Culture method for blueberry embryoids

A culture method and technology of embryoid body, which is applied in the field of rare fruit variety blueberry embryoid body culture, can solve the problems of reducing the number of effective seedlings, prone to browning, callus, etc., and achieve fast culture speed, less plant viruses, and improved Quantity effect

Inactive Publication Date: 2013-07-03
LUOYANG INST OF SCI & TECH
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  • Summary
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AI Technical Summary

Problems solved by technology

Blueberries use single-bud shoots for tissue differentiation and cluster buds. Although cluster buds are relatively easy to cultivate, it is very slow for mass cultivation of blueberry seedlings.
In the 5th issue of "Molecular Plant Breeding" in 2007, Ning Zhiyuan and others used the induction of blueberry cluster buds and plant regeneration, that is, the single bud shoots of young leaves to cultivate blueberry plants. When the concentration of ZT is too high, it will exceed 2.0 mg / L, and will form A large number of adventitious buds, some calluses and vitrified seedlings appear, reducing the number of effective seedlings; in the 9th issue of 2010 "Northern Horticulture", Yu Qiangbo et al. used blueberry semi-quality new slightly to cultivate blueberry plants, which is prone to brown change

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0017] A method for cultivating blueberry embryoids, using wild blueberries growing in Changbai Mountain in Northeast China as materials, and using leaves grown for 2-3 months as explants, after preparing a medium, sterilizing and inoculating the explants, callus Subculture, induce single-cell, single-cell differentiation, and culture embryoid bodies. The formulation of the prepared medium is: callus subculture medium formulation: WPM+ZT0.1mg / L+2,4-D 1mg / L+NAA 0.3mg / L+6-BA 1.0mg / L+sucrose 20g / L + PVP 0.5g / L + agar 5.2g / L, pH 5.2, temperature 25°C, dark culture; single cell culture formula: WPM + ZT0.1mg / L + 2,4-D 1mg / L + NAA0. 5mg / L + 6-BA 1.0mg / L + sucrose 20g / L + PVP 0.5g / L + agar 5.2g / L, pH 5.2, temperature 25°C, dark culture; single cell dedifferentiation formula: 1 / 2WPM + 6- BA 1.0mg / L + NAA0.5 mg / L + KT 0.1 mg / L + sucrose 15g / L + PVP 0.5g g / L, pH 5.2, temperature 25°C, dark culture; embryoid body culture formula: WPM + ZT 0.1 mg / L + IBA 1.0 mg / L + agar 5.2g / L, pH 5.2, ...

Embodiment 2

[0019] A method for cultivating blueberry embryoids, using wild blueberries growing in Changbai Mountain in Northeast China as materials, and using leaves grown for 2-3 months as explants, after preparing a medium, sterilizing and inoculating the explants, callus Subculture, induce single-cell, single-cell differentiation, and culture embryoid bodies. The formula of the prepared medium is: callus subculture medium formula: WPM+ZT0.5mg / L+2,4-D 4mg / L+NAA 1.4mg / L+6-BA 4.0mg / L+sucrose 20g / L + PVP 1g / L + agar 5.2g / L, pH 5.8, temperature 25°C, dark culture; single cell culture formula: WPM + ZT0.5mg / L + 2,4-D 4mg / L + NAA1.5mg / L +6-BA 4.0mg / L+sucrose 20g / L + PVP 1g / L + agar 5.2g / L, pH 5.8, temperature 25°C, culture in the dark; single cell dedifferentiation formula: 1 / 2WPM + 6-BA 3.0 mg / L + NAA1.6 mg / L + KT 1.5 mg / L + sucrose 40g / L + PVP 1 g / L, pH 5.8, temperature 25°C, dark culture; embryoid body culture formula: WPM + ZT 0.1~2mg / L + IBA 2.0 mg / L + agar 5.2g / L, pH 5.8, temperatu...

Embodiment 3

[0021]A method for cultivating blueberry embryoids, using wild blueberries growing in Changbai Mountain in Northeast China as materials, and using leaves grown for 2-3 months as explants, after preparing a medium, sterilizing and inoculating the explants, callus Subculture, induce single-cell, single-cell differentiation, and culture embryoid bodies. The formula of the prepared medium is: callus subculture medium formula: WPM+ZT0.1~0.5mg / L+2,4-D 1~4mg / L+NAA 0.3~1.4mg / L+6- BA 1.0~4.0mg / L + sucrose 20g / L + PVP 0.5~1g / L + agar 5.2g / L, pH 5.2~5.8, temperature 25°C, culture in the dark; single cell culture formula: WPM + ZT0.1~0.5 mg / L + 2,4-D 1~4mg / L + NAA0.5~1.5mg / L +6-BA 1.0~4.0mg / L+ sucrose 20g / L + PVP 0.5~1g / L + agar 5.2g / L , PH 5.2~5.8, temperature 25°C, dark culture; single cell dedifferentiation formula: 1 / 2WPM + 6-BA 1.0~3.0mg / L + NAA0.5~1.6 mg / L + KT 0.1~1.5 mg / L + Sucrose 15~40g / L + PVP 0.5g~1 g / L, pH 5.2-5.8, temperature 25°C, dark culture; embryoid body culture formu...

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Abstract

The invention discloses a culture method for blueberry embryoids. The culture method comprises the steps of preparing a culture medium by using blueberry with 2-3 months old leaves as an explant, preparing a culture medium, sterilizing the explant and inoculating, subculturing calluses, inducing single cells, differentiating the single cells and culturing the embryoids. The calluses of the blueberry are obtained by induction of the blueberry leaves, the calluses are cultured to obtain the single cells, the single cells are induced to the blueberry embryoids, and the blueberry embryoids are used for culturing the blueberry plants, so that the formation of adventitious buds and vitrified shoots can be reduced and the number of effective seedlings. The calluses can be prevented from browning by using PVP, which is very suitable for the calluses of the blueberry. A culture speed is fast, one hundred thousand of blueberry tissue cultured seedlings can be cultured within five months, and the cultured seedlings have similar growing states basically, high plant qualities and relatively small plant viruses.

Description

technical field [0001] The invention relates to a kind of rare fruit variety blueberry embryoid body culture, which belongs to the field of plant tissue culture. Background technique [0002] The use of leaves to induce callus is very common in tissue culture. Blueberries all use single-bud branch to carry out tissue differentiation cluster buds, although cluster buds are easier to cultivate, but this is very slow for mass cultivation of blueberry seedlings. In the 5th issue of "Molecular Plant Breeding" in 2007, Ning Zhiyuan and others used the induction of blueberry cluster buds and plant regeneration, that is, the single bud shoots of young leaves to cultivate blueberry plants. When the concentration of ZT is too high, it will exceed 2.0 mg / L, and will form A large number of adventitious buds, some calluses and vitrified seedlings appear, reducing the number of effective seedlings; in the 9th issue of 2010 "Northern Horticulture", Yu Qiangbo et al. used blueberry semi-qu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王安亭王祖华李永程关润伶黄向东
Owner LUOYANG INST OF SCI & TECH
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