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Method for obtaining distant hybridization offspring of common head cabbage and cabbage type rape

A technology for Brassica napus and Cabbage, which is applied in the directions of horticultural methods, botanical equipment and methods, plant genetic improvement, etc., can solve the problems of abortion of young embryos, inability to obtain hybrid plants, etc., achieves restoration of fertility, shortens induction time, the effect of improving the embryo rate

Inactive Publication Date: 2018-07-24
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the object of the present invention is to provide a method for obtaining distant hybrid offspring of head cabbage and Brassica napus, which overcomes the possibility of abortion of immature embryos during distant hybridization of cabbage and Brassica napus cultivars and makes it impossible to obtain hybrid plants To solve the problem, transfer the excellent recovery gene of rapeseed into the cytoplasmic male sterile cultivars of cabbage to restore the fertility of cabbage

Method used

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  • Method for obtaining distant hybridization offspring of common head cabbage and cabbage type rape
  • Method for obtaining distant hybridization offspring of common head cabbage and cabbage type rape
  • Method for obtaining distant hybridization offspring of common head cabbage and cabbage type rape

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] The female parent was selected from the cabbage Ogura cytoplasmic male sterile line cultivar Qingan 58 (2n=2x=18), and the male parent was selected from the excellent restorer line of rapeseed cultivar RFO-46 (2n=4x=38); selected disease-free Full parental seeds ready for planting, routine management;

[0057] Remove the flowers that have opened on the female parent's inflorescence 3 days before the hybridization at the initial flowering stage, tag the unopened flowers and bag them for isolation, take the anthers on the flowering day of the bagged male parent, and peel off the 4mm flower buds on the female parent plant for pollination ;

[0058] Cut the ovary 11 days after pollination, soak it with 70% ethanol for 30s, and then wash it with 0.1% HgCl 2 Disinfect for 15 minutes, then rinse with sterile water 4 times, 5 minutes each time;

[0059] The sterilized ovary was inoculated on the ovary induction medium under sterile conditions, and cultured for 20 days;

[00...

Embodiment 2

[0065] The female parent was selected from the cabbage Ogura cytoplasmic male sterile line cultivar Qingan 68 (2n=2x=18), and the male parent was selected from the excellent restorer line of rapeseed cultivar RFO-46 (2n=4x=38); selected disease-free Full parental seeds ready for planting, routine management;

[0066] Remove the bloomed flowers on the female parent's inflorescence 2 days before hybridization at the initial flowering stage, tag the unopened flowers and bag them for isolation, take the anthers on the flowering day of the bagged male parent, and peel off the 4mm flower buds on the female parent plant for pollination ;

[0067] Cut the ovary 13 days after pollination, soak it in 70% ethanol for 30s, and then wash it with 0.1% HgCl 2 Disinfect for 15 minutes, then rinse with sterile water 4 times, 5 minutes each time;

[0068] The sterilized ovary was inoculated on the ovary induction medium under sterile conditions, and cultured for 20 days;

[0069] The ovules ...

experiment example

[0074] The 10 hybrid plants that embodiment 1 and embodiment 2 obtain altogether carry out hybrid authenticity verification:

[0075] (1) Cytological identification: Sysmex CyFlow Cube8 flow cytometer was used to identify the ploidy of the hybrid offspring.

[0076] Prepare the solution: solution I is 4.2g citric acid, 0.5% Tween 20, dilute to 200mL, store at 4°C for later use; solution II is 28.65g sodium dihydrogen phosphate, PI20mg, dilute to 200mL.

[0077] Take 20mg of parent and hybrid F 1 The new leaves of the generation seedlings were placed in a plastic petri dish, and the leaves were chopped with a double-sided blade, and the cut samples were sucked up with a 1mL pipette gun and filtered into a 1.5mL Eppendorf tube through a 350-mesh nylon screen, and fresh I Suspend the sediment in 400 μL of the solution, add 800 μL of solution II to the sample when measuring the sample with a flow cytometer, and judge the hybrid F according to the abscissa position of the fluoresc...

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Abstract

The invention provides a method for obtaining a distant hybridization offspring of a common head cabbage and a cabbage type rape, and belongs to the field of biotechnological breeding. The method comprises the steps that (1) a common head cabbage cytoplasmic male sterility is selected to serve as a female parent, a cabbage type rape restorer serves as a male parent, and the female parent and the male parent are isolated after hand pollination is conducted; (2) an ovary after being pollinated for 18-25 days is cut and disinfected; (3) in-vitro culture is conducted for 18-25 days; (4) an ovule in the ovary which is subjected to in-vitro culture is peeled off, and then subjected to induction culture to germinate; (5) differentiation culture is conducted, and a F1 generation seedling is obtained; and (6) a lateral bud of the F1 generation seedling is separated and transplanted onto a rooting medium, and a complete hybridization seedling is obtained. The method utilizes an embryo rescue technique to overcome the problem that the hybridization seedling cannot be obtained due to abortion of a young embryo is prone to occurring in the distant hybridization process of the common head cabbage and the cabbage type rape, an excellent recovery gene of the rape is transferred into a common head cabbage cytoplasmic male sterile cultivated species, the fertility of cabbage is recovered, and the technical evidences are provided for innovation and application of germplasm resources of cabbage.

Description

technical field [0001] The invention belongs to the field of biotechnology breeding, in particular to a method for obtaining distant hybrid offspring of head cabbage and Brassica napus. Background technique [0002] Heading cabbage (Brassica oleracea L.var.capitata L., 2n=18), referred to as cabbage, belongs to Brassicaceae Brassica cabbage species. It is an important vegetable crop in China and is cultivated all over the country. The annual cultivation area reaches 9 ×10 5 hm 2 about. The heterosis of cabbage is more obvious. The materials of Ogura cytoplasmic male sterile line show maternal inheritance, pollen abortion and normal pistil, and are easy to restore fertility by the dominant nuclear restorer gene of the restorer material, but cannot carry out normal self-segregation and further innovation of germplasm resources . [0003] Both head cabbage and Brassica napus are important cultivated crops in the Brassicaceae. The rapeseed material is also a restorer line m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H4/00
CPCA01H1/02A01H4/001A01H4/008
Inventor 许忠民张恩慧王杏利姜娇李升娟郭佳马青山石汶汶吴晓凤董艺
Owner NORTHWEST A & F UNIV
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