Planting method for increasing number of Yunnan pleione bulbocodioides seedlings

Abstract

The invention provides a planting method for increasing the number of Yunnan pleione bulbocodioides seedlings, and relates to the technical field of south pleione bulbocodioides seedling breeding. The method comprises the following steps that Yunnan pleione bulbocodioides pseudobulbs are provided; planting and field management; when the pseudobulb of the pleione bulbocodioides in Yunnan province begins to have terminal buds, auxin is sprayed; earthing up the periphery of the terminal bud of the pseudobulb of the Pleione bulbocodioides Yunnan; performing water management after the pseudobulb of the pleione bulbocodioides in Yunnan province grows a terminal bud; collecting terminal buds, and culturing the terminal buds again to obtain Yunnan pleione bulbocodioides seedlings. By means of the pseudobulb terminal bud proliferation mode, the number of Yunnan pleione bulbocodioides seedlings can be increased, and compared with a pseudobulb lateral bud proliferation mode, the number of Yunnan pleione bulbocodioides seedlings is increased by 4-5 times.

Description

Technical Field

[0001] This application relates to the field of Yunnan single-flower orchid seedling propagation technology, and in particular to a planting method for increasing the number of Yunnan single-flower orchid seedlings. Background Technology

[0002] *Pleione yunnanensis*, belonging to the Orchidaceae family and the *Pleione* genus, is mainly distributed in Yunnan, Sichuan, Guizhou, and Tibet, China. It is listed in the *Chinese Pharmacopoeia* and possesses properties of clearing heat and detoxifying, relieving cough and phlegm, stopping bleeding and promoting tissue regeneration. It is primarily used to treat tuberculosis, pertussis, bronchitis, carbuncles, gastrointestinal bleeding, and traumatic bleeding. In recent years, due to its anti-tumor activity, the market demand for *Pleione yunnanensis* has been increasing. However, its seeds are small and rarely germinate and grow successfully in nature. Furthermore, wild resources have been severely damaged. Therefore, obtaining a large number of intact regenerated plants through relevant technologies is an important technical means and a major way to alleviate resource and environmental pressures.

[0003] Currently, the main propagation methods for *Cymbidium goeringii* in Yunnan are seed propagation and pseudobulb propagation. The fruit of *Cymbidium goeringii* is typically a capsule, 5-9 cm in length. The seeds are very small, dusty, and lack endosperm; under natural conditions, the germination rate of *Cymbidium goeringii* seeds is relatively low. The method of increasing seedlings through the lateral buds of the pseudobulbs is currently the most common method used by most growers and enterprises. This method increases the number of seedlings by 1-2, but the increase is still very low. Summary of the Invention

[0004] In view of this, the purpose of this application is to provide a method for increasing the number of *Orchidonia yunnanensis* seedlings, thereby solving the problem of low seedling quantity obtained in existing technologies. The technical solution adopted by this application to solve the above-mentioned technical problem is as follows:

[0005] Embodiments of the present invention provide a method for increasing the number of *Cymbidium goeringii* seedlings in Yunnan, comprising the following steps:

[0006] We provide pseudobulbs of *Gynostemma pentaphyllum* from Yunnan.

[0007] Planting and field management;

[0008] When the pseudobulbs of *Gynostemma pentaphyllum* in Yunnan begin to develop terminal buds, spray with growth hormone.

[0009] Mound soil around the terminal bud of the pseudobulb of *Gynostemma pentaphyllum* in Yunnan.

[0010] Water management is carried out on the pseudobulbs of *Cymbidium goeringii* in Yunnan after they produce terminal buds.

[0011] The terminal buds were collected and cultured again to obtain Yunnan single-garlic orchid seedlings.

[0012] In some embodiments, pseudobulbs of *Gynostemma pentaphyllum* from Yunnan are selected in November to December and soaked in a 1x solution of C1 mancozeb wettable powder for t1 time.

[0013] In some embodiments, C1 is 80%; and / or

[0014] x1 is 600 to 800 times; and / or

[0015] t1 is 15 min to 20 min; and / or

[0016] The pseudobulbs of *Gynostemma pentaphyllum* in Yunnan have a longitudinal and transverse diameter of 20mm–30mm and 20mm–30mm; and / or

[0017] The individual pseudobulbs of *Cymbidium goeringii* in Yunnan weigh between 10g and 25g.

[0018] In some embodiments, before planting, a planting area is selected. The planting area has an altitude of 2700m to 3000m, an air relative humidity of 60% to 80%, a soil moisture content of 20% to 30%, a pH value of 6.5 to 7.5, and is a well-drained, deep, loose and fertile farmland or dry land.

[0019] In some embodiments, the planting area is prepared by evenly spreading farmyard manure and leaf mold in the planting area before preparation, turning it over, and making ridges with a ridge width of 90cm-105cm, a furrow width of 20cm-30cm, and a ridge height of 25cm-35cm; and / or

[0020] The planting area is covered with shade netting that provides 80% shading, with the netting positioned 1.8m to 2.2m above the ground; and / or

[0021] Dig planting pits within the planting area for one *Cymbidium goeringii* pseudobulb per pit, with a row spacing of 12-15 cm; and / or

[0022] Foliar spraying with urea solution and potassium dihydrogen phosphate solution should be applied 70-80 days after planting the pseudobulbs of *Cymbidium goeringii* in Yunnan.

[0023] In some embodiments, the auxin includes gibberellin, the concentration of which is 0.5 mg / L to 5 mg / L; and / or

[0024] Auxins include 6-benzylaminopurine, with concentrations ranging from 2 mg / L to 16 mg / L.

[0025] In some embodiments, the soil covering time is 10 to 15 days after the terminal buds appear at the top of the pseudobulb of *Gynostemma pentaphyllum*; and / or

[0026] The soil should be mounded 8 to 10 times; and / or

[0027] The soil covering method involves completely covering the terminal buds of the pseudobulbs of *Gynostemma pentaphyllum* with a soil covering substrate, but not covering the terminal bud leaves. The soil covering substrate includes peat moss, leaf mold, and well-rotted manure.

[0028] In some embodiments, after the pseudobulbs of *Gynostemma pentaphyllum* produce terminal buds, the soil moisture is maintained at 70%–75%.

[0029] In some embodiments, after October, the terminal buds detached from the pseudobulbs are collected. The terminal buds are first soaked in a C1 concentration mancozeb wettable powder solution diluted 1 times for t1 time, and then soaked in a C2 concentration indole-naphthaleneacetic acid wettable powder solution diluted 2 times for t2 time. The soaked terminal buds are then cultured in seedling trays at a plant spacing of a. During the culture period, the seedlings are sprayed with a C2 concentration indole-naphthaleneacetic acid wettable powder solution diluted 3 times for y times every t3 time interval, with a spraying amount of b. The seedlings are cultured until the terminal buds grow new roots and lateral buds, thus obtaining Yunnan single-clove orchid seedlings.

[0030] In some embodiments, C1 is 80%; and / or

[0031] x1 is 600 to 800 times; and / or

[0032] t1 is 20 min to 30 min; and / or

[0033] C2 is 5%; and / or

[0034] x2 is 300 to 500 times; and / or

[0035] t2 is 50 min to 60 min; and / or

[0036] t3 is 15d to 20d; and / or

[0037] x3 is 1000 times; and / or

[0038] y is from the first to the third degree; and / or

[0039] a is 3cm to 5cm; and / or

[0040] b is 8ml / m 2 ~10ml / m 2 .

[0041] Compared with the prior art, the beneficial effects of the present invention are:

[0042] This application demonstrates a method for increasing the number of *Cymbidium goeringii* seedlings in Yunnan through pseudobulb apical bud propagation. Compared to pseudobulb lateral bud propagation, this method increases the number of seedlings by 4-5 times. Traditional pseudobulb lateral bud propagation, planting one acre of seedlings at the beginning of the year, yields approximately one acre of pseudobulbs by the end of the year. Using this method, planting the same area results in 4-5 acres of pseudobulbs by the end of the year. This significantly increases the number of pseudobulbs in *Cymbidium goeringii*, leading to a substantial increase in its economic value and providing strong support for the development of the *Cymbidium goeringii* industry in Yunnan. Attached Figure Description

[0043] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings of the embodiments will be briefly described below. Obviously, the drawings described below only relate to some embodiments of this application and are not intended to limit this application, wherein:

[0044] Figure 1 The image shows a high-quality Yunnan single-clove orchid pseudobulb.

[0045] Figure 2 These are images showing the effect of planting pseudobulbs of *Cymbidium goeringii* in Yunnan.

[0046] Figure 3 This is a diagram showing the initial formation of the terminal bud;

[0047] Figure 4 These are before and after spraying GA3 and 6-BA and hilling.

[0048] Figure 5 These are images showing the effects after spraying GA3 and 6-BA, as well as after hilling.

[0049] Figure 6 This is a diagram showing the growth of the terminal bud;

[0050] Figure 7 This is a picture of the collected apical buds;

[0051] Figure 8 This is a diagram of the terminal bud being cultured again;

[0052] Figure 9 This is a picture showing the effect after the terminal bud has been cultured again; Figure 10 This is a flowchart of a planting method to increase the number of *Gynostemma pentaphyllum* seedlings in Yunnan. Detailed Implementation

[0053] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application. Furthermore, it should be understood that the specific embodiments described herein are only for illustration and explanation of this application and are not intended to limit this application.

[0054] In this application, unless otherwise stated, directional terms such as "upper" and "lower" generally refer to the upper and lower states of the device in actual use or operation; in addition, in the description of this application, the term "comprising" means "including but not limited to".

[0055] In this application, "and / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, or B existing alone. A and B can be singular or plural.

[0056] In this application, "at least one" means one or more, and "more than one" means two or more. "One or more", "at least one of the following", or similar expressions refer to any combination of these items, including any combination of single or multiple items. For example, "at least one of a, b, or c", or "at least one of a, b, and c" can both mean: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can be single or multiple.

[0057] Various embodiments of this application may exist in the form of a range; it should be understood that the description in the form of a range is merely for convenience and brevity and should not be construed as a hard limitation on the scope of this application; therefore, it should be considered that the range description has specifically disclosed all possible sub-ranges and single numerical values ​​within that range. For example, it should be considered that the range description from 1 to 6 has specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers within the range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. Furthermore, whenever a numerical range is referred to herein, it means including any referenced number (fraction or integer) within the referred range.

[0058] Currently, tissue culture is used to improve the germination rate of *Cymbidium goeringii* seeds in Yunnan, which is also one way to obtain high-quality seedlings. The method of increasing seedlings by using lateral buds from the pseudobulbs of *Cymbidium goeringii* is currently the method used by most planting enterprises and growers, increasing the number of seedlings by 1-2.

[0059] Firstly, please refer to Figure 10 The present invention provides a method for increasing the number of *Gynostemma pentaphyllum* seedlings in Yunnan, comprising the following steps:

[0060] S01, Providing pseudobulbs of Yunnan single-clove orchid;

[0061] S02, Planting and Field Management;

[0062] S03. Spray auxin when the pseudobulbs of Yunnan single-clove orchid begin to have terminal buds.

[0063] S04. Mound soil around the terminal bud of the pseudobulb of *Cymbidium goeringii* in Yunnan.

[0064] S05. Water management should be carried out after the pseudobulbs of Yunnan Garlic Orchid have produced terminal buds.

[0065] S06. Collect the terminal buds, cultivate them again, and obtain Yunnan single-garlic orchid seedlings.

[0066] This application demonstrates a method for increasing the number of *Cymbidium goeringii* seedlings in Yunnan through pseudobulb apical bud propagation. Compared to pseudobulb lateral bud propagation, this method increases the number of seedlings by 4-5 times. Traditional pseudobulb lateral bud propagation, planting one acre of seedlings at the beginning of the year, yields approximately one acre of pseudobulbs by the end of the year. Using this method, planting the same area results in 4-5 acres of pseudobulbs by the end of the year. This significantly increases the number of pseudobulbs in *Cymbidium goeringii*, leading to a substantial increase in its economic value and providing strong support for the development of the *Cymbidium goeringii* industry in Yunnan.

[0067] In S01:

[0068] Please see Figure 1 In some embodiments, pseudobulbs of *Gynostemma pentaphyllum* from Yunnan were selected in November to December and soaked in a 1x solution of C1 mancozeb wettable powder for t1 time.

[0069] Furthermore, C1 is 80%;

[0070] x1 can be 600x to 800x, for example, it can be 600x, 650x, 700x, 750x, 600x, etc.

[0071] t1 is 15 min to 20 min, for example, it can be 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, etc.

[0072] Furthermore, the longitudinal and transverse diameters of the pseudobulbs of *Gynostemma pentaphyllum* in Yunnan are 20mm-30mm x 20mm-30mm, for example, they can be 20mm x 20mm, 20mm x 22mm, 20mm x 25mm, 20mm x 28mm, 20mm x 30mm, 25mm x 20mm, 25mm x 22mm, 25mm x 25mm, 25mm x 28mm, 25mm x 30mm, 28mm x 20mm, 28mm x 22mm, 28mm x 25mm, 28mm x 28mm, 28mm x 30mm, 30mm x 20mm, 30mm x 22mm, 30mm x 25mm, 30mm x 28mm, 30mm x 30mm, etc.

[0073] The individual pseudobulbs of Yunnan single garlic orchid weigh between 10g and 25g, for example, 10g, 12g, 15g, 16g, 17g, 18g, 20g, 22g, 24g, 25g, etc.

[0074] It is understandable that the selected Yunnan single-clove orchid pseudobulbs should be free from pests and diseases and have plump lateral buds. Weeds, withered leaves and other impurities should be removed from the selected Yunnan single-clove orchid pseudobulbs to prevent the growth of pathogens.

[0075] In S02:

[0076] In some embodiments, a planting area is selected before planting. The altitude of the planting area is 2700m to 3000m, for example, 2700m, 2800m, 2900m, 3000m, etc.; the relative humidity of the air is 60% to 80%, for example, 60%, 65%, 70%, 75%, 80%, etc.; the soil moisture content is 20% to 30%, for example, 22%, 24%, 25%, 26%, 28%, 30%, etc.; the pH value is 6.5 to 7.5, for example, 6.5, 6.6, 6.8, 7, 7.2, 7.3, 7.4, 7.5, etc., and the planting area is farmland or dry land with good drainage, deep soil layer, loose and fertile soil.

[0077] Furthermore, for land preparation in the planting area, before land preparation, farmyard manure and leaf mold are evenly spread in the planting area, turned over and buried, and ridges are made. The ridge width is 90cm to 105cm, for example, 90cm, 92cm, 95cm, 98cm, 100cm, 102cm, 105cm, etc.; the furrow width is 20cm to 30cm, for example, 20cm, 22cm, 25cm, 28cm, 30cm, etc.; the ridge height is 25cm to 35cm, for example, 25cm, 26cm, 27cm, 28cm, 29cm, 30cm, 32cm, 35cm, etc.

[0078] Preferably, the width of the raised bed is 100cm, the width of the furrow is 25cm, and the height of the raised bed surface is 30cm.

[0079] It's understandable that the root system of Yunnan single-clove orchid generally grows in the soil layer of 10cm to 15cm, so high ridges are made in conjunction with fertilization.

[0080] Furthermore, the planting area is covered with shade netting that provides 80% shading, positioned 1.8m to 2.2m above the ground; for example, the netting can be 1.8m, 1.9m, 2.0m, 2.1m, or 2.2m above the ground.

[0081] Furthermore, dig ponds within the planting area to plant one pseudobulb of *Cymbidium goeringii* per pond. The planting row spacing for *Cymbidium goeringii* pseudobulbs is 12cm to 15cm, for example, 12cm, 12cm, 12cm, etc., and the plant spacing is 8cm to 12cm, for example, 8cm, 9cm, 10cm, 11cm, 12cm, etc.

[0082] It is understandable that after planting the pseudobulbs of Yunnan single-clove orchid, sufficient water should be given to help the roots establish, and the bed surface should be covered with pine needles after planting to retain water and fertilizer.

[0083] In some embodiments, the air humidity during the seedling stage of Yunnan single-garlic orchid is maintained at 50% to 75%, for example, it can be 50%, 52%, 55%, 58%, 60%, 62%, 65%, 68%, 70%, 72%, 75%, etc.

[0084] It's understandable that people choose to spray water on sunny afternoons or evenings in summer.

[0085] Please see Figure 2 In some embodiments, the pseudobulbs of *Gynostemma pentaphyllum* in Yunnan are sprayed with urea solution and potassium dihydrogen phosphate solution 70 to 80 days after planting.

[0086] Furthermore, the concentration of the urea solution is 1% to 3%, for example, it can be 1%, 1.2%, 1.5%, 1.8%, 2%, 2.2%, 2.5%, 2.8%, 3%, etc., and the number of sprays is 1 to 4 times, for example, 1 time, 2 times, 3 times, 4 times, etc.; spray once every 7 days, and the total spraying amount is 18 kg / 667 m³. 2 ~20Kg / 667m 2 For example, it can be 18Kg / 667m 2 18.5Kg / 667m 2 19Kg / 667m 2 19.5Kg / 667m 2 20Kg / 667m 2 wait.

[0087] Preferably, the urea solution concentration is 2%, and the application is performed three times, once every seven days, with a total application rate of 19 kg / 667 m³.2 .

[0088] Furthermore, the concentration of the potassium dihydrogen phosphate solution is 0.1%–0.3%, for example, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, etc., and the number of sprayings is 1 to 4 times, for example, 1, 2, 3, 4 times, etc.; spraying once every 7 days, with a spraying amount of 5 kg / 667 m³. 2 ~6Kg / 667m 2 For example, it can be 5Kg / 667m 2 5.2Kg / 667m 2 5.5Kg / 667m 2 5.8Kg / 667m 2 6Kg / 667m 2 wait.

[0089] It is understandable that the high temperature and humidity from June to September make it easy for some fungal diseases to occur. Therefore, prevention should be the main focus, and a comprehensive green control method combining agricultural, physical, and chemical control techniques should be adopted.

[0090] In S03:

[0091] Please see Figure 4 and Figure 5 In some embodiments, the auxin includes gibberellin, with a concentration of 0.5 mg / L to 5 mg / L, for example, 0.5 mg / L, 1 mg / L, 1.5 mg / L, 2 mg / L, 2.5 mg / L, 3 mg / L, 3.5 mg / L, 4 mg / L, 4.5 mg / L, 5 mg / L, etc.; the number of sprays is 1 to 3 times, for example, 1 time, 2 times, 3 times, etc.; spraying once every 10 days; the spraying amount is 18 ml / m². 2 ~20ml / m 2 For example, it can be 18ml / m 2 18.5ml / m 2 19ml / m 2 19.5ml / m 2 20ml / m 2 wait.

[0092] In some embodiments, the auxin includes 6-benzylaminopurine, and the concentration of 6-benzylaminopurine is 2 mg / L to 16 mg / L, for example, 2 mg / L, 3 mg / L, 4 mg / L, 5 mg / L, 6 mg / L, 7 mg / L, 8 mg / L, 9 mg / L, 10 mg / L, 11 mg / L, 12 mg / L, 13 mg / L, 14 mg / L, 15 mg / L, or 16 mg / L; the number of sprayings is 1 to 3 times, for example, 1 time, 2 times, or 3 times; spraying is done once every 10 days; and the spraying volume is 10 ml / m². 2 ~12ml / m 2 For example, it can be 10ml / m 2 10.5ml / m 2 11ml / m 2 11.5ml / m 2 12ml / m 2 wait.

[0093] In S04:

[0094] Please see Figure 3 and Figure 6 In some embodiments, the soil covering time is 10 to 15 days after the terminal buds of the pseudobulb of *Gynostemma pentaphyllum* emerge, for example, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, etc.

[0095] The soil should be mounded 8 to 10 times, for example, 8, 9, or 10 times.

[0096] In some embodiments, the hilling method includes completely covering the terminal bud of the pseudobulb of *Gynostemma pentaphyllum* with a hilling substrate, without covering the terminal bud leaves.

[0097] Furthermore, the soil substrate includes peat moss, leaf mold, and well-rotted manure.

[0098] For example, the soil substrate, by weight percentage, comprises 60% peat moss, 35% leaf mold, and 5% well-rotted manure.

[0099] It is understandable that decomposed manure includes decomposed pig manure or decomposed sheep manure.

[0100] It is understood that the potting mix is ​​sterilized by conventional methods in the art, for example, at a sterilization temperature of 100°C for 1.5 hours.

[0101] It's understandable that the top bud leaves are not covered, as this is to encourage the top bud to grow new roots. Any part with a top bud needs to be covered with soil.

[0102] It is understandable that soil is mounded around the terminal bud of the pseudobulb of *Gynostemma pentaphyllum* in Yunnan to facilitate the growth and rooting of the terminal bud.

[0103] In S05:

[0104] In some embodiments, after the pseudobulbs of *Gynostemma pentaphyllum* produce terminal buds, the soil moisture is maintained at 70%–75%, for example, 70%, 71%, 72%, 73%, 74%, 75%, etc.

[0105] It's understandable that after mounding soil, it's important to maintain soil moisture, as low moisture levels are detrimental to the growth of terminal buds.

[0106] In S06:

[0107] Please see Figure 7 and Figure 8 In some embodiments, the terminal buds detached from the pseudobulbs are collected after October. The terminal buds are first soaked in a C1 concentration mancozeb wettable powder solution at a dilution of x1 for t1 time, and then soaked in a C2 concentration indole-naphthaleneacetic acid wettable powder solution at a dilution of x2 for t2 time. The soaked terminal buds are then cultured in seedling trays at a plant spacing of a. During the culture period, the seedlings are sprayed with a C2 concentration indole-naphthaleneacetic acid wettable powder solution at a dilution of x3 times every t3 time interval, with a spraying amount of b. The seedlings are cultured until the terminal buds grow new roots and lateral buds, thus obtaining Yunnan single-clove orchid seedlings.

[0108] Furthermore, C1 is 80%;

[0109] x1 can be 600x to 800x, for example, it can be 600x, 650x, 700x, 750x, 800x, etc.;

[0110] t1 is 20 min to 30 min, for example, it can be 20 min, 22 min, 25 min, 28 min, 30 min, etc.;

[0111] C2 is 5%;

[0112] x2 can be 300x to 500x, for example, it can be 300x, 350x, 400x, 450x, 500x, etc.;

[0113] t2 is 50 min to 60 min, for example, it can be 50 min, 52 min, 55 min, 58 min, 60 min, etc.;

[0114] t3 is 15d to 20d, for example, it can be 15d, 16d, 17d, 18d, 19d, 20d, etc.;

[0115] x3 is 1000 times;

[0116] y can be from the first to the third degree, for example, it can be the first, second, third degree, etc.;

[0117] a is 3cm to 5cm, for example, it can be 3cm, 3.2cm, 3.5cm, 3.8cm, 4cm, 4.2cm, 4.5cm, 4.8cm, 5cm, etc.

[0118] b is 8ml / m 2 ~10ml / m 2 For example, it can be 8ml / m 2 8.5ml / m 2 9ml / m 2 9.5ml / m 2 10ml / m 2 wait.

[0119] For example, after October, most of the terminal buds will detach from the pseudobulb. At this time, the terminal buds need to be collected and soaked in a 600-fold dilution of 80% mancozeb wettable powder for 20 minutes. Then soak them in a 300-fold dilution of 5% wettable powder indole-naphthaleneacetic acid for 60 minutes. Then, the terminal buds are cultured in seedling trays at a spacing of 5 cm. Every 15 days, they are sprayed twice with a 1000-fold dilution of 5% wettable powder indole-naphthaleneacetic acid at a rate of 9 ml / m2. After 90-100 days of culture, new roots and lateral buds will appear, with a rooting rate and lateral bud formation rate of over 95%.

[0120] Test case

[0121] The effects of spraying different concentrations of gibberellin GA3 on *Gnaphalium affine* in Yunnan are shown in Table 1.

[0122] Table 1. Effects of spraying different concentrations of gibberellin (GA3)

[0123]

[0124]

[0125] As shown in Table 1, by spraying different concentrations of GA3 on *Gynostemma pentaphyllum* in Yunnan, it was found that the number of apical buds did not increase significantly when the GA3 concentration was low, but the number of apical buds decreased when the GA3 concentration was high, thus inhibiting the growth of apical buds. The number of apical buds reached its highest when the GA3 concentration was 2 mg / L.

[0126] Different concentrations of 6-BA were sprayed on *Gynostemma pentaphyllum* in Yunnan. The results are shown in Table 2.

[0127] Table 2. Effects of spraying different concentrations of 6-benzylaminopurine (6-BA)

[0128]

[0129] Table 2 shows that spraying different concentrations of 6-BA on *Gynostemma pentaphyllum* in Yunnan revealed that: at lower 6-BA concentrations, the number of terminal buds was not significant; at higher 6-BA concentrations, the number of terminal buds decreased, thus inhibiting their growth. The highest number of terminal buds was observed at a 6-BA concentration of 10 mg / L.

[0130] Simultaneously spray different concentrations of gibberellin (GA3) and 6-benzylaminopurine (6-BA), each sprayed 3 times, with an interval of 10 days. The spraying amount of GA3 was 19 ml / m³. 2 The application rate of 6-BA is 11 ml / m³. 2 The spraying effect is shown in Table 3:

[0131] Table 3. Effects of simultaneous spraying of different concentrations of gibberellin (GA3) and 6-benzylaminopurine (6-BA).

[0132]

[0133] Table 3 shows that by simultaneously spraying different concentrations of gibberellin (GA3) and 6-benzylaminopurine (6-BA), and by adding soil, increasing humidity and moisture, the number of apical buds increased significantly under the combined effect of GA3 and 6-BA. The number of apical buds was highest when the concentration of GA3 was 2 mg / L and the concentration of 6-BA was 10 mg / L, indicating that the effect was good.

[0134] Table 4 shows the comparison of the number of seedlings obtained by planting using traditional methods and planting using the method of this invention.

[0135] Table 4 Comparison of seedling quantity (including lateral buds and apical buds) between traditional planting and this method

[0136]

[0137] As shown in Table 4, by spraying gibberellin (GA3) and 6-benzylaminopurine (6-BA) on *Gnaphalium affine* in Yunnan, and by combining this with measures such as hilling, increasing humidity, and watering, the number of seedlings increased by 4 to 5 times (average 4.67 times) compared to traditional planting. For example, the number of seedlings increased from 50 to 69 after one year of traditional planting, while the number of seedlings increased from 50 to 248 after using this method, thus increasing the number of seedlings.

[0138] The effects of using 5% wettable powder indole-naphthaleneacetic acid (IAA) on the re-culturing of apical buds are shown in Table 5.

[0139] Table 5 shows the effect of using 5% wettable powder indole-naphthaleneacetic acid on the re-culturing of apical buds.

[0140] Lateral bud formation rate (%) Rooting rate (%) Spray once 76 69 Spray twice 98 95

[0141] Table 5 shows that spraying with 5% wettable powder indole-naphthaleneacetic acid during the re-culturing of terminal buds is beneficial for rooting and the generation of lateral buds.

[0142] Only through the re-cultivation of the terminal bud can the entire process of seedling formation be completed, resulting in a true Yunnan single-stem orchid seedling. This is especially true for the two crucial criteria of lateral bud formation and rooting. If the terminal bud cannot form lateral buds and roots, it is meaningless for future field planting. This invention achieves the re-cultivation of the terminal bud, with lateral bud formation and rooting rates exceeding 95%, providing a new approach and method for the future cultivation of Yunnan single-stem orchid seedlings.

[0143] The basic concepts have been described above. Obviously, for those skilled in the art, the detailed disclosure above is merely illustrative and does not constitute a limitation of this application. Although not explicitly stated herein, those skilled in the art may make various modifications, improvements, and corrections to this application. Such modifications, improvements, and corrections are suggested in this application, and therefore remain within the spirit and scope of the exemplary embodiments of this application.

[0144] Furthermore, this application uses specific terms to describe embodiments of the application. For example, "an embodiment," "one embodiment," and / or "some embodiments" refer to a particular feature, structure, or characteristic associated with at least one embodiment of the application. Therefore, it should be emphasized and noted that "an embodiment," "one embodiment," or "an alternative embodiment" mentioned twice or more in different locations in this specification do not necessarily refer to the same embodiment. In addition, certain features, structures, or characteristics in one or more embodiments of the application can be appropriately combined.

Claims

1. A method for increasing the number of *Gynostemma pentaphyllum* seedlings in Yunnan, characterized in that, Includes the following steps: We provide pseudobulbs of *Gynostemma pentaphyllum* from Yunnan. Planting and field management; When the pseudobulbs of *Gynostemma pentaphyllum* in Yunnan begin to develop terminal buds, spray with growth hormone. Mound soil around the terminal bud of the pseudobulb of *Gynostemma pentaphyllum* in Yunnan. Water management is carried out on the pseudobulbs of *Cymbidium goeringii* in Yunnan after they produce terminal buds. The terminal buds were collected and cultured again to obtain Yunnan single-garlic orchid seedlings.

2. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, Select Yunnan single-flower orchid pseudobulbs in November and December, and soak them in a 1x solution of C1 mancozeb wettable powder for t1 time.

3. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 2, characterized in that, C1 is 80%; and / or x1 is 600 to 800 times; and / or t1 is 15 min to 20 min; and / or The pseudobulbs of *Gynostemma pentaphyllum* in Yunnan have a longitudinal and transverse diameter of 20mm–30mm and 20mm–30mm; and / or The individual pseudobulbs of *Cymbidium goeringii* in Yunnan weigh between 10g and 25g.

4. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, Before planting, select a planting area with an altitude of 2700m to 3000m, relative humidity of 60% to 80%, soil moisture content of 20% to 30%, pH value of 6.5 to 7.5, and good drainage, deep soil layer, loose and fertile farmland or dry land.

5. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 4, characterized in that, For land preparation in the planting area, before preparation, evenly spread farmyard manure and leaf mold in the planting area, turn them over and cover them, and make ridges. The ridges should be 90cm-105cm wide, the furrows 20cm-30cm wide, and the ridge surface 25cm-35cm high; and / or The planting area is covered with shade netting that provides 80% shading, with the netting positioned 1.8m to 2.2m above the ground; and / or Dig planting pits within the planting area for one *Cymbidium goeringii* pseudobulb per pit, with a row spacing of 12-15 cm; and / or Foliar spraying with urea solution and potassium dihydrogen phosphate solution should be applied 70-80 days after planting the pseudobulbs of *Cymbidium goeringii* in Yunnan.

6. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, Auxins include gibberellins, with concentrations ranging from 0.5 mg / L to 5 mg / L; and / or Auxins include 6-benzylaminopurine, with concentrations ranging from 2 mg / L to 16 mg / L.

7. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, The soil should be prepared 10-15 days after the terminal bud appears at the top of the pseudobulb of *Gynostemma pentaphyllum*; and / or The soil should be mounded 8 to 10 times; and / or The soil covering method involves completely covering the terminal buds of the pseudobulbs of *Gynostemma pentaphyllum* with a soil covering substrate, but not covering the terminal bud leaves. The soil covering substrate includes peat moss, leaf mold, and well-rotted manure.

8. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, After the pseudobulbs of *Gynostemma pentaphyllum* in Yunnan produce terminal buds, maintain soil moisture at 70%–75%.

9. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 1, characterized in that, After October, collect the terminal buds that have detached from the pseudobulbs. The terminal buds are first soaked in a C1 concentration mancozeb wettable powder solution diluted 1 times for t1 time, and then soaked in a C2 concentration indole-naphthaleneacetic acid wettable powder solution diluted 2 times for t2 time. The soaked terminal buds are then cultured in seedling trays at a plant spacing of a. During the culture period, every t3 time interval, spray with a C2 concentration indole-naphthaleneacetic acid wettable powder solution diluted 3 times y times, with a spraying amount of b. Culture until the terminal buds grow new roots and lateral buds to obtain Yunnan single garlic orchid seedlings.

10. The planting method for increasing the number of *Gynostemma pentaphyllum* seedlings according to claim 9, characterized in that, C1 is 80%; and / or x1 is 600 to 800 times; and / or t1 is 20 min to 30 min; and / or C2 is 5%; and / or x2 is 300 to 500 times; and / or t2 is 50 min to 60 min; and / or t3 is 15d to 20d; and / or x3 is 1000 times; and / or y is 1 to 3 times; and / or a is 3cm to 5cm; and / or b is 8ml / m 2 ~10ml / m 2 .

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