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Combined medium for in-vitro cucumber ovary regeneration

A technology of in vitro regeneration and rooting medium, applied in the field of in vitro regeneration of plants, can solve the problems of low regeneration frequency of regenerated plants, large genotype restriction, large workload, etc., and achieves saving seed usage, reducing cultivation steps, differentiation powerful effect

Inactive Publication Date: 2015-10-21
SICHUAN AAS HORTICULTURE RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many problems in this way of occurrence: ①The genotype is highly restricted, and some genotypes cannot be differentiated into plants through this way; ②The workload is heavy: seed sterilization-sterile seedling cultivation-cotyledon / cotyledon node cutting-inoculation of cotyledon / cotyledon node -Induction and differentiation-elongation culture of regenerated seedlings-rooting culture of regenerated seedlings, each step is indispensable; ③There are few differentiated seedlings obtained through this method, and a single cotyledon node / cotyledon can only differentiate 1-2 complete plants, and the regeneration frequency is low ;④ It takes a lot of seeds, if you need 1000 explants, you need to use 500-1000 plump and fresh seeds
There are some reports on using explants such as cucumber cotyledons, hypocotyls, leaves, and petioles to regenerate plants through callus, but there are generally problems such as large genotype constraints, low regeneration rates, and poor repeatability.
[0005] Also have to utilize ovary as explant in addition, as: Du Shengli etc. (cucumber gynogenesis and chromosomal ploidy identification and doubling research, Tianjin: Nankai University, 2002) utilize cucumber unpollinated ovary to establish the in vitro regeneration system, but The regenerated plants obtained by this method not only have a low regeneration frequency, but also are a mixture of haploid, diploid and polyploid. It is necessary to identify the ploidy of the regenerated plants to distinguish them, which greatly increases the workload.

Method used

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  • Combined medium for in-vitro cucumber ovary regeneration
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  • Combined medium for in-vitro cucumber ovary regeneration

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Embodiment 1 The preparation of the combination culture medium of the present invention

[0055] Combined medium includes callus induction medium, callus proliferation and differentiation medium, adventitious shoot elongation medium, rooting medium,

[0056] Callus induction medium: take NB medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 7g / L, TDZ at 1.5mg / L, and NAA at 0.2mg / L;

[0057] Callus proliferation and differentiation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 7g / L, 6-BA at 2.5mg / L, and NAA at 0.01mg / L;

[0058] Adventitious shoot elongation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 7g / L, and 6-BA at 0.5mg / L;

[0059] The rooting medium is: taking MS medium as basic medium, adding sucrose with a final concentration of 30g / L, agar with 7g / L, and IBA with 0.05mg / L.

[0060] The preparation method is as follows:

[0...

Embodiment 2

[0088] Embodiment 2 The preparation of the combination culture medium of the present invention

[0089] Combined medium includes callus induction medium, callus proliferation and differentiation medium, adventitious shoot elongation medium, rooting medium,

[0090] Callus induction medium: take NB medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 5g / L, TDZ at 1.2mg / L, and NAA at 0.5mg / L;

[0091] Callus proliferation and differentiation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 5g / L, 6-BA at 1.5mg / L, and NAA at 0.01mg / L;

[0092] Adventitious shoot elongation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 5g / L, and 6-BA at 0.3mg / L;

[0093] The rooting medium is: taking MS medium as basic medium, adding sucrose with a final concentration of 30g / L, agar with 7g / L, and IBA with 0.05mg / L.

[0094] The preparation method is as follows:

[009...

Embodiment 3

[0109] Embodiment 3 The preparation of the combination culture medium of the present invention

[0110] Combined medium includes callus induction medium, callus proliferation and differentiation medium, adventitious shoot elongation medium, rooting medium,

[0111] Callus induction medium: take NB medium as the basic medium, add sucrose at a final concentration of 40g / L, agar at 4g / L, TDZ at 2.0mg / L, and NAA at 0.5mg / L;

[0112] Callus proliferation and differentiation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 4g / L, 6-BA at 3.0mg / L, and NAA at 0.1mg / L;

[0113] Adventitious shoot elongation medium: use MS medium as the basic medium, add sucrose at a final concentration of 30g / L, agar at 5g / L, and 6-BA at 0.1mg / L;

[0114] The rooting medium is: taking MS medium as basic medium, adding sucrose with a final concentration of 30g / L, agar with 7g / L, and IBA with 0.05mg / L.

[0115] The preparation method is as follows:

[01...

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Abstract

The invention provides a combined medium for in-vitro cucumber ovary regeneration. The combined medium is characterized by comprising a callus induction medium, a callus proliferation and differential medium, an unstable seedling extension medium and a rooting medium. The invention also provides a preparation method of the combined medium. The combined medium is convenient to prepare, can be applied to the in-vitro cucumber ovary regeneration to obtain a great number of cucumber diploid regeneration plants, and is great in application prospect.

Description

technical field [0001] The invention belongs to the technical field of in vitro regeneration of plants, and in particular relates to a combined culture medium for in vitro regeneration of cucumber ovaries. Background technique [0002] Cucumber (Cucumis sativus L.) is one of the main vegetables in the world. It has a long history of cultivation. Cucumber is rich in cellulose, multivitamins and mineral elements. It has high nutritional and medicinal value and is deeply loved by consumers. Due to the limited resources within the species, it is difficult to develop new cucumber varieties through conventional breeding methods, and the use of genetic engineering technology will be an effective means of improving cucumber varieties in the future. Therefore, it is necessary to establish an efficient in vitro regeneration system of cucumber for further transgenic research. [0003] Since Masataka Sato obtained the first cucumber regenerated plant in 1979, cucumber tissue culture ha...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 杨宏李跃建刘小俊梁根云房超刘独臣
Owner SICHUAN AAS HORTICULTURE RES INST
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