A method for limiting stem cultivation of disease-resistant and high-yield Pinellia ternata strains

Abstract

The present invention discloses a method for stem-limited cultivation of disease-resistant and high-yield Pinellia ternata strains, comprising the following steps: S1, screening high-quality Pinellia ternata strains according to leaf shape; through screening, selecting Pinellia ternata with oval leaves as high-quality Pinellia ternata strains; S2, screening disease-resistant Pinellia ternata strains; S3, stem-limited cultivation of disease-resistant Pinellia ternata strains. The present invention reduces the occurrence of Pinellia ternata diseases by starting from internal strain screening and external planting condition optimization, and provides high-yield and stable yield of Pinellia ternata artificial cultivation; at the same time, reduces the cost of artificial cultivation and the use of chemical agents such as fungicides, improves the quality of Pinellia ternata, and further improves the benefits of artificial cultivation of Pinellia ternata.

Description

Technical Field

[0001] The invention belongs to the technical field of agricultural planting, and particularly relates to a stem-limited cultivation method for a disease-resistant and high-yield Pinellia ternata strain. Background Art

[0002] Pinellia ternata is a perennial herbaceous plant of the Araceae family. Its tubers are used as medicine. It has a mild smell, spicy taste, numbing tongue and piercing throat. It is a traditional Chinese herbal medicine used to relieve cough and asthma. Pinellia ternata is also widely used in Southeast Asian countries. It is a mainstream variety in the Chinese market and one of the important commodities for export and foreign exchange earnings. The pinellia ternata grown in Jingmen area is called Jingbanxia. The tubers are large and full, powdery and white in color. It has always been a high-quality authentic herbal medicine among pinellia ternata.

[0003] Due to the good quality of Pinellia ternata and the large market demand, the supply is in short supply. Pinellia ternata mainly comes from artificial cultivation, and its artificial cultivation depends on the oil sand soil in Jingmen. Therefore, the artificial cultivation area is limited; Pinellia ternata artificial cultivation adopts open-air planting, which is easily affected by weather factors such as rainfall and high temperature. Diseases are common and the yield per mu fluctuates greatly. The lowest yield per mu is 500-600 catties, and the highest can reach more than 2,000 catties. In addition to being affected by the weather, cultivation management, especially disease and weed management, is particularly important for high yield. The main measure to reduce diseases is to apply a large amount of various fungicides and herbicides, which not only damages the soil ecological environment, but also leads to problems such as pesticide residues in products. Therefore, Pinellia ternata relies on artificial open-air cultivation of oil sand soil, which not only has extremely unstable yields, but also has problems such as soil ecological environment damage and pesticide residues.

[0004] There are generally 3-5 different leaf shapes in wild or cultivated populations of Pinellia ternata. Pinellia ternata with different leaf shapes may differ in growth rate, active ingredient content, disease resistance, etc., which lays the foundation for obtaining high-yield, stable-yield, and stable-quality Pinellia ternata strains through leaf shape-based strain screening. Xiao Pinguo et al. (2008) studied the differences between the four leaf shape populations of Pinellia ternata and believed that the differences between populations were much greater than the differences within populations, indicating that it is feasible to distinguish strains by leaf shape; they then conducted a comprehensive comparison of the four different leaf shapes of Pinellia ternata in terms of botanical morphology, plant biology and physiological characteristics, stress resistance, yield, and quality. Wei Lixia et al. (2014) obtained the strain BY-1 by tissue culture asexual rapid propagation. The strain has consistent growth and its active ingredients meet the requirements of the Pharmacopoeia of the People's Republic of China.

[0005] Strain screening is a way to discover high-yield and high-quality pinellia from the perspective of genetic internal conditions, while optimization of cultivation conditions is a method to improve the yield and quality of pinellia from the perspective of external conditions. At present, the yield and quality of pinellia are mainly improved by controlling cultivation conditions such as shading, fertilization, planting density, and seed stem size. Mei Yan et al. (2012) studied the effects of planting tubers of different sizes on the yield of pinellia, net increase in tuber yield, and tuber input-output ratio, and proposed that using 1.0-1.5 cm tubers as seed stems is more effective. Wang Xing et al. (2008) found that shading promotes the growth of pinellia tubers. Although existing studies have made certain progress in germplasm resource screening and cultivation conditions, there has been no systematic study from the screening of pinellia strains to an effective cultivation system; at present, the artificial cultivation of pinellia is mainly based on traditional methods. Patent document CN106538222A provides a three-dimensional cultivation method of pinellia in the shape of a letter A, which achieves the improvement of pinellia yield and production efficiency by optimizing the cultivation method, but its planting method is relatively not suitable for large-scale promotion.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a stem-limited cultivation method for a disease-resistant and high-yield Pinellia ternata strain, which reduces the occurrence of diseases in Pinellia ternata and provides high and stable yields of artificial cultivation of Pinellia ternata by starting from internal strain screening and external planting condition optimization. At the same time, it reduces the cost of artificial cultivation and the use of chemical agents such as fungicides, improves the quality of Pinellia ternata, and thereby improves the efficiency of artificial cultivation of Pinellia ternata.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] A method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain comprises the following steps:

[0010] S1. Screening high-quality Pinellia ternata strains according to leaf shape; through screening, selecting Pinellia ternata with oval leaves as the high-quality Pinellia ternata strain;

[0011] S2, screening of disease-resistant Pinellia ternata strains;

[0012] S3, stem-limited cultivation of disease-resistant Pinellia ternata strains;

[0013] Wherein, in step S3, stem-limited cultivation comprises the following steps:

[0014] S31, leveling the land and forming ridges;

[0015] S32, spraying the ridge bed with a mineral source potassium humate solution containing Bacillus subtilis, and then flattening the ridge bed;

[0016] S33, placing a planting frame on a ridge bed, then placing the tubers of the disease-resistant Pinellia ternata strain obtained by screening on the planting frame, covering it with a compost-treated substrate, and then mixing it with a composite material;

[0017] S34, spraying the mineral source potassium humate solution containing Bacillus subtilis once more after the leaves of Pinellia ternata unfold;

[0018] S35, manage the water and light.

[0019] As a preferred embodiment of the technical solution of the present invention, the screening in step S2 includes the following steps:

[0020] S21, taking the oval-leafed petiole of Pinellia ternata screened in step S1, sterilizing it and inoculating it in a first culture medium for one-step culture to form seedlings; then taking the petiole of the obtained seedling and inoculating it in a second culture medium to obtain embryonic callus; inoculating the embryonic callus in a third culture medium for suspension dark culture to obtain a suspension culture solution;

[0021] S22, taking the suspension culture obtained in step S21, discarding the supernatant after centrifugation, adding it to a filtered and sterilized sodium azide solution, shaking it well, soaking it, and adding Tween dropwise; then washing away the sodium azide, inoculating the obtained suspension culture cells in a differentiation medium for cultivation, acclimatizing and transplanting them after the seedlings grow to obtain tubers;

[0022] S23, first obtain the pathogenic bacteria solution, then place the soil where Pinellia ternata has been cultivated and where the disease is more serious in a cultivation pot, then mix in the obtained pathogenic bacteria solution, put into an artificial climate chamber, and finally place the tubers obtained in step S22 in the cultivation pot for cultivation. After disease resistance screening in two growth stages, a disease-resistant Pinellia ternata strain is obtained.

[0023] As a preferred embodiment of the technical solution of the present invention, in step S21, the composition of the first culture medium is: MS+6-BA 2.0 mg / L+2,4-D 0.1 mg / L;

[0024] The composition of the second culture medium is: MS+2,4-D 2.0 mg / L+6-BA 0.5 mg / L+sucrose 60 g / L;

[0025] The composition of the third culture medium is: MS+2,4-D 1.0 mg / L+6-BA 0.3 mg / L+hydrolyzed casein 100 mg / L.

[0026] As a preferred embodiment of the technical solution of the present invention, in step S22, the soaking time is 1 to 3 hours;

[0027] The concentration of sodium azide is 0.02 to 1 mmol / L;

[0028] The amount of Tween added was 2 drops / L;

[0029] The composition of the differentiation medium is: MS+6-BA 1.0mg / L+NAA 0.2mg / L.

[0030] As a preferred embodiment of the technical solution of the present invention, in step S23, the method for obtaining the pathogenic bacteria liquid includes the following steps: sterilizing the fourth culture medium under high pressure and then packaging it into sterilized culture dishes, taking tubers infected with soft rot and stem rot respectively, disinfecting them in an ultra-clean workbench after washing, and then placing them on sterilized paper, drying them and cutting them, picking out the diseased tissues, putting them in sterile water, and then smearing them on a NA plate, placing them at a constant temperature, and picking them out after the growth of plaques and inoculating them in the fourth culture medium for culture, and then separating them to obtain the pathogenic bacteria liquid.

[0031] As a preferred embodiment of the technical solution of the present invention, in step S23, the volume ratio of the pathogenic bacteria liquid to the soil is 1:100;

[0032] The fourth culture medium is composed of: 10 g of peptone, 3 g of beef extract, 5 g of NaCl, 15-20 g of agar, 1000 mL of water, and pH 7.4-7.6.

[0033] As a preferred embodiment of the technical solution of the present invention, in step S23, the environmental conditions of the artificial climate box are: in the first 2 weeks, the light is maintained at 12h / d, the temperature is controlled at 25°C during light, the temperature is controlled at 18°C ​​in darkness, the soil moisture is maintained at 60%, and the air humidity is maintained at 60%; after 2 weeks, the light is maintained at 12h / d, the temperature is controlled at 30°C during light, the temperature is controlled at 25°C in darkness, the soil moisture is maintained at 80%, and the air humidity is maintained at 80%.

[0034] As a preferred embodiment of the technical solution of the present invention, in step S31 and step S34, the mineral-source potassium humate solution containing Bacillus subtilis is prepared by adding 1 kg of mineral-source potassium humate per 100 L.

[0035] As a preferred embodiment of the technical solution of the present invention, in step S33, the covering thickness of the compost-treated substrate is 5 to 10 cm;

[0036] The preparation of the composting substrate comprises the following steps: crushing the straw waste and stacking it, evenly applying a Bacillus subtilis aqueous solution, covering it with a film to make the straw pile airtight, and when the temperature of the straw pile reaches 70-80°C, it is ready.

[0037] As a preferred embodiment of the technical solution of the present invention, in step S33, the mixing amount of the composite material is 50 to 80 g / m 2 ;

[0038] The preparation of the composite material comprises the following steps:

[0039] (1) mixing rectorite with sodium hydroxide and subjecting it to high temperature treatment, and grinding it to obtain pre-modified rectorite powder for standby use;

[0040] (2) mixing the vermiculite and fly ash and placing them in a hydrochloric acid solution for treatment, filtering, washing, drying, and finally grinding to obtain a pre-modified mixed powder for standby use;

[0041] (3) treating the bentonite at high temperature to obtain pre-modified bentonite;

[0042] (4) mixing the pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) with peat in a mass ratio of 1-1.2:2-3:2-3:1 to obtain a composite material;

[0043] Wherein, in step (1), the mass ratio of rectorite to sodium hydroxide is 1 to 0.4 to 0.5, the high temperature treatment temperature is 780 to 800° C., and the treatment time is 0.5 to 2 h;

[0044] Wherein, in step (2), the mass ratio of vermiculite to fly ash is 1:0.5-2, the concentration of the hydrochloric acid solution is 0.6-1 mol / L, the amount ratio of vermiculite to hydrochloric acid solution is 1g:5-12mL; the treatment temperature is 85-95°C, and the treatment time is 0.5-4h;

[0045] Wherein, in step (3), the high temperature treatment temperature of bentonite is 480-510°C, and the treatment time is 0.5-2h.

[0046] Compared with the prior art, the present invention has the following advantages:

[0047] 1. The present invention provides a method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain, which greatly reduces the occurrence of Pinellia ternata diseases and increases the per mu yield of Pinellia ternata to more than 2,000 jin per mu through the screening of Pinellia ternata germplasm resources and strains and the regulation of the growth environment of Pinellia ternata tubers.

[0048] 2. The stem-limited cultivation method of a disease-resistant and high-yield Pinellia ternata strain provided by the present invention can significantly reduce the use of chemical agents such as fungicides and avoid harmful residues in the soil and medicinal materials compared with the traditional field Pinellia ternata production method.

[0049] 3. The stem-limited cultivation method of a disease-resistant and high-yield Pinellia ternata strain provided by the present invention can significantly reduce labor costs in weeding and harvesting compared with traditional field Pinellia ternata production methods, thereby improving the production efficiency of Pinellia ternata.

[0050] 4. The present invention provides a method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain. By screening Pinellia ternata with different leaf shapes, it is found that the oval leaf shape is the best planting strain.

[0051] 5. The present invention provides a method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain. By optimizing the cultivation process, a Pinellia ternata strain with good disease resistance is obtained, thereby controlling the occurrence of diseases from the source.

[0052] 6. The present invention provides a method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain, which is mainly based on limiting the stem cultivation, which is another major invention of the present invention. It is easy to plant on a large scale and improve economic benefits.

[0053] 7. The present invention provides a method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain, which adopts a composting treatment + composite material supply mode to improve the planting quality and yield of Pinellia ternata. Specifically, the composting treatment is mainly based on straw waste, and a low-cost planting matrix can be obtained after simple fermentation; the composite material uses bentonite, vermiculite, retort, fly ash and peat as main raw materials, and the amount of peat is small because the source of peat is limited and the cost is relatively high; bentonite has poor permeability and is easy to form lumps, so it is simply subjected to high-temperature treatment to improve its adsorption capacity, reduce its expansibility, and thus improve its permeability; retort itself does not provide external nutrients, but its porous structure can promote soil gas exchange and root respiration, so it is subjected to high-temperature alkali treatment during use to further enrich its pore structure, and its amount is controlled to be relatively small;

[0054] Vermiculite has excellent water retention capacity, can improve soil permeability, and can provide some trace elements, but it has weak nutrient fixation capacity and is easy to pulverize; fly ash is a type of industrial by-product, which has been used in some occasions to improve its added value, but its matrix structure is unstable and its degradability is relatively poor. In this regard, the present invention performs acid treatment on the mixture of the two, mainly considering the following aspects: 1) acid treatment can remove potential heavy metal ions in fly ash and impurities in vermiculite, and reduce the adverse effects of the use of the two on the soil; 2) the soil environment for planting pinellia is acidic or neutral, and the system of vermiculite and fly ash is alkaline, and acid treatment is beneficial to regulating the pH value of the system of the two, so as to facilitate the growth of pinellia; 3) acid treatment is beneficial to improving the pore structure and surface area of ​​vermiculite and fly ash, improving water retention capacity and nutrient slow control capacity, and promoting the growth of pinellia.

[0055] In summary, the present invention can improve soil planting conditions, enhance soil moisture retention capacity, and promote the growth quality and yield of Pinellia ternata to the greatest extent while ensuring the nutritional components by optimizing the types of composite materials and performing corresponding treatments. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 Schematic diagram of different leaf shapes of Pinellia ternata in the present invention.

[0057] Figure 2 This is a schematic diagram of the limited stem cultivation of Pinellia ternata of the present invention. DETAILED DESCRIPTION

[0058] In order to make the purpose, technical solution and advantages of the present invention more clearly explained, the present invention will be further explained in detail below in conjunction with the embodiments of the specification.

[0059] In the present invention, all substances are purchased from the market unless otherwise specified. Among them, the mineral source potassium humate solution containing Bacillus subtilis was purchased from Shandong Lvlong Biotechnology Co., Ltd., with an effective viable count of more than 200 million / g.

[0060] The present invention provides a method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain, comprising the following steps:

[0061] S1. Screening high-quality Pinellia ternata strains according to leaf shape; through screening, selecting Pinellia ternata with oval leaves as the high-quality Pinellia ternata strain;

[0062] S2, screening of disease-resistant Pinellia ternata strains;

[0063] S3, stem-limited cultivation of disease-resistant Pinellia ternata strains;

[0064] The screening in step S2 includes the following steps:

[0065] S21, taking the oval-leafed petiole of Pinellia ternata screened in step S1, sterilizing it and inoculating it in a first culture medium for one-step culture to form seedlings; then taking the petiole of the obtained seedling and inoculating it in a second culture medium to obtain embryonic callus; inoculating the embryonic callus in a third culture medium for suspension dark culture to obtain a suspension culture solution;

[0066] S22, taking the suspension culture obtained in step S21, discarding the supernatant after centrifugation, adding it to a filtered and sterilized sodium azide solution, shaking it well, soaking it, and adding Tween dropwise; then washing away the sodium azide, inoculating the obtained suspension culture cells in a differentiation medium for cultivation, acclimatizing and transplanting them after the seedlings grow to obtain tubers;

[0067] S23, first obtain the pathogenic bacteria solution, then place the soil where Pinellia ternata has been cultivated and where the disease is more serious in a cultivation pot, then mix in the obtained pathogenic bacteria solution, put into an artificial climate chamber, and finally place the tubers obtained in step S22 in the cultivation pot for cultivation. After disease resistance screening in two growth stages, a disease-resistant Pinellia ternata strain is obtained.

[0068] In the above technical scheme, it is obvious that the planting quality of Pinellia ternata is improved by screening the internal disease-resistant strains and optimizing the external planting conditions. It should be particularly emphasized that one-step seedling formation and acclimatization and transplanting after seedling formation are conventional planting techniques in the field, and the present invention does not describe or limit the specific details thereof, which shall be subject to the ability of those skilled in the art to implement.

[0069] In some embodiments, the specific screening process of step S1 is as follows:

[0070] See also Figure 1 , from left to right, four leaf shapes of Pinellia ternata with higher occurrence frequency were selected from the population of Pinellia ternata, which were recorded as oval leaves (LY), bamboo leaves (ZY), needle leaves (NY), and bamboo leaves + (ZY+). 0.9 kg of tubers with a diameter of about 1.0 cm were taken from each leaf shape of Pinellia ternata and planted in a 1.5*2.0 m plot. There were 3 replicates for each leaf shape. Planted in the oil sand soil of Yonglang, Jingshan, and managed according to conventional management methods. After growing for one year and two growing seasons, the yield, reproduction coefficient, moisture, ash, water-soluble extract, total acid, and total alkali were counted. The yield reflects the growth rate of Pinellia ternata, which is an important agronomic trait like the reproduction coefficient. Moisture, ash, water-soluble extract, and total acid are the indicators set by the "Pharmacopoeia" for the quality of Pinellia ternata. Total alkaloids (total alkaloids) are the sum of the contents of various alkaloids. Alkaloids are considered to be the active ingredients of Pinellia ternata in many reports. The test results are shown in Table 1. As can be seen from Table 1, the oval-leaf Pinellia ternata had no significant difference from the other leaf shapes in terms of total acid, total alkali, ash content, and reproduction coefficient, but had the highest yield, extract, and total acid content among the four leaf shapes. Therefore, the oval-leaf Pinellia ternata was selected as a high-quality strain.

[0071] Table 1 Comparison of yield and quality indexes of Pinellia ternata with different leaf shapes

[0072]

[0073]

[0074] In some embodiments, in step S21, the composition of the first culture medium is: MS+6-BA 2.0 mg / L+2,4-D 0.1 mg / L;

[0075] The composition of the second culture medium is: MS+2,4-D 2.0 mg / L+6-BA 0.5 mg / L+sucrose 60 g / L;

[0076] The composition of the third culture medium is: MS+2,4-D 1.0 mg / L+6-BA 0.3 mg / L+hydrolyzed casein 100 mg / L.

[0077] In step S21, the rotation speed during dark culture is preferably 100 r / min.

[0078] In some embodiments, in step S22, the rotation speed during the centrifugal treatment is 5000 r / min; the soaking treatment time is 1 to 3 hours; preferably, 2 hours;

[0079] The concentration of sodium azide is 0.02-1 mmol / L; preferably, 0.5 mmol / L;

[0080] The amount of Tween added was 2 drops / L;

[0081] The composition of the differentiation medium is: MS+6-BA 1.0mg / L+NAA0.2mg / L.

[0082] In some embodiments, in step S23, the method for obtaining the pathogenic bacteria liquid comprises the following steps: sterilizing the fourth culture medium by high pressure and then dispensing it into the sterilized culture dish, taking the tubers with soft rot and stem rot respectively, disinfecting them in the clean bench after washing, then placing them on sterilized paper, cutting them after drying, picking out the diseased tissues, putting them in sterile water, then smearing them on the NA plate, placing them at a constant temperature (such as spinning in a 30°C constant temperature box for 24 hours), picking them out and inoculating them in the fourth culture medium for culture (30°C, 100r / min) after the plaque grows, and then separating them to obtain the pathogenic bacteria liquid. The specific steps of the above disinfection are: soaking in 70% alcohol for 1 minute, washing with sterile water 3 times, soaking and disinfecting with 0.2% HgCl2 for 20 minutes, and washing with sterile water 5 times.

[0083] In some embodiments, in step S23, the volume ratio of the pathogenic bacteria liquid to the soil is 1:100;

[0084] The fourth culture medium is composed of: 10 g of peptone, 3 g of beef extract, 5 g of NaCl, 15-20 g of agar, 1000 mL of water, and pH 7.4-7.6.

[0085] In some embodiments, in step S23, the soil is oil sand soil, and the soil sampling range is 1 cm to 11 cm below the surface soil.

[0086] In some embodiments, in step S23, the environmental conditions of the artificial climate box are: for the first 2 weeks, maintain light at 12h / d, control the temperature at 25°C during light, control the temperature at 18°C ​​in darkness, maintain soil moisture at 60%, and maintain air humidity at 60%; after 2 weeks, maintain light at 12h / d, control the temperature at 30°C during light, control the temperature at 25°C in darkness, maintain soil moisture at 80%, and maintain air humidity at 80%.

[0087] In some embodiments, in step S23, the strains for the final stem-limited cultivation can be selected as follows: select the single plants with strong growth potential for tissue culture and rapid propagation to obtain disease-resistant clones, and detect the moisture, ash, water-soluble extract, total acid, and total alkali of each clone. Select the clones whose moisture, ash, and water-soluble extract meet the requirements of the pharmacopoeia for stem-limited cultivation.

[0088] In some embodiments, see Figure 2 In step S3, the stem-limited cultivation includes the following steps:

[0089] S31, leveling the land and forming ridges;

[0090] S32, spraying the ridge bed with a mineral source potassium humate solution containing Bacillus subtilis, and then flattening the ridge bed;

[0091] S33, placing a planting frame on a ridge bed, then placing the tubers of the disease-resistant Pinellia ternata strain obtained by screening on the planting frame, covering it with a compost-treated substrate, and then mixing it with a composite material;

[0092] S34, spray after the leaves of Pinellia ternata unfold (additional spraying amount: 1L / m 2 ) a mineral-derived potassium humate solution containing Bacillus subtilis;

[0093] S35, manage the water and light.

[0094] In the above technical scheme, the cultivation of Pinellia ternata adopts stem-limited cultivation, which can greatly promote the root development of Pinellia ternata and increase the yield of Pinellia ternata; at the same time, it is convenient to save land resources, facilitate management, and improve the planting efficiency of Pinellia ternata.

[0095] In some embodiments, in step S31, the ridge bed height is 40 cm.

[0096] In some embodiments, in step S33, the bottom of the planting frame has mesh holes, and the mesh hole size is 30 mesh, 40 mesh, or 50 mesh; preferably, 40 mesh.

[0097] In some embodiments, in step S31 and step S34, the mineral-source potassium humate solution containing Bacillus subtilis is prepared by adding 1 kg of mineral-source potassium humate per 100 L.

[0098] In some embodiments, in step S33, the compost-treated substrate has a covering thickness of 5 to 10 cm;

[0099] The preparation of the composting substrate comprises the following steps: crushing the straw waste and stacking it, evenly applying the Bacillus subtilis aqueous solution, covering it with a film to make the straw pile airtight, and when the temperature of the straw pile reaches 70-80° C. The straw waste is rice husk, rapeseed husk, rice, wheat or corn straw, etc., and the straw material needs to be crushed into 0.5 cm particles.

[0100] In some embodiments, in step S33, the mixing amount of the composite material is 50 to 80 g / m 2 ; Preferably, 60g / m 2 .

[0101] It should be particularly emphasized that in step S35, the planting links such as water and care management can refer to the general Pinellia planting technology, which is general knowledge mastered by those skilled in the art.

[0102] The preparation of the composite material comprises the following steps:

[0103] (1) mixing rectorite with sodium hydroxide and subjecting it to high temperature treatment, and grinding it to obtain pre-modified rectorite powder for standby use;

[0104] (2) mixing the vermiculite and fly ash and placing them in a hydrochloric acid solution for treatment, filtering, washing, drying, and finally grinding to obtain a pre-modified mixed powder for standby use;

[0105] (3) treating the bentonite at high temperature to obtain pre-modified bentonite;

[0106] (4) The pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) are mixed with peat in a mass ratio of 1-1.2:2-3:2-3:1 to obtain a composite material; preferably, the mass ratio of each substance is 1:2.5:3:1.

[0107] Wherein, in step (1), the mass ratio of rectorite to sodium hydroxide is 1 to 0.4 to 0.5, the high temperature treatment temperature is 780 to 800° C., and the treatment time is 0.5 to 2 h;

[0108] Wherein, in step (2), the mass ratio of vermiculite to fly ash is 1:0.5-2, the concentration of the hydrochloric acid solution is 0.6-1 mol / L, the amount ratio of vermiculite to hydrochloric acid solution is 1g:5-12mL; the treatment temperature is 85-95°C, and the treatment time is 0.5-4h;

[0109] Wherein, in step (3), the high temperature treatment temperature of bentonite is 480-510°C, and the treatment time is 0.5-2h.

[0110] Next, the technical concept of the present invention is described in detail in conjunction with specific embodiments.

[0111] Example 1

[0112] A method for preparing a composite material comprises the following steps:

[0113] (1) Rectorite and sodium hydroxide are mixed in a mass ratio of 1:0.45 and subjected to high temperature treatment (790°C, 0.5h), and then ground to obtain pre-modified rectorite powder for later use;

[0114] (2) Vermiculite and fly ash are mixed (mass ratio 1:1) and placed in a 0.8 mol / L hydrochloric acid solution at a dosage ratio of 1 g:10 mL for treatment (90°C, 1.5 h), filtered, washed, dried, and finally ground to obtain a pre-modified mixed powder for later use;

[0115] (3) treating the bentonite at high temperature (500° C., 40 min), and obtaining pre-modified bentonite after the treatment;

[0116] (4) The pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) are mixed with peat in a mass ratio of 1:2.5:3:1 to obtain a composite material.

[0117] Example 2

[0118] A method for preparing a composite material comprises the following steps:

[0119] (1) mixing rectorite and sodium hydroxide in a mass ratio of 1:0.5 and subjecting the mixture to high temperature treatment (780°C, 1 h), and grinding the mixture to obtain pre-modified rectorite powder for later use;

[0120] (2) Vermiculite and fly ash are mixed (mass ratio 1:1) and placed in a 0.8 mol / L hydrochloric acid solution at a dosage ratio of 1 g:10 mL for treatment (85°C, 2 h), filtered, washed, dried, and finally ground to obtain a pre-modified mixed powder for later use;

[0121] (3) treating the bentonite at high temperature (500° C., 30 min), and obtaining pre-modified bentonite after the treatment;

[0122] (4) The pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) are mixed with peat in a mass ratio of 1:2.5:3:1 to obtain a composite material.

[0123] Comparative Example 1

[0124] A method for preparing a composite material comprises the following steps:

[0125] (1) Take a rector stone, grind it and set it aside;

[0126] (2) Vermiculite and fly ash are mixed (mass ratio 1:1) and placed in a 0.8 mol / L hydrochloric acid solution at a dosage ratio of 1 g:10 mL for treatment (90°C, 1.5 h), filtered, washed, dried, and finally ground to obtain a pre-modified mixed powder for later use;

[0127] (3) treating the bentonite at high temperature (500° C., 40 min), and obtaining pre-modified bentonite after the treatment;

[0128] (4) The rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) are mixed with peat in a mass ratio of 1:2.5:3:1 to obtain a composite material.

[0129] Comparative Example 2

[0130] A method for preparing a composite material comprises the following steps:

[0131] (1) Rectorite and sodium hydroxide are mixed in a mass ratio of 1:0.45 and subjected to high temperature treatment (790°C, 0.5h), and then ground to obtain pre-modified rectorite powder for later use;

[0132] (2) Mix the vermiculite and fly ash (mass ratio 1:1) and grind them to obtain a mixture for later use;

[0133] (3) treating the bentonite at high temperature (500° C., 40 min), and obtaining pre-modified bentonite after the treatment;

[0134] (4) The pre-modified rectorite powder obtained in step (1), the mixed material obtained in step (2), and the pre-modified bentonite obtained in step (3) are mixed with peat in a mass ratio of 1:2.5:3:1 to obtain a composite material.

[0135] Comparative Example 3

[0136] A method for preparing a composite material comprises the following steps:

[0137] (1) Rectorite and sodium hydroxide are mixed in a mass ratio of 1:0.45 and subjected to high temperature treatment (790°C, 0.5h), and then ground to obtain pre-modified rectorite powder for later use;

[0138] (2) Vermiculite and fly ash are mixed (mass ratio 1:1) and placed in a 0.8 mol / L hydrochloric acid solution at a dosage ratio of 1 g:10 mL for treatment (90°C, 1.5 h), filtered, washed, dried, and finally ground to obtain a pre-modified mixed powder for later use;

[0139] (3) Take bentonite and set aside;

[0140] (4) The pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the bentonite and peat obtained in step (3) are mixed in a mass ratio of 1:2.5:3:1 to obtain a composite material.

[0141] Application Examples

[0142] The disease-resistant Pinellia ternata strains obtained by screening are subjected to stem-limited cultivation, and the specific method comprises the following steps:

[0143] S31, leveling the land and forming ridges;

[0144] S32, spraying the ridge bed with a mineral source potassium humate solution containing Bacillus subtilis, and then flattening the ridge bed;

[0145] S33, place a planting frame (nylon net) with a mesh size of 40 on the ridge bed, then place the tubers (size 1-1.5 cm) of the disease-resistant Pinellia ternata strain obtained by screening on the planting frame, with a planting spacing of 4*4 cm, and then cover it with a compost-treated substrate (rice straw, covering thickness 6 cm), and then 2 The amount of is mixed into the composite material (Example 1 and Comparative Examples 1 to 3);

[0146] S34, spraying the mineral source potassium humate solution containing Bacillus subtilis once more after the leaves of Pinellia ternata unfold;

[0147] S35. Manage water and light, and monitor growth regularly until it is mature and ready for harvest.

[0148] It should be noted that the above planting methods were carried out in the same place, and each experimental group was randomly divided into three plots of equal size.

[0149] The pinellia tuber was harvested and the yield of the pinellia tuber was counted. The results are shown in Table 2.

[0150] Table 2

[0151]

[0152] Note: 1. When the test results are relative ratios, they are all based on Example 1.

[0153] 2. Since there is no unified classification of the incidence of Pinellia ternata, the above incidence statistics are for the overall incidence of Pinellia ternata.

[0154] It can be seen from Table 1 that the stem-limited cultivation method provided by the embodiment of the present invention has a high germination rate, a low overall disease rate, a high yield, a good soil and water conservation rate, and considerable economic benefits.

[0155] The above description describes the preferred implementation of the present invention, which should not be regarded as limiting the scope of protection of the claims of the present invention. Any modification, equivalent replacement and improvement without departing from the principle and idea of ​​the present invention should be regarded as within the scope of protection of the claims of the present invention.

Claims

1. A method for limiting the stem cultivation of a disease-resistant and high-yield Pinellia ternata strain, characterized in that: The steps include: S1. Screening high-quality Pinellia ternata strains according to leaf shape; through screening, selecting Pinellia ternata with oval leaves as the high-quality Pinellia ternata strain; S2, screening of disease-resistant Pinellia ternata strains; S3, stem-limited cultivation of disease-resistant Pinellia ternata strains; Wherein, in step S3, stem-limited cultivation comprises the following steps: S31, leveling the land and forming ridges; S32, spraying the ridge bed with a mineral source potassium humate solution containing Bacillus subtilis, and then flattening the ridge bed; S33, placing a planting frame on a ridge bed, then placing the tubers of the disease-resistant Pinellia ternata strain obtained by screening on the planting frame, covering it with a compost-treated substrate, and then mixing it with a composite material; S34, spraying the mineral source potassium humate solution containing Bacillus subtilis once more after the leaves of Pinellia ternata unfold; S35, manage water and light; Wherein, in step S33, the covering thickness of the compost treated substrate is 5 to 10 cm; The preparation of the substrate for composting treatment comprises the following steps: crushing the straw waste and stacking it, evenly applying the Bacillus subtilis aqueous solution, covering it with a film to make the straw pile airtight, and when the temperature of the straw pile reaches 70-80°C; In step S33, the mixing amount of the composite material is 50-80 g / m 2 ; The preparation of the composite material comprises the following steps: (1) mixing rectorite with sodium hydroxide and subjecting it to high temperature treatment, and grinding it to obtain pre-modified rectorite powder for later use; (2) mixing the vermiculite and fly ash and placing them in a hydrochloric acid solution for treatment, filtering, washing, drying, and finally grinding to obtain a pre-modified mixed powder for later use; (3) treating the bentonite at high temperature to obtain pre-modified bentonite; (4) mixing the pre-modified rectorite powder obtained in step (1), the pre-modified mixed powder obtained in step (2), and the pre-modified bentonite obtained in step (3) with peat in a mass ratio of 1-1.2:2-3:2-3:1 to obtain a composite material; Wherein, in step (1), the mass ratio of rectorite to sodium hydroxide is 1-0.4-0.5, the high temperature treatment temperature is 780-800°C, and the treatment time is 0.5-2h; Wherein, in step (2), the mass ratio of vermiculite to fly ash is 1:0.5-2, the concentration of the hydrochloric acid solution is 0.6-1 mol / L, the dosage ratio of vermiculite to hydrochloric acid solution is 1 g:5-12 mL; the treatment temperature is 85-95°C, and the treatment time is 0.5-4 h; Wherein, in step (3), the high temperature treatment temperature of bentonite is 480-510°C, and the treatment time is 0.5-2h.

2. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 1, characterized in that: The screening in step S2 includes the following steps: S21, taking the oval-leafed petiole of Pinellia ternata screened in step S1, sterilizing it and inoculating it in a first culture medium for one-step culture to form seedlings; then taking the petiole of the obtained seedling and inoculating it in a second culture medium to obtain embryonic callus; inoculating the embryonic callus in a third culture medium for suspension dark culture to obtain a suspension culture solution; S22, taking the suspension culture obtained in step S21, discarding the supernatant after centrifugation, adding it to a filtered and sterilized sodium azide solution, shaking it well, soaking it, and adding Tween dropwise; then washing away the sodium azide, inoculating the obtained suspension culture cells in a differentiation medium for cultivation, acclimatizing and transplanting them after the seedlings grow to obtain tubers; S23, first obtain the pathogenic bacteria solution, then place the soil where Pinellia ternata has been cultivated and where the disease is more serious in a cultivation pot, then mix in the obtained pathogenic bacteria solution, put into an artificial climate chamber, and finally place the tubers obtained in step S22 in the cultivation pot for cultivation. After disease resistance screening in two growth stages, a disease-resistant Pinellia ternata strain is obtained.

3. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 2, characterized in that: In step S21, the composition of the first culture medium is: MS+6-BA 2.0 mg / L+2,4-D 0.1 mg / L; The composition of the second culture medium was: MS +2,4-D 2.0 mg / L + 6-BA 0.5 mg / L + sucrose 60 g / L; The composition of the third culture medium is: MS +2,4-D 1.0 mg / L + 6-BA 0.3 mg / L + hydrolyzed casein 100 mg / L.

4. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 1, characterized in that: In step S22, the soaking time is 1 to 3 hours; The concentration of sodium azide is 0.02~1mmol / L; The amount of Tween added was 2 drops / L; The composition of differentiation medium is: MS+6-BA 1.0 mg / L+NAA 0.2 mg / L.

5. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 2, characterized in that: In step S23, the method for obtaining the pathogenic bacteria liquid includes the following steps: sterilizing the fourth culture medium under high pressure and then packaging it into sterilized culture dishes, taking tubers infected with soft rot and stem rot respectively, disinfecting them in an ultra-clean workbench after washing, and then placing them on sterilized paper, drying them and cutting them, picking out the diseased tissues, putting them in sterile water, and then smearing them on a NA plate, placing them at a constant temperature, and picking them out after the growth of bacterial plaques and inoculating them in the fourth culture medium for culture, and then separating them to obtain the pathogenic bacteria liquid.

6. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 2, characterized in that: In step S23, the volume ratio of the pathogenic bacteria liquid to the soil is 1:100; The composition of the fourth culture medium is: 10 g peptone, 3 g beef extract, 5 g NaCl, 15-20 g agar, 1000 mL water, pH 7.4-7.

6.

7. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 2, characterized in that: In step S23, the environmental conditions of the artificial climate box are: for the first 2 weeks, the light is maintained at 12h / d, the temperature is controlled at 25°C during light, the temperature is controlled at 18°C ​​during darkness, the soil humidity is maintained at 60%, and the air humidity is maintained at 60%; after 2 weeks, the light is maintained at 12h / d, the temperature is controlled at 30°C during light, the temperature is controlled at 25°C during darkness, the soil humidity is maintained at 80%, and the air humidity is maintained at 80%.

8. The method for limiting stem cultivation of a disease-resistant and high-yield Pinellia ternata strain according to claim 1, characterized in that: In step S31 and step S34, the mineral-source potassium humate solution containing Bacillus subtilis is prepared by adding 1 kg of mineral-source potassium humate per 100 L.

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