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High effective tissue culture regeneration of cucumber, and method for transplanting of said seedlings with high viability

A technology for regenerating seedlings and cucumbers, applied in the biological field, can solve the problems of shortened internodes of regenerated cucumber plants, aging regenerated seedlings, difficulty in transplanting and survival, etc., achieve normal development, vigorous growth, and solve the effects of short internodes

Inactive Publication Date: 2007-07-25
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The object of the present invention is to provide a kind of high-efficiency cucumber tissue culture regeneration and its regenerated seedling transplanting and survival method, which can reduce the genotype's influence on cucumber regeneration ability, improve the regeneration frequency of cucumber, shorten the regeneration cycle, and solve the problem of cucumber regeneration plant. Internode shortening, flowering in the bottle, aging and regenerated seedlings are difficult to take root, and it is difficult to transplant and survive. Establish a stable and efficient regeneration system for cucumber gene transformation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Select the parental seeds Tm of the South China variety "Green Angel" with full grains and uniform size, soak them in tap water for 30 minutes, first treat them with 75% ethanol for 30 seconds, then sterilize the surface with 2% NaClO for 10 minutes, and wash them three times with sterile water , dispersed on water-soaked absorbent cotton / filter paper. After being treated in the dark at 30°C for one day, they were cultured in a cultivation room at 25°C, with a photoperiod of 12 hr / day and a light intensity of 1500 lux to obtain sterile seedlings.

[0027] Take the cotyledons of sterile cucumber seedlings 2 days after "Lubai", cut off 1 / 2 of the cotyledons near the petiole end, and connect them with MS+6-BA 1mg / L+ABA 1mg / L+AgNO with the surface facing up 3 0.5mg / L+Ades 20mg / L+3% sucrose+0.7% agar bud induction and differentiation medium; then the explant material was placed in the culture room under the same conditions for culture, and after 12 days a large number of Cl...

Embodiment 2

[0031] Select the seeds 007-71321 of the North China type strain "Xintai Mici" with full grain and uniform size, soak them in tap water for 30 minutes, then treat them with 75% ethanol for 30 seconds, and then sterilize them with 2% NaClO for 10 minutes. Wash 3 times with water and spread on water-soaked absorbent cotton / filter paper. After being treated in the dark at 30°C for one day, they were cultured in a cultivation room at 25°C, with a photoperiod of 12 hr / day and a light intensity of 1500 lux to obtain sterile seedlings.

[0032] Take the cotyledons of sterile cucumber seedlings 2 days after "Lubai", cut off 2 / 3 of the cotyledons near the petiole end, and connect them with MS+6-BA 1mg / L+ABA 1mg / L+AgNO 3 1mg / L+Ades 20mg / L+3% sucrose+0.7% agar on the bud induction and differentiation medium; then put the explant material in the culture room under the same conditions for culture, and after 15 days, a large number of clusters can be seen on the edge of the cut bud. At th...

Embodiment 3

[0036] Select the parental seeds HL7 of the European and American strain "Little Angel 2" with plump grains and the same size, soak them in tap water for 30 minutes, then treat them with 75% ethanol for 30 seconds, then disinfect the surface with 2% NaClO for 10 minutes, and wash them with sterile water for 3 minutes. Once, spread on water-soaked absorbent cotton / filter paper. After being treated in the dark at 30°C for one day, they were cultured in a cultivation room at 25°C, with a photoperiod of 12 hr / day and a light intensity of 1500 lux to obtain sterile seedlings.

[0037] Take the cotyledons of sterile cucumber seedlings 2 days after "Lubai", cut off 1 / 2 of the cotyledons near the petiole end, and connect them with MS+6-BA 1mg / L+ABA1mg / L+Ades 20mg / L+3% Sucrose + 0.7% agar on shoot induction and differentiation medium; then the explant material was cultured in the culture room under the same conditions, and after 12 days, a large number of clustered shoots could be seen...

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PUM

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Abstract

A method for the tissue culture and transplanting of cucumber includes such steps as taking the aseptic seedling of cucumber, choosing and inoculating explant, inducing regeneration bud, differentiating, growing, rooting culture, pratisizing seedlings and transplanting.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to the cucumber regeneration process, the hormone ratio of different mediums in the regeneration process, temperature control and a transplanting method for regenerated seedlings. Background technique: [0002] Cucumber (cucumis sativus.L) breeding mainly adopts conventional methods at present. Due to the lack of germplasm resources and narrow genetic background, it is difficult to breed good varieties with strong stress resistance by existing resources and conventional breeding methods. The use of biotechnology to assist traditional breeding and the improvement of cucumber quality and stress resistance through genetic engineering have attracted widespread attention. Establishing an efficient and rapid cucumber regeneration system is the premise of using genetic engineering technology. There have been many reports on plant regeneration of cucumbers, such as: [0003] 1) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01C1/00A01G1/00C12N5/04
Inventor 张卫华孙小镭王志峰曹齐卫刘文宝
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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