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A kind of high-efficiency cultivation method of rapeseed microspores

A culture method and technology of microspores, applied in the field of high-efficiency culture of rapeseed microspores, can solve the problems of low doubling rate and low embryo emergence rate

Inactive Publication Date: 2021-04-02
WUHAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some genotypes with low embryonic rate in microspore culture and low doubling rate after seedling growth.

Method used

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  • A kind of high-efficiency cultivation method of rapeseed microspores
  • A kind of high-efficiency cultivation method of rapeseed microspores
  • A kind of high-efficiency cultivation method of rapeseed microspores

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] This embodiment provides a method for high-efficiency cultivation of rapeseed microspores, the steps comprising:

[0028] 1. Take materials

[0029] The sampling time is between 10:00 a.m. when the weather is fine, and the buds of the main inflorescence are selected as the site for sampling. The inflorescence has the following characteristics:

[0030] 1) It is best to choose the main branch. The inflorescence of the main branch is very important for the embryo emergence rate (the pollen vitality of the main inflorescence is strong, and the development period of the microspores is more consistent). The inflorescence on the selected main branch has bloomed, and the number of blooms is between 1-10, and the bud emergence rate of the main inflorescence with 3-5 flowers is relatively high.

[0031] 2) When selecting flower buds, do not choose those with exposed stigmas and cracked flower buds to avoid incomplete sterilization;

[0032] 3) If there is no suitable main infl...

Embodiment 2

[0097] This embodiment provides a method for high-efficiency cultivation of rapeseed microspores, the steps of which are basically the same as in Example 1, the difference being:

[0098] The doubling process is as follows:

[0099] 1) Dilute the precipitated pollen with NLN-16 liquid medium containing 16mg / L colchicine (note: the dilution should not exceed 10ml, because after doubling, it will be diluted and centrifuged again in a 10ml centrifuge tube), and poured into a triangle The bottle was cultured in the dark at 31.5°C for 60 hours;

[0100] 2) After 60 hours, check whether there is any contamination. If there is no contamination, separate the dishes and continue to cultivate;

[0101] The embryo induction culture process is as follows:

[0102] 1) Pour the doubled non-contaminated NLN-16 pollen suspension into a centrifuge tube again and centrifuge at 800 rpm for 5 minutes, pour off the NLN-16 liquid, and leave the precipitated pollen;

[0103] 2) Dilute the precipi...

Embodiment 3

[0107] This embodiment provides a method for high-efficiency cultivation of rapeseed microspores, the steps of which are basically the same as in Example 1, the difference being:

[0108] The doubling process is as follows:

[0109] 1) Dilute the precipitated pollen with NLN-16 liquid medium containing 14mg / L colchicine (note: the dilution should not exceed 10ml, because after doubling, it will be diluted and centrifuged again in a 10ml centrifuge tube), and poured into a triangle The bottle was cultured in the dark at 32.5°C for 55 hours;

[0110] 2) After 55 hours, check whether there is any contamination. If there is no contamination, separate the dishes and continue to cultivate;

[0111] The embryo induction culture process is as follows:

[0112] 1) Pour the doubled non-contaminated NLN-16 pollen suspension into a centrifuge tube again and centrifuge at 800 rpm for 5 minutes, pour off the NLN-16 liquid, and leave the precipitated pollen;

[0113] 2) Dilute the precipi...

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Abstract

The invention relates to a method for efficiently cultivating microspores of oilseed rape, comprising the following steps: S1, material selection and bud selection; S2, double culture: pollen is diluted with a NLN-16 liquid medium containing 14-16 mg / L of colchicines for dark culture; S3, embryo induced culture: a uncontaminated NLN-16 pollen suspension obtained after double culture is centrifuged, and the precipitated pollen is diluted with a NLN-13 medium containing 7-8 mg / L of colchicine for dark culture; S4, regeneration seedling cultivation; and S5, seedling transplanting. A high-efficiency technical system for cultivation of oilseed rape microspores is established. By the technique, the embryogenic efficiency, the primary seedling rate and the doubling rate of existing genotype materials which are difficult in embryogenesis can be improved effectively.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a high-efficiency culture method for rapeseed microspores. Background technique [0002] Microspore culture is to separate and cultivate the microspores in the middle and late stages of mononuclei from the anthers to form haploid embryos, and then form normal diploid plants after doubling. In the 1980s, after German researchers obtained plants from rapeseed microspore culture, they continued to study the physiological state of the donor plant, the developmental stage of the mononuclear microspore, the components of the medium, the genotype of the recipient material, the method of isolating the microspore, and The environmental conditions of microspore culture were studied in detail, and finally a set of microspore regeneration system with wide adaptability was obtained (Lichter, 1982; Choung and Beversdorf, 1985). Domestic researchers have also conducted in-depth st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H1/08A01H4/00
CPCA01H1/08A01H4/001A01H4/008
Inventor 万丽丽王转茸洪登峰杨光圣辛强
Owner WUHAN ACADEMY OF AGRI SCI
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