Unlock instant, AI-driven research and patent intelligence for your innovation.

Orchid gene transformation method

A technology of gene transformation and orchid, applied in biochemical equipment and methods, horticultural methods, genetic engineering and other directions, can solve the problems of long transformation period and low transformation efficiency, and achieve the effect of short transformation period, high transformation efficiency and low cost

Pending Publication Date: 2019-01-04
ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the Agrobacterium-mediated method has the characteristics of strong feasibility, simple operation, low cost, and good repeatability, but there are still shortcomings such as too long transformation cycle and low transformation efficiency. Seedlings take 2-3 years
Therefore, the lack of an efficient and rapid orchid genetic transformation system is still a key constraint for the study of gene function in orchid molecular biology and molecular breeding

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Orchid gene transformation method
  • Orchid gene transformation method
  • Orchid gene transformation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The establishment of embodiment 1 bamboo leaf orchid multiplication and rapid propagation system

[0030] (1) Selection of orchid explants

[0031] In tissue culture experiments, the selection of explants is very important, and the explants selected by different plants are also different. The experimental results showed that when the leaves or roots of Bambooleaf orchid were used as explants, the roots or leaves were easy to brown and could not induce protocorms, while the stems and buds could successfully induce protocorms in a suitable medium, and the buds The induction effect is good (Table 1). Inoculate the buds or stem segments into the culture medium, and after 30 days of dark treatment, small light green protocorms can be seen at the bud cuts and stem nodes. Then transferred to normal photoperiod conditions and cultured for 30 days, the protocorms turned green and became larger. Protocorms induced from stems are larger than those induced from buds, about 0.3-0...

Embodiment 2

[0038] Influencing factors of embodiment 2 bamboo leaf orchid transformation efficiency

[0039] (1) Effect of different pre-cultivation time on transformation efficiency

[0040]Pre-cultivation keeps the recipient material in a state of cell division, making it easier to integrate foreign genes and improve the efficiency of genetic transformation. The pre-culture time of the receptor has a great relationship with the transformation efficiency. Each receptor has an optimal pre-culture time. If the pre-culture time is too long, the cutting wound will heal and the transformation efficiency will be reduced. In the present invention, the protocorm of Bambooleaf Orchid is cut into 0.5cm size and pre-cultured for 1-3 days, and the culture condition is that the light intensity is 40 μmol m -2 ·s -1 , photoperiod 16h / d, ambient temperature 24±1°C, relative humidity 50-60%, pre-medium contains Huabao No. 1 3g, NAA 0.2mg, 6-BA 2mg per liter, and the balance is water. Then soak in OD ...

Embodiment 3

[0052] Embodiment 3 Agrobacterium transformation method transfers green fluorescent protein into bamboo leaf orchid

[0053] Inoculate the buds of Bambooleaf orchids in the protocorm induction medium (containing 3g of Huabao No. 1, 0.2mg of NAA, 0.2mg of 6BA per liter, and the balance is water). After 30 days of dark treatment, it can be seen everywhere in the bud cut Light green small protocorms. Move to a photoperiod of 16h / d, and keep the light intensity at 40μmo1·m -2 ·s -1 After 30 days of cultivation under the condition, the protocorm turns green and becomes larger, and the protocorm with a diameter of 0.5 cm is used as the acceptor material. Protocorms with a diameter of 0.5 cm were pre-cultured at a light intensity of 40 μmo1 m -2 ·s -1 , cycle 16h / d, ambient temperature 24±1°C, relative humidity 55%, pre-medium contains Huabao No. 1 3g, NAA 0.2mg, 6-BA 2mg per liter, the balance is water, pre-cultivation after 1 day , soaked in OD 600 Infect in the 0.5 Agrobacte...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an orchid gene transformation method. According to the method, buds or stem segments of orchids are adopted as explants for protocorm induction until the diameter of protocormsis 0.2-0.5 cm; the obtained protocorms are pre-cultured, soaked in an agrobacterium solution with OD600 of 0.4 to 0.6 and then transferred to a co-culture medium for dark culture; after excess agrobacteria are removed, the co-cultured protocorms are transferred to a proliferation culture medium for culture to obtain resistant protocorms; the resistant protocorms are transferred to a bud inductionculture medium for culture until dark green seedlings which are 5-6 cm high are obtained, and seedling hardening and transplanting are conducted. The orchid gene transformation method has the advantages of being easy to operate, short in transformation period, high in transformation efficiency and low in cost and is particularly suitable for in-vivo verification of gene functions of the orchids.A basis is laid for obtaining arundina graminifolia and other orchid plants with disease resistance, stress resistance, an early flowering period, changed flower colors, changed flower shapes and changed leaf colors.

Description

technical field [0001] The invention relates to a method for gene transformation in the field of bioengineering, in particular to an orchid gene transformation method. Background technique [0002] Orchidaceae (commonly known as orchids) is the largest family of monocots, with 801 genera and nearly 3,000 species. Most orchid species have high ornamental and medicinal value, so orchids play an important role in the flower industry. At present, the annual sales of orchids in the world have exceeded 7 billion US dollars, becoming an important new industry. The pursuit of novelty is the biggest feature of the flower industry, and it is also an important factor to promote the development of the industry. However, most orchid varieties grow slowly, and it takes a long time for the seeds to mature. The traditional hybrid breeding method has a long cycle, and the progeny traits are difficult to predict, which requires high investment. Compared with traditional breeding methods, t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/62A01H4/00
CPCA01H4/008C12N15/8205
Inventor 杨凤玺金建鹏朱根发郭阿瑾
Owner ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI