Method for obtaining haplobiont through inducing sporidia of eustoma grandiflorum

A technology of microspores and eustoma eustoma, applied in the field of haploid plants induced by free eustoma eustoma, can solve the problems of difficult to obtain parent plants of inbred lines, dependence on import of eustoma hybrid seeds, high production cost of fresh cut flowers, etc., to achieve The effect of huge potential commercial value

Inactive Publication Date: 2014-01-15
YUNNAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a kind of efficient and practical eustoma single seed for the serious separation of offspring of eustoma hybrid seed and the technical problem that it is difficult to obtain inbred parent plants The ploidy breeding method provides excellent parental inbred line materials for eustoma variety improvement and new variety breeding, and solves the problems of eustoma hybrid seeds (F1) relying on imports and high production costs of fresh cut flowers. This method can quickly and stably obtain a large number of haploid plants

Method used

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  • Method for obtaining haplobiont through inducing sporidia of eustoma grandiflorum
  • Method for obtaining haplobiont through inducing sporidia of eustoma grandiflorum
  • Method for obtaining haplobiont through inducing sporidia of eustoma grandiflorum

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The method for obtaining haploid plants by induction of free microspores of Shelley (champagne color) is as follows:

[0028] (1) Determining the material: Select the hybrid first-generation Eustoma Shelley (champagne color) with comprehensive and excellent properties as the test material.

[0029] (2) Bud selection, pretreatment, and disinfection: In the early stage of flowering, select flower buds in the single-nucleus marginal stage under a microscope by fluorescent staining, put them in a plastic bag and place them in a refrigerator at 4°C for 1 to 3 days, and wash them with detergent. Clean the surface of the flower buds and rinse them with clean water; disinfect with 75% alcohol for 60 sec, then disinfect with 0.1% mercury chloride for 10-15 minutes, and finally rinse with sterile water for 3 times and dry them in the air.

[0030] (3) Separation of free microspores: Grind the sterilized flower buds in a mortar containing B5 medium, filter the mixture through ...

Embodiment 2

[0042] The method of inducing haploid plants from Senna (purple) free microspores is as follows:

[0043] (1) Determine the material: select the first-generation hybrid eustoma Senna (purple) with comprehensive and excellent properties as the test material;

[0044] (2) Bud selection, pretreatment, disinfection: the same as in Example 1

[0045] (3) Separation of free microspores: the same as in Example 1

[0046] (4) Microspore callus induction: same as Example 1, while solid layer: MS + 0.8% agar powder + 3% sucrose, liquid layer: MS + 2 mg / L KT + 2 mg / L 2,4- D+3% sucrose;

[0047] (5) Adventitious bud induction: same as Example 1, but adventitious bud induction medium MS+2mg / L KT+0.5mg / L IBA;

[0048] (6) Haploid plants are obtained the same as in Example 1

[0049] (7) Ploidy identification of regenerated plants is the same as in Example 1. But the control plant of first-generation eustoma imported from abroad is Saina (purple).

Embodiment 3

[0051] The method of obtaining haploid plants induced by free microspores of myth (purple fringe) is as follows:

[0052] (1) Determining the material: Select the hybrid first-generation eustoma (purple fringe) with comprehensive and excellent traits as the test material;

[0053] (2) Bud selection, pretreatment, disinfection: the same as in Example 1

[0054] (3) Separation of free microspores: the same as in Example 1.

[0055] (4) Microspore callus induction: same as Example 1, while solid layer: MS + 0.8% agar powder + 3% sucrose; liquid layer: MS + 1 mg / L KT + 2 mg / L 2,4-D +3% sucrose;

[0056] (5) Adventitious bud induction: same as Example 1, but adventitious bud induction medium MS+1mg / L KT+0.2mg / L IBA;

[0057] (6) Haploid plants are obtained the same as in Example 1

[0058] (7) Ploidy identification of regenerated plants is the same as in Example 1. But the control plant of the first-generation eustoma imported from abroad is a myth (purple fringe).

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Abstract

The invention provides a method for obtaining haplobiont through inducing sporidia of eustoma grandiflorum. The method comprises the following steps: (1) determining materials; (2) selecting and pretreating flower buds; (3) separating free sporidia; (4) inducing sporidia calluses; (5), inducing adventitious buds; (6), obtaining haplobiont; and (7) identifying regeneration plant ploidy to and determined that the haplobiont is obtained. Through the method, the interference from the diploid regeneration plant, which is caused by anther culture, can be avoided, a large quantity of haploid embryoids can be obtained, the haplobiont rate can reach 100%, so that a large enough selection group is formed, and the selection group can serve as a homozygous inbred line after being subjected to colchicine treatment. A powerful mean is provided for breed improvement and new breed selection of eustoma grandiflorum through obtaining the homozygous inbred line of eustoma grandiflorum, a good first-filial generation (F1) can be screened quickly when the method is used for hybrid crossing, the production problem of seeds of eustoma grandiflorum is solved, and the method has a huge potential commercial value.

Description

technical field [0001] The invention relates to a method for inducing free microspores of eustoma eustoma to obtain haploid plants, and belongs to the technical field of flower breeding. Background technique [0002] eustoma ( Eustoma grandiflorum ), also known as grassland gentian, is a plant of the Gentianaceae grassland Gentiana. Oily green, with high ornamental value and economic value, is one of the most popular high-end cut flowers in the world, and has now ranked among the top ten cut flowers in the world. [0003] Eustoma is a cross-pollinated plant, easy to pollinate, with many seeds, and there is a decline in self-fertilization, but hybrids between inbred lines (F 1 ) showed strong heterosis, therefore, the imported eustoma cultivar F 1 The offspring of the first-generation hybrid seeds are seriously separated and cannot be reserved for planting. In my country, it is necessary to import a large number of eustoma seedlings from abroad every year, resulting in hi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 和凤美朱永平王小巧赵兴富
Owner YUNNAN AGRICULTURAL UNIVERSITY
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