Micro-grafting method for chamomile and artemisia annua
By using Artemisia annua and chamomile seedlings of matching age and diameter, combined with silicone tube fixation, the problem of low grafting survival rate of chamomile was solved, achieving high aphid resistance and simple grafting operation, thus promoting the industrial application of chamomile.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-03-04
- Publication Date
- 2026-04-07
AI Technical Summary
The existing cleft grafting method for chamomile and artemisia has problems with rootstock lignification and hollowing, resulting in a low healing rate. In addition, traditional grafting methods are prone to physical damage and low survival rate, making it difficult to promote on a large scale.
Artemisia annua seedlings aged 28 days were used as rootstocks, and Chrysanthemum morifolium seedlings aged 35 days were used as scions. The grafting interface was fixed with a silicone tube with an inner diameter similar to that of the scion, and the grafting was performed under sterile conditions with controlled light and humidity.
It improved the grafting survival rate, reduced the mechanical damage rate and adventitious root rate, enhanced the aphid resistance of chamomile, simplified the operation process, and has scientific research value and promotion potential.
Smart Images
Figure CN121795244A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of plant micrografting technology, specifically relating to a micrografting method for chamomile and artemisia. Background Technology
[0002] Chamomile is an important ornamental flower with high economic, medicinal, and cultural value, and is widely used in landscaping, cut flower production, and medicinal processing. However, aphids and other piercing-sucking pests cause serious damage during chamomile production, leading to stunted plant growth, curled leaves, deformed flower buds, and even the spread of various viral diseases, greatly affecting the ornamental quality and commercial value of chamomile. Currently, the control of chamomile aphids mainly relies on chemical pesticides, but long-term use can easily lead to environmental pollution, increased pesticide resistance in pests, and excessive pesticide residues, which is detrimental to the development of a green and sustainable flower industry.
[0003] Artemisia annua, a plant of the Asteraceae family, possesses significant medicinal and ecological value due to its rich content of artemisinin and other secondary metabolites. Studies have shown that Artemisia annua plants exhibit strong natural resistance to pests such as aphids, and its root exudates contain various insecticidal compounds that effectively inhibit pest reproduction. Therefore, using Artemisia annua as rootstock and grafting it onto chamomile to impart aphid resistance is a biological control approach that combines ecological safety with effective pest control.
[0004] Currently, chamomile and Artemisia annua are mainly grafted using the cleft grafting method. However, the selected Artemisia annua rootstocks often have issues such as being older (over 45 days old) or taller (over 45 cm). At this age, the pith of the rootstock has already lignified and become hollow, causing a sharp drop in the proportion of thin-walled cells at the cleft grafting site, making it difficult for the cambium layers to align, and reducing the healing rate from over 90% to below 50%. Furthermore, the cleft grafting method is prone to physical damage during the grafting process, causing the scion to fall off during growth, or its activity to decrease due to prolonged direct exposure to air. This results in low grafting survival rates, poor rootstock compatibility, and high operational difficulty, hindering large-scale promotion and industrial application. Summary of the Invention
[0005] To address the above technical problems, this invention provides a micrografting method for chamomile and artemisia.
[0006] To achieve the above objectives, the technical solution of the present invention is as follows: A micrografting method for chamomile and artemisia, comprising the following steps: Under sterile conditions, Artemisia annua seedlings aged 28 days were used as rootstocks, and Chrysanthemum morifolium shoots aged 35 days were used as scions. The rootstock and scion were tightly grafted using a flat grafting method, and the graft union was secured with a silicone tube with an inner diameter similar to that of the scion and cut on the side. After grafting, the grafted seedlings were placed on the surface of the culture medium, and the culture dishes were placed vertically in a plant culture room, where the light intensity was controlled at 8000 lux and the humidity at 60%.
[0007] Preferably, the rootstock is obtained by selecting Artemisia annua seedlings that have been cultured in a culture medium for 28 days, cutting them horizontally at the junction of the root and stem with a blade, and using the remaining Artemisia annua roots as rootstock.
[0008] Preferably, the chamomile tissue culture seedlings are propagated by subculture for 35 days, and 5mm chamomile terminal shoots are selected as scions.
[0009] Preferably, the culture medium is 1 / 2 MS sugar-free medium, which is prepared by adding 0.2203g of MS medium (including vitamins) powder and 0.8g of agar to every 100mL of water and adjusting the pH to 5.8.
[0010] Preferably, the diameter of the cut surface of the chamomile tip is 1.0mm ± 0.1mm; and the diameter of the cut surface of the Artemisia annua root is 1.0mm ± 0.1mm.
[0011] Preferably, the silicone tube has a length of 3 mm and an inner diameter of 1 mm.
[0012] Preferably, during the grafting process, the cut surfaces of the chamomile tip and the Artemisia root are kept perpendicular to the axial direction of the silicone tube.
[0013] Preferably, the seeds of Artemisia annua seedlings are sterilized by adding 75% ethanol, shaking for 1 minute, pouring out the ethanol, adding 5% sodium hypochlorite and shaking for 20 minutes, rinsing three times with sterile water, and after sterilization, evenly placing them on the surface of the culture medium, vernalizing them for 3 days under shaded conditions at 4°C, and then transferring them to light for 28 days of cultivation.
[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The grafting method of the present invention is simple to operate, has a short material preparation cycle, fast grafting interface healing, and high survival rate.
[0015] 2. The grafting method of the present invention performs grafting during the seedling stage, enabling chamomile to have aphid resistance from the seedling stage, reducing the risk of aphid damage during the seedling stage.
[0016] 3. The grafting method of this invention not only provides an efficient technical solution for the grafting research of chamomile and artemisia annua, but also provides a reliable reference for the micro-grafting technology and rootstock-scion interaction mechanism research of other plants, and has important scientific research value and potential for technology promotion. Attached Figure Description
[0017] Figure 1 This is a schematic diagram illustrating the implementation process of the micrografting method for Artemisia annua in this invention.
[0018] Figure 2 This is a comparative analysis chart showing the average number of aphids per plant in Example 1 and in ungrafted chamomile seedlings of the same age.
[0019] Figure 3 This is a statistical analysis of the phenotypes of silicone tubes of different specifications grafted 10 days after the invention's embodiments and comparative examples.
[0020] Figure 4 The micro-grafted seedlings of Chrysanthemum indicum and Artemisia annua were grafted on the same day in this embodiment of the invention.
[0021] Figure 5 This is a diagram showing the adventitious roots generated on the scion above the grafting interface of the grafted plant in an embodiment of the present invention.
[0022] Figure 6 This is a grafting point diagram of the grafted plant 14 days after transplanting it into the soil, as shown in this embodiment of the invention.
[0023] Figure 7 This is a picture of a grafted plant that has been transplanted into the soil and grown for 14 days in an embodiment of the present invention.
[0024] Figure 8 This is a grafting point diagram of the grafted plant 60 days after transplanting, as shown in this embodiment of the invention.
[0025] Figure 9 This is a diagram showing the grafted plant's growth status 60 days after transplanting, as described in an embodiment of the present invention. Detailed Implementation
[0026] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0027] Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0028] Currently, the main grafting technique for chamomile and Artemisia annua is cleft grafting. However, since the selected Artemisia annua rootstock is taller than 45cm, it is prone to pith lignification and hollowing, which causes a sharp drop in the proportion of thin-walled cells at the cleft grafting site, making it difficult for the cambium layers to align and resulting in a decrease in the healing rate.
[0029] Furthermore, the traditional PE film used in grafting suffers from insufficient elasticity, leading to physical damage and a graft union rot rate of 5-8%. The grafting clips used lack sufficient mechanical strength, causing them to detach during the growth of the grafted plant. Exposed scions are prone to wilting due to water loss. The lack of precise control over the geometric tolerances of the scion's wedge-shaped surface and the rootstock's cleft can easily cause misalignment of the cambium layers of the rootstock and scion, hindering the reconnection of vascular bundles. These problems result in low grafting survival rates and high operational difficulty, hindering large-scale promotion and industrial application.
[0030] The technical solution of the present invention will be further described below through specific embodiments.
[0031] In the following embodiments, unless otherwise specified, the methods described are conventional methods; and unless otherwise specified, the reagents and materials described are commercially available.
[0032] Example 1 A micrografting method for chamomile and artemisia, comprising the following steps: S1. Cultivation of Artemisia annua rootstock: Preparation of 1 / 2 MS sugar-free medium: Add 0.2203g of MS medium (including vitamins) powder and 0.8g of agar to every 100mL of water, and adjust the pH to 5.8.
[0033] Sterilization of Artemisia annua seeds: Add 75% ethanol, shake for 1 min, pour out the ethanol, add 5% sodium hypochlorite and shake for 20 min, rinse three times with sterile water, and sow evenly on the surface of 1 / 2 MS medium.
[0034] After sowing, Artemisia annua seeds were vernalized for 3 days under shaded conditions at 4℃, and then transferred to light for cultivation for 28 days to be used as rootstock. During the cultivation process, the materials were observed for contamination.
[0035] S2. Cultivation of Chamomile: Select chamomile tissue culture seedlings for propagation. The specific operation is as follows: Prepare 1 / 2 MS medium, sterilize tools such as tweezers and scissors by autoclaving, operate in a clean bench, cut off the leaves, cut the stem segments into small segments, each segment containing at least one bud, insert the cut stem segments into new medium, and cultivate for 35 days.
[0036] S3. Grafting implementation: Under sterile conditions, select 28-day-old Artemisia annua seedlings with a diameter of 1mm, and cut them horizontally with a blade at the junction of the rootstock and stem, using the remaining Artemisia annua root as the rootstock; use the top shoot to the bottom 5mm of chamomile seedlings with a diameter of 1mm at 35 days as the scion.
[0037] Select a 3mm long, 1mm inner diameter silicone tube with a side cut, and use a flat grafting method to tightly attach the rootstock and scion of Artemisia annua and Chrysanthemum morifolium. Place the fixed grafted seedling on the surface of the culture medium, with the culture dish placed vertically to avoid contact between the grafting point and the culture medium.
[0038] S4. Post-grafting cultivation: After 10 days, the graft union will heal. Prepare sterile water and immerse the lower end of the rootstock in the water, avoiding contact between the graft union and water. Three days later, transplant into a substrate with a peat moss:vermiculite volume ratio of 3:1, ensuring the graft union is far from the substrate to prevent adventitious root formation. Observe promptly after transplanting and remove any adventitious roots that appear. Water appropriately after transplanting, controlling light intensity to 8000 lux and humidity to 60%. The plants will grow vigorously approximately two months after transplanting. Calculate the graft union healing rate, damage rate, and survival rate 10 days after grafting; the results are shown in Table 1. Calculate the aphid resistance rate of the grafted seedlings 60 days after grafting; the results are shown in Table 1. Figure 2 As shown.
[0039] Comparative Example 1 A micrografting method for chamomile and artemisia, comprising the following steps: S1. Cultivation of Artemisia annua rootstock: Preparation of 1 / 2 MS sugar-free medium: Add 0.2203g of MS medium (including vitamins) powder and 0.8g of agar to every 100mL of water, and adjust the pH to 5.8.
[0040] Sterilization of Artemisia annua seeds: Add 75% ethanol, shake for 1 min, pour out the ethanol, add 5% sodium hypochlorite and shake for 20 min, rinse three times with sterile water, and sow evenly on the surface of 1 / 2 MS medium.
[0041] After sowing, Artemisia annua seeds were vernalized for 3 days under shaded conditions at 4℃, and then transferred to light for cultivation for 28 days to be used as rootstock. During the cultivation process, it was observed in time whether there was any contamination.
[0042] S2. Cultivation of Chamomile: Select chamomile tissue culture seedlings for propagation. The specific operation is as follows: Prepare 1 / 2 MS medium, sterilize tools such as tweezers and scissors by autoclaving, operate in a clean bench, cut off the leaves, cut the stem segments into small segments, each segment containing at least one bud, insert the cut stem segments into new medium, and cultivate for 35 days.
[0043] S3, S3, Grafting Implementation: Under aseptic conditions, select 28-day-old Artemisia annua seedlings with a diameter of 1mm, and cut them horizontally with a blade at the junction of the rootstock and stem, using the remaining Artemisia annua root as the rootstock; use the top shoot to the bottom 5mm of chamomile seedlings with a diameter of 1mm and a diameter of 35 days as the scion.
[0044] Select a 3mm long silicone tube with a 0.9mm inner diameter and cut on the side. Use a flat grafting method to tightly attach the rootstock and scion of Artemisia annua and Chrysanthemum morifolium. Place the fixed grafted seedling on the surface of the culture medium, with the culture dish placed vertically to avoid contact between the grafting point and the culture medium.
[0045] S4. Post-grafting cultivation: After 10 days, the graft union will heal. Prepare sterile water and immerse the lower end of the rootstock in the water, avoiding contact between the graft union and water. Three days later, transplant into a substrate with a peat moss:vermiculite volume ratio of 3:1, ensuring the graft union is far from the substrate to prevent adventitious root formation. Observe promptly after transplanting and remove any adventitious roots that appear. Water appropriately after transplanting, controlling light intensity to 8000 lux and humidity to 60%. The plants will grow vigorously approximately two months after transplanting. Calculate the graft union healing rate, damage rate, and survival rate 10 days after grafting; the results are shown in Table 1. Calculate the aphid resistance rate of the grafted seedlings 60 days after grafting; the results are shown in Table 1. Figure 2 As shown.
[0046] Comparative Example 2 A micrografting method for chamomile and artemisia, comprising the following steps: S1. Cultivation of Artemisia annua rootstock: Preparation of 1 / 2 MS sugar-free medium: Add 0.2203g of MS medium (including vitamins) powder and 0.8g of agar to every 100mL of water, and adjust the pH to 5.8.
[0047] Sterilization of Artemisia annua seeds: Add 75% ethanol, shake for 1 min, pour out the ethanol, add 5% sodium hypochlorite and shake for 20 min, rinse three times with sterile water, and sow evenly on the surface of 1 / 2 MS medium.
[0048] After sowing, Artemisia annua seeds were vernalized for 3 days under shaded conditions at 4℃, and then transferred to light for cultivation for 28 days to be used as rootstock. During the cultivation process, it was observed in time whether there was any contamination.
[0049] S2. Cultivation of Chamomile: Select chamomile tissue culture seedlings for propagation. The specific operation is as follows: Prepare 1 / 2 MS medium, sterilize tools such as tweezers and scissors by autoclaving, operate in a clean bench, cut off the leaves, cut the stem segments into small segments, each segment containing at least one bud, insert the cut stem segments into new medium, and cultivate for 35 days.
[0050] S3, S3, Grafting Implementation: Under aseptic conditions, select 28-day-old Artemisia annua seedlings with a diameter of 1mm, and cut them horizontally with a blade at the junction of the rootstock and stem, using the remaining Artemisia annua root as the rootstock; use the top shoot to the bottom 5mm of chamomile seedlings with a diameter of 1mm and a diameter of 35 days as the scion.
[0051] Select a 3mm long silicone tube with an inner diameter of 1.1mm and cut on the side. Use a flat grafting method to tightly attach the rootstock and scion of Artemisia annua and Chrysanthemum morifolium. Place the fixed grafted seedling on the surface of the culture medium, with the culture dish placed vertically to avoid contact between the grafting point and the culture medium.
[0052] S4. Post-grafting care: After 10 days, once the graft union has healed, immerse the lower end of the rootstock in sterile water, avoiding direct contact between the graft union and water. Three days later, transplant into a substrate with a peat moss:vermiculite volume ratio of 3:1, ensuring the graft union is far from the substrate to prevent adventitious root formation. Observe the plant promptly after transplanting; remove any adventitious roots that appear. Water appropriately after transplanting, controlling light intensity to 8000 lux and humidity to 60%. Vigorous plant growth is expected approximately two months after transplanting. The graft union healing rate, damage rate, and survival rate were calculated 10 days after grafting, as shown in Table 1. The aphid resistance rate of the grafted seedlings was calculated 60 days after grafting, as shown in Table 1. Figure 2 As shown.
[0053] Table 1. Comparative Analysis of Technical Indicators of Example 1 and Comparative Examples 1-2 Table 1 presents the statistical data of Example 1, Comparative Example 1, and Comparative Example 2. The data shows that the management method for micro-grafted seedlings of *Artemisia annua* exhibits significant advantages in several key indicators. The mechanical damage rate of Example 1 was 7%, a decrease of 91.3% compared to Comparative Example 1; in terms of the rate of adventitious root formation, Example 1 was 13%, a decrease of 83.8% compared to Comparative Example 2. It is evident that the improved survival rate of the grafted seedlings is mainly due to the appropriate selection of silicone tubes. These data fully demonstrate the comprehensive advantages of this invention in improving grafting quality and reducing production costs.
[0054] Figure 2 Statistical analysis was conducted on the average aphid count per plant of *Chrysanthemum indicum* (Example 1) and ungrafted chamomile seedlings of the same age. The data shows that the management method for micro-grafted *Chrysanthemum indicum* seedlings exhibits a significant advantage in aphid resistance compared to ungrafted chamomile. The average aphid count per plant in Example 1 was 6.46, while the average aphid count per plant in Example 2 was 33.6. The aphid resistance rate of Example 1 was increased by 77.7% compared to ungrafted chamomile. This demonstrates that grafting significantly improves the aphid resistance of chamomile, and these data fully prove the comprehensive advantages of this invention in improving grafting quality and reducing production costs.
[0055] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A method for micrografting chamomile and artemisia, characterized in that, Includes the following steps: Under sterile conditions, Artemisia annua seedlings aged 28 days were used as rootstocks and Chrysanthemum morifolium shoots aged 35 days were used as scions. The rootstock and scion were tightly attached using a flat grafting method, and the grafting interface was fixed with a silicone tube with an inner diameter similar to that of the scion and cut on the side. After grafting, the grafted seedlings were placed on the surface of the culture medium, and the culture dish was placed vertically in the plant culture room for cultivation.
2. The micrografting method for Artemisia annua according to claim 1, characterized in that, The method for obtaining the rootstock is as follows: select seedlings of Artemisia annua that have been cultivated in a culture medium for 28 days, cut them horizontally with a blade at the junction of the root and stem, and use the remaining roots of Artemisia annua as the rootstock.
3. The micrografting method for Artemisia annua according to claim 1, characterized in that, The method for obtaining the scion is as follows: chamomile tissue culture seedlings are propagated by subculture for 35 days, and 5mm long chamomile top shoots are selected as scions.
4. The micrografting method for *Artemisia annua* according to claim 1, characterized in that, The culture medium was 1 / 2 MS sugar-free solid medium; The method for preparing the 1 / 2 MS sugar-free medium is as follows: add 0.2203g of MS Medium powder and 0.8g of agar to every 100mL of water, and adjust the pH to 5.
8.
5. The micrografting method for *Artemisia annua* according to claim 1, characterized in that, The diameter of the cut surface of the chamomile tip is 1.0mm ± 0.1mm; the diameter of the cut surface of the Artemisia annua root is 1.0mm ± 0.1mm.
6. The micrografting method for *Artemisia annua* according to claim 1, characterized in that, The silicone tube is 3mm long and has an inner diameter of 1mm.