A kind of hard leaf orchid tissue culture breeding method

Abstract

A sclerophyll tissue culture breeding method, comprising the steps of: (1) taking newly germinated shoots of sclerophyll healthy plants as explants for disinfection; (2) placing the sterilized explants in MS basic culture (3) placing the sterile test-tube plantlets in MS propagation medium for rapid propagation of test-tube plantlets to obtain clustered buds. The multiplication coefficient of the clustered buds of the sclerophyllum obtained by adopting the cultivation method of the invention reaches 20-30 times, can provide robust high-quality sclerophyllum seedlings in a short period of time, and effectively solves the problem of large-scale seedling cultivation of the sclerophyllia.

Description

technical field

[0001] The invention relates to a plant propagation method, in particular to a sclerophylla tissue culture propagation method. Background technique

[0002] Cymbidium bicolor Lindl. is a medicinal plant of Orchidaceae. The whole herb is used as medicine. In addition, sclerophylla is also of great ornamental value. However, the seeds of Sclerophyllum sclerophyllum are very small, with only immature embryos, which cannot store nutrients. Under natural conditions, the germination stage of its seeds completely depends on mycorrhizal fungi to provide nutrients for them. The natural reproduction rate is low and the time is long. Therefore, wild Sclerophyllum is very rare. Due to long-term over-exploitation and destruction of the ecological environment, Sclerophyllum is in an endangered state. The seeds and seedlings are scarce, and resource renewal is very difficult. Contents of the invention

[0003] The purpose of the present invention is to provide a method ...

Examples

Embodiment 1

[0013] An example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0014] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0015] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and cultu...

Embodiment 2

[0018] Another example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0019] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0020] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and ...

Embodiment 3

[0023] Another example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0024] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0025] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and ...

technical field

[0001] The invention relates to a plant propagation method, in particular to a sclerophylla tissue culture propagation method. Background technique

[0002] Cymbidium bicolor Lindl. is a medicinal plant of Orchidaceae. The whole herb is used as medicine. In addition, sclerophylla is also of great ornamental value. However, the seeds of Sclerophyllum sclerophyllum are very small, with only immature embryos, which cannot store nutrients. Under natural conditions, the germination stage of its seeds completely depends on mycorrhizal fungi to provide nutrients for them. The natural reproduction rate is low and the time is long. Therefore, wild Sclerophyllum is very rare. Due to long-term over-exploitation and destruction of the ecological environment, Sclerophyllum is in an endangered state. The seeds and seedlings are scarce, and resource renewal is very difficult. Contents of the invention

[0003] The purpose of the present invention is to provide a method ...

Examples

Embodiment 1

[0013] An example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0014] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0015] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and cultu...

Embodiment 2

[0018] Another example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0019] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0020] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and ...

Embodiment 3

[0023] Another example of the sclerophylla tissue culture breeding method of the present invention comprises the following steps:

[0024] (1) Selection and disinfection of explants: take the newly germinated shoots of healthy plants of Sclerophyllum sclerophyllum as explants, peel off the outer wrapping leaves, soak them in 2v / v% aqueous solution of detergent for 5min, and linearly Rinse with tap water for 15-30min, add 2-3 drops of Tween-20 to 100ml of 0.1v / v% mercuric chloride for disinfection for 8-10min, rinse with sterile water for 3-5 times, and finally use sterile filter paper to remove surface moisture to obtain the Implants, wherein the sterile water is distilled water sterilized by high temperature and high pressure;

[0025] (2) First-generation induction to obtain aseptic test-tube plantlets: the explants obtained in step (1) are placed under a dissecting microscope, and the shoot tip meristem of 0.2-0.3mm is stripped off, inoculated into MS induction medium, and ...