic rate: measured using a photosynthetic Rate meter model Lcpro +. The measurement is carried out in three times, namely three days after the spraying. In each measurement, five radix astragali is measured by adopting a five-point sampling method, and the leaf of the sixth branch from bottom to top is measured.
Determination of astragalus chlorophyll
Chlorophyll: measured using a chlorophyll meter. The measurement is carried out in three times, namely three days after the spraying. In each measurement, five radix astragali is measured by adopting a five-point sampling method, and the leaf of the sixth branch from bottom to top is measured.
Determination of astragalus malondialdehyde content
The malondialdehyde content was determined with reference to the method of plum arylene in the design of the plant malondialdehyde content determination assay.
(1) Extraction of malonaldehyde: weighing 1g of astragalus membranaceus leaf material, shearing, adding 2mL of 5% trichloroacetic acid and a small amount of quartz sand, grinding to obtain homogenate, adding 8mL of trichloroacetic acid, further grinding, centrifuging the homogenate at 4000r/min for 10min, and taking the supernatant as a sample extracting solution.
(2) Color reaction and measurement: 2mL of the centrifuged supernatant (control 2mL of distilled water) was aspirated, and 2mL of 0.6% thiobarbituric acid solution was added and shaken. Placing the test tube in boiling water bath, boiling for 10min (counting time from small bubbles appearing in the solution in the test tube), taking out the test tube, cooling, centrifuging at 3000r/min for 15min, sucking supernatant, and measuring absorbance values at 532nm, 600nm and 450nm with control as blank.
(3) Calculation of results
C(μmol/L)=6.45(A532-A600)-0.56A450
MDA content [ mu mol/g]＝CVTV1/1000V2W
Wherein, the concentration of C-MDA is mu mol/L;
VT-total volume of sample extract, mL;
V1-total reaction night volume, mL, of sample extract and TBA solution;
V2-volume of sample extract reacted with TBA, mL;
w-fresh sample mass, g.
Determination of Astragalus root yield
And (4) adopting a five-point sampling method, removing soil after harvesting, weighing and recording.
Determination of astragalus polysaccharide content
The determination is carried out by referring to the extraction and content determination research methods of astragalus polysaccharide of Liuruisheng and Wangbuyi.
Drawing of glucose standard curve
(1) 100mg of glucose dried to constant weight at 105 ℃ is taken, dissolved in water in a 100mL volumetric flask, diluted to the scale and shaken up. Precisely measuring 10mL, putting the solution into a 100mL volumetric flask, adding water to the scale, shaking up, and preparing a glucose standard solution of 0.1 mg/mL.
(2) Accurately sucking 6 parts of glucose standard solution, namely 0.0mL, 1.0mL, 2.0mL, 3.0mL, 4.0mL, 5.0mL and adding distilled water to make up to the scale mark in a 10mL volumetric flask respectively.
(3) Precisely sucking 2mL of each solution, placing in a test tube with a plug, adding 1.0mL of 5% phenol solution, shaking, quickly dropwise adding 5.0mL of concentrated sulfuric acid, shaking, standing for 5min, heating in 80 deg.C water bath for 15min, taking out, and quickly cooling to room temperature. A blank was prepared by equilibrating 2.0mL of distilled water as above and measuring the absorbance at 490 nm.
(4) And (3) performing linear regression by taking the concentration (X) of the anhydrous glucose as an abscissa and the absorbance (Y) as an ordinate to obtain a linear equation.
Extraction and content determination of polysaccharide
Weighing radix astragali powder 3g, adding 7 times of distilled water, boiling for 1.5 hr, filtering with 4 layers of gauze, and boiling the residue for 3 times by the same method. Mixing filtrates, heating and concentrating, centrifuging the concentrated solution at 2000 r/min for 10min, adding 3 times of 95% ethanol into the supernatant, and precipitating. Centrifuging at 2000 r/min for 10min in a centrifuge, removing supernatant, adding 95% ethanol into the precipitate to reach concentration of 80%, standing, and decanting the supernatant. The filter residue is washed with acetone for 2 times and filtered. And putting the filter residue into a drying oven, and drying at 80 ℃ to obtain the astragalus polysaccharide.
Taking 25 mg of astragalus polysaccharide which is extracted and dried at 80 ℃ to constant weight, placing the astragalus polysaccharide in a 25mL volumetric flask, adding distilled water to dissolve the astragalus polysaccharide to a scale, shaking up, accurately sucking 1 mL, placing the astragalus polysaccharide in a 10mL volumetric flask, adding distilled water to dilute the astragalus polysaccharide to the scale, shaking up to prepare a polysaccharide solution of 0.1 mg/mL. Sucking 0.5 mL of the solution, placing the solution in a 25mL volumetric flask, adding distilled water to supplement the solution to 25mL, measuring the absorbance value, substituting the absorbance value into a regression equation to calculate the polysaccharide concentration, and obtaining the astragalus polysaccharide content through conversion.
Determination of Total Flavonoids content
And (4) determining the content of the total flavonoids in the astragalus by referring to a plum spring red ultraviolet spectrophotometry.
Drawing standard curve of total flavone
(1) Accurately weighing rutin 20mg, placing in a 100mL volumetric flask, adding 70% ethanol, heating in water bath for dissolving, cooling, diluting with 70% ethanol to scale, shaking, and making into 0.2mg/mL reference solution.
(2) Precisely sucking the solution 0.0mL, 1.6mL, 1.8mL, 2.0mL, 2.2mL and 2.4.mL into a 10mL volumetric flask, adding 5% sodium nitrite 0.5 mL, shaking up, standing for 6min, adding 10% aluminum nitrate 0.5 mL, shaking up, standing for 6min, adding 4% sodium hydroxide 4 mL, diluting the solution with 70% ethanol solution to constant volume, shaking up, and standing for 15 min. The first tube solution was blanked at 425nm wavelength. And measuring absorbance values under different concentrations, and performing regression by taking the absorbance values as a vertical coordinate (Y) and the concentrations as a horizontal coordinate (X) to draw a standard curve.
Extraction and content determination of total flavone
Taking 1g of astragalus crude powder into a round-bottom flask, repeating each group for three times, adding 10 times of 95% ethanol, soaking overnight, leaching for 1h in a 75 ℃ water bath, cooling, filtering, collecting filtrate, and fixing the volume in a 100mL volumetric flask for later use. The solution was diluted 10-fold and the absorbance value was measured at a wavelength of 425nm according to the procedure in (2) above, and brought into the standard curve.
Results and analysis
3.1 Effect of different spraying times on the photosynthetic Rate of Astragalus
3.1.1 Effect of one-time spraying on photosynthesis rate of radix astragali
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between various classes of growth regulators, between various concentrations, and between indoleacetic acid and paclobutrazol interactions with concentrations are therefore also needed.
TABLE 1 multiple comparison Table of the Effect of one spray application on the photosynthetic Rate of Astragalus (unit: mg CO)2/10cm2/hr）
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 18.26 a A
Paclobutrazol 200mg/L 15.68 b B
Indolylacetic acid 200mg/L 15.12 b B
Paclobutrazol 100mg/L 13.54 c C
Indolylacetic acid 400mg/L 13.43 c C
Indolylacetic acid 100mg/L 13.23 c C
Blank control 8.43 d D
As can be seen from table 1, the photosynthetic rate was higher than that of the blank control in the treatment of spraying indolacetic acid and paclobutrazol, and the photosynthetic rate was significantly different from that of the blank control. The strongest photosynthetic rate of the treated astragalus is spraying paclobutrazol with the concentration of 400mg/L, the weakest photosynthetic rate is indoleacetic acid with the concentration of 100mg/L, and the two have very obvious difference. In the treatment of spraying indoleacetic acid, the photosynthetic rate is strongest at the concentration of 200mg/L, and the difference is very obvious compared with the other two concentrations; there was no significant difference between the other two concentrations in the treatment with paclobutrazol spray. During the treatment of astragalus by paclobutrazol, the photosynthesis of astragalus is continuously strengthened along with the increase of the concentration of the growth regulator, and the concentrations are very obviously different.
Influence of spraying twice on photosynthesis rate of astragalus membranaceus
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between various classes of growth regulators, between various concentrations, and between indoleacetic acid and paclobutrazol interactions with concentrations are therefore also needed.
TABLE 2 multiple comparison Table of the Effect of spraying twice on the photosynthetic Rate of Astragalus (unit: mg) CO2/10cm2/hr）
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 22.28 a A
Indolylacetic acid 200mg/L 20.57 b B
Paclobutrazol 200mg/L 19.65 c B
Indolylacetic acid 100mg/L 16.62 d C
Indolylacetic acid 400mg/L 16.49 d C
Paclobutrazol 100mg/L 16.37 d C
Blank control 10.65 e D
As can be seen from table 2, there was a very significant difference in both treatments with the two growth regulators sprayed, compared to the blank control. The astragalus root has the strongest photosynthetic rate of spraying paclobutrazol with the concentration of 400mg/L, and the astragalus root has the weakest photosynthetic rate of spraying paclobutrazol with the concentration of 100 mg/L. In the indoleacetic acid treatment group, the photosynthesizing is strongest at the concentration of 200mg/L, and has very obvious difference with other two concentrations, and the photosynthesizing is weakest at 400mg/L and has no obvious difference with 100 mg/L. In the paclobutrazol treatment group, the photosynthetic rate increased with increasing concentration.
Influence of spraying for three times on photosynthesis rate of astragalus membranaceus
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between various classes of growth regulators, between different concentrations, and between indoleacetic acid and paclobutrazol interactions with concentrations are therefore also needed.
TABLE 3 multiple comparison Table of the Effect of three spray applications on the photosynthetic Rate of Astragalus (unit: mg CO)2/10cm2/hr）
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 18.76 a A
Paclobutrazol 200mg/L 15.39 b B
Indolylacetic acid 200mg/L 14.11 c C
Paclobutrazol 100mg/L 13.76 c C
Indolylacetic acid 400mg/L 12.41 d D
Indolylacetic acid 100mg/L 12.37 d D
Blank control 9.72 e E
As can be seen from table 3, there was a very significant difference in both treatments with the two growth regulators sprayed, compared to the blank control. The astragalus root has the strongest photosynthetic rate of spraying paclobutrazol with the concentration of 400mg/L, and the astragalus root has the weakest photosynthetic rate of spraying indoleacetic acid with the concentration of 100 mg/L. In the indoleacetic acid treatment group, the photosynthesizing is strongest at the concentration of 200mg/L, and has very obvious difference with other two concentrations, and the photosynthesizing is weakest at 100mg/L and has no obvious difference with 400 mg/L. In the paclobutrazol treatment group, the photosynthetic rate increased with increasing concentration.
As can be seen from tables 1, 2 and 3, the high concentration of indoleacetic acid is not conducive to photosynthesis of astragalus membranaceus, and in the treatment of paclobutrazol, it is conducive to photosynthesis as the paclobutrazol concentration increases.
Influence of different spraying times on chlorophyll content of radix astragali
3.2.1 Effect of spraying once on chlorophyll content of Astragalus membranaceus
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01, multiple comparisons between various species of growth regulators, between different concentrations, and between concentration interactions of indole acetic acid and paclobutrazol are also needed.
TABLE 4 multiple comparison Table of the effect of spraying once on the chlorophyll content of Astragalus membranaceus (Unit: SPAD)
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 21.47 a A
Indolylacetic acid 200mg/L 17.93 b AB
Paclobutrazol 200mg/L 17.03 bc BC
Paclobutrazol 100mg/L 15.06 cd BC
Indolylacetic acid 400mg/L 14.03 d C
Indolylacetic acid 100mg/L 14.02 d C
Blank control 14 d C
As can be seen from Table 4, the treated radix astragali has the highest chlorophyll content of paclobutrazol with the spraying concentration of 400mg/L, and has obvious difference compared with the blank control, and the content of the paclobutrazol is 1.534 times of that of the blank control; the chlorophyll content was the least of indoleacetic acid sprayed at a concentration of 100mg/L, 1.001 times that of the blank, but with no significant difference.
Influence of spraying twice on chlorophyll content of radix astragali
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01, multiple comparisons between various species of growth regulators, between different concentrations, and between concentration interactions of indole acetic acid and paclobutrazol are also needed.
TABLE 5 multiple comparison Table (Unit: SPAD) of the influence of spraying twice on the chlorophyll content of Astragalus membranaceus
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 27.76 a A
Indolylacetic acid 200mg/L 24.97 b AB
Paclobutrazol 200mg/L 22.63 b B
Indolylacetic acid 100mg/L 18.43 c C
Paclobutrazol 100mg/L 18.00 c C
Indolylacetic acid 400mg/L 17.43 c C
Blank control 16.44 c C
As can be seen from Table 5, the treated Astragalus membranaceus has the largest chlorophyll content, and is sprayed with paclobutrazol with the concentration of 400mg/L, and the difference is obvious compared with the blank control, 1.689 times of the blank control, and is obvious compared with other groups. The lowest chlorophyll content was indoleacetic acid sprayed at a concentration of 400mg/L, which was 1.060 times greater than the blank, but was not significantly different from the blank.
Influence of spraying for three times on chlorophyll content of radix astragali
From SPSS analysis, F between various classes of growth regulators>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01, multiple comparisons between various species of growth regulators, between different concentrations, and between concentration interactions of indole acetic acid and paclobutrazol are also needed.
TABLE 6 multiple comparison Table of the influence of three spray applications on the chlorophyll content of Astragalus membranaceus (Unit: SPAD)
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 34.03 a A
Paclobutrazol 200mg/L 28.90 b B
Indolylacetic acid 200mg/L 28.60 b B
Paclobutrazol 100mg/L 22.93 c C
Indolylacetic acid 100mg/L 21.90 c C
Indolylacetic acid 400mg/L 20.20 c C
Blank control 20.13 c C
As can be seen from Table 6, the treated Astragalus membranaceus has a chlorophyll content of at most 400mg/L as sprayed paclobutrazol, and has a very significant difference compared with the blank control, which is 1.691 times that of the blank control; the minimal chlorophyll content was paclobutrazol at a spray concentration of 100mg/L, which was 1.003 times that of the blank, but no significant difference compared to the blank.
As can be seen from tables 4, 5 and 6, the content of astragaloside chlorophyll is in an increasing state in three times of measurement, which indicates that both indoleacetic acid and paclobutrazol can promote the increase of the content of chlorophyll. In the treatment of the indoleacetic acid, the chlorophyll content is highest when the content is 200mg/L, and the chlorophyll content is lowest when the content is 400mg/L, and the indoleacetic acid with high concentration is not suitable for the accumulation of the chlorophyll content of the astragalus. In the treatment with paclobutrazol, the chlorophyll content increases with increasing concentration.
Influence of different spraying times on content of malondialdehyde in radix astragali
3.3.1 Effect of one-time spraying on the content of malondialdehyde in radix astragali
From SPSS analysis, the concentration of F>F0.01（3，16）、P<0.01, so multiple comparisons were performed.
TABLE 7 multiple comparison Table of the Effect of one spray application on the malondialdehyde content of Astragalus membranaceus (Unit:. mu. mol/L)
Treatment of Mean value 5% level 1% level
Indolylacetic acid 400mg/L 4.38 a A
Indolylacetic acid 100mg/L 3.70 b B
Paclobutrazol 100mg/L 3.55 b B
Paclobutrazol 200mg/L 3.52 b B
Indolylacetic acid 200mg/L 3.44 b B
Paclobutrazol 400mg/L 3.37 b B
Blank control 3.27 b B
As can be seen from Table 7, the indole acetic acid malondialdehyde concentration of 400mg/L was the highest, and was significantly different from the blank control and the other treatment groups, and was not significantly different from the blank control in any of the other groups.
Influence of spraying twice on the content of malondialdehyde in radix astragali
From SPSS analysis, the concentration of F>F0.01（3，16）、P<0.01, so multiple comparisons were performed.
TABLE 8 multiple comparison Table of the Effect of spraying twice on the malondialdehyde content of Astragalus membranaceus (Unit: μmol/L)
Treatment of Mean value 5% level 1% level
Indolylacetic acid 400mg/L 4.34 a A
Indolylacetic acid 100mg/L 3.86 b B
Paclobutrazol 200mg/L 3.75 b B
Paclobutrazol 100mg/L 3.73 b B
Indolylacetic acid 200mg/L 3.50 b B
Paclobutrazol 400mg/L 3.44 b B
Blank control 3.33 b B
As can be seen from Table 8, the indole acetic acid malondialdehyde concentration of 400mg/L was the highest, and was significantly different from the blank control and the other treatment groups, and was not significantly different from the blank control in any of the other groups.
Influence of spraying for three times on content of malondialdehyde in radix astragali
From SPSS analysis, F>F0.01（3，16）、P<0.01, so multiple comparisons were performed.
TABLE 9 multiple comparison Table of the Effect of three spray applications on the malondialdehyde content of Astragalus membranaceus (Unit: μmol/L)
Treatment of Mean value 5% level 1% level
Indolylacetic acid 400mg/L 4.40 a A
Indolylacetic acid 100mg/L 3.66 b B
Paclobutrazol 200mg/L 3.65 b B
Paclobutrazol 100mg/L 3.53 b B
Indolylacetic acid 200mg/L 3.49 b B
Paclobutrazol 400mg/L 3.44 b B
Blank control 3.30 b B
As can be seen from Table 9, the indole acetic acid malondialdehyde concentration of 400mg/L was the highest, and was significantly different from the blank control and the other treatment groups, and was not significantly different from the blank control in any of the other groups.
As can be seen from tables 7, 8 and 9, the content of high-concentration indole acetic acid malondialdehyde is the largest, which indicates that the membrane peroxidation degree of the astragalus membranaceus leaf cell is high, the cell membrane is seriously damaged, and the astragalus membranaceus leaf has stress damage.
Effect of different plant growth regulators on the yield of Astragalus
Growth regulators of the various classes F as determined by SPSS analysis>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between different growth regulator species, different concentrations, different growth regulators and concentration interactions are therefore also required.
TABLE 10 multiple comparison of Astragalus yields (unit: g)
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 71.33 a A
Paclobutrazol 200mg/L 59.33 b B
Indolylacetic acid 400mg/L 53.33 c C
Indolylacetic acid 200mg/L 52.03 c C
Paclobutrazol 100 mg/L 50.00 c C
Indolylacetic acid 100mg/L 43.33 d D
Blank control 43.23 d D
As can be seen from Table 10, the highest yield of the radix astragali is paclobutrazol with the spraying concentration of 400mg/L, which is 1.653 times of that of the blank control, and has very significant difference with other groups; the astragalus yield is the least that of indoleacetic acid with the spraying concentration of 100mg/L, which is almost equal to that of blank control and has no obvious difference.
Under the treatment of the indole acetic acid, the yield is increased along with the increase of the concentration of the indole acetic acid, and no significant difference exists between 400mg/L and 200mg/L, but the difference between the two concentrations and 100mg/L is very significant; under the treatment with paclobutrazol, the yield increases with increasing paclobutrazol concentration, and there are very significant differences between all three concentrations. Under the concentration of 400mg/L, the yield of the astragalus sprayed with paclobutrazol is 1.202 times higher than that of the astragalus sprayed with indoleacetic acid, and the difference is very obvious. Therefore, the treatment of spraying paclobutrazol is better than the treatment of indoleacetic acid, and the high-concentration paclobutrazol is beneficial to improving the yield of astragalus.
Effect of different plant growth regulators on Astragalus polysaccharides content
TABLE 11 glucose Standard Curve concentration and Absorbance
Concentration of 0 0.010 0.020 0.030 0.040 0.050
Absorbance of the solution 0 0.073 0.146 0.223 0.295 0.378
After conversion, the growth regulators of the various types F were analyzed by SPSS software>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between different growth regulator species, different concentrations, different growth regulators and concentration interactions are therefore also required.
TABLE 12 Astragalus polysaccharides multiple comparison table (unit: mg/g)
Treatment of Mean value 5% significant level 1% very significant level
Paclobutrazol 400mg/L 29.33 a A
Indolylacetic acid 200mg/L 26.02 b B
Paclobutrazol 200mg/L 25.33 b B
Indolylacetic acid 400mg/L 22.21 c C
Paclobutrazol 100mg/L 20.33 c C
Indolylacetic acid 100mg/L 20.65 c C
Blank control 16.67 d D
As can be seen from Table 12, indole acetic acids of 100mgl/L, 200mg/L, 400mg/L and blank control all have significant differences, and the concentrations also have significant differences; obvious differences exist among paclobutrazol 100mg/L, paclobutrazol 200mg/L and paclobutrazol 400mg/L and blank control, and obvious differences also exist among various concentrations; the applied paclobutrazol has the highest content of 400mg/L and has obvious difference with each group; the content of the indole acetic acid administered is the lowest, and has no obvious difference with the indole acetic acid administered which is 400mg/L and paclobutrazol administered which is 100 mg/L; there was no significant difference between the administration of 200mg/L indole acetic acid and 200mg/L paclobutrazol.
In the comparison of the two growth regulators, the highest content of astragalus polysaccharide was 400mg/L of paclobutrazol, which is 1.7 times of the blank control, and the lowest content of astragalus polysaccharide was 100mg/L of indoleacetic acid, which is 1.238 times of the blank control. The two growth regulators have promoting effect on the accumulation of polysaccharide content in radix astragali. In the comparison of the same growth regulator, the content of the astragalus root polysaccharide applied with the indoleacetic acid is in an ascending trend between 100mg/L and 200mg/L, is in a descending trend between 200mg/L and 400mg/L, has the maximum content at 200mg/L and is 1.535 times of that of a blank control, and the effect is best; the radix astragali polysaccharide applied with paclobutrazol increases along with the increase of the concentration, and the high concentration of paclobutrazol can promote the accumulation of the polysaccharide content. The effect of applying paclobutrazol is better than that of applying indoleacetic acid.
3.6 Effect of different plant growth regulators on the content of Total Flavonoids in Astragalus
TABLE 13 concentration and Absorbance of rutin Standard Curve
Concentration of 0.032 0.036 0.040 0.044 0.048
Absorbance of the solution 0.17 0.212 0.235 0.263 0.295
After conversion, the growth regulator F is analyzed by SPSS software>F0.01（1，16）、p<0.01, concentration of F>F0.01（3，16）、P<0.01, F of interaction>F0.01（3，16）、p<0.01; multiple comparisons between different growth regulator species, different concentrations, different growth regulators and concentration interactions are therefore also required.
TABLE 14 multiple comparison of total flavonoids in Astragalus membranaceus (unit: mg/g)
Treatment of Mean value 5% significant level 1% significant level
Paclobutrazol 400mg/L 33.13 a A
Paclobutrazol 200mg/L 30.88 b B
Indolylacetic acid 200mg/L 29.40 c C
Indolylacetic acid 400mg/L 28.75 c CD
Indolylacetic acid 100mg/L 28.45 cd D
Paclobutrazol 100mg/L 28.20 d D
Blank control 27.08 e E
From FIG. 2, Table 14 shows that the total flavone content in radix astragali at 100mg/L, 200mg/L and 400mg/L of indoleacetic acid was significantly different from that of the blank control, but the difference between the concentrations was not significant; the total flavone content of astragalus roots, which is 100mg/L, 200mg/L and 400mg/L of paclobutrazol, is very obviously different from that of a blank control, and the concentrations are also very obviously different; the paclobutrazol with the highest total flavone content and the application concentration of 400mg/L has obvious difference with each group, the paclobutrazol with the lowest total flavone content and the application concentration of 100mg/L has no obvious difference with the indoleacetic acid with 100mg/L and has obvious difference with other treatments.
In the comparison of the two growth regulators, the paclobutrazol with the concentration of 400mg/L is applied to the root of the astragalus root with the most accumulated content and is about 1.2 times of that of the blank control, the paclobutrazol with the concentration of 100mg/L is applied to the root of the astragalus root with the least accumulated content and is about 1.04 times of that of the blank control, and the contents of the two growth regulators are improved compared with that of the blank control, so that the two growth regulators have promotion effects on the total flavone content of the astragalus root. In the treatment of spraying the paclobutrazol growth regulator, the flavone content is increased along with the increase of the concentration, the maximum value is reached when the concentration is 400mg/L, and the effect is best; in the treatment of spraying the indoleacetic acid growth regulator, the flavone content is in an ascending trend between 100mg/L and 200mg/L, and in a descending trend between 200mg/L and 400mg/L, and reaches a maximum value at 200mg/L, which is 1.08 times of that of a blank control, so that the effect is best. The high-concentration paclobutrazol can promote the accumulation of the total flavone content in the root of the astragalus, and the effect of applying the high-concentration paclobutrazol is better than that of indoleacetic acid.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes or modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention. The embodiments and features of the embodiments of the present application may be combined with each other arbitrarily without conflict.

Claims (4)

1. A method for improving the active ingredients of polysaccharide and total flavonoids in astragalus is characterized by comprising the following steps: indoleacetic acid and/or paclobutrazol are administered.
2. The method for improving the polysaccharides and the total flavonoids as the effective components of astragalus according to claim 1, which comprises the following steps: indolylacetic acid was administered at 200 mg/L.
3. The method for improving the polysaccharides and the total flavonoids as the effective components of astragalus according to claim 1, which comprises the following steps: 400mg/L paclobutrazol was administered.
4. The method for improving the polysaccharides and the total flavonoids as the effective components of astragalus according to claim 1, which comprises the following steps: 200mg/L of indole acetic acid and 400mg/L of paclobutrazol were administered.
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