Propagation method for improving cutting rooting rate of pueraria lobata

Abstract

The invention belongs to the technical field of plant propagation, and particularly relates to a propagation method for improving the cutting rooting rate of pueraria lobata, which comprises the following steps: pruning mother branches of pueraria lobata to obtain cutting slips, and disinfecting the cutting slips to obtain disinfected cutting slips; the disinfected cutting slips are treated with a growth regulator, and the cutting slips treated with the growth regulator are obtained; and inserting the cutting slips treated by the growth regulator into a seedbed matrix, and carrying out cuttage culture to obtain pueraria lobata breeding seedlings. According to the method, stems of leguminous plant pueraria lobata are used as raw materials, a cuttage vegetative propagation mode is adopted, and the rooting rate of pueraria lobata cuttage propagation is increased by selecting a plurality of key parameters such as proper types and concentrations of growth regulators and controlling treatment time. Results show that the rooting rate of pueraria lobata cutting propagation can be remarkably increased by using the mixed growth regulator for treatment. The method is simple, convenient, feasible and environment-friendly, can effectively promote the large-scale breeding of the pueraria lobata excellent germplasm material, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of plant propagation technology, specifically relating to a propagation method for improving the rooting rate of wild kudzu cuttings. Background Technology

[0002] Wild kudzu (Pueraria lobata (Willd.) Ohwi) is a perennial herbaceous vine belonging to the genus Pueraria in the legume family. Its dried root is included in the 2025 edition of the Chinese Pharmacopoeia. As a medicinal resource, it has the effects of promoting body fluid production and quenching thirst, relieving muscle tension and reducing fever, promoting blood circulation and relieving stagnation, invigorating yang and quenching thirst, and detoxifying alcohol. It is known as "Northern Ginseng and Southern Kudzu". With the development and application of kudzu root-related products, wild resources can no longer meet market demand, so there is an urgent need to develop artificial cultivation to improve its yield and quality.

[0003] In recent years, many regions have attempted to introduce and cultivate wild kudzu, but due to low propagation rates and unstable yields, industrialization and large-scale production have not yet been achieved. Seed propagation faces numerous limitations, including limited seed sources, long growth cycles, and low germination rates. Some varieties even fail to flower in the same year of planting, restricting its application in practical production. In contrast, cutting propagation, with its high propagation coefficient, short cycle, and lack of seasonal limitations, has become an important means of rapid propagation of wild kudzu. Cutting propagation not only effectively maintains the superior traits of the parent plant and achieves high genetic gain, but also allows for the propagation of high-quality germplasm materials in a relatively short time. Currently, wild kudzu cutting propagation technology faces bottlenecks such as low rooting rates and low propagation efficiency, hindering its large-scale application. Therefore, developing a propagation method that can significantly improve the rooting rate of wild kudzu cuttings is crucial. Summary of the Invention

[0004] The purpose of this invention is to provide a propagation method that improves the rooting rate of kudzu cuttings. This invention uses a mixed growth regulator treatment, which can significantly improve the rooting rate of kudzu cuttings. This method is simple, easy to implement, and environmentally friendly, and can effectively promote the large-scale breeding of superior kudzu germplasm materials, and has broad application prospects.

[0005] This invention provides a propagation method for improving the rooting rate of kudzu cuttings, comprising the following steps:

[0006] Cuttings are obtained by pruning the mother branches of wild kudzu. The cuttings are then disinfected to obtain disinfected cuttings.

[0007] The disinfected cuttings were treated with a growth regulator to obtain growth regulator-treated cuttings.

[0008] Cuttings treated with growth regulators were inserted into the seedbed substrate for propagation to obtain wild kudzu seedlings.

[0009] The growth regulator comprises a combination of at least two of IAA, IBA, and NAA, wherein the concentration of IAA is 200–500 mg·L⁻¹. -1 The concentration of IBA is 200–500 mg·L⁻¹. -1 The concentration of NAA is 200–500 mg·L⁻¹. -1 ;

[0010] The treatment time with the growth regulator is 0.5 to 5 hours.

[0011] As a preferred embodiment, the pruning method is as follows: the cutting is 10-12cm long and 0.8cm or more in diameter, retains one bud, the upper morphological cut is flat and 2-2.5cm away from the bud, and the lower morphological cut is at a 45° angle and 8-9cm away from the bud.

[0012] As a preferred embodiment, the disinfectant solution is a 0.1% carbendazim solution, and the disinfection time is 10-15 minutes.

[0013] As a preferred method, when treating with growth regulators, the lower 1.5–2.4 cm of the disinfected cuttings is inserted into the growth regulator.

[0014] As a preferred embodiment, the seedbed substrate includes river sand + vermiculite or loam + river sand.

[0015] As a preferred option, when the seedbed substrate is river sand + vermiculite, the volume ratio of river sand + vermiculite is 1:1;

[0016] When the seedbed substrate is loam + river sand, the volume ratio of loam + river sand is 1:1.

[0017] As a preferred embodiment, the seedbed substrate is disinfected with a 0.5% KMnO4 solution 4 days before cutting, and then covered with a film for 3 days.

[0018] As a preferred embodiment, the plant spacing during cutting is 5cm and the row spacing is 10cm.

[0019] As a preferred embodiment, the cuttings are inclined at 60° to the seedbed substrate during propagation, with the buds of the cuttings just protruding from the substrate surface.

[0020] As a preferred method, the temperature for cutting culture is 15–25℃, and the substrate moisture content is 60%–80%.

[0021] Beneficial Effects: This invention provides a propagation method for improving the rooting rate of kudzu cuttings, comprising the following steps: pruning kudzu mother branches to obtain cuttings; disinfecting the cuttings to obtain disinfected cuttings; treating the disinfected cuttings with a growth regulator to obtain growth regulator-treated cuttings; inserting the growth regulator-treated cuttings into a seedbed substrate for propagation to obtain kudzu seedlings. This invention uses stem segments of the legume kudzu as raw material and employs asexual propagation by cuttings. By selecting suitable types and concentrations of growth regulators and controlling treatment time, among other key parameters, the rooting rate of kudzu cutting propagation is improved. Results show that treatment with mixed growth regulators significantly improves the rooting rate of kudzu cutting propagation. This method is simple, easy to implement, and environmentally friendly, effectively promoting the large-scale propagation of superior kudzu germplasm materials and has broad application prospects.

[0022] The present invention also has the following advantages:

[0023] 1. The wild kudzu propagation method of the present invention can significantly improve the rooting rate of wild kudzu cutting propagation and improve the problems of low rooting rate and low propagation efficiency in traditional wild kudzu seedling cultivation methods.

[0024] 2. This invention uses a combination of growth regulators to treat kudzu cuttings, which can effectively overcome the rooting inhibition of rooting agents (such as tannins, resins, organic acids, etc.) in the plant, thereby greatly improving the reliability and efficiency of seedling cultivation.

[0025] 3. The wild kudzu propagation method adopted in this invention is low in cost, simple and feasible to operate, environmentally friendly, and has broad application prospects and great economic value. Attached Figure Description

[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0027] Figure 1 The morphology of kudzu cuttings at different stages of rooting;

[0028] Figure 2 The condition of a kudzu cutting 60 days after cutting. Detailed Implementation

[0029] This invention provides a propagation method for improving the rooting rate of kudzu cuttings, comprising the following steps:

[0030] Cuttings are obtained by pruning the mother branches of wild kudzu. The cuttings are then disinfected to obtain disinfected cuttings.

[0031] The disinfected cuttings were treated with a growth regulator to obtain growth regulator-treated cuttings.

[0032] Cuttings treated with growth regulators were inserted into the seedbed substrate for propagation to obtain wild kudzu seedlings.

[0033] This invention first prunes the mother branches of wild kudzu to obtain cuttings, which are then disinfected to obtain disinfected cuttings. The pruning method described in this invention can be as follows: select one-year-old, two-year-old, or multi-year-old wild kudzu branches, cut them to a length of 10-12cm and a diameter of at least 0.8cm, retaining one plump bud without leaves. The upper morphological cut is flat, 2-2.5cm from the bud, and the lower morphological cut is at a 45° angle, 8-9cm from the bud. The cuts should be smooth and free from mechanical damage. As a specific embodiment, the cutting length can be any one of 10cm, 11cm, and 12cm, or any value between two values; the diameter can be any one of 0.8cm, 0.9cm, 1.0cm, and 1.1cm, or any value greater than 0.8cm. The distance between the upper cut and the bud can be any one of 2.0cm, 2.1cm, 2.2cm, 2.3cm, and 2.5cm, or any value between two of these. If the distance is too short, sufficient water cannot be stored to support the bud's germination and initial growth due to transpiration; if it is too long, the rapid growth of the new shoot will consume nutrients and affect root growth, resulting in a "false survival" phenomenon. As a specific implementation method, the distance between the lower cut and the bud can be any one of 8.0cm, 8.1cm, 8.2cm, 8.3cm, 8.4cm, 8.5cm, 8.6cm, 8.7cm, 8.8cm, 8.9cm, and 9.0cm, or any value between two of these. If the lower cut is too short, nutrient storage will be insufficient; if it is too long, water cannot be effectively replenished before root formation, and excessive water loss will cause the cutting to wilt. Furthermore, if inserted too deeply into the substrate, anaerobic bacteria can easily proliferate, causing root rot. The cutting length of this invention provides an appropriate amount of nutrients for early growth, and the nutrient transport path is of suitable length, reducing losses and ensuring that nutrients are efficiently and timely delivered to various organs to meet their growth and development needs, thus ensuring the balanced growth of the cutting.

[0034] The disinfection solution of the present invention can be a carbendazim solution with a mass percentage of 0.1%, and the disinfection time can be 10 to 15 minutes. As a specific embodiment, the disinfection time can be any one of 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes and 15 minutes, or any value between two values.

[0035] Then, the disinfected cuttings are treated with a growth regulator to obtain growth regulator-treated cuttings. The growth regulator of this invention comprises a combination of at least two of IAA, IBA, and NAA, wherein the concentration of IAA is 200–500 mg·L⁻¹.-1 In one specific embodiment, the concentration of the IAA can be 200 mg·L⁻¹. -1 300mg·L -1 400 mg·L -1 and 500 mg·L -1 Any one of the values ​​and any value between the two. The concentration of IBA is 200–500 mg·L⁻¹. -1 In one specific embodiment, the concentration of IBA can be 200 mg·L⁻¹. -1 300mg·L -1 400 mg·L -1 and 500 mg·L -1 Any one of the values ​​and any value between the two. The concentration of NAA is 200–500 mg·L⁻¹. -1 In one specific embodiment, the concentration of NAA can be 200 mg·L⁻¹. -1 300mg·L -1 400 mg·L -1 and 500 mg·L -1 Any one of the values ​​and any value between the two values. In a specific embodiment of the present invention, the growth regulator can be 300 mg·L⁻¹. -1 IAA + 300mg·L -1 IBA combination solution, 300 mg·L -1 IAA + 300mg·L -1 NAA combination solution, 300 mg·L -1 IBA + 300mg / L -1 NAA combination solution and 300 mg·L -1 IAA + 300mg·L -1 IBA + 300mg / L -1 The NAA combination solution. The wild kudzu mother branches described in this invention may include one-year-old tender branches, two-year-old or perennial hard branches; the treatment time of the growth regulator described in this invention is 0.5 to 5 hours. As a specific embodiment, the treatment time of the growth regulator can be any one of 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours and 5 hours, or any value between two values.

[0036] When treating the cuttings with the growth regulator, the lower end of the sterilized cuttings can be inserted into the growth regulator at a depth of 1.5–2.4 cm. If the insertion length is too long, excessive hormone absorption will inhibit rooting and increase the risk of rot; if it is too short, the effective treatment area will be insufficient. As a specific embodiment, the length of the lower end of the cutting inserted into the growth regulator can be any one of 1.5 cm, 1.6 cm, 1.7 cm, 1.8 cm, 1.9 cm, 2.0 cm, 2.1 cm, 2.2 cm, 2.3 cm, and 2.4 cm, or any value between two of these two values.

[0037] The embodiments of the present invention show that when using kudzu branches of different ages as materials, and employing 300 mg·L⁻¹... -1 IAA + 300mg·L -1 After treating the cuttings with an IBA solution for 3 hours, statistical analysis was performed on the surviving plants on day 45. One-year-old cuttings showed the best growth, with a rooting rate of 91.67%. The average number of roots per cutting was 10.26 ± 0.57, the average total root length was 14.48 ± 2.28 cm, and the average maximum root length was 2.52 ± 0.22 cm, indicating a well-developed root system. When using kudzu branches of different ages as material, treated with 300 mg·L⁻¹ solution… -1 IAA + 300mg·L -1 When treated with NAA combined solution for 3 hours, the growth of one-year-old and two-year-old cuttings was similar, with one-year-old cuttings having a slightly higher rooting rate. The latter, however, showed significantly better average number of roots per cutting, average total root length, and average maximum root length compared to the former. When using kudzu branches of different ages as material, treated with 300 mg·L⁻¹ NAA combined solution... -1 IBA + 300mg / L -1 When treated with NAA combined solution for 3 hours, the two-year-old cuttings showed good growth, with a rooting rate of 55.38%, an average of 9.17±1.49 roots per cutting, an average total root length of 10.51±1.99 cm, and an average maximum root length of 2.03±0.06 cm. When using kudzu branches of different ages as material, treated with 300 mg·L⁻¹ NAA combined solution... -1 IAA + 300mg·L -1 IBA + 300mg / L -1When treated with the NAA combination solution for 3 hours, the rooting rate of perennial cuttings reached 81.58%, with an average of 12.47±0.71 roots per cutting, an average total root length of 14.41±0.88 cm, and an average maximum root length of 1.97±0.09 cm. In summary, the propagation effect of kudzu cuttings is closely related to the age of the cuttings and the matching degree of the plant growth regulator combination. One-year-old cuttings showed significant advantages with the IAA+IBA combination, two-year-old cuttings were more suitable for the IAA+NAA or IBA+NAA combination, while perennial cuttings performed best when all three regulators were used in combination. These experimental results provide precise technical references for the propagation of kudzu cuttings of different ages, clarifying the optimal regulator combination schemes for cuttings of various ages, and helping to improve the efficiency and quality of kudzu cutting propagation.

[0038] Finally, the cuttings treated with growth regulators were inserted into the seedbed substrate for propagation to obtain wild kudzu seedlings.

[0039] The seedbed substrate described in this invention may include river sand + vermiculite or loam + river sand. As a specific embodiment, when the seedbed substrate is river sand + vermiculite or loam + river sand, the volume ratio of river sand + vermiculite or loam + river sand is 1:1. This invention, through statistical analysis of the growth of one-year-old kudzu cuttings in different substrates, found that using river sand + vermiculite (1:1, v / v) as a substrate can balance aeration and moisture retention, which is beneficial for the rooting of kudzu cuttings. The rooting rate was 88.18%, the average number of roots per cutting was 8.19±0.52, the average total root length was 10.04±2.93 cm, and the average maximum root length was 2.21±1.00 cm. The second best result was achieved with loam mixed with river sand (1:1, v / v), which had a rooting rate of 80.00%, an average of 5.45±0.55 roots per cutting, an average total root length of 4.69±1.01cm, and an average maximum root length of 1.23±0.20cm.

[0040] The seedbed substrate of the present invention is disinfected by spraying with a KMnO4 solution of 0.5% by mass 4 days before cutting, and the volume ratio of substrate to disinfectant solution is 80:1. After disinfection, the substrate is covered with film for 3 days.

[0041] The cuttings of this invention can be spaced 5cm apart and 10cm apart in rows. The cuttings are placed at a 60° angle to the seedbed substrate, with the buds just protruding from the substrate surface. The temperature for cultivation is 15–25℃, and the substrate humidity is 60%–80%. As a specific embodiment, the cultivation temperature can be any one of 15℃, 16℃, 17℃, 18℃, 19℃, 20℃, 21℃, 22℃, 23℃, 24℃, and 25℃, or any value between two of these. In one specific implementation, the humidity of the substrate for the cutting culture can be any one of 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, and 80%, or any value between two values.

[0042] To further illustrate the present invention, the following detailed description of a propagation method for improving the rooting rate of wild kudzu cuttings, in conjunction with embodiments, is provided by the present invention, but should not be construed as limiting the scope of protection of the present invention.

[0043] Unless otherwise specified, the present invention does not have special requirements for the raw materials used in the preparation, and commercially available products well known to those skilled in the art can be used.

[0044] The reagents used in this invention are shown in Table 1.

[0045] Table 1 Reagent Instructions

[0046] name Manufacturer batch number Specification 3-Indoleacetic acid (IAA) Hangzhou Mumu Biotechnology Co., Ltd. 20230501 5g 3-Indolebutyric acid (IBA) Hangzhou Mumu Biotechnology Co., Ltd. 20230508 5g Naphthaleneacetic acid (NAA) Hangzhou Mumu Biotechnology Co., Ltd. 20230612 50g potassium permanganate Kunming Ketewei Biotechnology Co., Ltd. 24071201 500g / bottle Carbendazim Sichuan Runer Technology Co., Ltd. 230530002 50g

[0047] Example 1

[0048] (1) Seedbed preparation: land preparation, ridging, and substrate preparation. Four days before cutting, the substrate was disinfected with a 0.5% KMnO4 solution, and then covered with plastic film. The film was removed one day before cutting. No other fertilizer was applied.

[0049] (2) Material selection: Based on the degree of lignification of the mother branch, select semi-lignified tender branches (one-year-old) and fully lignified hard branches. The latter are further subdivided into two-year-old and multi-year-old hard branches according to the bark color, hardness and cross-sectional structural characteristics. They should be healthy, free from diseases, pests and damage.

[0050] (3) Cutting preparation: Select one-year-old, two-year-old, and multi-year-old kudzu branches for batch experiments. Each treatment contains 40 cuttings, with three replicates and one replicate. Cut the stems into segments 10-12cm long and 0.8cm or more in diameter, retaining one plump bud without leaves. The upper morphological end is cut flat, 2-2.5cm from the bud, and the lower morphological end is cut at a 45° angle, 8-9cm from the bud. The cuts are smooth and free of mechanical damage.

[0051] (4) Cutting treatment: Bundle the cuttings neatly, immerse them completely in a 0.1% carbendazim solution for 10 minutes for disinfection, and then air dry them. Apply a 300 mg·L⁻¹ solution to the lower 2 cm of each cutting. -1 IAA + 300mg·L -1 The IBA combination solution was used for treatment for 3 hours.

[0052] (5) Cutting method: First, use a stick of the same thickness as the cutting to make holes in the substrate (river sand + vermiculite, volume ratio of 1:1), with a plant spacing of 5cm, a row spacing of 10cm, and a depth of 8cm, at a 60° angle to the substrate surface. The depth of the hole should be such that the bud just protrudes from the substrate surface. Place the cutting in the substrate so that the substrate just covers the bud. Compact the substrate on both sides appropriately and water thoroughly.

[0053] (6) Seedbed management: Control the nighttime temperature in the greenhouse to 17±2℃ and the daytime temperature to 22±2℃. Maintain the substrate moisture at around 70%. Allow the seedbed to dry out between wet and dry periods. Water the seedbed promptly when the substrate below 2cm is dry. Spray the seedbed surface with a 0.1% carbendazim solution every 7 days for disinfection.

[0054] After 45 days of propagation by cuttings, a statistical analysis was conducted on the surviving plants. The morphology of kudzu cuttings at each growth stage is shown in the figure. Figure 1 The condition of the entire plant after 60 days of kudzu cuttings is shown in the figure. Figure 2 The growth of cuttings at different ages is shown in Table 2. One-year-old cuttings achieved a rooting rate of 91.67%, with an average of 10.26±0.57 roots per cutting, an average total root length of 14.48±2.28 cm, and an average maximum root length of 2.52±0.22 cm, indicating a well-developed root system. Two-year-old cuttings followed, with a rooting rate of 53.73%, an average of 6.60±1.41 roots per cutting, an average total root length of 12.54±3.24 cm, and an average maximum root length of 2.10±0.24 cm.

[0055] Table 2. Statistical table of growth of cuttings of three different ages in Example 1 (Mean±SD, n=3)

[0056]

[0057] Example 2

[0058] Wild kudzu branches of one-year, two-year, and several-year-old age were selected for batch experiments. Each treatment contained 40 cuttings, with three replicates and one replicate. Cuttings were prepared into stem segments 10–12 cm long and at least 0.8 cm in diameter, retaining one plump bud without leaves. The upper morphological cut was made horizontally, 2–2.5 cm from the bud, and the lower morphological cut was made at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of the cutting was then treated with a 300 mg / L solution. -1 IAA + 300mg·L -1 The cuttings were treated with a NAA solution for 3 hours. Afterward, holes were made in the substrate (river sand + vermiculite, volume ratio 1:1) using a wooden stick, with a plant spacing of 5cm, a row spacing of 10cm, and a depth of 8cm. The cuttings were then placed in the substrate, ensuring it just covered the buds, and the substrate was compacted appropriately. The remaining steps were the same as in Example 1.

[0059] After 45 days of propagation, the surviving plants were statistically analyzed, and the growth of cuttings at different ages is shown in Table 3. One-year-old and two-year-old cuttings showed similar growth, with one-year-old cuttings having a slightly higher rooting rate. However, the latter showed significantly better growth in terms of the average number of roots per cutting, average total root length, and average maximum root length. Overall, two-year-old cuttings exhibited superior growth.

[0060] Table 3. Statistical table of growth of cuttings of three different ages in Example 2 (Mean±SD, n=3)

[0061]

[0062] Example 3

[0063] Wild kudzu branches of one-year, two-year, and several-year-old age were selected for batch experiments. Each treatment contained 40 cuttings, with three replicates and one replicate. Cuttings were prepared into stem segments 10–12 cm long and at least 0.8 cm in diameter, retaining one plump bud without leaves. The upper morphological cut was made horizontally, 2–2.5 cm from the bud, and the lower morphological cut was made at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of the cutting was then treated with a 300 mg / L solution. -1 IBA + 300mg / L -1 The cuttings were treated with a NAA solution for 3 hours. Afterward, holes were made in the substrate (river sand + vermiculite, volume ratio 1:1) using a wooden stick, with a plant spacing of 5cm, a row spacing of 10cm, and a depth of 8cm. The cuttings were then placed in the substrate, ensuring it just covered the buds, and the substrate was compacted appropriately. The remaining steps were the same as in Example 1.

[0064] After 45 days of propagation, the surviving plants were statistically analyzed, and the growth of cuttings of different ages is shown in Table 4. Two-year-old cuttings showed better growth, with a rooting rate of 55.38%, an average of 9.17±1.49 roots per cutting, an average total root length of 10.51±1.99 cm, and an average maximum root length of 2.03±0.06 cm.

[0065] Table 4. Statistical table of growth of cuttings of three different ages in Example 3 (Mean±SD, n=3)

[0066]

[0067]

[0068] Example 4

[0069] Wild kudzu branches of one-year, two-year, and several-year-old age were selected for batch experiments. Each treatment contained 40 cuttings, with three replicates and one replicate. Cuttings were prepared into stem segments 10–12 cm long and at least 0.8 cm in diameter, retaining one plump bud without leaves. The upper morphological cut was made horizontally, 2–2.5 cm from the bud, and the lower morphological cut was made at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of the cutting was then treated with a 300 mg / L solution. -1 IAA + 300mg·L -1 IBA + 300mg / L -1 The cuttings were treated with a NAA solution for 3 hours. Afterward, holes were made in the substrate (river sand + vermiculite, volume ratio 1:1) using a wooden stick, with a plant spacing of 5cm, a row spacing of 10cm, and a depth of 8cm. The cuttings were then placed in the substrate, ensuring it just covered the buds, and the substrate was compacted appropriately. The remaining steps were the same as in Example 1.

[0070] After 45 days of propagation, the surviving plants were statistically analyzed, and the growth of cuttings at different ages is shown in Table 5. The rooting rate of perennial cuttings reached 81.58%, with an average of 12.47±0.71 roots per cutting, an average total root length of 14.41±0.88 cm, and an average maximum root length of 1.97±0.09 cm. Two-year-old cuttings were the next best, with a rooting rate of 56.25%, an average of 11.84±0.99 roots per cutting, an average total root length of 13.32±0.85 cm, and an average maximum root length of 2.18±0.17 cm.

[0071] Table 5. Statistical table of growth of cuttings of three different ages in Example 4 (Mean±SD, n=3)

[0072]

[0073] Comparative Example 1

[0074] Wild kudzu branches of one-year, two-year, and multi-year age were selected for batch experiments. Each treatment contained 40 cuttings, with three replicates and one destroyed. Stem segments 10–12 cm long and at least 0.8 cm in diameter were cut, retaining one plump bud without leaves. The upper morphological end was cut flat, 2–2.5 cm from the bud, and the lower morphological end was cut at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of the cuttings was treated with clean water as a control for 3 hours. The cuttings were then removed, and holes were punched in the substrate (river sand + vermiculite, 1:1 volume ratio) with a wooden stick, spaced 5 cm apart at plant spacing, 10 cm apart at row spacing, and 8 cm deep. The cuttings were placed in the substrate, ensuring it just covered the bud, and the soil was compacted appropriately and thoroughly watered. Other operating steps were the same as in Example 1.

[0075] After 45 days of propagation, the surviving plants were statistically analyzed. The growth of the cuttings in the water control group is shown in Table 1. All three types of cuttings showed poor growth and low rooting rates.

[0076] Table 6. Statistical table of growth of cuttings of three different ages in Comparative Example 1 (Mean±SD, n=3)

[0077]

[0078] Comparative Example 2

[0079] One-year-old kudzu branches were selected for batch experiments, with each treatment containing 40 cuttings. Three replicates were set up, with one replicate destroyed. Stem segments 10–12 cm long and at least 0.8 cm in diameter were cut, retaining one plump bud without leaves. The upper morphological cut was flat, 2–2.5 cm from the bud, and the lower morphological cut was at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of each cutting was then treated with a hormone (100–1000 mg / L). -1 IAA or 100-1000 mg / L -1 IBA or 100-1000 mg / L -1 The NAA (Natural Acid) was applied for 3 hours. The cuttings were then removed, and holes were punched in the substrate (river sand + vermiculite, volume ratio 1:1) with a wooden stick. The spacing between plants was 5cm, the row spacing 10cm, and the depth 8cm. The cuttings were then placed in the substrate, ensuring it just covered the buds. The substrate was compacted appropriately, and the cuttings were thoroughly watered. Other operating steps were the same as in Example 1.

[0080] After 45 days of cutting, the surviving plants were statistically analyzed. The growth of one-year-old kudzu cuttings treated with different concentrations of a single growth regulator is shown in Table 7.

[0081] Concentrations of 100, 300, 500, 700, and 1000 mg·L were used. -1 The rooting rates of cuttings treated with a single IAA growth regulator were 3.51%, 23.17%, 21.00%, 29.37%, and 25.89%, respectively; and the concentrations used were 100, 300, 500, 700, and 1000 mg·L⁻¹. -1 The rooting rates of cuttings treated with IBA single growth regulator were 17.54%, 23.88%, 22.50%, 33.33%, and 37.34%, respectively; the concentrations used were 100, 300, 500, 700, and 1000 mg·L⁻¹. -1 The rooting rates of cuttings treated with NAA single growth regulators were 7.35%, 36.17%, 23.29%, 17.83%, and 26.50%, respectively.

[0082] Overall, the rooting rate of kudzu cuttings treated with a single growth regulator was not as good as that treated with a combination of hormones.

[0083] Table 7. Statistical table of growth of one-year-old kudzu cuttings treated with different concentrations of a single growth regulator (Mean±SD, n=3)

[0084]

[0085] Example 5

[0086] One-year-old kudzu branches were selected for batch experiments, with each treatment containing 40 cuttings. Three replicates were set up, with one replicate destroyed. Cuttings were prepared into stem segments 10–12 cm long and at least 0.8 cm in diameter, retaining one plump bud without leaves. The upper morphological cut was made horizontally, 2–2.5 cm from the bud, and the lower morphological cut was made at a 45° angle, 8–9 cm from the bud, with smooth cuts and no mechanical damage. The cuttings were completely disinfected by soaking in a 0.1% carbendazim solution for 10 minutes, then air-dried. The lower 2 cm of the cutting was then treated with a 300 mg / L solution. -1 IAA + 300mg·L -1 The cuttings were treated with an IBA combination solution for 3 hours. After treatment, holes were made in the substrate using a wooden stick (substrate could be river sand, loam, loam + river sand (1:1, v / v), loam + vermiculite (1:1, v / v), or river sand + vermiculite (1:1, v / v)). The spacing between plants was 5 cm, the row spacing was 10 cm, and the depth was 8 cm. The cuttings were then placed in the substrate, ensuring it just covered the buds. The substrate was compacted appropriately, and the cuttings were thoroughly watered. Other operating steps were the same as in Example 1.

[0087] Table 8 shows the growth of one-year-old kudzu cuttings in different substrates after 45 days of propagation, based on statistical analysis of the surviving plants. The best substrate combination was river sand + vermiculite (1:1, v / v), which balanced aeration and moisture retention, promoting rooting of the kudzu cuttings. The rooting rate was 88.18%, with an average of 8.19 ± 0.52 roots per cutting, an average total root length of 10.04 ± 2.93 cm, and an average maximum root length of 2.21 ± 1.00 cm. The second best substrate combination was loam + river sand (1:1, v / v), with a rooting rate of 80.00%, an average of 5.45 ± 0.55 roots per cutting, an average total root length of 4.69 ± 1.01 cm, and an average maximum root length of 1.23 ± 0.20 cm.

[0088] Table 8. Statistical table of growth of one-year-old kudzu cuttings in different substrates (Mean±SD, n=3)

[0089]

[0090] Experimental Example 6

[0091] The rooting rates of cuttings in Examples 1-4 and Comparative Example 1 (15 treatment groups) were statistically analyzed. The M4 treatment group showed the highest rooting rate at 91.67%. The results indicate that selecting one-year-old kudzu branches and using a 300 mg·L⁻¹ treatment is effective. -1 IAA + 300mg·L -1 Treatment with an IBA solution for 3 hours, using a 1:1 volume ratio of river sand and vermiculite as the seedbed substrate, yielded the best propagation results for wild kudzu cuttings. The treatment conditions of M4 provide a scientific and efficient technical reference for the large-scale cultivation of high-quality wild kudzu seedlings.

[0092] Therefore, this invention uses stem segments of the legume *Pueraria lobata* as raw material and employs asexual propagation via cuttings. By optimizing several key parameters, such as the age of the cuttings, selecting suitable types and concentrations of growth regulators, and controlling the treatment time, the rooting rate of *Pueraria lobata* cuttings is improved. Results show that treatment with a mixed growth regulator significantly increases the rooting rate of *Pueraria lobata* cuttings. This method is simple, easy to implement, and environmentally friendly, and can effectively promote the large-scale propagation of superior *Pueraria lobata* germplasm, showing broad application prospects.

[0093] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A propagation method for increasing the rooting rate of Pueraria lobata cuttings, characterized by, The method comprises the following steps: The mother branch of the kudzu vine is cut to obtain a cutting, the cutting is disinfected to obtain a disinfected cutting; The disinfected cutting is treated with a growth regulator to obtain a growth regulator treated cutting; The growth regulator treated cutting is inserted into a seedbed substrate to carry out cutting culture, and a kudzu vine seedling is obtained. The growth regulator includes a combination of at least two of IAA, IBA and NAA, the concentration of the IAA is 200-500 mg·L -1 , the concentration of the IBA is 200-500 mg·L -1 , and the concentration of the NAA is 200-500 mg·L -1 . The growth regulator treatment time is 0.5-5 hours.

2. The propagation method according to claim 1, characterized in that, The cutting method is as follows: The length of the cutting is 10-12 cm, the diameter is more than 0.8 cm, one bud eye is reserved, the upper end is cut flat, 2-2.5 cm from the bud eye, and the lower end is cut at an angle of 45°, 8-9 cm from the bud eye.

3. The propagation method according to claim 1, characterized in that, The disinfecting solution is a 0.1% carbendazim solution, and the disinfecting time is 10-15 minutes.

4. The propagation method according to claim 1, characterized in that, When the cutting is treated with the growth regulator, the lower end of the disinfected cutting is inserted into the growth regulator for 1.5-2.4 cm.

5. The propagation method according to claim 1, characterized in that, The seedbed substrate comprises river sand+vermiculite or loam soil+river sand.

6. The propagation method according to claim 5, characterized in that, When the seedbed substrate is river sand+vermiculite, the volume ratio of the river sand+vermiculite is 1:

1. When the seedbed substrate is loam soil+river sand, the volume ratio of the loam soil+river sand is 1:

1.

7. The propagation method according to claim 1, characterized in that, The seedbed substrate is disinfected with a 0.5% KMnO4 solution 4 days before cutting, and is covered with a film after disinfection, and the film covering time is 3 days.

8. The propagation method according to claim 1, characterized in that, The cutting is carried out with a plant spacing of 5 cm and a row spacing of 10 cm.

9. The propagation method according to claim 1, characterized in that, The cutting is carried out with the cutting and the seedbed substrate being inclined at an angle of 60°, and the bud eye of the cutting just protruding out of the substrate surface.

10. The propagation method according to claim 1, characterized in that, The cutting culture temperature is 15-25°C, and the substrate humidity is 60%-80%.

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