Field non-in-vitro regeneration method for rubus plants

Abstract

The invention discloses a field non-in-vitro regeneration method for rubus plants, and belongs to the technical field of rubus plant breeding. The method comprises the steps of girdling a target branch, spraying a plant growth regulator, wrapping moss and covering soil. The method has the advantages of being environmentally friendly and efficient (in-vitro culture is not needed, field resources are directly utilized, and use of chemical agents is reduced), genetic stability is kept (good characters of stock plants are reserved through asexual propagation), adaptability is high (the rooting process can be completed in the natural environment, the rooting rate exceeds 90%, and the transplanting survival rate reaches 90% or above), population can be rapidly expanded, and the method is suitable for large-scale popularization and application. And regional biodiversity is maintained. Therefore, a beneficial method is provided for biodiversity protection of rubus plants, and a reference method is also provided for field breeding of other rare and endangered species.

Description

Technical Field

[0001] The present invention relates to the technical field of Rubus plant breeding, and particularly to a method for in-situ regeneration of Rubus plants in the wild. Background Art

[0002] As an important part of the forest ecosystem, Rubus spp. plants have functions such as soil and water conservation, species symbiosis, and fruit economic value (such as Rubus idaeus, Rubus parvifolius, Rubus corchorifolius, Rubus fruticosus, etc.). However, traditional breeding techniques have significant ecological risks: (1) Destruction of wild resources: A large number of wild branches need to be collected for traditional division or cutting, resulting in damage or even death of the mother plants, threatening the continuation of the population; (2) Gene simplification: Seed propagation is vulnerable to environmental screening pressure, weakening the genetic diversity of natural populations; (3) Disturbance of the ecological chain: In vitro propagation requires cultivation in other places, and the transplantation of seedlings is prone to the risk of alien species invasion; (4) Chemical pollution: Tissue culture relies on hormones and antibiotics, which have a negative impact on the soil microbial community.

[0003] The existing techniques related to Rubus breeding are generally cutting, tissue culture, root sucker propagation, etc., which are all in vitro regeneration. Due to seasonal limitations, environmental regulation generally needs to be carried out manually, and it is mostly applicable to commercial asexual reproduction. Compared with other breeding methods, there is no report on the in-situ regeneration technology for wild Rubus plants. Especially, there is still a blank in the method for achieving asexual propagation without destroying the integrity of the mother plants and maintaining the natural structure of the population.

[0004] Based on this, a method for in-situ regeneration of Rubus plants in the wild with simple operation and environmental friendliness is provided, which can break through the limitations of traditional breeding and promote the protection of biodiversity. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for in-situ regeneration of Rubus plants in the wild to solve the problems existing in the above-mentioned prior art. The present invention not only provides a beneficial method for the protection of Rubus plant biodiversity, but also provides a reference method for the in-situ breeding of other rare and endangered species.

[0006] To achieve the above purpose, the present invention provides the following scheme:

[0007] The present invention provides a method for in-situ regeneration of Rubus plants in the wild, which includes the steps of girdling the target branches, spraying a plant growth regulator solution, wrapping moss, and covering with soil;

[0008] The width of the girdling is 0.5 - 1.5 cm.

[0009] Optionally, it includes the following steps:

[0010] Select one-year-old or two-year-old branches and conduct girdling at the part of the branch 15 - 30 cm above the ground. The width of the girdling is 0.5 - 1.5 cm.

[0011] Spray 200 - 1000 mg / L of plant growth regulator on the girdled part and then wrap it with moss. Cover the moss with 1 - 1.5 cm thick soil and cover the outermost layer with a breathable film.

[0012] After taking root, transplantation can be carried out.

[0013] Preferably, the width of the girdling is 0.5 cm.

[0014] Preferably, the concentration of the plant growth regulator is 800 mg / L.

[0015] Preferably, the plant growth regulator includes IBA.

[0016] Optionally, the length of the moss wrapping is 5 - 8 cm; the water content of the moss is 60 - 70%.

[0017] Optionally, the time required for rooting is 4 - 8 weeks.

[0018] Optionally, the Rubus plants include Rubus idaeus.

[0019] The present invention discloses the following technical effects:

[0020] 1. Environmentally friendly and efficient: It does not require in vitro culture, directly utilizes field resources, and reduces the use of chemical agents.

[0021] 2. Maintain genetic stability: Retain the excellent traits of the mother plant through asexual reproduction.

[0022] 3. Strong adaptability: The rooting process can be completed in the natural environment, with a rooting rate exceeding 90% and a transplantation survival rate reaching 90% and above.

[0023] 4. Promote ecological restoration: Rapidly expand the population and maintain regional biodiversity.

[0024] In summary, the present invention provides a method for non-in vitro regeneration of Rubus plants in the wild, which not only provides a beneficial method for the protection of Rubus plant biodiversity, but also provides a method reference for the field cultivation of other rare and endangered species. Detailed implementation manners

[0025] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0026] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0027] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0028] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the present invention specification are obvious to those skilled in the art. The present invention specification and examples are merely exemplary.

[0029] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0030] The technical solution of the present invention is a method for in-situ regeneration of Rubus plants in the wild, comprising the following steps:

[0031] 1. Branch selection: Select one-year-old or two-year-old branches that are robust and free from diseases and pests on the mother plant of Rubus plants; prune and remove the redundant and weak branches on the branches.

[0032] 2. Propagation by girdling and rooting: Perform girdling on the branch at a position 15 - 30 cm above the ground. The width of the girdling is 1 / 3 - 1 / 2 of the branch diameter, but try not to exceed 2 cm; it is required that the cut is aligned without rough edges and the width is consistent to facilitate wound healing. The cambium of the cut should be removed completely to avoid forming too much callus to fill the wound and delaying root formation; at the same time, the girdling should not be too deep and should not damage the xylem.

[0033] Spray a rooting agent containing 200 - 1000 mg / L indole - 3 - butyric acid (IBA) or 200 - 1000 mg / L naphthaleneacetic acid (NAA) on the girdled part, and wrap a 5 - 8 cm long stem segment at the girdled part with moss having a water content of 60 - 70%. Try to place the cut at the middle part of the stem segment. Then wrap the outer layer of the moss with 1 - 1.5 cm thick humus soil or the soil from the roots of the plant, with the water content of the soil being 65 - 80%. Cover the outermost layer with a breathable film or other waterproof and breathable materials; after wrapping, tie both ends tightly to prevent rapid water evaporation.

[0034] According to the length of the branch, multiple parts of the same branch can be girdled, generally with an interval of 5 - 10 cm.

[0035] This method can be used throughout the year.

[0036] 3. Rooting and transplanting management: After the complete roots are formed at the girdled part (about 4 - 8 weeks), cut the rooted branch from the mother plant and transplant it to the target area. Generally, roots can form in 15 - 20 days, and after 25 - 30 days, cut off the connection between the branch and the mother body, leaving 3 - 4 leaves. Then water and fertilize according to the weather and the growth condition of the seedlings. In the spring of the second year, dig out the seedlings, grade them, and directly plant them in the field or transplant them to the nursery for cultivation.

[0037] Example 1: In - situ regeneration method of Rubus chingii Hu in the wild

[0038] Taking Rubus chingii Hu, which belongs to the genus Rubus and blooms in spring, as an example, its in - situ regeneration in the wild is carried out as follows:

[0039] 1. Select biennial branches with a thickness of 1 cm germinated from the base of the mother plant in spring;

[0040] 2. Girdle the branches at 30 cm above the ground, with the girdling widths being 0.25 cm, 0.5 cm, 1 cm, 1.5 cm, 2 cm, and 3 cm respectively, 10 in a group, and repeat 3 times.

[0041] 3. Spray 500 mg / L IBA solution on all girdled parts, wrap with moist sphagnum moss, then cover with 1 cm rhizosphere soil, and tie the outer layer tightly with a polyethylene film;

[0042] 4. After 6 weeks, roots are observed to form. Cut the branches and transplant them to the nursery. The callus induction rate, rooting rate, and survival rate of different girdling widths are shown in Table 1. It can be seen from Table 1 that when the girdling width is 0.5 cm or 1 cm, the callus induction rate can reach the highest 93.33 ± 5.77%, and the rooting rate can also reach the highest 93.33 ± 5.77%. Under conventional management, when the girdling width is 0.5 cm - 1.5 cm, the survival rate can reach 87.72 ± 1.52 or above. Based on this result, the optimal girdling width is selected as 0.5 cm.

[0043] Table 1 Results of in - vitro regeneration of Rubus idaeus with different girdling widths

[0044] Width of girdling / cm Callus induction rate / % Rooting rate / % Survival rate / % 0.25 86.67±5.77 16.67±5.77 66.67±7.22 0.5 93.33±5.77 93.33±5.77 97.78±2.22 1 93.33±5.77 93.33±5.77 94.70±1.31 1.5 83.33±5.77 90.00±0.00 87.72±1.52 2 73.33±5.77 76.67±5.77 74.44±1.92 3 36.67±5.77 33.33±5.77 61.33±2.31

[0045] Example 2 Method for in - vitro regeneration of Rubus idaeus in the wild

[0046] Optimize the concentration of IBA.

[0047] 1. Select biennial branches with a thickness of 1 cm germinated from the base of the mother plant in spring;

[0048] 2. Girdle the branches at 30 cm above the ground, with a girdling width of 0.5 cm, 10 branches in a group, and repeat 3 times.

[0049] 3. Spray 100mg / L, 200mg / L, 400mg / L, 800mg / L, 1000mg / L, 1200mg / L IBA solutions on the girdled parts respectively, wrap with wet sphagnum moss and then cover with 1 cm of rhizosphere soil, and tie the outer layer tightly with a polyethylene film;

[0050] 4. After 6 weeks, when roots are observed to form, cut the branches and transplant them to the nursery. The callus induction rate, rooting rate and survival rate with different IBA concentrations are shown in Table 2. It can be seen from Table 2 that when the IBA concentration is 800mg / L, the callus induction rate can reach the highest 100.00±0.00%, and the rooting rate can reach the highest 96.67±5.77%. Under conventional management, when the IBA concentration is 400mg / L, 800mg / L, 1000mg / L and 1200mg / L, the survival rate can reach 90% and above. Among them, when the IBA concentration is 800mg / L, the survival rate can reach the highest 100.00±0.00%.

[0051] Table 2 Results of in - vitro regeneration of Rubus idaeus with different IBA concentrations

[0052] IBA concentration / (mg / L) Callus induction rate / % Rooting rate / % Survival rate / % 100 76.67±5.77 23.33±5.77 57.58±5.29 200 83.33±5.77 53.33±5.77 77.33±2.31 400 100.00±0.00 76.67±5.77 91.43±2.86 800 100.00±0.00 96.67±5.77 100.00±0.00 1000 93.33±5.77 96.67±5.77 96.30±1.28 1200 43.33±5.77 46.67±5.77 90.00±5.00

[0053] The above - described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for in vitro regeneration of Rubus plants in the field, characterized in that: The method comprises the steps of ring-barking target branches, spraying plant growth regulators, wrapping with moss, and covering with soil; The width of the ring stripping is 0.5-1.5 cm.

2. The method according to claim 1, characterized in that The steps include: Select one-year-old or two-year-old branches and perform ring peeling at a position 15-30 cm above the ground, with a ring peeling width of 0.5-1.5 cm; Spray 200-1000mg / L plant growth regulator on the ring-barking part and wrap the moss. Cover the outer layer of the moss with 1-1.5cm thick soil and the outermost layer with a breathable film. After roots have taken shape, transplanting can be carried out.

3. The method according to claim 2, characterized in that The ring stripping width is 0.5 cm.

4. The method according to claim 2, characterized in that The concentration of the plant growth regulator is 800 mg / L.

5. The method according to claim 2, characterized in that The plant growth regulator includes IBA.

6. The method according to claim 2, characterized in that The length of the wrapped moss is 5-8 cm; the water content of the moss is 60-70%.

7. The method according to claim 2, characterized in that The time required for rooting is 4-8 weeks.

8. The method according to claim 1, characterized in that The Rubus plant includes raspberry.