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A method for rapid propagation of Paphiopedilum high-quality seedling tissue culture through leaf cluster buds

A tissue culture and seedling technology, applied in the field of plant biology, can solve the problem of not passing patent applications, and achieve the effect of consistent traits

Active Publication Date: 2020-12-01
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no paper report on the rapid propagation of Paphiopedilum tissue culture through the phyllophyte approach at home and abroad, and there is no patent application through the phyllophyll approach.

Method used

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  • A method for rapid propagation of Paphiopedilum high-quality seedling tissue culture through leaf cluster buds
  • A method for rapid propagation of Paphiopedilum high-quality seedling tissue culture through leaf cluster buds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022]Embodiment 1: the tissue culture of green Modi pure Paphiopedilum (Paphiopedilum SCBG Purity)

[0023] 1. Obtaining of sterile adventitious buds

[0024] Use the lateral buds germinated from the mother plant of Paphiopedilum SCBG Purity cultivated in the greenhouse as explants, and use 80mg / L streptomycin sulfate solution to irrigate the roots once two weeks before taking the explants. Irrigate the roots once with 80mg / L streptomycin sulfate solution, cut off the lateral buds, then soak the cut lateral buds with 75% alcohol aqueous solution for 30 seconds on the ultra-clean workbench, cut off the leaves, and then use the volume fraction Soak in 75% alcohol aqueous solution for 60 seconds, rinse with sterile water for 3 times, then sterilize with 0.1% mercuric chloride aqueous solution for 5 minutes, rinse with sterile water for 4 times, inoculate into adventitious bud induction medium, and then add with 3 0.75g of cefuroxime sodium for injection dissolved in 1 ml of ste...

Embodiment 2

[0033] Embodiment 2: the tissue culture of red Modi gift Paphiopedilum (Paphiopedilum SCBG Souvenir)

[0034] 1. Obtaining of sterile adventitious buds

[0035] Take the lateral buds germinated from the tissue culture mother plant of Paphiopedilum SCBG Souvenir cultivated in the greenhouse as explants, and water the roots once with 90mg / L streptomycin sulfate solution two weeks before getting the explants, and the interval After 7 days, irrigate the roots once with 90mg / L streptomycin sulfate solution, cut off the side buds, then soak the cut side buds with 75% alcohol solution by volume fraction for 30 seconds on the ultra-clean workbench, cut off the leaves, and then Soak in 75% alcohol aqueous solution for 60 seconds, rinse 3 times with sterile water, then sterilize with 0.1% mercuric chloride aqueous solution for 8 minutes, rinse 4 times with sterile water, inoculate into adventitious bud induction medium, and then Add 0.75g cefuroxime sodium for injection dissolved in 5 ...

Embodiment 3

[0044] Example 3: Tissue Culture of Red Modi Returning Paphiopedilum (Paphiopedilum SCBG Returnee)

[0045] 1. Obtaining of sterile adventitious buds

[0046] Use the lateral buds germinated from the mother plant of Paphiopedilum SCBG Returnee cultivated in the greenhouse as explants, and use 100 mg / L streptomycin sulfate solution to irrigate the roots once two weeks before taking the explants, and then after 7 days Irrigate the roots once with 100mg / L streptomycin sulfate solution, cut off the lateral buds, then soak the cut lateral buds with 80% alcohol aqueous solution for 60 seconds on the ultra-clean workbench, cut off the leaves, and then use the volume fraction Soak in 80% alcohol aqueous solution for 30 seconds, rinse with sterile water for 5 times, then sterilize with 0.2% mercuric chloride aqueous solution for 10 minutes, rinse with sterile water for 5 times, inoculate into adventitious bud induction medium, and then add 6 0.75g of cefuroxime sodium for injection di...

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Abstract

The invention discloses a method for culturing and rapidly breeding high-quality paphiopedilum seedling tissues by leafage buds. According to the method, lateral buds of paphiopedilum serve as explants, the leafage buds are induced, proliferated and disintegrated into normal adventitious buds by the aid of unique cutting of the explants and a culture medium, rooting and strong seedling culture isimplemented to obtain a lot of uniform tissue culture seedlings. According to the method for culturing and rapidly breeding the high-quality paphiopedilum seedling tissues by the leafage buds, the leafage buds can be successfully induced, proliferated and reset into the normal adventitious buds, each cluster of leafage buds acquire adventitious buds of 89-122 pieces within half year, rooting and strong seedling culture of 6-8 months of the acquired leafage buds is implemented and can be transplanted, and large-scale and commercialization production of the paphiopedilum tissue culture seedlingsis achieved. The culture method and components of the used culture medium are unique, simple and practical, a culture system is stable, a technique is operable, high in application value, and an effective path is provided for improvement of breeding efficiency and market requirement meeting of the paphiopedilum.

Description

Technical field: [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a rapid propagation method for tissue culture of Paphiopedilum high-quality seedlings through leaf cluster buds. Background technique: [0002] Paphiopedilum is a rare orchid with high ornamental value, great market potential and expensive seedlings. The hybrid progeny of Paphiopedilum hybrids tend to segregate in traits after aseptic sowing, which affects their commercial production. Tissue culture asexual propagation of Paphiopedilum hybrids is an effective method to obtain Paphiopedilum seedlings with stable and consistent traits. However, when propagating Paphiopedilum seedlings through tissue culture by common adventitious bud proliferation, its large-scale production is affected due to reasons such as slow growth, long subculture cycle, and low multiplication factor. Tissue culture of high-quality Paphiopedilum seedlings through leaf cluster buds can efficie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 毛创学吴坤林曾宋君郑枫房林李琳周依清
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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