Method for efficiently inhibiting growth of bulbil of tulipa gesneriana and application thereof
By using tebufenozide as an inhibitor in field cultivation and in vitro culture of Lilium tigrinum, and configuring a specific concentration in combination with topping treatment, the problem of disordered bulbil growth was solved, achieving a synergistic effect of bulbil inhibition and bulb yield increase. A stable regulatory system was established, which is suitable for the industrial production and research of Lilium tigrinum.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- INSTITUTE OF VEGETABLES & FLOWERS CHINESE ACADEMY OF AGRICULTURAL SCIENCES
- Filing Date
- 2026-04-10
- Publication Date
- 2026-07-31
AI Technical Summary
Existing technologies are insufficient for efficiently and specifically inhibiting the growth of bulbils in Lilium tigrinum. Furthermore, traditional methods and general growth regulators are ineffective in field cultivation and in vitro culture, failing to meet production needs and potentially affecting bulb growth.
Using terbufos as the core inhibitor, a specific effective concentration was prepared for field cultivation and in vitro culture. In field cultivation, terbufos was applied by dripping terbufos solution onto the top of the plant, and in in vitro culture, terbufos was added to the culture medium to target and inhibit the key developmental stage of bulbil formation.
It achieves targeted and efficient inhibition of bulbil growth, synergistically promotes bulb growth, improves the yield and quality of underground bulbs, and establishes a stable regulatory system suitable for large-scale field cultivation and scientific research of Lilium tigrinum.
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Figure CN122477932A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the technical field of methods for inhibiting the growth of bulbils in Lilium tigrinum, and particularly relates to a method and application for efficiently inhibiting the growth of bulbils in Lilium tigrinum. Background Technology
[0002] The tiger lily (Lilium tigrinum) is a lily variety with edible, medicinal, and ornamental value. Its underground bulb is the core economically utilized part. However, the abundant natural germination and growth of bulbils competes with the bulb for nutrients, water, and photosynthetic products, hindering bulb enlargement and significantly reducing yield and quality. This has become a key issue restricting the improvement of the economic benefits of large-scale field cultivation of tiger lilies. Traditional production often uses physical methods such as topping to try to inhibit bulbil growth, but this method only achieves partial inhibition, has limited effect, and cannot block the bulbil formation process at the developmental level, failing to meet the actual needs of efficient and targeted bulbil inhibition in production.
[0003] While some growth regulators have been attempted to inhibit bulbils in lilies, and reagents such as flubendiamide and pendimethalin have been preliminarily tested, these reagents are mostly general-purpose and lack specific adaptability to bulbil growth in *Lilium tigrinum*. They do not differentiate between the needs of field cultivation and in vitro culture, nor do they clarify the effective concentration and the target bulbil development stage, resulting in inconsistent inhibitory effects. Furthermore, current technologies have not achieved synergistic development of bulbil inhibition and bulb yield increase. Some reagents may even have adverse effects on bulb growth, and there is a lack of stable and controllable methods for inducing and inhibiting bulbils in in vitro culture systems. This limits the industrial production of *Lilium tigrinum* and hinders in-depth basic research related to bulbil development. Summary of the Invention
[0004] In view of the aforementioned problems, and in conjunction with the first aspect of the present invention, the present invention provides a method for efficiently inhibiting the growth of bulbils of Lilium tigrinum, including at least one implementation method of field cultivation bulbil inhibition and in vitro culture bulbil induction inhibition, using tebufenozide as the core inhibitory reagent, configuring corresponding effective concentrations of tebufenozide for different implementation methods, and achieving targeted inhibition of Lilium tigrinum bulbil growth through reagent application; In the field cultivation implementation, topping treatment can be used to inhibit bulbil growth and increase the yield of underground bulbs of Lilium tigrinum. In the in vitro culture implementation, terbutaline is added to the culture medium to inhibit bulbil induction, and terbutaline can target and inhibit the banded ridge stage of bulbil formation in Lilium tigrinum.
[0005] Preferably, the specific steps for inhibiting bulbils in field cultivation are as follows: prepare an aqueous solution of 50 mg / L terbinafine, and apply the reagent to the field-grown Lilium tigrin plants by top dripping. The Lilium tigrin plants can be topped first before applying the terbinafine aqueous solution. The reagent application is set to be repeated 3 times, and the sample size for each treatment is not less than 30 plants.
[0006] Preferably, the field-cultivated lily of tiger lily is grown in a mountainous area at an altitude of 1300-1400m. After the reagent is applied, the growth of bulbils is statistically analyzed at 15 days and 30 days. The bulbil inhibition rate is calculated with the untopped water treatment group as the control.
[0007] Preferably, the specific steps for in vitro culture of bulbil induction inhibition are as follows: after vernalization treatment, stem segments are cut from Lilium tigrin bulbs, and after surface sterilization, small stem segments containing single leaf axils are cut and inoculated into in vitro culture medium supplemented with tebufenozide for culture, wherein the effective concentration of tebufenozide in the in vitro culture medium is 15 mg / L.
[0008] Preferably, the surface sterilization is performed sequentially using 75% alcohol and 10% NaClO; the in vitro culture medium is formulated as follows: Sucrose + 6 g / L agar, culture conditions: 25℃, 16 / 8 h light / dark cycle, light intensity... .
[0009] Preferably, the vernalization treatment conditions for the *Lilium tigrinum* bulbs are low-temperature vernalization at 4℃, and the stem segments are cut and inoculated when they elongate to about 10cm after vernalization.
[0010] Preferably, the specific stage at which the tebufenozide targets and inhibits the formation of bulbils in Lilium tigrin is the 12-day ridge stage of bulbil development. After treatment with 15 mg / L tebufenozide, the cell division activity in the leaf axils of Lilium tigrin decreases, and by the 16-day ridge of bulbil development, it is still only in a slightly ridged state and cannot form bulbil growth points and scale differentiation.
[0011] Preferably, sec-butylamine can be used in combination with one or two of flubendiamide and pendimethalin. Flubendiamide and pendimethalin are used in the form of aqueous solutions in field cultivation and are added directly to the in vitro culture medium in in vitro culture. When used in combination, the concentration of each reagent is the effective concentration when used alone.
[0012] Preferably, after treatment with 50 mg / L tebufenozide during field cultivation, the diameter, fresh weight, and dry weight of the underground bulbs of Lilium tigrin were significantly increased compared with the water treatment group, achieving a synergistic effect of bulbil inhibition and bulb yield increase.
[0013] Based on the above, this invention uses tebufenozide as the core inhibitory agent and configures specific effective concentrations for both field cultivation and in vitro culture of Lilium tigrinum, achieving targeted and efficient inhibition of bulbil growth. In field cultivation, topping treatment can be used to further enhance the inhibitory effect. Furthermore, tebufenozide can precisely target the key developmental stage of bulbil formation in Lilium tigrinum, blocking the formation and growth process of bulbils from the source of development. Compared with inhibitors such as flubendiamide and pendimethalin, its inhibitory effect on bulbils is more targeted and stable, effectively solving the problem of disordered bulbil growth in Lilium tigrinum.
[0014] This invention achieves efficient bulbil inhibition while synergistically promoting the growth of underground bulbs in Lilium tigrinum, significantly improving bulb growth and realizing the dual effects of bulbil inhibition and bulb quality improvement and yield enhancement, thus significantly increasing the economic value of large-scale field cultivation of Lilium tigrinum. Simultaneously, the established in vitro culture method for bulbil inhibition provides a stable and controllable regulatory system for related research on bulbil development in Lilium tigrinum, and also provides a replicable and referable technical solution for the directional regulation of bulbil growth in other plants of the Lilium genus, possessing both practical production application value and scientific research reference value. Attached Figure Description
[0015] Figure 1 The topping method provided in this embodiment of the invention has an impact on the underground bulbs of *Tilia tigrinosa*. A: Phenotypic observation of underground bulbs after topping and control (CK), scale bar = 10cm. B: Effect of topping on the fresh and dry weight of *Tilia tigrinosa* underground bulbs.
[0016] Figure 2 This invention relates to the effects of terbufotoxin treatment on the phenotype and underground bulbs of *Tilia tigrin* plants, as provided in this embodiment. A: Phenotypic observation of T4 plants after topping, scale bar = 20cm. B: Phenotypic observation of T4 underground bulbs after topping, scale bar = 10cm. Figure 3 This invention provides an embodiment of the observation of leaf axil morphology during the formation of *Tilia tigrinosa* bulbils under in vitro conditions. A→G: Observation of leaf axil morphology during the formation of bulbils of *Lysimachia christinae* under in vitro conditions (A→G are 0d, 2d, 4d, 6d, 8d, 10d, 12d respectively), scale bar = 1mm; Figure 4 This is an observation of leaf axil morphology during bulbil formation in the 15 mg / L sec-butylamine treatment group provided in this embodiment of the invention; A→G: Observation of leaf axillary morphology during bulbil formation in the 15mg / L sec-butyridine treatment group (A→G were 0d, 2d, 4d, 6d, 8d, 10d, 12d respectively), scale bar = 1mm; Figure 5 This is a histological observation of the bulbil formation process in the 15 mg / L sec-butylamine treatment group provided in the embodiments of the present invention; AC: Morphological observation during bulbil induction (A→C at 8d, 12d, and 16d respectively). ac: Anatomical observation during bulbil induction (a→c were 8d, 12d, and 6d respectively); DF: Morphological observation during bulbil induction treated with Butralin 15 mg / L (D→F were 8d, 12d, and 16d respectively). df: Anatomical observation during bulbil induction treated with Butralin 15mg / L (d→f were 8d, 12d, and 16d respectively). The scales for A, B, D, E, and F are 1 mm, and the scale for C is 0.5 mm. The scale bars for A, B, D, E, and F are 200 μm, and the scale bar for C is 400 μm. Detailed Implementation
[0017] The invention will now be described in detail with reference to the accompanying drawings, including a preliminary screening of field inhibition of bulbils in *Lilium tigrinum*. Experimental Methods: Location: Huawu Weicheng Lily Base, Weicheng Town, Qingzhen City, Guiyang City, Guizhou Province, 26°44'34"N, 106°22'58"E, altitude 1347 m. Top drip irrigation was used, n=30, 3 replicates.
[0018] Inhibition rate = (Number of bulbils in experimental group - Number of bulbils in control group) / Number of bulbils in control group × 100% Table 1: Effects of different inhibitors on *Lilium tigrinum* bulbils;
[0019] It was found that, compared with not topping, the number of bulbils was significantly reduced after 30 days, with an inhibition rate of 72.41%.
[0020] It was found that three reagents, namely flubendiamide, sec-butylamine, and pendimethalin, could inhibit the growth of *Tilia tigrin* bulbils to varying degrees. Among them, sec-butylamine was the most effective, and the inhibition effect of sec-butylamine at 50 mg / L was the best, reaching 94.21% compared with no topping and 79.01% compared with topping.
[0021] Experimental method: The bulbs of *Ligustrum lucidum* located at the Huawu Weicheng Lily Base in Weicheng Town, Qingzhen City, Guiyang City, Guizhou Province were harvested on August 14, 2025.
[0022] Bulb diameter measurement: Measure the diameter of the longest single bulb.
[0023] Individual bulb weight: 30 samples were randomly selected from the experimental area as a group, and their weight was measured using a balance, n=30, with 3 replicates. Dry weight: The bulbs were dried in an oven until their weight no longer changed.
[0024] Increase in yield = (Experimental group yield - Control group yield) / Experimental group yield × 100%.
[0025] Table 2: Effects of different inhibitors on the yield of *Tiger lily bulbils*;
[0026] Further explanation Figure 1 The impact of topping on the underground bulbs of *Tiger lily*; A: Phenotypic observation of underground bulbs after topping and control (CK), scale bar = 10cm. B: Effect of topping on the fresh and dry weight of *Tilia tigrinosa* underground bulbs; Figure 2 The effects of cypermethrin treatment on the phenotype and underground bulbs of *Tilia tigrin* plants; A: Phenotypic observation of T4 plants after topping, scale bar = 20cm. B: Phenotypic observation of T4 underground bulbs after topping, scale bar = 10cm. Upon harvesting, it was found that the diameter, weight, fresh weight, and dry weight of individual underground bulbs treated with 50 mg / L terbufotazone increased. This indicates that treatment with 50 mg / L terbufotazone significantly increased the yield of underground bulbs.
[0027] Verification of the inhibitory effect of gentian on bulbils of *Tiger lily* In vitro induction culture of *Lysimachia christinae* bulbils Experimental Methods: Triploid *Lilium lancifolium* bulbs preserved in the Lily Research Group of the Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, were used as experimental materials. In August 2024, bulbs of uniform size were selected for embedding and vernalization (4℃). An in vitro bulbil induction system was used. After vernalization, when the stem segments of the *Lilium lancifolium* bulbs elongated to approximately 10 cm, the stem segments were cut, the bottom and outer leaves were removed, and the surfaces were sterilized with 75% alcohol and 10% NaClO. Small stem segments containing single leaf axils were then inoculated onto a culture medium formulated as MS + 30 g / L sucrose + 6 g / L agar. The culture conditions were: 25℃, 16 / 8 h light / dark cycle. Light intensity.
[0028] growth status of bulbils 0-12 days as follows Figure 3 As shown: A→G: Observation of leaf axil morphology during bulbil formation in *Lysimachia christinae* under in vitro conditions (A→G are 0d, 2d, 4d, 6d, 8d, 10d, 12d respectively), scale bar = 1mm.
[0029] Figure 3 Observation of leaf axil morphology during bulbil formation in *Lysimachia christinae* under in vitro conditions; Concentration screening of cyprodinium for inhibiting the growth of *Tilia tigrin* bulbils; Table 3; Concentration screening of methylphenidate for inhibition of in vitro culture of Tilanthus tigrin;
[0030] In terms of treatment concentration, 15 mg / L of tebufenozide showed the best inhibitory effect on bulbils of Tilia tiglium, reaching 51.57%.
[0031] A→G: Observation of leaf axillary morphology during bulbil formation in the 15mg / L tebufenozide treatment group (A→G were 0d, 2d, 4d, 6d, 8d, 10d, and 12d, respectively), scale bar = 1mm.
[0032] Figure 4 Observation of leaf axillary morphology during bulbil formation in the 15mg / L terbufos treatment group; Effects of three bulbil inhibitors on *Lysimachia nummularia* bulbils under in vitro conditions; Table 4; Screening of different inhibitors on the inhibitory effect of different inhibitors on in vitro culture of *Lysimachia nummularia* bulbils;
[0033] The results were largely similar to those in the field trials, with the treatment with cyprodinil achieving the best results, resulting in a bulbil induction rate of only 53.33%.
[0034] Paraffin sections; AC: Morphological observation during bulbil induction (A→C at 8d, 12d, and 16d respectively). ac: Anatomical observation during bulbil induction (a→c were 8d, 12d, and 6d respectively); DF: Morphological observation during bulbil induction treated with Butralin 15 mg / L (D→F were 8d, 12d, and 16d respectively). df: Anatomical observation during bulbil induction treated with Butralin 15 mg / L (d→f were 8d, 12d, and 16d respectively). The scales for A, B, D, E, and F are 1 mm, and the scale for C is 0.5 mm. The scale bars for A, B, D, E, and F are 200 μm, and the scale bar for C is 400 μm. Figure 5 Histological observation of bulbil formation in the 15 mg / L sec-butylamine treatment group; Histological results from paraffin sections showed that in the CK group, ridges began to appear at 8 days, bulbils emerged at 12 days, and the bulbils had differentiated into multiple layers of scales by 16 days. In contrast, no axillary organ ridges were observed in the 15 mg / L Butralin group at 8 days; only slight ridges began to appear at 12 days, and the ridges remained at the ridge stage by 16 days. We also observed that the staining in the leaf axils was darker at 8 days, gradually lightening by 12 and 16 days in the treatment groups, indicating less vigorous cell division.
[0035] 5. Through long-term observation, we found that the bulbils in the CK group grew normally, while those in the 15 mg / L Butralin group did not grow. We found that Butralin inhibited the growth of bulbils at the 12-day banded ridge stage, indicating that it inhibited the occurrence of bulbils in *Lysimachia christinae*.
[0036] When the bulbils prolong in the 15 mg / L sec-butylamine treatment group.
[0037] The embodiments of this application have been described above with reference to the accompanying drawings. Unless otherwise specified, the embodiments and features in the embodiments of this application can be combined with each other. This application is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this application.
Claims
1. A method of efficiently inhibiting growth of a Japanese Show Lily bulb, comprising: Includes the following steps: The invention includes at least one of field cultivation bulbil inhibition and in vitro culture bulbil induction inhibition, with tebufenozide as the core inhibitory agent. The effective concentration of tebufenozide is configured for different implementation methods, and the targeted inhibition of bulbil growth of Lilium tigrin is achieved by applying the agent. In the field cultivation implementation, topping treatment can be used to inhibit bulbil growth and increase the yield of underground bulbs of Lilium tigrinum. In the in vitro culture implementation, terbutaline is added to the culture medium to inhibit bulbil induction, and terbutaline can target and inhibit the banded ridge stage of bulbil formation in Lilium tigrinum.
2. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: The specific steps for inhibiting bulbils in field cultivation are as follows: Prepare a 50 mg / L solution of terbufotoxin and apply the reagent to the field-grown Lilium tigrin plants by top dripping. The Lilium tigrin plants can be topped first before applying the terbufotoxin solution. The reagent application is set up to be repeated 3 times, and the sample size for each treatment is no less than 30 plants.
3. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: The field-grown *Lilium tigrinum* was planted in a mountainous area at an altitude of 1300-1400m. After the reagent was applied, the growth of bulbils was statistically analyzed at 15 and 30 days. The bulbil inhibition rate was calculated using the un-topped water treatment group as a control.
4. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: The specific steps for in vitro culture of bulbil induction inhibition are as follows: after vernalization treatment, stem segments are cut from Lilium tigrinum bulbs, and after surface sterilization, small stem segments containing single leaf axils are cut and inoculated into in vitro culture medium supplemented with tebuconazole for culture. The effective concentration of tebuconazole in the in vitro culture medium is 15 mg / L.
5. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 4, characterized in that: The surface sterilization was performed sequentially using 75% alcohol and 10% NaClO; the in vitro culture medium was formulated as MS + 30 g / L sucrose + 6 g / L agar, and the culture conditions were 25℃, 16 / 8h light / dark cycle, and light intensity [not specified]. .
6. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 4, characterized in that: The vernalization treatment conditions for the *Lilium tigrinum* bulbs were 4℃ low-temperature vernalization. After vernalization, the stem segments were cut and inoculated when they reached about 10cm in length.
7. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: The specific stage at which tebufenozide targets and inhibits bulbil formation in Lilium tigrin is the 12-day ridge stage of bulbil development. After treatment with 15 mg / L tebufenozide, the cell division activity in the leaf axils of Lilium tigrin decreases, and by the 16-day ridge of bulbil development, it is still only in a slightly ridged state and cannot form bulbil growth points and scale differentiation.
8. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: It is also possible to use sec-butylamine in combination with one or two of flubendiamide and pendimethalin. Flubendiamide and pendimethalin are used in the form of aqueous solution in field cultivation, and are added directly to the in vitro culture medium in in vitro culture. When used in combination, the concentration of each reagent is the effective concentration when used alone.
9. The method for efficiently inhibiting the growth of bulbils in Lilium tigrinum according to claim 1, characterized in that: In the field cultivation, after treatment with 50 mg / L tebufenozide, the diameter, fresh weight, and dry weight of the underground bulbs of Lilium tigrin were significantly increased compared with the water treatment group, achieving a synergistic effect of bulbil inhibition and bulb yield increase.