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Method for direction development and plant regeneration of somatic embryos of ilex crenata

A technology of Ilex serrata and somatic embryos, which is applied in the field of tissue culture, and can solve the problems that hinder the large-scale application of Ilex serrata industrial seedlings, the low rate of callus induction, and the low rate of somatic embryo induction

Active Publication Date: 2020-05-12
YANGTZE UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, at home and abroad, the cotyledons, hypocotyls, leaves, buds (tender buds and dormant buds) are used as explants to obtain test-tube plantlets through the organogenesis pathway, but there are generally low callus induction rates and somatic cells. A series of technical problems such as low embryo induction rate have hindered the large-scale application of industrial seedlings of Ilex serrata and the production of excellent clones

Method used

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  • Method for direction development and plant regeneration of somatic embryos of ilex crenata
  • Method for direction development and plant regeneration of somatic embryos of ilex crenata
  • Method for direction development and plant regeneration of somatic embryos of ilex crenata

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Embodiment 1

[0022] A method for the direct generation of somatic embryos and plant regeneration of Ilex serrata provided by the present embodiment, comprising:

[0023] (1) Rinse the immature seeds of holly with running water for 20 minutes to remove surface dirt. Transfer to an ultra-clean workbench, put into a sterile beaker, and rinse once with sterile water. The surface disinfection of seeds is first soaked in 75% alcohol for 5 minutes, then soaked in 8% sodium hypochlorite for 15 minutes, and finally rinsed with sterile water for 5-6 times, and put into sterile water for later use;

[0024] (2) Peel off the immature embryos, inoculate them on the medium of 1 / 4MS+0.1mg / L 6-BA+1.5mg / L 2,4-D+60g / L sucrose+6.5mg / L agar, culture in dark for 30 Days later, somatic embryos were obtained;

[0025] (3) Transfer the somatic embryos to germinate on MS medium without any plant growth regulator, and transfer to light conditions for 4 weeks after 2 weeks;

[0026] (4) Wash the seedlings and tra...

Embodiment 2

[0028] A method for the direct generation of somatic embryos and plant regeneration of Ilex serrata provided by the present embodiment, comprising:

[0029] (1) Rinse the immature seeds of holly with running water for 20 minutes to remove surface dirt. Transfer to an ultra-clean workbench, put into a sterile beaker, and rinse once with sterile water. The surface disinfection of seeds is first soaked in 75% alcohol for 5 minutes, then soaked in 8% sodium hypochlorite for 15 minutes, and finally rinsed with sterile water for 5-6 times, and put into sterile water for later use;

[0030] (2) Peel off the immature embryos, inoculate them on the medium of 1 / 4MS+0.1mg / L 6-BA+1.5mg / L 2,4-D+50g / L sucrose+6.5mg / L agar, culture in dark for 30 Days later, somatic embryos were obtained;

[0031] (3) Transfer the somatic embryos to germinate on the MS medium without any plant growth regulator, and transfer to light conditions for 26 days after 12 days;

[0032] (4) Wash the seedlings and...

Embodiment 3

[0034] A method for the direct generation of somatic embryos and plant regeneration of Ilex serrata provided by the present embodiment, comprising:

[0035] (1) Rinse the immature seeds of holly with running water for 20 minutes to remove surface dirt. Transfer to an ultra-clean workbench, put into a sterile beaker, and rinse once with sterile water. The surface disinfection of seeds is first soaked in 75% alcohol for 5 minutes, then soaked in 8% sodium hypochlorite for 15 minutes, and finally rinsed with sterile water for 5-6 times, and put into sterile water for later use;

[0036] (2) Peel off the immature embryos, inoculate them on the medium of 1 / 4MS+0.1mg / L 6-BA+1.5mg / L 2,4-D+70g / L sucrose+6.5mg / L agar, culture in dark for 30 Days later, somatic embryos were obtained;

[0037] (3) Transfer the somatic embryos to germinate on MS medium without any plant growth regulator, and transfer to light conditions for 30 days after 16 days;

[0038] (4) Wash the seedlings and tra...

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Abstract

The invention discloses a method for direction development and plant regeneration of somatic embryos of ilex crenata. The method comprises the following steps: 1, collecting immature seeds of ilex crenata, cleaning and sterilizing the seeds, and taking out young embryos; 2, inoculating the obtained young embryos with a somatic embryo induction culture medium, and performing dark culture so as to obtain somatic embryos, wherein the somatic embryo culture medium comprises 1 / 4MS, 0.05-0.2mg / L 6-BA, 1.3- / 1.7mg / L 2,4-D, 50-70g / L cane sugar and 6.2-6.7mg / L agar; 3, transferring the somatic embryos to an MS (Murashige&Skoog) culture medium, performing germination culture, transferring buds to a lighting condition, and performing culture to form complete plants with grown roots; and 4, transplanting the plants in a cultivation medium, and performing culture. According to the method, the young embryos of the ilex crenata are adopted as explants, a plant regeneration rapid propagation system issuccessfully obtained through direct development of the somatic embryos, and both the frequency of callus induction and the frequency of somatic embryo induction are high.

Description

technical field [0001] The invention relates to the technical field of tissue culture, in particular to a method for direct generation of somatic embryos and plant regeneration of Ilex serrata. Background technique [0002] Ilex serrata is a multi-branched evergreen shrub, up to 5 meters high; the leaves are leathery, obovate, elliptic or oblong-elliptic, 1-7 male flowers are arranged in cymes, 4 bases, white; calyx disc-shaped , petals 4, broadly elliptic, corolla diameter about 6 mm, petals ovate, ovary ovoid, fruit spherical, black when mature; endocarp leathery. The flowering period is from May to June, and the fruiting period is from August to October. Born in hills, mountain forests or bushes at an altitude of 700-2100 meters. Distributed in China, Japan and Korea. It is often cultivated as an ornamental tree species in gardens, and it is also cultivated in Europe and America. The fruit of Ilex serrata belongs to the seeds of plants, and it is very difficult to ger...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005A01H4/008
Inventor 杨玉洁郝明灼邹义萍耿芳
Owner YANGTZE UNIVERSITY
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