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Induction of peony embryoid

An embryoid and peony technology, which is applied in horticultural methods, botanical equipment and methods, plant cells, etc., can solve the problem that the embryoid seedlings are not strong enough for domestication, obtain embryoid and embryoid seedlings, and have no problems. Complete problems such as organ differentiation, and achieve the effects of improving breeding methods, shortening breeding cycles, and increasing proliferation coefficients

Inactive Publication Date: 2008-12-17
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The fact that Paeoniaceae plants have multinucleated or multicellular pollen (embryo) indicates that they have a relatively obvious potential for embryoid body formation. It has been reported that embryoid bodies can be obtained from anthers, pollen, and zygotic embryo culture in Paeoniae officinalis (Zenkteler M et al. 1975), but the embryoid seedlings were not strong enough to be domesticated in vases (Brukhin V.B et al.1994); and a small amount of pollen embryos obtained from peony and tree peony were only composed of more than 30 cells and were always coated with The outer wall of anthers has not completed organ differentiation, so far there is no report of obtaining complete embryoid bodies and embryoid body seedlings from peony

Method used

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  • Induction of peony embryoid
  • Induction of peony embryoid
  • Induction of peony embryoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The acquisition of embodiment 1 explant

[0033] Peony carpels collected from robust plants were taken back to the laboratory, rinsed with running water for 24 hours, thoroughly scrubbed and cleaned, stripped of ovules, and disinfected on an ultra-clean bench. The ovules were first treated with 70% ethanol for 3 minutes, washed with sterilized distilled water for 3 times, then treated with 0.2% NaClO for 15 minutes, washed with sterilized distilled water for 3 times, peeled off the seed coat, broke open the endosperm, and picked out the seed embryo with a dissecting needle for inoculation .

Embodiment 2

[0035] (1) After the isolated embryos were cultured on the starting medium Q-1 for 30 days, the rooted seedlings were subcultured (I) to J-2, and cultured for 35 days. After the embryoid body completed organ differentiation, it had two obvious cotyledons time division;

[0036] (2) Inoculate the divided embryoid bodies onto E-2, and after culturing for 30 days, subculture again (1) onto E-2;

[0037] (3) After 35 days, the embryoid body seedlings were subcultured (II) onto E'-1;

[0038] (4) Put the E'-1 seedlings in the refrigerator for 60 days at 4°C;

[0039] (5) Take it back to the cultivation room for cultivation;

[0040] (6) One month later, a few embryoid seedlings germinated, took root and became seedlings.

[0041] (7) Pick out the embryoid body seedlings with good roots and leaves and start seedling hardening.

[0042] Culture conditions: culture temperature 25±1°C, light 16h / d, light intensity 1600Lx.

Embodiment 3

[0044] (1) After the isolated embryos were cultured on the starting medium Q-2 for 40 days, the rooted seedlings were subcultured (I) to J-1, and cultured for 35 days. After the embryoid body completed organ differentiation, it had two obvious cotyledons time division;

[0045] (2) Inoculate the divided embryoid body onto E-2, and after culturing for 35 days, subculture again (1) onto E-2;

[0046] (3) After 35 days, the embryoid body seedlings were subcultured (II) onto E'-1;

[0047] (4) Put the E'-1 seedlings in the refrigerator for 90 days at 4°C;

[0048] (5) Take it back to the cultivation room for cultivation;

[0049] (6) One month later, a few embryoid seedlings germinated, took root and became seedlings.

[0050] (7) Pick out the embryoid body seedlings with good roots and leaves and start seedling hardening.

[0051] Culture conditions: culture temperature 25±1℃, light 20h / d, light intensity 2000Lx.

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Abstract

The invention relates to a method for inducing embryoid bodies of tree peony: inoculate the explants into the starting medium for 30-40 days after cleaning and disinfection, and then transfer them to the embryoid body differentiation medium for 15-70 days. Embryoid bodies are produced on the axis. After the embryoid body differentiation is completed, divide it quickly, inoculate it into the embryoid body inoculation medium and culture it for 30-40 days, then transfer it to the embryoid body subculture medium, and refrigerate the rooted embryoid body seedlings at 4°C for 60-90 days , and then cultivate normally in the cultivation room. When the roots and leaves are well developed and the embryoid seedlings are strong, the seedlings are hardened, potted and transplanted. The invention provides a reliable method for successfully inducing somatic embryogenesis and cultivating seedlings of peony, which not only provides a new way for peony breeding, but also provides an alternative system for studying the embryogenesis of peony, and reveals cell differentiation, morphology of peony for research. Mechanisms of genesis and ontogenetic major theoretical questions opened up a new field.

Description

technical field [0001] The invention relates to tissue culture technology, in particular to a method for inducing embryoid body of peony. Background technique [0002] Since Steward and Reinert first discovered in the 1950s that somatic embryos (embryoid bodies) could be induced to form regenerated plants from somatic carrot cultures in vitro, people have been involved in a large number of plant tissue cultures, single-cell suspension cultures, and protoplasts. Somatic embryogenesis or pollen embryogenesis has been observed in both somatic and pollen cultures. According to incomplete statistics, more than 150 species of plants in more than 40 families have been able to induce embryoid body formation from plant tissue culture through the somatic embryogenesis pathway (Zhu Zhiqing edited. Plant Cell Engineering. 2003, Beijing: Chemical Industry Press) , which includes almost all important families of angiosperms and some gymnosperms. More and more studies have shown that in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12N5/04A01H3/00
Inventor 成仿云何贵梅
Owner BEIJING FORESTRY UNIVERSITY
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