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Tissue culture method for broccoli

A technology of tissue culture and broccoli, which is applied in the field of plant tissue culture, can solve the problems of long breeding time, etc., and achieve the effects of convenient operation, improvement of culture efficiency, and promotion of broccoli microspore embryogenesis

Inactive Publication Date: 2013-11-20
ZHENJIANG SUIHAN AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Chinese invention patent with the patent number ZL201110112536.9 mainly adopts the method of co-cultivating the microspores of the stubborn genotype of broccoli and the microspores of rapeseed that are easy to emerge embryos, and achieves the purpose of promoting the emergence of microspores of broccoli. Co-cultivation, the embryoids obtained may be broccoli embryoids or rapeseed embryoids, which can only be judged after planting in the field, and the selection time is relatively long

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The cultivation method is carried out as follows.

[0038] Culture medium preparation: including the culture medium of different culture stages of microspores, its components and the weight of each component contained in each liter of medium are:

[0039] NLN-13 liquid medium: NLN liquid medium 1L + sucrose 130g / L, pH6.0, filter sterilized;

[0040] NLN liquid medium, in 1L, consists of: KNO 3 125mg, Ca(NO 3 ) 2 4H 2 O500mg, MgSO 4 ·7H 2O125mg, KH 2 PO 4 125 mg, H 3 BO 3 6.2 mg, MnSO 4 ·H 2 O18.95mg, ZnSO 4 ·7H 2 O8.6mg, Na 2 MoO 4 2H 2 O0.25mg, CuSO 4 ·5H 2 O0.025mg, CoCl 2 ·6H 2 O0.025mg, vitamin B10.5mg, vitamin B60.5mg, biotin 0.05mg, folic acid 0.5mg, Na 2 EDTA37.3mg, FeSO 4 ·7H 2 O27.8mg, inositol 100mg, glycine 2mg, niacin 5mg, L-glutamine 800mg, glutathione 30mg, vitamin B55mg, serine 100mg and the rest sterile water.

[0041] Embryoid body differentiation medium: B5 medium + sucrose 30g / L, agar 9g / L, pH6.0, high temperature and high pres...

Embodiment 2

[0057] Medium preparation:

[0058] NLN-13 induction medium: NLN-13 medium + sucrose 130 g / L, pH 6.2, filtered and sterilized, the components of the NLN liquid medium are the same as in Example 1.

[0059] Embryoid differentiation medium: B5 medium + 20 g / L sucrose, 10 g / L agar, pH 6.1, sterilized under high temperature and high pressure; the composition of B5 medium is the same as that in Example 1.

[0060] Rooting medium: composed of 1 / 2 MS medium 1L+0.2mg NAA+0.1mg IBA, 20g sucrose and 7g agar, pH 6.0, sterilized by high temperature and high pressure; MS medium components are the same as in Example 1.

[0061] The method for improving microspore embryogenesis of the stubborn genotype of broccoli:

[0062] 1) Selection of donor materials: take broccoli inflorescences with a petal-to-anther length ratio of 1.0, healthy flower buds free from diseases and insect pests, and wheat ears 2 days before fertilization as the test donor materials;

[0063] 3) Put 14 sterile broccoli...

Embodiment 3

[0069] Medium preparation:

[0070] NLN-13 induction medium: NLN-13 medium + sucrose 130g / L, pH6.1, filter sterilized; NLN liquid medium components are the same as in Example 1;

[0071] Embryoid body differentiation medium: B5 medium + sucrose 20g / L, agar 10g / L, pH6.0, high temperature and high pressure sterilization; B5 medium components are the same as in Example 1;

[0072] Rooting medium: composed of 1 / 2 MS medium 1L+0.2mg NAA+0.2mg IBA, 25g sucrose and 7.5g agar, pH 5.9, sterilized by high temperature and high pressure; MS medium components are the same as in Example 1.

[0073] The method for improving microspore embryogenesis of the stubborn genotype of broccoli:

[0074] 1) Selection of donor materials: flower buds and wheat ears with a petal-to-anther length ratio of 1.2, healthy oil, and no pests and diseases on broccoli inflorescences were used as donor materials for microspore culture;

[0075] 3) Put 15 sterile broccoli flower buds in a sterile beaker on a clean ...

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Abstract

The invention discloses a tissue culture method for broccoli. The tissue culture method comprises the following steps of (1) taking flower buds of the broccolis and wheat ovaries, and sterilizing; (2) preparing the sterilized flower buds into a microspore suspension solution; putting the microspore suspension solution and the wheat ovaries into culturing liquid to be mixed and cultured; inducing to obtain cotyledon-shaped embryoid; (3) inoculating the cotyledon-shaped embryoid into a differential culture medium and culturing to obtain a regenerated plant; and (4) cutting regenerated buds from the regenerated plant and inoculating the regenerated buds into a rooting culture medium to obtain a complete plant. According to the tissue culture method for the broccolis disclosed by the invention, obstinate gene type free microspores of the broccolis and the wheat ovaries are proportioned according to a certain number and are commonly cultured so as to accelerate embryogenesis of the obstinate gene type microspores of the broccolis.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a tissue culture method for promoting microspore embryogenesis of the stubborn genotype of broccoli. Background technique [0002] Broccoli (Brassica oleracea L.var.italica), also known as broccoli, green cauliflower, stem broccoli, etc., is a primary or secondary herbaceous plant with green curds as product organs in Brassicaceae Brassica species. The green flower ball formed on the top of the main stem and side branches is used as the product. It is rich in nutrition, good in color, aroma and taste, and is a famous and special vegetable that is very popular in the international market. At present, most of the main varieties of broccoli in my country come from abroad, and the varieties with our independent intellectual property rights are scarce. The reason is that there are few excellent breeding resources of broccoli in China, and no good materials can be prepared...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张振超潘跃平戴忠良毛忠良吴国平姚悦梅秦文斌肖燕潘永飞王建华马志虎孙春青
Owner ZHENJIANG SUIHAN AGRI
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