Polyploidy cell breeding technique for red winged euonymus leaves

A polyploidy, red-leaf technique, applied in gardening, botanical equipment and methods, gardening tools/equipment, etc.

Active Publication Date: 2017-01-04
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there is no report on the use of flow cytometry for plant cell ploidy sorting...

Method used

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  • Polyploidy cell breeding technique for red winged euonymus leaves
  • Polyploidy cell breeding technique for red winged euonymus leaves
  • Polyploidy cell breeding technique for red winged euonymus leaves

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Callus induced by endosperm of Euonymus euonymus

[0021] Red leaf euonymus ( Euonymus alatus 'Compactus'), mature seeds were collected in late November, arils were removed, rinsed with water for 30 min, soaked in 70% ethanol for 60 s, and placed in a disinfectant solution containing 1% Tween-80 and sodium dichloroisocyanurate ( The effective chlorine concentration is 1.0%) and the shock treatment is 20 min. The seeds were rinsed with sterile water 5 times, 3 min each time. Inoculate the sterilized seeds in test tubes containing semi-solidified agar medium. After 7 days, the sterile seeds were screened, and the seed coat was removed with a sterilized scalpel and forceps under sterile conditions (121°C, 0.11 MPa, 20 min, the same below), and the embryos and endosperms were inoculated together in the callus induction medium cultured in the dark at 24°C and subcultured every two weeks. The embryos were removed in the 4th week, and the endosperm was continued to be cul...

Embodiment 2

[0023] Free callus protoplasts

[0024]Red leaf euonymus ( Euonymus alatus 'Compactus') endosperm callus, select the light yellow endosperm callus, and treat it in the dark at 4°C for 24 hours, slice the callus with sterilized forceps and a scalpel under sterile conditions, and weigh it on an electronic balance Callus 0.5g. The callus was placed in a 50 ml centrifuge tube containing 10 ml of enzymatic hydrolysis solution, sealed with parafilm, treated in the dark at 27 °C for 4 h, and mixed gently by inversion every hour. After enzyme treatment, filter with a 300-mesh cell sieve, put the filtrate into a new centrifuge tube and seal it with parafilm, and centrifuge in a low-temperature centrifuge with a centrifugal force of 100 g for 3 minutes. Aspirate the supernatant, add the protoplast washing solution to resuspend, and repeat the aspiration-resuspension three times. Dilute the suspension to 500 μl, take a little suspension, and stain with trypan blue. The concentration...

Embodiment 3

[0026] Protoplast ploidy sorting and plant regeneration

[0027] Red leaf euonymus ( Euonymus alatus 'Compactus') protoplasts were stained with Hoechst 33342 staining solution. The concentration of the staining solution was 5 μg / ml, and the staining time was 5 min. Centrifuge at 100 g for 3 min, discard the staining solution, and resuspend with protoplast medium, repeat three times. Protoplasts were collected, flow cytometric sorting was performed under sterile conditions, and triploid protoplasts were sorted according to the instrument manual.

[0028] Mix the sorted triploid protoplasts with low-melting point agarose (freezing point: 26-30°C) and protoplast culture medium at a temperature of 30-40°C, and pour them into a petri dish with a diameter of 5 cm. After solidification, cut into four equal parts with a sterilized scalpel, transfer to a culture bottle with a diameter of 9 cm containing the protoplast culture solution, and culture with gentle shaking. After the cal...

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Abstract

The invention relates to a polyploidy cell breeding technique for red winged euonymus leaves, belongs to the field of plant ploidy breeding and relates to a method for cultivating a triploid plant by inducing the cells of red winged euonymus leaves. The method comprises the processes of adopting the endosperms of the red winged euonymus leaves for inducing callus and free protoplast and sorting the triploid protoplast under an axenic cultivation condition and inducing and cultivating the triploid protoplast into a triploid plant. The method further comprises the process of eliminating the apical bud dormancy of the triploid red winged euonymus leaves plant. According to the invention, the rich triploid cells are sorted from abundant diploid cells of diploid wild plant seed endosperms and cultivated into the triploid plant and the growth of the triploid plant is boosted.

Description

technical field [0001] The invention belongs to the field of plant ploidy breeding and relates to a method for engineering breeding of Euonymus euonymus triploid cells. It involves the processes and methods of inducing callus formation from endosperm cells of wild Euonymus euonymus seeds; dissociating protoplasts, staining and ploidy sorting triploid protoplasts; protoplast-induced regeneration of plantlets; terminal bud dormancy relief of plantlets and other processes and methods. Background technique [0002] Red leaf euonymus ( Euonymus alatus 'Compactus') is a deciduous shrub of the genus Euonymus in the family Euonymus, with green, glabrous branches when young and corky wings on older branches. Flowers light red or light yellow. The leaves are dark green in spring and summer, and turn purple or fiery red in autumn, so it is called "flaming euonymus". As a garden flower and tree, it is planted alone, in rows, or in groups in scenic spots, campuses, courtyards, lakesid...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 王华芳卜祥潘王蒙蒙邓舒雨修宇唐文思
Owner BEIJING FORESTRY UNIVERSITY
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