A kind of efficient propagation and seedling raising method of ginger flower

Abstract

The present invention discloses a method for efficiently breeding and raising seedlings of ginger flower, comprising the following steps: leaf stem selection; stem segment processing: cutting the leaf stem and soaking the stem segment in a growth regulator solution; stem segment cultivation: burying the stem segment in perlite and placing it in a shade shed to moisturize and cultivate until the hidden buds germinate into small rhizomes; small rhizome transplanting: cutting the stem segment into segments with small rhizomes according to the nodes and planting them in batches; strong seedling cultivation: using an aqueous solution containing a plant growth regulator to irrigate the substrate, and when the average height of the plant reaches the standard, it can be transplanted or out of the nursery. The advantages of the present invention include: being suitable for large-scale seedling cultivation of ginger flower in conventional production, and having the advantages of high breeding efficiency, short seedling cycle, and low production cost, and being able to quickly and in large quantities carry out efficient breeding of high-quality ginger flower seedlings to meet the strong demand of the market such as garden landscape and flower gardening.

Description

Technical Field

[0001] The invention relates to the technical field of horticultural seedling propagation, in particular to a propagation technology of ginger flower. Background Art

[0002] Hedychium coronarium is a perennial herbaceous plant in the Zingiberaceae family. Its stems are 1-2 meters tall, with alternate, narrow leaves and terminal spikes. Its bracts are arranged in an imbricate pattern, each containing two to three flowers. During flowering, the flowers resemble a fluttering flock of beautiful butterflies. The fragrant white flowers bloom from July to November.

[0003] Ginger flower, a specialty flower in South China, is an excellent choice for landscape gardening, potted plants, and cut flowers. It is often cultivated for ornamental purposes, has a wide range of applications, and boasts high economic value, with seedlings often in short supply. Sexual propagation of ginger flower is rarely used due to its low natural fruit set rate and inconsistent flowering quality of seedlings. Asexual propagation, which includes tissue culture and division, requires high sterile facilities and operating techniques, and is costly. Therefore, rhizome division is often used for asexual propagation. Division of ginger flower is fast and has a high seedling success rate, but the number of seed stems is limited and the reproduction coefficient is low, making it difficult to effectively produce high-quality ginger flower seedlings.

[0004] In addition, Chen Shiren and others have also developed a ginger flower breeding method based on the promotion of leaf stem latent buds, publication number CN110915446A. This method is a breeding method using the leaf stem latent buds of 'Hanyue' ginger flower as the material source, and the seedling rate is high and stable. However, this method requires indoor sterile conditions to control the size of the sealed bag and the number of segments placed in the sealed bag, and the temperature, humidity and thickness of the leaf stem segments in the sealed bag must be strictly controlled to achieve effective germination of the leaf stem latent buds. Its operation is similar to the aseptic culture in tissue culture, and has very high requirements for sterile environment and aseptic operation. In addition, the utilization rate of peeled leaf stem segments with a diameter of less than 10 mm is not high, making it difficult to carry out large-scale seedling cultivation in conventional production. Summary of the Invention

[0005] The purpose of the present invention is to provide an efficient propagation and seedling raising method for ginger flower, so as to solve the problem in the prior art that the number of ginger flower seed stems is limited and the propagation coefficient is low, which makes it difficult to effectively meet the needs of efficient propagation of high-quality ginger flower seedlings.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for efficiently breeding and raising seedlings of ginger flower comprises the following steps:

[0008] (1) Leaf and stem selection;

[0009] (2) Stem segment treatment: Cut the leaf stems and soak the stem segments in a growth regulator solution;

[0010] (3) Stem segment culture: bury the stem segment in perlite and place it in a shade shed to keep it moist until the latent buds germinate into small rhizomes;

[0011] (4) Transplanting small rhizomes: Cut the stem segments into segments with small rhizomes and transplant them in batches;

[0012] (5) Cultivation of strong seedlings: Use an aqueous solution containing plant growth regulators to irrigate the substrate. When the average height of the plants reaches the standard, they can be transplanted or taken out of the field.

[0013] Furthermore, the detailed operation of step (1) includes: from August to December each year, selecting strong leaf stems that have bloomed, cutting them off from the base and removing the residual inflorescences; carefully peeling the leaf sheaths of the leaf stems from bottom to top, layer by layer, to the top, exposing the hidden buds on the nodes.

[0014] Furthermore, the detailed operation of step (2) includes: cutting the leaf stem from the middle to divide it into two bud-bearing stem segments of approximately equal length, each containing 4 to 6 hidden buds; then soaking the upper and lower stem segments in a solution containing a growth regulator for 10 to 20 minutes;

[0015] In the solution containing the growth regulator, the ratio of chlorfenapyr to 3-indolebutyric acid is 2:1.

[0016] Furthermore, the solution containing the growth regulator is as follows: per 1L of solution, it contains 40-60 mg of chlorfenuron (CPPU), 20-30 mg of 3-indolebutyric acid (IBA) and the balance of water;

[0017] The lower stem segment is thick and the soaking time is 15 to 20 minutes, while the upper stem segment is thin and the soaking time is 10 to 15 minutes.

[0018] Furthermore, the detailed process of step (3) includes: first spreading perlite on the bottom of the plug tray, placing the stem segments flat on different plug trays, covering the surface of the stem segments with a layer of perlite, and placing them in a shade shed with automatic spray, the shade shed has a shading rate of 40% to 60%; using spraying to moisturize, depending on the dryness and wetness of the weather, spraying mist water once in the morning, noon and evening every day, each spraying time is 2 to 4 minutes; culturing the stem segments for 3 to 5 weeks.

[0019] Furthermore, the detailed process of step (3) includes: first spreading perlite at the bottom of the plug tray with a thickness of 2 to 4 cm; placing the stem segments flat on different plug trays, and covering the surface of the stem segments with a layer of perlite with a thickness of 1 to 2 cm; placing all the plug trays in a shade shed with automatic spraying, and the shade shed has a shading rate of 40% to 60%; using a spray method to moisturize the plug trays, and spraying mist water once in the morning, noon and evening every day depending on the dryness and wetness of the weather, and each spraying time is 2 to 4 minutes; during the stem segment cultivation process, it is strictly forbidden to use fungicides for disinfection; the lower thick stem segments are cultivated for 3 to 4 weeks, and the upper thin stem segments are cultivated for 4 to 5 weeks.

[0020] Furthermore, the detailed process of step (4) includes: cutting the stem segments into segments with small rhizomes according to the nodes and planting them in batches; placing the segments flat on the surface of the substrate, and continuously filling the mixed substrate to completely cover the small rhizomes. Depending on the dryness and wetness of the weather, spraying with mist water once in the morning, noon and evening, each spraying lasting 3 to 6 minutes, and drenching with 2000 to 3000 times of 30% hydroxychloroquine solution once every 15 days to prevent and control fungal diseases.

[0021] Furthermore, the detailed process of step (4) includes: cutting the upper and lower stem segments into segments with small rhizomes according to the nodes and planting them in batches, each segment is 2 to 3 cm long; placing the segments flat on the substrate surface of the flower pot or nutrient pot, placing one segment per pot, and then filling the mixed substrate to completely cover the small rhizomes. Depending on the weather, spray mist water once in the morning, noon and evening every day, each spraying lasting 3 to 6 minutes, and drench once every 15 days with 2000 to 3000 times of 30% hydroxychloroquine solution to prevent fungal diseases.

[0022] Furthermore, the detailed process of step (5) includes: 3 to 5 days after planting, in order to accelerate the root development of the rhizomes and the growth of the seedlings, the substrate is irrigated once with an aqueous solution containing a plant growth regulator; when the ginger flower seedlings unfold their leaves, in order to promote the healthy development of the seedlings, water-soluble fertilizer is sprayed on the leaves; if the temperature continues to be low in winter, fertilization is stopped; spraying is continued several times, and the plants can be transplanted or out of the field when the average height exceeds 30 cm.

[0023] Furthermore, in step (5), each 1L of the aqueous solution containing the plant growth regulator contains 50mg of naphthaleneacetic acid (NAA), 25mg of 6-benzylaminopurine (6-BA) and the balance of clean water;

[0024] When ginger flower seedlings have 3 to 4 leaves, spray the leaves with a 0.2% to 0.3% water-soluble fertilizer every 7 to 10 days to promote healthy growth. The N:P:K ratio should be 20:20:20. If temperatures remain below 15°C in winter, stop fertilizing. Continue spraying 3 to 5 times until the plants reach an average height of more than 30 cm before transplanting or outplanting.

[0025] Compared with the prior art, the present invention has the following advantages:

[0026] 1. The present invention uses mature leaf stems after ginger flower blooming to promote the development of latent buds, which greatly expands the asexual reproduction mode and material source of ginger flower. In particular, the segmented treatment of leaf stems can make full use of stem segments of different thicknesses, so that the utilization rate of each mature leaf stem is as high as 100%, which doubles the reproduction coefficient.

[0027] 2. The present invention utilizes a combination of growth regulator solutions with varying concentrations to soak segmented stems, effectively promoting the differentiation and expansion of latent buds and inducing adventitious roots, thus laying a good foundation for the subsequent growth and development of rhizomes. By applying different soaking and incubation times to account for the varying thickness and maturity of the upper and lower stem segments, individual differences in rhizome development can be minimized, ensuring consistency in the resulting seedlings from each batch.

[0028] 3. This method uses pure perlite for layering of budded stem segments. Compared to traditional layering, this method eliminates the need for tedious soil preparation and sterilization, making it highly convenient and efficient, making it suitable for large-scale cultivation. Pure perlite is free of pathogens and insect eggs, is fluffy and breathable, and does not accumulate water, facilitating the preservation of budded stem segments and the initiation of latent buds. The overall stem segment preservation rate can reach over 95%, and it effectively reduces stem segment infection and black rot.

[0029] 4. The chlorfenapyr selected by the present invention is a plant growth regulator with high cytokinin activity, which can affect the development of plant buds, accelerate cell mitosis, and promote cell enlargement and differentiation. Through appropriate growth regulator treatment and layering culture in a suitable environment, after culturing the lower thick stem segment for 3 to 4 weeks, the hidden bud germination rate reaches 93.5%, and the hidden buds germinate into small rhizomes with a diameter of 7 to 12 mm and 1 to 4 roots. The average diameter of the rhizomes is 9.35 mm, the average number of roots is 2.6, and 4 to 6 small rhizomes can be obtained from each stem segment. After culturing the upper thin stem segment for 4 to 5 weeks, the hidden bud germination rate reaches 87.1%, and the hidden buds germinate into small rhizomes with a diameter of 6 to 10 mm and 1 to 3 roots. The average diameter of the rhizomes is 8.29 mm, the average number of roots is 2.2, and 3 to 5 small rhizomes can be obtained from each stem segment.

[0030] 5. The mixed matrix used in the present invention can effectively preserve rhizomes and promote root growth and elongation. After 30 days of planting segments with rhizomes in the mixed matrix, the transplant preservation rate of rhizomes can reach 100%, with no infection, black rot, etc. The present invention separates the differentiation and induction of leaf stem crypt buds from the rooting and development of rhizomes, combining the advantages of conventional layering and cutting propagation, but is simpler to operate than conventional layering and has a faster rooting speed than cuttings. The propagation coefficient of each leaf stem is as high as 8.8, and the seedling establishment speed is also significantly improved.

[0031] 6. The seedling cultivation method adopted by the present invention includes root induction and foliar fertilization. The induction of growth regulators can effectively promote the root growth of small rhizomes, significantly increase the number of tender roots, and significantly elongate the roots. A dense root mass can be formed after 15 days of growth. On this basis, the use of appropriate water-soluble fertilizer foliar spraying can accelerate the growth of seedlings. After 3 to 5 consecutive fertilizations, the plants have well-developed root systems, thick leaf stems, and bright green leaves, and the seedling cultivation effect is significant.

[0032] The method of the present invention expands the asexual propagation mode and material source of ginger flower, and has the advantages of both layering propagation and cutting propagation. It can well promote the germination of hidden buds of ginger flower leaf stems, the differentiation of root systems of rhizomes and the rapid development of seedlings. The innovative layering culture method is suitable for batch culture of mature ginger flower stem segments. Through the differentiation of hidden buds of leaf stems and the induction of root systems of rhizomes, the propagation coefficient and seedling speed are significantly improved, so that the utilization rate of mature leaf stems reaches 100%, the overall germination rate of leaf stems reaches 87.1% to 93.5%, and the transplanting seedling rate of rhizomes reaches 100%. The method of the present invention is suitable for large-scale seedling cultivation of ginger flower in conventional production, and has the advantages of high propagation efficiency, short seedling cycle and low production cost. It can quickly and efficiently breed high-quality ginger flower seedlings in large quantities to meet the strong demand of the market such as garden landscape and flower gardening. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The drawings described herein are used to provide a further understanding of the present invention, constitute a part of this application, and do not constitute an improper limitation of the present invention. In the drawings:

[0034] Figure 1 : Experimental photos of peeling off the leaf sheath of mature leaf stems to reveal the hidden buds on the nodes;

[0035] Figure 2 : Experimental photos of segmented culture of leaf stems using pure perlite;

[0036] Figure 3 : Experimental photos of the latent buds of the stem segments germinating and swelling;

[0037] Figure 4 : Experimental photos of latent buds germinating into rhizomes with root systems;

[0038] Figure 5 : Experimental photos comparing the bud germination of upper and lower stem segments;

[0039] Figure 6 : Experimental photos of rhizomes ready for transplantation after stem segment cutting;

[0040] Figure 7 : Experimental photos of continued differentiation and growth of rhizomes;

[0041] Figure 8: Experimental photos comparing the root system of rhizomes before and after induction;

[0042] Figure 9 : Experimental photos of cultivating strong seedlings with small rhizomes into high-quality large seedlings. DETAILED DESCRIPTION

[0043] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. The exemplary embodiments and descriptions of the present invention are used to explain the present invention but are not intended to limit the present invention.

[0044] Example 1 Ginger Flower Propagation and Seedling Raising Method

[0045] 1. Leaf stem selection: From August to December each year, select healthy leaf stems that have already bloomed, cut them off from the base and remove the residual inflorescence; carefully peel off the leaf sheaths of the leaf stems from bottom to top, layer by layer, to the top, exposing the hidden buds on the nodes, such as Figure 1 shown.

[0046] 2. Stem Segment Processing

[0047] Cut the leaf stem in the middle and divide it into two bud-bearing stem segments of nearly equal length, each containing 4 to 6 hidden buds; then soak the upper and lower stem segments in a solution containing a growth regulator for 10 to 20 minutes. The solution containing the growth regulator is as follows: each 1L of solution contains 40 to 60 mg of chlorpyrifos (CPPU), 20 to 30 mg of 3-indolebutyric acid (IBA) and the remainder of clean water.

[0048] Among them, the lower stem segment is thick and the soaking time is 15 to 20 minutes, while the upper stem segment is thin and the soaking time is 10 to 15 minutes; in the growth regulator solution, the ratio of chlorpyrifos to 3-indolebutyric acid is 2:1.

[0049] 3. Stem Segment Culture

[0050] (1) Culture container and substrate: The above-mentioned stem segments with buds were cultured in a common gardening flat-bottomed plug tray with holes at the bottom for drainage. The culture substrate was pure perlite.

[0051] (2) Culture method: First, spread perlite on the bottom of the hole tray with a thickness of 2 to 4 cm; Figure 2 As shown, place the upper and lower stem segments flat on different plug trays, and cover the surface of the stem segments with a layer of perlite with a thickness of 1 to 2 cm.

[0052] (3) Cultivation environment: All plug trays are placed in a shade shed equipped with an automatic sprayer, with a shading rate of 40% to 60%. Moisturize the plug trays by spraying them once a day in the morning, noon, and evening, depending on the weather. Each spraying time is 2 to 4 minutes. During the cultivation of stem segments, the use of fungicides is strictly prohibited.

[0053] (4) Culture period: After culturing the lower thick stem segment for 3 to 4 weeks, the latent bud germination rate reached 93.5%. Figure 3 As shown, the latent buds germinated into rhizomes with a diameter of 7 to 12 mm and 1 to 4 roots, and 4 to 6 rhizomes were obtained from each stem segment. After culturing the upper thin stem segment for 4 to 5 weeks, the latent bud germination rate reached 87.1%, and the latent buds germinated into rhizomes with a diameter of 6 to 10 mm and 1 to 3 roots, and 3 to 5 rhizomes were obtained from each stem segment. Figure 4-5 shown.

[0054] 4. Transplanting small rhizomes

[0055] (1) Transplanting container and substrate: Use plastic flower pots or nutrient pots with a diameter of 8 to 10 cm to culture the above-mentioned small rhizomes. The culture substrate is general nutrient soil and perlite. Mix them evenly in a volume ratio of 7:3, fill 2 / 3 of the flower pots or nutrient pots, and place them neatly on the seedbed in the shade.

[0056] (2) Root segment planting: Cut the upper and lower stem segments into segments with small rhizomes and plant them in batches. Each segment is 2 to 3 cm long. Figure 6 As shown; Place the segments flat on the substrate surface of the flower pot or nutrient pot, place one per pot, and continue to fill with mixed substrate to completely cover the rhizomes. Depending on the weather, spray mist water once every morning, noon and evening, each spray lasting 3 to 6 minutes. Spray once every 15 days with 2000 to 3000 times the amount of 30% hydroxychloroquine solution to prevent fungal diseases. Figure 7 As shown, the rhizomes continue to differentiate and grow.

[0057] 5. Cultivation of strong seedlings

[0058] (1) 3 to 5 days after planting, in order to accelerate the root development of rhizomes and the growth of seedlings, water the substrate once with an aqueous solution containing plant growth regulators. Each liter of aqueous solution containing plant growth regulators contains 50 mg of naphthaleneacetic acid (NAA), 25 mg of 6-benzylaminopurine (6-BA) and the balance of water. Figure 8 Shown is the comparison before and after root induction.

[0059] (2) When the ginger flower seedlings have 3 to 4 leaves, spray the leaves with 0.2% to 0.3% water-soluble fertilizer every 7 to 10 days to promote the healthy development of the seedlings. The N:P:K mass ratio of the water-soluble fertilizer should be 20:20:20. If the temperature is below 15℃ in winter, stop fertilizing. Spray 3 to 5 times continuously. When the average height of the plants exceeds 30 cm, they can be transplanted or taken out of the nursery. Figure 9 shown.

[0060] Example 2 Effect of ginger flower leaf and stem maturity on layering latent bud germination

[0061] A layering experiment was conducted using ginger flower stems of two different maturities. Treatment 1 consisted of stems without an inflorescence at the top, and Treatment 2 consisted of stems with wilted terminal inflorescences. In both treatments, the leaf sheaths were carefully peeled from bottom to top, layer by layer, to the top, exposing the hidden buds at the nodes. The stems were then divided into two segments of approximately equal length, one above and one below, for conventional layering. The culture medium used was ordinary garden soil, covering each segment with a thickness of approximately 2 cm. The soil in the seedbed was watered regularly to maintain moisture, depending on the soil's dryness. Sterilization was performed every 7 to 10 days using a 1000-fold solution of 50% carbendazim.

[0062] Sixty stem segments were cultured for each treatment, replicated three times, with each stem segment containing 4 to 6 latent buds. After one month of culture, the germination rate was calculated, with a latent bud diameter greater than 3 mm as the germination standard, and significance analysis was performed using SPSS 21.0. The results are as follows:

[0063]

[0064] Different letters within treatments indicate significant differences at the 0.05 level, the same below.

[0065] The experiment showed that in Treatment 1, the overall germination rate of latent buds on the leaf stems that had not yet produced inflorescences was low, with germination rates of 17.7% on the lower, thicker stem segments and only 3.8% on the upper, thinner stem segments. In Treatment 2, the germination rate of latent buds on the leaf stems that had already flowered was slightly higher, reaching 39.4% and 21.3% on the lower and upper stem segments, respectively. In both treatments, latent bud germination in the upper, thinner stem segments was slower than in the lower, thicker stem segments. When conventionally layered using standard garden soil, both stem segments showed obvious signs of infection and black rot.

[0066] Comparison of the two treatments revealed that the latent bud germination rate of ginger flower stems was closely related to the maturity of the stems. Stems without inflorescences were relatively tender and had high water content, resulting in small or inconspicuous latent buds that were difficult to differentiate and a low stem preservation rate. Stems with wilted terminal inflorescences were more mature, had more pronounced latent buds, and were more likely to differentiate into rhizomes. The germination rates of both upper and lower stem segments were significantly higher than those in Treatment 1. Therefore, it can be concluded that robust, flowering stems are the most suitable propagation material for layering, and stem segments of varying thickness can be used for asexual propagation.

[0067] Example 3 Effect of Layering Culture Method on Hidden Bud Germination

[0068] In order to improve the layering preservation rate and latent bud germination rate of stem segments, layering experiments with different substrates were carried out on mature stem segments that had already bloomed (treatment 3). The leaf stems with leaf sheaths removed were divided into two bud-bearing stem segments of nearly equal length, and the culture substrates were B1: general nutrient soil, B2: plain river sand, and B3: perlite.

[0069] Layering culture is carried out in a shade shed with a shading rate of 40% to 60%. The shed is equipped with automatic spraying facilities. First, a layer of culture matrix is ​​spread on the independent seedbed in the shade shed, and the stem segments are placed on different matrixes respectively. The surface of the stem segments is covered with a layer of matrix of 1 to 2 cm. Depending on the dryness and wetness of the weather, spray mist water once in the morning, noon and evening every day, and each spraying time is 2 to 4 minutes. Sterilize once every 7 to 10 days with 1000 times solution of 50% carbendazim.

[0070] For each treatment, 60 stem segments (30 upper and 30 lower stem segments) were cultured, replicated three times, and each stem segment contained 4 to 6 latent buds. After one month of culture, the germination standard was a latent bud diameter greater than 3 mm. The final overall germination rate was calculated and analyzed for significance using SPSS 21.0. The results are as follows:

[0071]

[0072] The test showed that compared with the garden soil used in Example 2, the use of general nutrient soil, plain river sand, and perlite for simple facility cultivation can effectively improve the preservation rate and latent bud germination rate of mature leaf stems. The latent bud germination rate of each treatment group is above 55%, and the stem segments cultured with perlite have the highest latent bud germination rate, with an overall germination rate of 71.1%. This is probably related to the fact that perlite does not contain pathogens or insect eggs, is fluffy and breathable, and does not accumulate water. Pure perlite cultivation is more conducive to the preservation of stem segments with buds and latent bud germination.

[0073] It should be noted that during this treatment, all excess stem segments with buds in each group were cultured with perlite and no carbendazim solution was used during subsequent maintenance. Surprisingly, infection and black rot in each stem segment were significantly reduced, and the stem segment preservation rate and germination rate were higher than those in the groups in Treatment 3. Subsequently, the inventors repeated the above culture using "pure perlite" without using any fungicide (Treatment 4). The final statistics showed that the overall latent bud germination rate in each repeated group was above 80%, indicating that strictly prohibiting the use of fungicides for disinfection during the stem segment culture process has a better implementation effect.

[0074] Example 4: Induction test of cryptogenic bud differentiation by growth regulators

[0075] 4.1 Materials and Methods

[0076] (1) Effects of growth regulator types on cryptic bud differentiation

[0077] To promote cryptic bud differentiation in leaf stems, different cytokinins were used to induce cryptic bud differentiation in mature stem segments (Treatment 5). The leaf stems, stripped of their leaf sheaths, were divided into two bud-bearing segments, upper and lower, of approximately equal length. These segments were then immersed for 15 minutes in solutions of different cytokinins: 6-benzylaminopurine (6-BA), kinetin (KT), and chlorfenapyr (CPPU), all at a concentration of 20 mg / L. Water was added as a control. The cells were then cultured in a simple facility using pure perlite as the culture medium. See Section 4.2 for Statistics and Analysis for the results.

[0078] (2) Effects of growth regulator concentration on cryptic bud differentiation

[0079] Based on Treatment 5, to investigate the effects of different growth regulator concentrations on cryptic bud differentiation and development, the sheathed stems were divided into two bud-bearing segments of approximately equal length, upper and lower. These segments were then immersed in solutions of chlorfenapyr (CPPU) at different concentrations for 15 minutes (Treatment 6), with concentrations of 20 mg / L, 40 mg / L, and 60 mg / L, respectively. The buds were then cultured in a simple facility using pure perlite as the culture medium. The results are shown in Section 4.2: Statistics and Analysis.

[0080] (3) Effects of growth regulator combinations on cryptic bud differentiation and rhizome development

[0081] To explore the effects of growth regulators on rhizome development after cryptic bud differentiation, treatment 7 (treatment 7) added varying concentrations of auxin to co-regulate rhizome growth and development. The auxin selected was 3-indolebutyric acid (IBA). Three concentration combinations were set: 40 mg / L CPPU + 20 mg / L IBA, 40 mg / L CPPU + 40 mg / L IBA, and 40 mg / L CPPU + 80 mg / L IBA. The lower stem segments were thicker and soaked for 15–20 minutes, while the upper stem segments were thinner and soaked for 10–15 minutes. The plants were then cultured in a simple facility on pure perlite. See Section 4.2 for statistics and analysis.

[0082] All cultivations were conducted in a shaded shed with a shading ratio of 40% to 60%. The shed was equipped with an automatic spray system, and mist water was applied once daily, morning, noon, and evening, depending on the weather conditions. Each spray lasted 2 to 4 minutes. Sixty stem segments (30 from the upper and lower stem segments) were cultured for each treatment, with three replicates. After 3 to 5 weeks of cultivation, the germination rate, rhizome diameter, and rooting were calculated, with a bud diameter greater than 3 mm as the germination standard. Significance analysis was performed using SPSS 21.0.

[0083] 4.2 Statistics and Analysis

[0084] (1) Analysis of the effects of growth regulator types on cryptic bud differentiation

[0085]

[0086] Statistics show that compared with the control group, treatment with different growth regulators can effectively increase the overall germination rate of cryptic buds and the average diameter of rhizomes. In terms of overall germination rate, 6-BA and KT have basically the same effect, but there is a significant difference with CPPU. CPPU has the highest overall germination rate, reaching 82.6%, which is 20.7% higher than the control group. In terms of the average diameter of rhizomes, there are significant differences among the treatment groups. Among them, the average diameter of rhizomes treated with CPPU is the largest, reaching 8.67mm, almost twice the diameter of the control group. Overall, the cytokinin chlorpyrifos (CPPU) has the most significant effect on the differentiation of stem cryptic buds and is the most suitable plant growth regulator for promoting the differentiation of leaf and stem cryptic buds.

[0087] (2) Effects of growth regulator concentration on cryptic bud differentiation

[0088]

[0089] Statistical data show that, compared with the control group, treatment with different concentrations of chlorfenapyr (CPPU) effectively increased the overall germination rate of latent buds, the average diameter of rhizomes, and the average number of roots. In terms of germination rate, the latent bud germination rate showed an initial increase followed by a decrease with increasing CPPU concentration. At a CPPU concentration of 40 mg / L, the latent bud germination rate reached 91.6%, which was not significantly different from the CPPU concentration of 60 mg / L. In terms of average diameter, the average diameter of rhizomes showed a continuous increase with increasing CPPU concentration, but the difference between CPPU concentrations of 40 mg / L and 60 mg / L was not significant. The average diameters of both treatments were above 9 mm and approximately twice that of the control group. In terms of average root number, the average number of rhizomes showed a continuous increase with increasing CPPU concentration, but the difference between CPPU concentrations of 40 mg / L and 60 mg / L was not significant. The average number of roots in both treatments was above 2, which was more than 2.6 times that of the control group. In summary, when the concentration of chlorfenapyr (CPPU) is between 40 and 60 mg / L, the differentiation effect on leaf and stem cryptic buds and rhizomes is most significant.

[0090] (3) Effects of growth regulator combinations on cryptic bud differentiation and rhizome development

[0091]

[0092] The statistical results showed that different growth regulator combinations had varying effects on cryptic bud differentiation and rhizome development, with the most significant effect on the average diameter of rhizomes, followed by the cryptic bud germination rate, and the effect on the average root number being less significant. Specifically, a 2:1 CPPU to IBA concentration ratio was most conducive to cryptic bud differentiation and rhizome development. The cryptic bud germination rate, average rhizome diameter, and average root number of the lower thick stem segments reached their maximum values, as did the cryptic bud germination rate and average rhizome diameter of the upper thin stem segments. When the CPPU to IBA concentration ratio was 1:1, all other indicators were lower than those of Group 1, except for the slightly higher average root number of the upper thin stem segments. However, there were no significant differences between the two treatment groups in terms of the cryptic bud germination rate and average root number of rhizomes in the lower thick stem segments, or the average diameter and average root number of rhizomes in the upper thin stem segments. When the concentration ratio of CPPU to IBA was 1:2, the values ​​of various indicators of the upper and lower stem segments decreased significantly, which was significantly different from that of Group 1 and Group 2.

[0093] In summary, the combined use of CPPU and IBA can play a good regulatory role in the differentiation of ginger flower hidden buds and the development of rhizomes. When the concentration ratio of CPPU to IBA is ≥1, the regulatory effect on the plant is more obvious. In this embodiment, "CPPU 40mg / L+IBA 20 mg / L" was the optimal treatment combination, achieving an overall stem segment preservation rate exceeding 95%. After 3-4 weeks of cultivation, the lower, thicker stem segments achieved a latent bud germination rate of 93.5%. These latent buds gave rise to small rhizomes with a diameter of 7-12 mm and 1-4 roots. The average diameter of the rhizomes was 9.35 mm, and the average number of roots reached 2.6. Four to six rhizomes were obtained per stem segment. After 4-5 weeks of cultivation, the upper, thinner stem segments achieved a latent bud germination rate of 87.1%. These latent buds gave rise to small rhizomes with a diameter of 6-10 mm and 1-3 roots. The average diameter of the rhizomes was 8.29 mm, and the average number of roots reached 2.2. Three to five rhizomes were obtained per stem segment. Through the rational cultivation of both upper and lower stem segments, the utilization rate of each mature leaf stem reached 100%, and the overall latent bud germination rate reached 87.1% to 93.5%.

[0094] Example 5 Experiment on the influence of different transplanting media and planting methods on the seedling formation of small rhizomes

[0095] In order to explore the effects of different transplanting substrates and planting methods on the transplanting and seedling formation of small rhizomes, this example tested the combination treatments of two substrate formulas and two planting methods (treatments 8 and 9); the substrate formulas were: ① garden soil and plain river sand were evenly mixed in a volume ratio of 7:3, and ② nutrient soil and perlite were evenly mixed in a volume ratio of 7:3; the two planting methods were: ① the upper and lower stem segments were cut into small rhizomes with segments according to the nodes, each segment was 2 to 3 cm long, and ② the small rhizomes were carefully cut off from the upper and lower stem segments and divided into independent small rhizomes without segments.

[0096] Then, fill the flower pots or nutrient pots 2 / 3 full with the above-mentioned matrix and arrange them neatly on the seedbed in the shade. Place the rhizomes flat on the matrix surface of the flower pots or nutrient pots, one per pot, and continue to fill with the mixed matrix until the rhizomes are completely covered. The above cultivation is carried out in a shaded shed with a shading rate of 40% to 60%. The shed is equipped with an automatic spraying device. Depending on the weather, mist water is sprayed once in the morning, noon and evening every day, and each spraying lasts for 3 to 6 minutes.

[0097] 90 rhizomes were cultured for each treatment, with three replicates. After 30 days of culture, the transplant survival rate and seedling height of the rhizomes were calculated and analyzed for significance using SPSS 21.0. The results are as follows:

[0098]

[0099] The results showed that different transplanting substrates and planting methods had significant effects on the transplanting seedlings of small rhizomes. From the perspective of transplanting substrate, the transplanting survival rate of treatment 8 was 76.7% to 88.9%, which was significantly lower than the 91.1% to 100% of treatment 9. The seedling height was only 9.4cm to 10.5cm, which was also significantly shorter than the 12.3cm to 13.7cm of treatment 9. Therefore, it can be determined that the transplanting effect of the mixed substrate of nutrient soil and perlite is better than the substrate combination of garden soil and plain river sand.

[0100] From the perspective of transplanting methods, the transplant survival rate and seedling height of segmented rhizomes in treatment 8 were better than those of independent rhizomes. The transplant seedling effect of segmented rhizomes in treatment 9 was also significantly better than that of independent rhizomes. Therefore, it can be determined that the most suitable transplanting method for rhizomes is to cut the upper and lower stem segments into segments with rhizomes according to the nodes and ensure that each segment is 2 to 3 cm long.

[0101] In summary, small rhizomes with segments are most suitable for transplanting using a mixed matrix of nutrient soil and perlite (mixed evenly in a volume ratio of 7:3). After 30 days of transplanting, the transplant survival rate reached 100%, and the average plant height reached 13.8 cm.

[0102] Example 6: Test on the Effect of Seedling Culture on Seedling Growth

[0103] (1) Effects of growth regulator induction on root growth

[0104] To promote root growth and seedling development of small rhizome transplants, an aqueous solution containing plant growth regulators was used to irrigate the substrate. This example tested a combination treatment of two auxins and two cytokinins (Treatment 10), namely: (1) IBA 50 mg / L + 6-BA 25 mg / L, (2) IBA 50 mg / L + CPPU 25 mg / L, (3) NAA 50 mg / L + 6-BA 25 mg / L, and (4) NAA 50 mg / L + CPPU 25 mg / L.

[0105] All cultivation was conducted in a shaded shed with a shading ratio of 40% to 60%. The shed was equipped with an automatic misting system. Depending on the weather, mist water was applied once daily, morning, noon, and evening, for 3 to 6 minutes each time. Sixty rhizomes were cultured for each treatment, with three replicates. After 15 days, root development was analyzed for each treatment, and significance was analyzed using SPSS 21.0. The results are presented below.

[0106] (2) Effect of fertilization methods on seedling growth

[0107] In order to promote the healthy growth of seedlings, this embodiment carried out three kinds of topdressing experiments (treatment 11) based on root induction, namely:

[0108] F1: When the seedlings have 3 to 4 leaves, evenly apply 1g of quick-acting compound fertilizer (N:P:K mass ratio of 16:16:16) on the surface of the substrate every 10 to 15 days, and apply fertilizer 2 to 3 times in succession.

[0109] F2: When the seedlings have 3 to 4 leaves, evenly apply 0.3 to 0.5% urea solution on the surface of the substrate once every 10 days, and apply fertilizer continuously for 3 to 4 times.

[0110] F3: When the seedlings have 3 to 4 leaves, spray the leaves with 0.2% to 0.3% water-soluble fertilizer (N:P:K mass ratio of 20:20:20) once every 7 to 10 days, and spray continuously for 3 to 5 times.

[0111] All cultivation was conducted in a shaded shed with a shading ratio of 40% to 60%. The shed was equipped with an automatic misting system. Depending on the weather, mist water was applied once daily, morning, noon, and evening, with each misting period lasting 3 to 6 minutes. Sixty rhizomes were cultured for each treatment, with three replicates. Fertilization was discontinued during winter when temperatures remained below 15°C. After fertilization and maintenance, plant growth and development were analyzed for each treatment using SPSS 21.0. The results are presented below.

[0112] Here are the results:

[0113] (1) Effects of growth regulator induction on root growth

[0114]

[0115] Statistics show that different growth regulator combinations have different effects on the root differentiation of rhizomes, with the most significant effect on the average root length of rhizomes, followed by the average number of roots. Specifically, "NAA 50mg / L+6-BA 25mg / L" and "NAA 50mg / L+CPPU 25mg / L" have the most significant effects on the average number of roots, but the difference between the two is not significant; "NAA 50mg / L+6-BA25 mg / L" has the most significant effect on the average root length, followed by "NAA 50mg / L+CPPU 25mg / L", and the difference between the two is significant. Therefore, it can be determined that "NAA 50mg / L+6-BA25 mg / L" is the most suitable combination treatment, and its effect on root growth is the most significant. After 15 days of induction culture, the average number of roots of rhizomes can reach 6.2, and the average root length is 6.59cm.

[0116] (2) Effect of fertilization methods on seedling growth

[0117]

[0118] Statistics show that different fertilization methods have different effects on seedling growth. Based on root induction, the seedling rate of all three fertilization methods was 100%, but the effects of the three treatments on average plant height were very significant, with F3 (36.2cm) > F2 (32.7cm) > F1 (27.5cm), indicating that foliar spraying of water-soluble compound fertilizer is most conducive to increasing seedling height. In terms of seedling quality, F3 had the best seedling quality, with thick stems and green leaves, followed by F1. F2 had the worst seedling quality, with thin stems and many leaves withered and yellowed leaves. This may be related to the single urea fertilizer applied to F2, which did not improve the ginger flower seedlings' resistance to low winter temperatures as much as the compound fertilizer applied to F1 and F3. Therefore, F3 can be determined to be the most suitable fertilization method for strong seedlings.

[0119] Example 7 Comparison of the present invention and similar inventions

[0120] To illustrate the innovative effect of the present invention, this embodiment adopts the optimal processing combination method of each of the above embodiments and compares it with the existing similar invention method (CN110915446A), and is implemented as follows:

[0121] Treatment 13: "A ginger flower breeding method based on leaf and stem latent bud promotion" (CN110915446A), specifically:

[0122] (1) Material selection: Select ginger flower stems with inflorescences at the top and past flowering period, with a stem diameter of 10-11 mm (in conjunction with the embodiment, this is the stem diameter after removing the leaf sheath).

[0123] (2) Aseptic treatment of materials: peel off the leaf sheath of the leaf stem layer by layer to the last layer, and cut it into a segment of 4 to 6 cm in a sterile environment; then soak it in 75% alcohol for 1 min, take it out and dry it, then peel off the last layer of leaf sheath to obtain the segment with the hidden bud exposed.

[0124] (3) Hormone treatment: Soak the segments with exposed cryptic buds in a sterile solution of benzylaminopurine (6-BA 50 mg / L) and phenylurea derivatives (TDZ 10-20 mg / L) for 30 minutes, then take them out and dry them in the air.

[0125] (4) Bagging and culturing: Place the air-dried segments in a sterile sealed bag, control the humidity in the sealed bag to 90-95%, and the culture temperature to 20-30°C. Cultivate for 30 days to obtain segments with rhizomes (the average diameter of the rhizomes is 8-10 mm); the sterile treatment of the sealed bag is to sterilize the sealed bag with 75% alcohol for 30 seconds and then dry it under sterile conditions.

[0126] (5) Segment transplanting: Transplant the segments with small rhizomes in the seedbed with a spacing of 10×10 cm; the seedbed matrix is ​​a mixture of peat, river sand and garden soil in a mass ratio of 1:1:1, and 20 to 40 g of organic compound fertilizer is applied per m2 of the seedbed.

[0127] Process 14: The most effective combined method of the present invention

[0128] The inventors strictly repeated the experiment according to the steps of process 13 and compared the results with the final implementation results of the present invention.

[0129] Thirty mature leaf stems were cultured for each treatment, replicated three times. When the average plant height exceeded 30 cm, the leaf stem utilization rate, latent bud germination rate, average reproduction coefficient, seedling rate, seedling cycle, and implementation process evaluation were calculated for each treatment. The results are as follows:

[0130]

[0131]

[0132] Statistics show that although the invention embodiment of CN110915446A of treatment 13 does not mention the utilization rate of the leaf stem stripping segments, the invention requires the segment diameter to be 10-11 mm, and clearly states in Test Example 5 that "the thickness of the segment has an important influence on the germination of latent buds, and not all leaf stems after flowering are suitable for promoting latent buds." This leads to the fact that the utilization rate of the leaf stem segments with a diameter of less than 10 mm is not high. According to the repeated experiments of the inventor, the utilization rate of the leaf stem segments is only 40%, while the present invention can make full use of all leaf stem segments, and the material utilization rate reaches 100%.

[0133] The latent bud germination rate of treatment 13 was 90% to 95%, and the inventor's experiment was repeated to reach 70% to 90%, which was not significantly different from the germination rate of 87.1% to 93.5% of the present invention. This shows that in conventional production, the leaf-peeled stem segments can have a good germination rate without undergoing strict and complicated sterilization, which further verifies the advanced nature of the present invention.

[0134] Although the invention embodiment of treatment 13 does not mention the reproduction coefficient, combined with its utilization rate of leaf stems and the inventor's experimental repetitions, its reproduction coefficient is only 3.6, while the reproduction coefficient of the present invention is 8.8, which is 2.44 times that of the former. The segmented treatment of leaf stems by the present invention can not only make full use of stem segments of different thicknesses, but also multiply the reproduction coefficient, ensuring that under the premise of a limited number of seed stem divisions, the efficient reproduction of ginger flowers can be achieved through leaf stems as much as possible.

[0135] The invention example of treatment 13 shows that the survival rate of small rhizomes after transplantation can reach 100%. After repeated experiments, the inventors found that the survival rate was difficult to exceed 80%. The inventors believe that after a series of sterile treatments and sterile bagging culture, the segments with small rhizomes obtained are also sterile. When they are transplanted into natural materials containing river sand, garden soil, etc., they will inevitably become infected and black rot due to fungi and bacteria in the river sand and garden soil, or they will wilt and die due to difficulty adapting to the natural environment of the seedbed in a short period of time, thereby reducing the ultimate survival rate of the small rhizomes. In contrast, the present invention cultivates stem segments in the natural environment of a shade shed and obtains small rhizomes with root systems. They have strong adaptability to the natural environment, and the transplanting mixed medium used contains almost no fungi and bacteria, so the transplanting survival rate can reach 100%.

[0136] The invention embodiment of treatment 13 focuses on the study of various influencing factors on the initiation of latent buds, and does not involve rooting and seedling cultivation after segment transplantation. After the inventors repeated the experiment according to the transplanting method of the invention, they found that it takes 12 to 14 weeks from segment cultivation to when the average height of the plant exceeds 30 cm, while the present invention as a whole only takes 9 to 13 weeks, which can save 1 to 3 weeks; among them, the lower thick stem segment of the present invention only needs to be cultured for 9 to 10 weeks to become a seedling, which can save 2 to 5 weeks compared with treatment 13, and the seedling cycle is significantly shortened.

[0137] From the implementation process, process 13 needs to be under indoor aseptic conditions, control the size of the sealed bag and the number of segments put into the sealed bag, and the temperature, humidity and the stem thickness of the leaf stem in the sealed bag need to be strictly controlled to achieve the effective germination of the leaf stem hidden bud, and its operation is similar to the aseptic culture in tissue culture, and it is very demanding on the sterile environment and aseptic operation, and aseptic or sterilization operations are cumbersome, strict and complicated, and it is difficult to carry out large-scale seedling cultivation in conventional production. The inventive method is simple to operate, does not require complex sterilization, and even uses sterilizing agents to disinfect during the stem segment culture process, which can also reduce the stem segment preservation rate and germination rate. The inventive method is suitable for carrying out large-scale seedling cultivation of ginger flower in conventional production, and has the advantages of high breeding efficiency, short seedling cycle and low production cost, and can quickly and in large quantities carry out efficient breeding of ginger flower high-quality seedlings, meeting the strong demand in the markets such as landscape and flower gardening.

[0138] The technical solutions provided by the embodiments of the present invention are introduced in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the embodiments of the present invention. The description of the above embodiments is only applicable to help understand the principles of the embodiments of the present invention. At the same time, for those skilled in the art, according to the embodiments of the present invention, there may be changes in the specific implementation methods and application scopes. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. A method for efficiently breeding and raising seedlings of ginger flower, characterized by: The steps include: (1) Leaf stem selection: From August to December each year, select healthy leaf stems that have already bloomed, cut them off at the base and remove the remaining inflorescences; carefully peel off the leaf sheaths of the leaf stems from bottom to top, layer by layer, to the top, exposing the hidden buds on the nodes; (2) Stem segment treatment: Cut the leaf stem in the middle and divide it into two bud-bearing stem segments of approximately equal length, each containing 4 to 6 hidden buds; then soak the upper and lower stem segments in a solution containing a growth regulator for 10 to 20 minutes; The growth regulator solution comprises chlorfenuron and 3-indolebutyric acid in a ratio of 2:

1. The growth regulator solution comprises 40-60 mg of chlorfenuron (CPPU), 20-30 mg of 3-indolebutyric acid (IBA), and the balance of water per liter of solution. The lower stem segment is thick and needs to be soaked for 15 to 20 minutes, while the upper stem segment is thin and needs to be soaked for 10 to 15 minutes. (3) Stem segment culture: First, spread perlite at the bottom of the plug tray with a thickness of 2 to 4 cm; Place the upper and lower stem segments on different plug trays respectively, and cover the surface of the stem segments with a layer of perlite with a thickness of 1 to 2 cm; Place all plug trays in a shade shed with automatic spray, and the shade shed has a shading rate of 40% to 60%; Use spraying to moisturize the plug trays. Depending on the dryness and wetness of the weather, spray mist water once in the morning, noon and evening every day, and each spraying time is 2 to 4 minutes; During the stem segment culture process, it is strictly forbidden to use fungicides for disinfection; Cultivate the lower thick stem segment for 3 to 4 weeks, and the upper thin stem segment for 4 to 5 weeks; (4) Transplanting small rhizomes: Cut the stem segments into segments with small rhizomes and transplant them in batches; (5) Cultivation of strong seedlings: Use an aqueous solution containing plant growth regulators to irrigate the substrate. When the average height of the plants reaches the standard, they can be transplanted or taken out of the field.

2. The method for efficiently breeding and raising seedlings of ginger flower according to claim 1, wherein: The detailed process of step (4) includes: cutting the stem segments into segments with small rhizomes according to the nodes and planting them in batches; placing the segments flat on the substrate surface, and continuing to fill the mixed substrate to completely cover the small rhizomes. Depending on the weather conditions, spray mist water once in the morning, noon and evening, each spraying lasting 3 to 6 minutes. Spray once every 15 days with 2000 to 3000 times of 30% carbendazim solution to prevent fungal diseases.

3. The efficient breeding and seedling raising method of ginger flower according to claim 2, characterized in that: The detailed process of step (4) includes: cutting the upper and lower stem segments into segments with small rhizomes according to the nodes and planting them in batches, each segment is 2 to 3 cm long; placing the segments flat on the substrate surface of the flower pot or nutrient pot, placing one segment per pot, and continuing to fill the mixed substrate to completely cover the small rhizomes. Depending on the dryness and wetness of the weather, spray mist water once in the morning, noon and evening every day, each spraying lasting 3 to 6 minutes, and drench once every 15 days with 2000 to 3000 times of 30% carbendazim solution to prevent and control fungal diseases.

4. The efficient breeding and seedling raising method of ginger flower according to claim 1, characterized in that: The detailed process of step (5) includes: 3 to 5 days after planting, in order to accelerate the root development of the rhizomes and the growth of the seedlings, the substrate is irrigated once with an aqueous solution containing plant growth regulators; when the ginger flower seedlings unfold their leaves, in order to promote the healthy development of the seedlings, water-soluble fertilizer is sprayed on the leaves; if the temperature continues to be low in winter, fertilization is stopped; spraying is continued several times, and the plants can be transplanted or out of the field when the average height exceeds 30 cm.

5. The efficient breeding and seedling raising method of ginger flower according to claim 4, characterized in that: In step (5), each 1 L of the aqueous solution containing the plant growth regulator contains 50 mg of naphthaleneacetic acid (NAA), 25 mg of 6-benzylaminopurine (6-BA), and the balance of water; When the ginger flower seedlings have 3 to 4 leaves, in order to promote the healthy development of the seedlings, spray the leaves with 0.2% to 0.3% water-soluble fertilizer once every 7 to 10 days. The N:P:K mass ratio of the water-soluble fertilizer is 20:20:

20. If the temperature remains below 15°C in winter, stop fertilizing and spray continuously for 3 to 5 times. When the average height of the plants exceeds 30 cm, they can be transplanted or planted out.

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