Breeding method of poplar haploid

A cultivation method and haploid technology, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of low callus induction rate, low callus organ differentiation rate, etc., to achieve robust growth, The effect of high reproduction coefficient

Active Publication Date: 2012-07-11
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, there have always been problems such as low callus induction rate and low callus organ differentiation rate in the cultivation process of poplar haploids selected from poplar anthers as explants, resulting in the existing poplar haploids. The breeding method is far from meeting the needs of research and production practice, and needs to be improved urgently

Method used

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  • Breeding method of poplar haploid
  • Breeding method of poplar haploid
  • Breeding method of poplar haploid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1. Test materials

[0028] 1. Test material: Populus beijingensis;

[0029] 2. Plant growth regulator

[0030] The plant growth regulating substance used in the present invention adopts domestic naphthaleneacetic acid (NAA), 6-benzylaminoadenine (BAP), 6-furfurylaminopurine (KT), gibberellin (GA 3 ), 2,4-dichlorophenoxyacetic acid (2,4-D), indole butyric acid (IBA).

[0031] 3. Preparation of culture medium:

[0032] (1) Composition and preparation method of "MS basic medium":

[0033] Table 1 MS medium (Murashige and Skoog, 1962)

[0034]

[0035] The consumption of above-mentioned various nutrient components, except that mother liquor I is 20 times of concentrated solution, all the other are 200 times of concentrated solution. After the above MS medium mother solution was prepared, it was stored in a refrigerator at 4°C until use. According to the total amount of the prepared medium, weigh the required agar and sucrose, pour it into distilled water with 3 / 4 o...

Embodiment 2

[0049] 1. The appropriate development period of anthers and microspores cultured in vitro

[0050] Cut the poplar branches with full flower buds and culture them in water at room temperature 25°C for several days. Poplar flower buds were placed on glass slides, anthers were peeled off, stained with acetic acid carmine, and microspore development stages were observed under a microscope to determine the pollen development stages. In this experiment, the anthers whose microspores were at the uninucleated marginal stage were selected for inoculation, and the research results showed that this is the best time for anther culture.

[0051] 2. Disinfection of explants

[0052] Remove the flower buds that are in the single-nucleus marginal stage from the catkins, first use a brush to lightly brush the surface of the Beijing poplar flower bud scales, remove the dirt and bacteria on the surface, and improve the disinfection effect, and then rinse with sterile water for 5 times , filter...

Embodiment 3

[0095] 1. Preparation of sterile anthers

[0096] Take the poplar flower buds at the single-nucleus stage, rinse the flower buds with sterile water for 3-5 times, blot the water with filter paper, soak the flower buds with 70.0% alcohol by volume for 30 seconds, and then use 7.0% sodium hypochlorite The solution was sterilized for 5 minutes, and then the flower buds were washed 3-5 times with sterile water. Peel off the scales of the flower buds on an ultra-clean workbench, and take out the anthers in the middle of the inflorescence.

[0097] 2. Callus induction culture

[0098] The anthers were inoculated in the callus induction medium for dark culture, culture conditions: under dark conditions, the culture temperature was 25±1°C, cultured for 120 days, and the statistical anther callus induction rate was 30.0%. Among them, the callus The induction medium is: H+NAA 1.0mg / L+KT 1.0mg / L+sucrose 30g / L+agar 5.5g / L, pH5.5.

[0099] 3. Adventitious bud differentiation culture

...

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Abstract

The invention discloses a breeding method of poplar haploid and belongs to the field of selective breeding of poplar haploid. The breeding method comprises the following steps: (1) inoculating sterilized poplar anthers to a callus induction culture medium for callus induction culture to obtain poplar calli; (2) inoculating the poplar calli to an adventitious bud differentiation culture medium for differentiation culture of adventitious buds to obtain poplar adventitious buds; (3) inoculating the poplar adventitious buds to a rooting culture medium for rooting culture to obtain test-tube plantlets; and (4) performing ploidy identification to obtain poplar haploid plants. The breeding method of poplar haploid solves the problems occurring in poplar haploid breeding, has the advantages of high callus induction rate, high callus organ differentiation rate, high breeding rate and simple induced breeding operation, can produce high-quality poplar test-tube plantlets with high yield during a short time, achieves the purpose of large-scale industrial production, and provides a material basis for breeding, quality improvement and genetic research of poplar.

Description

technical field [0001] The invention relates to a method for cultivating haploid plants, in particular to a method for cultivating poplar haploids by plant tissue culture, and belongs to the field of poplar haploid breeding. Background technique [0002] Beijing poplar (Populus beijingensis) is a hybrid offspring of Qingyang and Zhuantian poplar. It is the first new poplar variety bred by artificial hybrid breeding in the forestry industry after the founding of New China. It has the characteristics of cold resistance, water and fertilizer, and beautiful tree shape. The timber forests, shelter forests, street trees and "surrounding" greening tree species are mainly distributed in North China and Northwest China, and are deeply loved by the masses. [0003] Populus plants are dioecious, and self-pollination is basically impossible. Obtaining homozygous lines through conventional cross breeding requires a long breeding period. In order to obtain homozygous poplar lines, resea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01H5/04A01H6/00
CPCA01H1/08C12N5/04A01H5/04A01H4/005A01H4/00A01H1/04A01H6/00
Inventor 安新民李英
Owner BEIJING FORESTRY UNIVERSITY
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