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Method and culture medium for culturing peony cotyledons to directly form somatic embryos

A technology of somatic embryos and medium, which is applied in the field of a method and medium for directly forming somatic embryos from cotyledons of peony, can solve problems such as difficulties in the formation of somatic embryos, and reduce the difficulty of culturing embryogenic callus in peony , Improving the rate of direct somatic embryo formation and improving the effect of application efficiency

Inactive Publication Date: 2022-03-29
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the above-mentioned defects existing in the prior art, the present invention provides a method and culture medium for directly forming somatic embryos from peony cotyledons. The method uses cotyledons of aseptic seedlings of peony as explants, and optimizes tissue culture method, to solve the key problem of the difficulty in the formation of somatic embryos in the establishment of the peony regeneration system, and to improve the process of establishing the peony regeneration system

Method used

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  • Method and culture medium for culturing peony cotyledons to directly form somatic embryos
  • Method and culture medium for culturing peony cotyledons to directly form somatic embryos
  • Method and culture medium for culturing peony cotyledons to directly form somatic embryos

Examples

Experimental program
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Effect test

Embodiment 1

[0054] (1) Select the after-ripened 'Fengdan' peony seeds newly harvested in August of that year, peel off the seed coats of healthy mature seeds with full grains, soak in distilled water for 37 hours, and set aside;

[0055] (2) Bring the soaked peeled seeds into the ultra-clean workbench, irradiate them with an ultraviolet lamp (rated power 18W; working voltage: 220V / 50Hz) for 25 minutes, ventilate them for 8 minutes, and soak them in medical alcohol with a volume concentration of 75%. After 20s, rinse with sterile water twice, then soak in mercuric chloride with a volume concentration of 0.1% for 7 minutes, then rinse with sterile water for 5 times, and set aside;

[0056] (3) The peony seed after above-mentioned processing is cut out seed embryo, is inoculated in the tree peony seedling culture medium, and described tree peony seedling culture medium is basal medium with MS culture medium, comprises the component of following concentration: sucrose 25g / L, agar 8g / L, NAA0....

Embodiment 2

[0061] (1) Select the after-ripened 'Fengdan' peony seeds newly harvested in August of that year, peel off the seed coats of healthy mature seeds with full grains, soak in distilled water for 37 hours, and set aside;

[0062] (2) Bring the soaked peeled seeds into the ultra-clean workbench, irradiate them with an ultraviolet lamp (rated power 18W; working voltage: 220V / 50Hz) for 25 minutes, ventilate them for 8 minutes, and soak them in medical alcohol with a volume concentration of 75%. After 20s, rinse with sterile water twice, then soak in mercuric chloride with a volume concentration of 0.1% for 7 minutes, then rinse with sterile water for 5 times, and set aside;

[0063] (3) The peony seed after above-mentioned processing is cut out seed embryo, is inoculated in the tree peony seedling culture medium, and described tree peony seedling culture medium is basal medium with MS culture medium, comprises the component of following concentration: sucrose 25g / L, agar 8g / L, NAA0....

Embodiment 3

[0068](1) Select the after-ripened 'Fengdan' peony seeds newly harvested in August of that year, peel off the seed coats of healthy mature seeds with full grains, soak in distilled water for 24 hours, and set aside;

[0069] (2) Bring the soaked peeled seeds into the ultra-clean workbench, irradiate them with an ultraviolet lamp (rated power 18W; working voltage: 220V / 50Hz) for 20 minutes, ventilate for 5 minutes, and then soak them in medical alcohol with a volume concentration of 70%. After 10 seconds, rinse with sterile water twice, then soak with 0.05% mercuric chloride for 5 minutes, then rinse with sterile water for 3 times, and set aside;

[0070] (3) The peony seed after above-mentioned processing is cut out seed embryo, is inoculated in the peony aseptic seedling culture medium, and described peony aseptic seedling culture medium is the basal medium with MS culture medium, comprises the component of following concentration: sucrose 20g / L, agar 5g / L, NAA0.5mg·L -1 , ...

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Abstract

The invention belongs to the technical field of plant tissue culture, and provides a method and a culture medium for culturing peony cotyledons to directly form somatic embryos. The method comprises the following steps: inoculating embryos of peony seeds into a peony aseptic seedling culture medium, sequentially carrying out dark culture and light culture to obtain peony aseptic seedlings, then inoculating cotyledons of the peony aseptic seedlings into a peony somatic embryo formation culture medium, and sequentially carrying out dark culture and light culture to obtain the peony somatic embryos. The method provided by the invention solves the problems of difficult survival of tissue culture materials and lack of effective test materials in subculture proliferation due to high pollution rate in the peony tissue culture process, and improves the application efficiency of the peony tissue culture technology.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method and a culture medium for directly forming somatic embryos from cotyledons of peony. Background technique [0002] Peony (Paeonia suffruticosa Andr.) is a perennial deciduous subshrub of the Paeoniaceae family (Paeoniaeeae) and is a famous ornamental, edible and medicinal plant. 'Fengdan' tree peony (P.ostii 'FengDan') has become one of the emerging woody oil plants because of its high quality seed oil. The traditional breeding method of peony has a long cycle and cannot meet the market demand. Therefore, speeding up the research on tissue culture technology of peony is beneficial to the rapid seedling growth of peony, preserving the excellent traits of the mother plant, and providing a basis for molecular biology breeding research. [0003] Currently, peony tissue culture has not yet established a mature and stable regeneration system. The main bottlenecks...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01C1/00A01C1/08C12N5/04
CPCA01H4/002A01H4/005A01C1/00A01C1/08C12N5/0025C12N5/04
Inventor 侯小改张婉青张红晓郭丽丽宋程威郭琪张利霞李昱莹
Owner HENAN UNIV OF SCI & TECH
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