A tissue culture method of mangrove plant Sonneratia

Abstract

The present invention discloses a tissue culture method of a mangrove plant, Sonneratia salsa, and belongs to the field of plant tissue culture. The method includes: (1) explant selection and disinfection: using the stem segment of Sonneratia salsa as an explant for disinfection treatment; (2) cluster bud induction: inoculating the disinfected stem segment onto a bud induction medium for cluster bud induction; (3) cluster bud subculture: when the cluster buds are induced to an average height of 1-2 cm, a single cluster bud is transferred to a subculture medium for subculture; (4) rooting induction: transferring the mature buds after subculture to a rooting medium for rooting culture. The present invention can quickly obtain a large number of Sonneratia salsa tissue culture seedlings through tissue culture, and the rooting rate is as high as 100%.

Description

Technical Field

[0001] The invention relates to the technical field of plant tissue culture, and in particular to a tissue culture method of a mangrove plant Sonneratia salsa. Background Art

[0002] Sonneratia is a pioneer mangrove plant species that settles on the frontier bare beach. It has the characteristics of tall tree body, rapid growth, high fruiting rate, and deep water resistance. It has become a mangrove afforestation tree species with great development prospects. It has grown into a forest along the coast of Shenzhen Bay and is growing well. However, at present, Sonneratia plants mainly rely on seed propagation, which limits their application. Summary of the invention

[0003] The object of the present invention is to provide a tissue culture method of a mangrove plant Sonneratia salsa to solve the problems existing in the above-mentioned prior art, and a large number of Sonneratia salsa tissue culture seedlings can be quickly obtained through tissue culture.

[0004] To achieve the above object, the present invention provides a method for tissue culture of Sonneratia salsa, comprising the following steps:

[0005] (1) Seed treatment: Collect mature fruits of Sonneratia salsa, remove the pulp, take out the seeds from the fruits and soak them in a 100 mg / L gibberellin solution for 2 hours. Sow the soaked seeds on the seedbed and water the seedbed with diluted sea salt water every day to keep it moist until the seeds grow into 8-10 cm seedlings.

[0006] (2) Selection and disinfection of explants: Take the seedlings growing on the seedbed, cut them into stem segments of 0.3-0.5 mm, wash them with tap water, soak them in 75% alcohol for 30 seconds, rinse them with sterile water for 3-5 times, then soak them in 0.1% mercuric chloride for 8-10 minutes, and rinse them with sterile water for 3-5 times.

[0007] (3) Cluster bud induction: The disinfected stem segments were inoculated onto a bud induction medium to induce cluster buds. The bud induction medium was: MS + 0.4 mg / L 6-BA + 0.2 mg / L NAA + 0.1 mg / L KT + 7 g / L agar + 25 g / L sucrose.

[0008] (4) Subculture of cluster buds: When the cluster buds are induced to an average height of 1-2 cm, individual cluster buds are transferred to a subculture medium and cultured to an average height of 3-4 cm. The subculture medium is: MS + 0.6 mg / L 6-BA + 0.5 mg / L NAA + 7 g / L agar + 30 g / L sucrose.

[0009] (5) Rooting culture: The mature buds after subculture are transferred to a rooting medium for rooting culture. The rooting medium is: 1 / 2MS + 0.6 mg / L NAA + 0.5 mg / L IBA + 7 g / L agar + 30 g / L sucrose.

[0010] The invention provides a mangrove plant tissue culture method. A large number of Sonneratia morsiana tissue culture seedlings can be quickly obtained by adopting the method. Specific implementation plan

[0011] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0012] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0013] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation 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 associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0014] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0015] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0016] Example 1: Collect the mature fruits of Sonneratia salsa, remove the pulp, take out the seeds from the fruits and soak them in a 100 mg / L gibberellin solution for 2 hours, sow the soaked seeds on the seedbed, and water the seedbed with diluted sea salt water every day to keep the seedbed moist until the seeds grow into 8-10 cm seedlings. Take the seedlings growing on the seedbed, cut them into 0.3-0.5 mm stem segments, wash them with tap water, soak them in 75% alcohol for 30 seconds, rinse them with sterile water for 3 to 5 times, then soak them in 0.1% mercuric chloride for 8-10 minutes, and rinse them with sterile water for 3 to 5 times. The disinfected stem segments are inoculated onto the bud induction medium to induce clustered buds. The induction effects of the bud induction medium and different culture media are as follows:

[0017]

[0018]

[0019] It can be seen that the most suitable medium for bud induction using Sonneratia salsa stem segments as explants is: MS+0.4mg / L6-BA+0.2mg / LNAA+0.1mg / LKT+7g / L agar+25g / L sucrose. Therefore, this medium was used as the bud induction medium for bud induction and subsequent experiments.

[0020] Example 2: When the clustered buds were induced to an average height of 1-2 cm, the individual clustered buds were transferred to a subculture medium and cultured to an average height of 3-4 cm. The subculture medium and subculture proliferation effects were as follows:

[0021]

[0022] It can be seen that the most suitable subculture proliferation medium is: MS+0.6mg / L6-BA+0.5mg / LNAA+7g / L agar+30g / L sucrose. Therefore, this medium was used as the subculture medium for subculture and subsequent experiments.

[0023] Example 3: The mature buds after subculture are transferred to a rooting medium for rooting culture. The rooting medium and the rooting effect are as follows:

[0024]

[0025] It can be seen that the most suitable rooting medium is 1 / 2MS+0.6mg / L NAA+0.5mg / L IBA+7g / L agar+30g / L sucrose. Therefore, this medium was used as the rooting medium.

[0026] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A tissue culture method of a mangrove plant Sonneratia salsa, comprising the following steps: (1) Seed treatment: collect mature fruits of Sonneratia salsa, remove the pulp, take out the seeds from the fruits and soak them in a 100 mg / L gibberellin solution for 2 hours, sow the soaked seeds on the seedbed, and water the seedbed with diluted sea salt water every day to keep it moist until the seeds grow into 8-10 cm seedlings; (2) Selection and disinfection of explants: Take seedlings grown on the seedbed, cut them into 0.3-0.5 mm stem segments, wash them with tap water, soak them in 75% alcohol for 30 seconds, rinse them with sterile water 3-5 times, then soak them in 0.1% mercuric chloride for 8-10 minutes, and rinse them with sterile water 3-5 times; (3) Cluster bud induction: The sterilized stem segments were inoculated into a bud induction medium to induce cluster buds. The bud induction medium was: MS + 0.4 mg / L 6-BA + 0.2 mg / L NAA + 0.1 mg / L KT + 7 g / L agar + 25 g / L sucrose; (4) Subculture of clustered buds: When the clustered buds are induced to an average height of 1-2 cm, individual clustered buds are transferred to a subculture medium and cultured to an average height of 3-4 cm. The subculture medium is: MS + 0.6 mg / L 6-BA + 0.5 mg / L NAA + 7 g / L agar + 30 g / L sucrose; (5) Rooting culture: The mature buds after subculture are transferred to a rooting medium for rooting culture. The rooting medium is: 1 / 2MS + 0.6 mg / L NAA + 0.5 mg / L IBA + 7 g / L agar + 30 g / L sucrose.

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