Cultivation method for cabbage type rape Isolated microspore plant strain

A technology of Brassica napus and its cultivation method, which is applied in the field of cultivating free microspore plants of Brassica napus, can solve the problems of costing more manpower, material resources and financial resources, low induction rate of microspore embryos, and induction rate of microspore embryoid bodies in rapeseed. Issues such as low widespread application

Inactive Publication Date: 2008-08-20
贵州省生物技术研究所
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

[0004] At present, there are many reports on rapeseed microspore culture technology, but the low induction rate of rapeseed microspore embryoid bodies has always been a problem that limits the wide application of this technology
The method commonly used in the prior art is to induce and cultivate all the isolated microspores. Because the induction rate of microspore embryos is low, it will cost more manpower, material and financial resources in the initial stage of microspore induction and culture. Screening out microspores with embryo potential for culture will greatly improve induction efficiency and save time and resources

Method used

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  • Cultivation method for cabbage type rape Isolated microspore plant strain
  • Cultivation method for cabbage type rape Isolated microspore plant strain

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1: In the initial stage of flowering of rapeseed in spring, in the morning of fine weather (8:00~11:00), select the inflorescence of flowering 3~7 days in the field, insert water and preserve in 4 ℃ of refrigerators (1~5 days all It will not affect the production of embryos), then select the flower buds (the length of the flower buds is about 3.5mm) in the uninucleate stage and the early stage of dinucleate in the microspore development period, put them into the filter cloth, and use 70% alcohol under aseptic conditions Disinfect for about 1 min, then disinfect with 0.1% mercury liter for 10 min under aseptic conditions, and rinse with sterile water for 3 to 5 times.

[0033] Put the washed and sterilized flower buds into the B solution with a pH of 5.8-6.0 and 13% sucrose. 5 Gently extrude the pollen from the hormone-free liquid medium with a glass rod, filter it through a 300-mesh nylon membrane into a centrifuge tube, centrifuge at 800rpm for 8min, and rem...

Embodiment 2

[0036] Embodiment 2: In the initial stage of flowering of rapeseed, cut the inflorescences that have bloomed for 3 to 7 days at 8:00 to 10:00 in the morning, select flower buds with a size of 3.5 mm, place them in 4% sodium hypochlorite solution for disinfection for 10 minutes, and wash them with sterile water for 3 to 7 days. 4 times, join B 5 Culture medium, gently squeeze the flower buds with a glass rod to free the microspores into the medium, sieve to remove the bud tissue residues, and collect the microspores by centrifugation; add 55mg / l colchicine full filter sterilized NLN-13 Hormone-free liquid medium (13% sucrose, pH 5.9), cultured in dark at 32°C for 2 days at high temperature, select materials with a microspore expansion rate greater than 50%, add colchicine-free filter-sterilized NLN- 13 Hormone-free liquid medium was centrifuged and washed twice to remove colchicine, and then added to fully filtered and sterilized NLN-13 hormone-free liquid medium without colchi...

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Abstract

The invention discloses a cultivating method of the isolated microspores of brassica napus, which aims to solve the drawback of embryo index unavailable in prior art that, the production of microspore embryos can not be judged in the early stage. The invention is characterized in that, a whole filtered and sterilized NLN-13 hormone-free liquid culture medium comprising 45 to 55 mg/L colchicine is added to the prepared isolated microspores of brassica napus; the microspores are cultured in dark under a high temperature raging from 30 deg C to 35 deg C for two days; the materials having the microspore swell ratio large than 50% are chosen for subsequent induction culture. The cultivating method has the advantages that, the microspores having the microspore swell ratio large than 50% are chosen for subsequent induction culture, so the early rejection of the microspores that cannot produce embryos is facilitated; the workload is greatly reduced; the induction efficiency is improved, and the waste of time and resources is avoided.

Description

technical field [0001] The invention relates to plant regeneration through tissue culture technology, in particular to a method for cultivating free microspore plants of Brassica napus. Background technique [0002] The culture of free microspores is an important way to obtain the oily naploid. The induction of haploid plants successfully speeds up the homozygous speed of breeding parents, accelerates the breeding and breeding process of near-isogenic lines, greatly shortens the breeding cycle, and can shorten the breeding time by 3-5 years compared with traditional hybrid breeding methods. Rapeseed microspores are good recipients for introducing foreign genes; microspore cultures to induce double haploid (DH) populations are good systems for genetic research and mutation breeding research. [0003] The use of microspore culture technology in the breeding of new varieties of rapeseed has become one of the breeding methods, such as being used to screen high, low erucic acid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 黄先群毛堂芬徐涵刘永翔李丽董颖苹
Owner 贵州省生物技术研究所
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