A method for rapid propagation of Saxifraga cordifolia

Abstract

The present invention relates to the field of biotechnology, and provides a method for rapid propagation of saxifrage with egg heart leaves. By collecting leaves for asexual propagation and controlling physical conditions to induce the generation of new plants, the propagation problem of saxifrage with egg heart leaves is effectively solved, and effective technical support is provided for the introduction, conservation and development and utilization of the wild plant resources. The leaves of saxifrage with egg heart leaves are clustered and compact, and have strong ornamental value. As a shade-tolerant potted plant, it can also be used as a shade-bearing ground cover plant under the forest. However, due to its narrow seed collection window period and poor germination performance, inducing leaves to generate asexual propagules and constructing seedlings will provide new insights and ideas for the protection of the wild plant resources.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and more particularly to a method for rapid propagation of Saxifraga cordifolia. Background Art

[0002] Saxifraga ovata Saxifraga epiphylla ) belongs to the Saxifragaceae family, Saxifragaceae genus, a perennial herb, mainly distributed in northeastern Guangdong, northeastern Guangxi, western Sichuan, southeastern to northwestern Yunnan, growing under forests and in rocky crevices at an altitude of 800-3800 meters, with good shade and barrenness tolerance. At the same time, its leaves are clustered and compact, with strong ornamental value, and can be used as an excellent shade-tolerant potted plant and ground cover plant.

[0003] At present, there are no reports on the development of germination technology for Saxifraga seeds because the seeds of Saxifraga are usually small, difficult to collect, and the seed germination rate is very low. The research on Saxifraga mainly focuses on chemical composition, phylogeny, and tissue culture of Saxifraga. There are almost no reports on seed sowing and division propagation. For division propagation, the new buds developed on the side of the root of the mother bead are usually used for propagation. There is currently no relevant research and report on the rapid propagation of Saxifraga cordifolia.

[0004] Therefore, there is an urgent need for a method for rapid propagation of Saxifrage ovalifolia to provide a technical basis for its cultivation and planting, and also to provide new insights and ideas for the protection of this wild plant resource. Summary of the invention

[0005] In order to solve or partially solve the problems existing in the related art, the present invention provides a method for rapid propagation of Saxifraga cordifolia, comprising the following steps:

[0006] S1: Collect mature leaves of Saxifraga ovata together with petioles, and dip the base of petioles into wood ash 2-3 times;

[0007] S2: Place the leaves in S1 in a petri dish covered with absorbent paper, add sterile water until the absorbent paper is completely wet, then cover and move to a dark environment at 4°C for 7 days;

[0008] S3: After 7 days, take out the culture dish in S2, keep the absorbent paper moist, and place it in the dark light conditions of 8℃ for 12h and 18℃ for 12h to induce, until 1-1.5mm granular swelling structures are formed at the base of the leaves; the light intensity of the dark light is 100-200lux;

[0009] S4: Lay the leaves of the expanded structure in S3 on the planting medium for planting until the seedlings grow 2-3 leaves and take root;

[0010] S5: Transplant the entire plant in S4 into the cultivation medium and place it in an environment with 50% shade and 50%-70% air humidity until it matures into a seedling.

[0011] Furthermore, when laying the blades in S2, ensure that the base of the blades is in full contact with the absorbent paper.

[0012] Furthermore, the planting matrix in S4 is a 2-cm thick layer of limestone gravel with a particle size of 0.5-1 cm at the bottom, and a mixed matrix of coconut bran and perlite is used on the upper layer, with a volume ratio of coconut bran to perlite of 3:1;

[0013] Furthermore, the air humidity for planting in S4 is 50%-80%; the shading degree is 70%; and the planting medium is kept moist.

[0014] Furthermore, the cultivation medium in S5 is mountain soil that has passed through a 4-mesh sieve.

[0015] Furthermore, the culture dish in S2 is sterilized and clean.

[0016] Beneficial effects:

[0017] 1. The invention provides a method for rapid propagation of saxifrage with egg heart leaves, which effectively solves the propagation problem of saxifrage with egg heart leaves by directly picking leaves and controlling physical conditions to induce the generation of new plants. The saxifrage with egg heart leaves has many leaves and is easy to obtain. The leaf propagation method reduces the difficulty of obtaining propagation materials, and provides effective technical support for the introduction, conservation and development and utilization of the wild plant resources. At present, most of the propagation methods of saxifrage are mainly tissue culture, stolon division propagation and seed propagation. These methods either have high technical thresholds, or the plants do not have runners and cannot be propagated by division, or the seed collection is difficult. Therefore, the method of propagating saxifrage with egg heart leaves by using leaves provided by the invention greatly reduces the propagation cost and technical threshold, and the technology is more easily promoted effectively, and has greater applicability.

[0018] 2. The present application provides a method for rapid propagation of Saxifraga ovata, and provides conditions for hardening the seedlings, which effectively improves its propagation coefficient (multiplier). The field collection window of Saxifraga ovata seeds is narrow, seed collection is difficult, and the seed germination rate is low. In addition, Saxifraga ovata does not have runners, and the method of propagating by division like other Saxifraga is not feasible. At present, there is no research and technical report on the breeding of this species. Therefore, the present invention is a domestic first, and also provides new insights and ideas for the protection of this wild plant resource. At the same time, the method reduces the difficulty of new propagation of Saxifraga ovata, improves the survival rate of new plants, and provides propagation technology support for its commercial development. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The leaves of a single plant of Saxifraga ovata are clustered together. DETAILED DESCRIPTION

[0020] The optional embodiments of the present invention will be described in more detail below. Although the optional embodiments of the present invention are described, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0021] For the purpose of greater clarity, the invention is described in detail through the following examples.

[0022] Example

[0023] A method for rapid propagation of Saxifraga cordifolia, comprising the following steps:

[0024] (1) In June 2023 and June 2024, leaves were collected together with petioles at the National Field Scientific Observation and Research Station for Forest Biodiversity in Lijiang, Yunnan. The back of the leaves was off-white, the front of the leaves was dark green, and the glandular hairs of the leaves were erect and hard. Ten leaves were selected from each plant, and a total of 5 plants were selected as one group. Three groups were selected for each experiment. The base of the petiole of the collected leaves was dipped in plant ash 2-3 times for disinfection. The so-called base refers to the truncated part of the leaf with part of the petiole (about 1 cm).

[0025] (2) Use a 90 mm diameter glass culture dish and lay absorbent paper at the bottom of the dish. Place the mature leaves of Saxifraga ovata in the sterilized clean dish and add sterile water until the absorbent paper is completely moistened. Keep the base of the leaves in full contact with the absorbent paper. Then cover the dish and move it to a dark environment at 4°C for 7 days for low-temperature induction.

[0026] (3) Take out the culture dish and, while keeping the absorbent paper moist, place it in a cycle of 8°C for 12 h, 18°C ​​for 12 h, and full light. Keep the light intensity at 100-200 lux and induce it under dark light conditions at different temperatures until a 1-1.5 mm granular bulge structure is formed at the base of the leaves.

[0027] (4) Lay the swollen leaves at the base on the planting substrate. The planting substrate is a layer of limestone gravel with a thickness of 2 cm and a particle size of 0.5-1 cm at the bottom, and a mixed substrate of coconut bran and perlite is used on the upper layer. The volume ratio of coconut bran to perlite is 3:1. The planting environment condition is under a shade net with a shade degree of 70%. Spray the substrate daily to keep it moist, and maintain the air humidity at 50%-80%. Until the seedlings grow 2-3 leaves and take root; transplant the whole plant with leaves into the cultivation substrate. The cultivation substrate is mountain base soil that has passed through a 4-mesh sieve and is placed under a shade net with a shade degree of 50%. Maintain the air humidity at 50%-70% until the seedlings are established and the seedling rate is calculated.

[0028] Comparative Example 1

[0029] According to the method for rapid propagation of Saxifraga ovata provided in the embodiment, young leaves and mature leaves with petioles of about 1 cm are collected, and then steps (2) to (4) of the embodiment are implemented.

[0030] Comparative Example 2

[0031] According to the method for rapid propagation of Saxifraga ovata provided in the embodiment, after the implementation of step (1) of the embodiment, without low-temperature induction (not placed in a 4°C constant temperature refrigerator, but kept at room temperature), the method is directly implemented according to steps (3) and (4) of the embodiment.

[0032] Comparative Example 3

[0033] According to the method for rapid propagation of Saxifraga ovata provided in the embodiment, after implementing steps (1) and (2) of the embodiment, the culture conditions of step (3) are changed to 8°C, 12h dark culture, 18°C, 12h normal light culture (light intensity 800-1200lux), and after step (4) of the embodiment, the seedling rate is counted.

[0034] Comparative Example 4

[0035] According to the method for rapid propagation of Saxifraga ovata provided in the embodiment, after implementing steps (1) and (2) of the embodiment, the culture conditions of step (3) are changed to 8°C for 12 h and 18°C ​​for 12 h in complete darkness. After step (4) of the embodiment, the seedling rate is counted.

[0036] Seedling rate = single leaf seedling formation / number of leaves;

[0037] Reproduction coefficient = final number of surviving seedlings / leaves;

[0038] Table 1 Effects of different treatments on the seedling rate of Saxifraga ovata leaves

[0039]

[0040] Note: The values ​​in the table are mean ± standard error. The difference in lowercase letters indicates significant difference in the column (p < 0.05)

[0041] As can be seen from Table 1, in the comparative example and comparative example 1, when carrying out rapid propagation for the egg heart leaf saxifrage, mature leaves with grayish white on the back, dark green on the front, and erect and hard glandular hairs should be selected. Young leaves cannot be used for rapid propagation of this species. In the comparative example and comparative example 2, although the overall seedling rate cannot be significantly increased through one week of low temperature induction, the reproduction coefficient of a single leaf of the egg heart leaf saxifrage can be significantly improved (i.e., multiple individuals can be produced through one leaf), which is mainly due to the induction of glycoprotein synthesis in the leaves through low temperature stimulation, thereby providing more effective nutrition for asexual propagules when growth is restored. In the comparative example and comparative example 3, during the establishment of the seedlings of the egg heart leaf saxifrage, normal or strong light conditions are unnecessary. When the light is too strong, its overall seedling rate and reproduction coefficient will be significantly reduced. The establishment of the seedlings of the egg heart leaf saxifrage needs to be carried out under dark light conditions. To further illustrate this point, by comparing the examples and control example 4, in a completely dark environment, the establishment process of the Saxifraga cordifolia seedlings was affected to a certain extent and was significantly reduced, further illustrating that the construction of the Saxifraga cordifolia seedlings by asexual propagules should be carried out under dark light conditions.

[0042] In summary, the method for rapid propagation of saxifrage provided by the present invention is simple to operate, and asexual propagation is performed by collecting leaves, and physical conditions are controlled to induce the generation of new plants, which effectively solves the propagation problem of saxifrage, and provides effective technical support for the introduction, conservation and development and utilization of the wild plant resources. The leaves of saxifrage are clustered and compact, and have strong ornamental value. As a shade-tolerant potted plant, it can also be used as a shade-bearing ground cover plant under the forest. However, due to its narrow seed collection window period, germination is poor. By inducing the leaves to generate asexual propagules and constructing seedlings, new insights and ideas will be provided for the protection of the wild plant resources.

Claims

1. A method for rapid propagation of Saxifraga cordifolia, characterized in that: The method comprises the following steps: S1: Collect mature leaves of Saxifraga ovata together with petioles, and dip the base of the petioles into wood ash 2-3 times; S2: Place the leaves in S1 in a culture dish covered with absorbent paper, add sterile water until the absorbent paper is completely wet, then cover and move to a dark environment at 4°C for 7 days; S3: After 7 days, the culture dish in S2 was taken out, and the dish was placed in a dark light condition of 8°C for 12 h and 18°C ​​for 12 h for induction, with the absorbent paper kept moist, and the culture was cycled until a 1-1.5 mm granular swelling structure was generated at the base of the leaves; the light intensity of the dark light was 100-200 lux; S4: laying the leaves of the expanded structure in S3 on a planting substrate for planting until the seedlings grow 2-3 leaves and take root; S5: Transplant the entire plant in S4 into a cultivation medium and place it in an environment with a 50% shade and an air humidity of 50%-70% until it becomes a seedling.

2. The method according to claim 1, characterized in that When laying the blades in S2, ensure that the base of the blades is in full contact with the absorbent paper.

3. The method according to claim 1, characterized in that: The planting matrix in S4 is a layer of limestone gravel at the bottom; a mixed matrix consisting of coconut husk and perlite is laid on the upper layer; the volume ratio of coconut husk to perlite is 3:1; the thickness of the limestone gravel is 2 cm, and the particle size is 0.5-1 cm.

4. The method according to claim 1, characterized in that The air humidity for planting in S4 is 50%-80%; the shading degree is 70%; and the planting substrate is kept moist.

5. The method according to claim 1, characterized in that The cultivation medium in S5 is mountain soil that has passed through a 4-mesh sieve.

6. The method according to claim 1, characterized in that The culture dish in S2 is sterilized and clean.

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