CN110622808A - Planting method for promoting growth of bletilla striata and accumulation of effective components - Google Patents
Planting method for promoting growth of bletilla striata and accumulation of effective components Download PDFInfo
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Classifications
-
- 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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
-
- 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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/60—Flowers; Ornamental plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention relates to a planting method for promoting growth of bletilla striata and accumulation of effective components, and belongs to the technical field of bletilla striata planting. The planting method comprises the following steps: keeping the relative water content of the soil of 75-85% after the bletilla striata seedlings germinate to the mature period, and adjusting the relative water content of the soil to 20-30% before harvesting; the method for calculating the relative water content of the soil comprises the following steps: the invention obviously improves the yield and the content of effective components of bletilla striata by controlling the relative water content of the soil.
Description
Technical Field
The invention relates to the technical field of bletilla striata planting, and particularly relates to a planting method for promoting growth of bletilla striata and accumulation of effective ingredients.
Background
Bletilla striata (Bletilla striata) also known as Nepeta stramonium, Gansu root, Baiyao, is a perennial warm-zone terrestrial orchid. Bletilla striata is not only an ornamental horticultural plant, but also a traditional Chinese medicinal material in China, and has important medicinal value. Recorded in Ben Cao gang mu, bletilla striata is bitter, mild and nontoxic in smell (root). According to the record of pharmacopoeia, bletilla has the functions of astringing to stop bleeding, eliminating swelling and promoting granulation, and is mainly used for treating hemoptysis, hematemesis, epistaxis, traumatic hemorrhage, sore and ulcer swelling and pain, unhealed ulcer and other diseases. At present, the bletilla striata polysaccharide is considered as the main effective component in bletilla striata, and the content can reach 35%. Other main active ingredients also include dihydrophenanthrene compounds, bibenzyl compounds, triterpenes and saponins thereof, steroids and saponins thereof, and the like.
Bletilla striata is a national secondary protection plant and a protected species endangered in the world, wild bletilla striata resources are collected and dug without restriction in recent years, the quantity of the resources is reduced sharply, medicinal raw materials are seriously deficient, and the bletilla striata is taken in from red famous records of Chinese species and is also a protection species of International trade Convention (CITES) for endangered wild animal and plant species. With the increase of demand for bletilla striata, the raw material requirements of pharmaceutical enterprises and other enterprises cannot be met only by digging wild bletilla striata. At present, due to the lack of systematic knowledge on the biological characteristics of bletilla striata, the influence of environmental factors on the growth and effective component accumulation of bletilla striata and the like, efficient production and planting rules of bletilla striata cannot be established in a targeted manner, and stable yield and quality are difficult to obtain.
Disclosure of Invention
The invention aims to provide a planting method for promoting growth of bletilla striata and accumulation of effective components. The invention obviously improves the yield and the content of effective components of the bletilla striata by controlling the relative water content of the soil.
The invention provides a planting method for promoting growth of bletilla striata and accumulation of effective components, which comprises the following steps:
keeping the relative water content of the soil of 75-85% after the bletilla striata seedlings germinate to the mature period, and adjusting the relative water content of the soil to 20-30% before harvesting;
the method for calculating the relative water content of the soil comprises the following steps:
the relative water content of the soil is equal to the water content of the soil/field water capacity multiplied by 100 percent.
Preferably, the germination rate of the bletilla striata reaches more than 95%.
Preferably, the period before harvesting is 4-6 weeks before harvesting the bletilla striata.
Preferably, the bletilla striata seedlings are selected from biennial bletilla striata pseudobulbs.
Preferably, the diameter of the biennial bletilla striata pseudobulb is 0.8-1.5 cm.
Preferably, the maximum light for said planting is 50% of full light.
Preferably, the planting temperature is 20-32 ℃.
Preferably, the relative humidity of the planting is 50-70%.
Preferably, the planting substrate comprises a mixture of leaf mold and red soil.
Preferably, the volume ratio of the leaf mold to the red soil is (0.8-1.2): 1.
the invention provides a planting method for promoting growth of bletilla striata and accumulation of effective components. In the production and planting process, higher water supply is kept from the germination stage to the mature stage of bletilla striata seedlings, so that the yield and polysaccharide content of bletilla striata are improved; the accumulation of secondary metabolites is promoted and the quality of the bletilla striata is improved by controlling water before harvesting, so that the aim of synchronously improving the yield and the quality is fulfilled. Test results show that the relative water content of the soil has obvious influence on the physiology, growth and content of substances in bletilla striata, and high-water-treated bletilla striata plants have higher leaf area, light saturation point, stomatal conductance, chlorophyll content and CO2Assimilation rate, biomass accumulation of bletilla striata and pseudo bulb polysaccharide content increase; the content of total phenols and dactylorhin A in the bletilla striata new pseudobulb treated by low water is obviously improved. In addition, the accumulation of bletilla dactylorhin A is promoted by controlling water, so that the effect of inhibiting the growth of peripheral weeds can be achieved to a certain extent, the use of herbicides is reduced, and the production cost is reduced.
Drawings
FIG. 1 is a graph showing the response of bletilla photosynthetic rate (A) and stomatal conductance (B) to optical intensity (PAR) under different moisture treatments provided by the present invention;
FIG. 2 is a plant height frequency distribution diagram of bletilla striata under different moisture treatments provided by the present invention;
FIG. 3 shows the total amount of polysaccharides, total phenols, militarin, and dactylorhin A in the bletilla striata new pseudobulb under different moisture treatments as provided by the present invention.
Detailed Description
The invention provides a planting method for promoting growth of bletilla striata and accumulation of effective components, which comprises the following steps:
keeping the relative water content of the soil of 75-85% after the bletilla striata seedlings germinate to the mature period, and adjusting the relative water content of the soil to 20-30% before harvesting;
the method for calculating the relative water content of the soil comprises the following steps:
the relative water content of the soil is equal to the water content of the soil/field water capacity multiplied by 100 percent.
In the invention, the relative water content of the soil is preferably kept at 80% after the bletilla striata seedlings germinate to the mature period, and the relative water content of the soil is preferably adjusted to 20% before harvesting. Keeping the relative water content of the soil of 75-85% after the bletilla striata seedlings germinate to the mature period can obviously improve the photosynthetic rate of the bletilla striata seedlings, so that the large biomass and polysaccharide content can be obtained, and the relative water content of the soil is adjusted to 20-30% before harvesting, so that the accumulation of secondary metabolites of the bletilla striata seedlings can be promoted, the purpose of synchronously improving the yield and quality of the bletilla striata is achieved, and good benefits can be brought to large-scale planting and efficient production of the bletilla striata. In the invention, the bletilla striata germchit is selected from biennial bletilla striata pseudobulb. In the invention, the diameter of the biennial bletilla striata pseudobulb is 0.8-1.5 cm. In the invention, the germination rate of bletilla striata is preferably more than 95%. In the invention, before harvesting, the period of time before harvesting is 4-6 weeks, more preferably 1 month, which is beneficial to accumulation of effective ingredients. In the invention, the maximum illumination of the planting is 50% of the full illumination, and the planting is suitable for the growth of bletilla striata. In the invention, the planting temperature is 20-32 ℃, more preferably 26 ℃, and bletilla striata growth is facilitated. In the invention, the relative humidity of planting is 50-70%, preferably 60%, and the planting method is suitable for growth of bletilla striata. In the present invention, the planting substrate preferably includes a mixture of leaf mold and red soil, and more preferably includes a mixture of yellow soil, red soil and leaf mold. In the invention, when the planting substrate is leaf mold and red soil, the volume ratio of the leaf mold to the red soil is (0.8-1.2): 1, preferably 1: 1.
In the invention, the field water capacity refers to the maximum value of water capacity which can be maintained without the influence of underground water on soil, and the measuring method comprises the following steps: taking an undisturbed soil sample, fully soaking the undisturbed soil sample in water, taking out the undisturbed soil sample, weighing m1 on the upper layer of the soil sample after the water infiltration process is carried out for 8 hours, putting the soil sample into a constant-temperature oven at 105 ℃ for drying until the constant weight is reached, taking out the soil sample, putting the soil sample into a dryer for cooling to the room temperature, and weighing m 2.
And the field water capacity X is (m1-m2)/m 2.
The water content (weight percent) of the soil is (original soil weight-dried soil weight)/dried soil weight multiplied by 100 percent is water weight/dried soil weight multiplied by 100 percent.
The planting method for promoting the growth of bletilla striata and the accumulation of active ingredients is further described in detail with reference to the following specific examples, and the technical scheme of the invention includes but is not limited to the following examples.
Example 1
Selecting two-year-old bletilla pseudobulb (diameter 0.8-1.5cm) with consistent specification, planting in mixed matrix (volume ratio 1: 1) of leaf mold and red soil, and planting 1 plant per pot. After planting, placing the plants in a greenhouse, and keeping the maximum illumination at 50 percent of full illumination, the temperature at 20-32 ℃ and the relative humidity at 50-70 percent.
The bletilla pseudobulb germinates about 1 month after being planted, and when the germination rate reaches more than 95%, the water treatment is started. The moisture gradient is set by controlling the relative water content of the soil: high water treatment (HW) 80%; middle-water treatment (MW) is 50%; the low water treatment (low-water, LW) was 20%. The relative water content of the soil was calculated as: the relative water content (%) of the soil is equal to the water content of the soil/field water capacity x 100%. During the experiment, the relative water content of the soil was maintained by weighing the entire pot. After the water treatment for 90 days, healthy mature leaves are selected for test determination.
Statistical analysis of the data was performed using the software Spss 16.0 for Windows (SPSS Inc, Chicago, USA), univariate analysis of variance was used for comparison between different moisture treatments, and Tukey test was used for significance test. All statistical plots were plotted using prism6.01(GraphPad Software Inc, La Jolla, USA).
First, the influence of the moisture condition on the photosynthetic gas exchange of bletilla striata
The photoresponse curves were measured with a LI-6400 photosynthesizer (Li-Cor, USA) using a 6400-40 fluorescent leaf cell. When the photoresponse curve is measured, the temperature of the leaf chamber is maintained at 25 ℃, the relative humidity is 60-80 percent, and CO is added2The concentration is maintained at 400. mu. mol-1. The light intensity gradient was set at 1000, 800, 600, 400, 300, 200, 100, 50, 0. mu. mol m-2s-1. Before starting the measurement, at 400. mu. mol-1CO2At a concentration of 600. mu. mol m-2s-1The saturation light intensity is induced for about 20min until the stomatal conductance and the photosynthetic rate reach steady states.
The instantaneous Water Use Efficiency (WUEi) was calculated as: pn/Tr，TrIs the transpiration rate. In this study, the light intensity of 600 μmol m in the photoresponse curve was selected-2s-1Time corresponding transpiration rate (T)r) And photosynthetic Rate (P)n) The value is obtained.
The chlorophyll content determination method comprises the following steps: about 0.07g of leaves were weighed, cut into 0.5 cm. times.0.5 cm pieces, put into a 10ml volumetric flask, subjected to constant volume with N, N-dimethylformamide, placed in the dark to extract a pigment for 48 hours, and then subjected to measurement of absorbance values at 647nm and 664.5nm with an ultraviolet spectrophotometer UV-2550(Shimadzu, Japan). Chlorophyll content per unit mass (mg g)-1) And the area of the leaf per fresh weight, and the chlorophyll content (μ g cm) based on the area was calculated-2)。
The soil moisture conditions had a significant effect on the photosynthesis of bletilla striata (fig. 1), the maximum photosynthetic rate (P) of the leaves treated with low water (P)max) Apparent Quantum Efficiency (AQE) and Light Saturation Point (LSP) were all lower than for high water and medium water treatments (table 1). By observing the light response curve, the photosynthetic rate of the plants under the low water treatment is 200 mu mol m in light intensity-2s-1The water is saturated, while the high water and the medium water have the light intensity of 600 mu mol m-2s-1The rhizoma bletilla can adapt to stronger illumination when the water content is sufficient. Although the light response curves of the high-water and medium-water treatment blades are similarHowever, the high water treatment blade has a higher air porosity than the medium water treatment blade. Photosynthetic Rate (P) for Low Water treatmentn) Harmonize the conductance of the stoma (g)s) At all light intensities, significantly lower than high water and medium water treatments (fig. 1). The low water treatment chlorophyll a content, chlorophyll b content, and the ratio of chlorophyll a to chlorophyll b were all lower than the high water treatment (table 1).
FIG. 1 shows the response of photosynthetic rate (A) and stomatal conductance (B) of bletilla striata to light intensity (PAR) under different moisture treatments. Wherein, high water treatment (HW, ● - ●), medium water treatment (MW, □ - □) and low water treatment (LW, a-solidup). Error bars are the standard error of the mean (SE), n for high and medium water treatment is 7 and n for low water treatment is 4.
TABLE 1 comparison of photosynthetic parameters of bletilla under different moisture treatments
Note: LSP, light saturation point intensity; AQE, apparent quantum efficiency; rd, dark breathing rate; pmax, maximum photosynthetic; gs, gas hole conductivity; WUEi, instantaneous water use efficiency; chla, chlorophyll a content per unit area; chlb, chlorophyll b content per unit area; chl (a + b), total chlorophyll per unit area; Chla/Chlb, ratio of chlorophyll a to chlorophyll b. HW, high water treatment; MW, treating reclaimed water; LW, low water treatment. The data in the table are mean ± sem (n is 6-8), and the same row of data shows significant difference in different letters on the right side (P < 0.05).
Secondly, influence of moisture conditions on biomass of bletilla striata
The water treatment had a significant effect on the plant height of bletilla striata (fig. 2), leaf area, biomass and biomass distribution (table 2). The high water treated plants were taller, while the low water treated plants were generally shorter. Also, low water treatment resulted in individual mortality of 13.92%. Biomass also corroborates this phenomenon, with higher and medium water treated plants exhibiting significantly higher biomass gain than low water and with a negative increase in low water treated biomass, indicating that the organic matter of bletilla striata plants is more depleted than accumulated under low water treatment.
FIG. 2 is a graph showing the plant height frequency distribution of bletilla striata under different moisture treatments. HW, high water treatment; MW, treating reclaimed water; LW, low water treatment.
TABLE 2 Effect of different moisture treatments on leaf area and dry weight
Note: LA, total leaf area of individual plants; Δ DW, individual plant dry weight increment; DW of leaf, dry weight of individual plant leaves; DW of root, dry root weight of individual plants; DW of the new pseudobulbil, dry weight of new pseudobulb; DW of the old pseudobulb dry weight. HW, high water treatment; MW, treating reclaimed water; LW, low water treatment. The data in the table are mean ± sem (n ═ 9), the same row of data, with different letters on the right indicating significant differences (P < 0.05).
Thirdly, the influence of the water treatment on the content of the effective components of the pseudobulb of bletilla striata
The polysaccharide content is determined by phenol-sulfuric acid method, the pseudobulb is dried at 70 ℃, ground into powder and sieved by a 50-mesh sieve. Adding 0.3g of sample into 200mL of distilled water, refluxing in a boiling water bath for 2h, adding absolute ethyl alcohol into filtrate for precipitation, centrifuging, dissolving the precipitate in water to prepare a sample solution, measuring absorbance at 488nm by using an ultraviolet-visible spectrophotometer, and calculating the polysaccharide content in the sample by using a glucose standard curve.
The method for measuring the total phenol content comprises the following steps: drying pseudobulb, pulverizing, and sieving with 50 mesh sieve. Accurately weighing about 1g of powder, adding 200ml of 80% ethanol, refluxing in 80 ℃ water bath for 2h, filtering, placing in a flask with the weighed weight, performing rotary evaporation concentration, adding 70% ethanol into an extract sample, and performing ultrasonic dissolution to prepare a sample solution. Adding the welan color developing solution into a sample to be tested, standing for 10min in the dark, and then adding Na2CO3(20%) solution, 50 deg.C water bath 10min, ultraviolet visible spectrophotometer at 760nm to determine the absorbance, through gallic acid standard curve calculation sample in total phenol content.
Measuring the contents of mileanine and dactylorhin A by HPLC, drying pseudobulb, pulverizing, and sieving with 50 mesh sieve. Accurately weighing about 1g of powder, adding 200ml of 80% ethanol, refluxing in 80 ℃ water bath for 2h, filtering, placing in a flask with the weighed weight, performing rotary evaporation concentration, adding 70% ethanol into an extract sample, and performing ultrasonic dissolution to prepare a sample solution. And simultaneously preparing a reference solution, injecting samples respectively, recording the chromatogram, measuring the peak area and calculating the content of the medicinal materials.
The polysaccharide is not only the main effective component of bletilla striata, but also the energy storage substance of plants. In the three treated new pseudobulbs, the polysaccharide content increased significantly with increasing soil moisture content (fig. 3), indicating that moisture contributes to the accumulation of bletilla striata polysaccharides and has a significant effect.
FIG. 3 shows the total amount of polysaccharides, total phenols, militarinin content, and dactylorhin A content in the fresh pseudobulbs of bletilla striata under different moisture treatments. HW, high water treatment; MW, treating reclaimed water; LW, low water treatment. Error bars are standard errors of mean (SE), n is 3, different letters indicate significant difference (P < 0.05).
Total phenols are a generic term for a class of substances containing phenolic hydroxyl groups, and represent to some extent a large class of secondary metabolites. The total phenols in rhizoma Bletillae include bibenzyl, phenanthrene, eugenol, gallic acid, etc. The total phenolic content of the new pseudobulb in the low water treatment group was significantly higher than in the other two treatments (fig. 3), indicating that water stress can induce secondary metabolite accumulation.
Militarine and dactylorhin A are two benzyl ester compounds with similar structures, and are also secondary metabolites of bletilla striata. There was no significant difference in the amount of mileanine between the new pseudobulbs from the different moisture treatments, whereas the dactylorhin a from the low moisture treatment was significantly higher than the other two treatments (fig. 3), with a similar trend to the total phenol content. Researches show that the milettine and dactylorhin A are stored in the whole plant of the bletilla striata, belong to xenobiotic substances and are related to the weed inhibition effect of the bletilla striata. When the environment changes, they may be released into the surrounding environment to inhibit the growth of other plants nearby. It can be speculated that the increase of the content of dactylorhin A under the low water condition is probably the increase of secondary metabolites caused by stress stimulation, and the aim is to inhibit the growth of surrounding plants so as to ensure the superiority of the plants in water resource robbery.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A planting method for promoting growth of bletilla striata and accumulation of effective components comprises the following steps:
keeping the relative water content of the soil of 75-85% after the bletilla striata seedlings germinate to the mature period, and adjusting the relative water content of the soil to 20-30% before harvesting;
the method for calculating the relative water content of the soil comprises the following steps:
the relative water content of the soil is equal to the water content of the soil/field water capacity multiplied by 100 percent.
2. The planting method according to claim 1, wherein the germination is that the germination rate of bletilla striata reaches more than 95%.
3. The planting method according to claim 1, wherein the period before harvesting is 4-6 weeks before harvesting bletilla striata.
4. The planting method of claim 1, wherein the bletilla striata seedlings are selected from biennial bletilla striata pseudobulbs.
5. The planting method of claim 4, wherein the diameter of the biennial bletilla pseudobulb is 0.8-1.5 cm.
6. The growing method of claim 1, wherein said growing maximum light is 50% of full light.
7. The planting method of claim 1, wherein the planting temperature is 20-32 ℃.
8. The planting method of claim 1, wherein the relative humidity of the planting is 50-70%.
9. The planting method of claim 1, wherein the planting substrate comprises a mixture of leaf mold and red soil.
10. The planting method according to claim 9, wherein the volume ratio of the leaf mold to the red soil is (0.8-1.2): 1.
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