In Vitro Regeneration Method of Cucumber Ovary

An in vitro regeneration and ovary technology, which is applied in plant regeneration, horticultural methods, botany equipment and methods, etc., can solve the problems of low regeneration frequency of regenerated plants, large genotype constraints, heavy workload, etc., and achieve seed saving Quantity, reduced culture steps, strong differentiation ability

Inactive Publication Date: 2017-03-15
SICHUAN AAS HORTICULTURE RES INST
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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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  • In Vitro Regeneration Method of Cucumber Ovary
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  • In Vitro Regeneration Method of Cucumber Ovary

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1 The in vitro regeneration method of cucumber ovary of the present invention

[0062] 1. Experimental materials

[0063] South China-type hybrid cucumber variety 'Baisitiao';

[0064] Planted in the plastic greenhouse of the Institute of Horticulture, Sichuan Academy of Agricultural Sciences, the row spacing, water and fertilizer management are managed according to the general level.

[0065] 2. Experimental culture medium

[0066] 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;

[0067] 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;

[0068] Adventitious seedling elongation medium: take 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;

[0069] The r...

Embodiment 2

[0109] Embodiment 2 The in vitro regeneration method of cucumber ovary of the present invention

[0110] 1. Experimental materials

[0111] With embodiment 1.

[0112] 2. Experimental culture medium

[0113] 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;

[0114] 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;

[0115] 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;

[0116] 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.

[0117] 3. Experimental method

[0118] (1) Material handling

[0119] With embodiment 1.

[0120] (2) I...

Embodiment 3

[0132] Embodiment 3 The in vitro regeneration method of cucumber ovary of the present invention

[0133] 1. Experimental materials

[0134] Cucumber variety 'Chuanlu No. 1', a local variety in South China;

[0135] Planted in the plastic greenhouse of the Institute of Horticulture, Sichuan Academy of Agricultural Sciences, the row spacing, water and fertilizer management are managed according to the general level.

[0136] 2. Experimental culture medium

[0137] With embodiment 1.

[0138] 3. Experimental method

[0139] (1) Material handling

[0140] With embodiment 1.

[0141] (2) Induction of callus

[0142] With embodiment 1.

[0143] (3) Callus proliferation and differentiation

[0144] The light yellow callus was transferred to the callus proliferation and differentiation medium for culture, and the callus area increased, gradually changing from light yellow to yellow green. Make the area larger than 3cm 2 The callus was then cut to a size of 0.5 cm 2 Small ca...

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Abstract

The invention provides an in-vitro regeneration method for a cucumber ovary. The method comprises the following steps of: 1) callus induction: sterilizing the cucumber ovary, inoculating the cucumber ovary into a callus induction medium, performing heat-shock treatment, and performing culture to obtain calluses; 2) callus proliferation and differentiation: transferring the calluses to a callus proliferation and differentiation medium, and performing culture to obtain cluster bud seedlings; 3) adventitious seedling elongation: transferring the cluster bud seedlings to an adventitious seedling elongation medium, and performing culture for seedling elongation; and 4) rooting culture: transferring adventitious seedlings to a rooting medium, and performing culture to obtain in-vitro regeneration plants. With the adoption of the in-vitro regeneration method for the cucumber ovary, efficient in-vitro regeneration of cucumbers can be realized and a large amount of diploid regeneration plants of the cucumbers are obtained, so that the application prospect is good.

Description

technical field [0001] The invention belongs to the technical field of in vitro regeneration of plants, in particular to a method 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 has gradually developed. Up to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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