Method for establishing genetic transformation system of dendrobium nobile

Dendrobium nobile, transgenic technology, applied in the field of genetic transformation

Inactive Publication Date: 2020-10-27
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The improvement of its ornamental traits or medicinal ingredients and content is still in the exploratory stage, especially there is no report on its genetic improvement by means of molecular biology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Induction of Dendrobium nobile callus: Transfer the aseptic seedlings of Dendrobium nobile to sucrose 30g / L+agar 7.5g / L+1 / 2MS+1.4mg / L 6-BA+0.2mg / L NAA medium . After 3 weeks, green protrusions appeared around the rhizoids, which was the germination of callus tissue. After 4 weeks, green protrusions were seen to expand to form green callus tissue. At this time, subculture was carried out again to induce proliferation of callus. The culture temperature is 25±1°C, and the culture is dark.

[0079] Screening of Spectinomycin resistance of Dendrobium nobile callus: inoculate the callus of Dendrobium nobile into medium supplemented with different concentrations of spectinomycin, the inoculation amount of each concentration is not less than 10, every 5 days The survival rate of the callus was counted, and the counting was stopped after 30 days. Medium formula: sucrose 30g / L+agar 7.5g / L+1 / 2MS+1.0mg / L 6-BA+0.1mg / L NAA. Spectinomycin concentrations were 50, 100, 150, 200, 250...

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PUM

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Abstract

The invention provides a method for establishing a genetic transformation system of dendrobium nobile and relates to the technical field of genetic transformation. The method provided by the inventioncomprises the following steps: inoculating an induction culture medium with aseptic seedlings of the dendrobium nobile, so as to obtain calli; inoculating a subculture medium with the calli, so as toobtain subculture calli; transferring a target gene into the subculture calli, so as to obtain infected calli; inoculating a 1 / 2MS plate culture medium with the infected calli; inoculating a differentiation culture medium with the co-cultured infected calli, so as to obtain clustered buds; inoculating a bud strengthening culture medium with the clustered buds, so as to obtain strengthened buds; and inoculating a rooting and seedling strengthening co-culture medium with the strengthened buds, thereby obtaining transgenic dendrobium nobile. According to the method, the system is established based on a tissue culture rapid-propagation system, and a technical foundation is settled for carrying out genetic improvement on ornamental properties and medicinal characteristics of the dendrobium nobile by using molecular biology ways.

Description

technical field [0001] The invention belongs to the technical field of genetic transformation, and in particular relates to a method for establishing a genetic transformation system of Dendrobium nobile. Background technique [0002] Dendrobium nobile Dendrobium orchidaceae Dendrobium, also known as Wanzhangxu, Nobile Stone, Flat Nobile, Flat Yellow Grass, Flat Grass, etc., is a perennial herb, named for its shape like the hairpin on the ancient head. The stems of Dendrobium nobile are clustered, the upper part is slightly flattened and slightly curved upward, 10-60cm high and 1.4cm thick. The raceme is issued from the upper part of the old stem, the flowers are large, white with lavender apex, sometimes all lavender or white except for a purple spot on the lip disc; the flower shape is peculiar, gray and elegant , is an excellent ornamental plant species. [0003] Dendrobium nobile is one of the authentic products of the medicinal material Dendrobium in the 2015 edition o...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 韩福贵傅力顾永华杨如同郑玉红杨军
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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