Method for high frequency plant regenerating of tallow tree tissue culture adventitious bud

A high-frequency, Chinese tallow tree technology, applied in the field of high-frequency plant regeneration of Chinese tallow tree tissue culture adventitious buds, can solve the problems of uneven seed maturity, long reproductive cycle, and species degradation, so as to increase the frequency of reproduction, accelerate the cycle of reproduction, The effect of increasing the multiplication factor

Inactive Publication Date: 2007-10-24
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is: for Chinese tallow tree with long natural reproduction cycle, long-term repeated cross-pollination process, easy to affect variety quality, and natural flowering or artificial induction of its flowering, all due to different seed maturity, easy shattering and extremely high germination rate. However, the survival rate of the vegetative propagation method using seed stems is low, and the long-term cutting propagation is likely to cause the accumulation of fungi and

Method used

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  • Method for high frequency plant regenerating of tallow tree tissue culture adventitious bud
  • Method for high frequency plant regenerating of tallow tree tissue culture adventitious bud
  • Method for high frequency plant regenerating of tallow tree tissue culture adventitious bud

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Effect test

Embodiment 1

[0027] Cultivation of explant material:

[0028] Chinese tallow tree species: Chinese tallow tree from Nanjing, China introduced from the Jiangsu Institute of Botany in 2006.

[0029] Seed germination medium: using WPM, adding 3% sucrose and 0.8% agar strips by mass ratio to the WPM, and adjusting the pH value to 5.8 with potassium hydroxide before autoclaving.

[0030] Remove the seed coat from the mature fruit of Chinese tallow tree, sterilize the surface with 75% ethanol for 1min, and then use 10% H 2 o2 Disinfect with hydrogen peroxide for 30 minutes, then rinse with sterile water three times, inoculate into the seed germination medium, put 1 seed in every 50ml triangular flask, the temperature of the cultivation room is 26°C-28°C, and the light is 16h per day. Under culture, the light intensity is 2000lux. After 16-20 days, about 38% of the cotyledons of Chinese tallow tree seeds have unfolded, and there are 3-4 terminal buds of unexpanded true leaves, 2-3cm long hypocot...

Embodiment 2

[0033] Induction culture of adventitious buds from different explants of Chinese tallow tree A

[0034] Basic medium: WPM, 3% by mass of sucrose, 0.8% by mass of agar strips, pH5.8;

[0035] Cotyledon adventitious bud induction medium: WPM+0.2mg / L IBA+1.0mg / L KT+2.0mg / L6-BA.

[0036] Terminal bud, hypocotyl and root adventitious bud induction medium: WPM+0.1mg / L IBA+0.1mg / LIAA+0.2mg / L 6-BA.

[0037] Each of the above-mentioned media needs to be adjusted to pH 5.8 with potassium hydroxide before autoclaving.

[0038] Culture room environment: temperature: 26°C-28°C, scattered light for 16 hours a day, light intensity 2000lux.

[0039] Induction culture of adventitious buds:

[0040] Under aseptic conditions, the sterile Chinese tallow tree seedlings were divided into two longitudinally to obtain cotyledon blocks, terminal buds containing unexpanded true leaves, and transversely to obtain 5 mm long hypocotyl sections and root sections. , they were inoculated on the correspon...

Embodiment 3

[0043] Induction culture of adventitious buds from different explants of Chinese tallow tree B

[0044] The difference between this embodiment and Example 2 is only that in the process of implementing the high-frequency plant regeneration of adventitious buds for the hypocotyls and root cut sections of aseptic Chinese tallow tree seedlings, different adventitious bud induction media were selected to achieve Adventitious bud clusters were induced directly without callus.

[0045] Hypocotyl adventitious bud induction medium: WPM+0.5mg / L IBA+2.0mg / L KT+3.0mg / L6-BA.

[0046] Adventitious shoot induction medium: WPM+0.5mg / L NAA+3.0mg / L 6-BA.

[0047] All of the above media need to be adjusted to pH 5.8 with potassium hydroxide before autoclaving.

[0048] The 5 mm long hypocotyl section was placed in this hypocotyl adventitious bud induction medium, and the hypocotyl was directly differentiated into adventitious bud clusters at both ends of the wound, and the induction rate was a...

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Abstract

The invention relates to a method for high-frequency regenerating Chinese sapium bark through tissue culturing adventitious bud. The invention is characterized in that it comprises following steps: removing seed coat from Chinese sapium bark's seed, disinfecting surface with hydrogen dioxide solution and inoculating seed germinating culture medium, and getting intact Chinese sapium bark seed seedling; cutting the cotyledon of seed seedling, apical bud containing undeveloped true leaf and hypocotyl and root, and then inoculating them into adventitious bud inducing culture medium suitable for their growth, inducing adventitious bud directly or inducing with callus and getting light green adventitious bud cluster; replanting it into adventitious bud lengthening culture medium, getting lengthened adventitious bud seedling; replanting into seedling breeding culture medium for enlarged breeding, and replanting it into rooting culture medium, getting strong tube Chinese sapium bark with developed true leaf; finally replanting it out of room for natural growth and getting strong Chinese sapium bark seedling.

Description

technical field [0001] The invention relates to a technical method for obtaining high-frequency adventitious bud plant regeneration through tissue culture of Chinese tallow tree, belongs to the field of biotechnology, is specially used for micropropagation of Chinese tallow tree seedlings in test tubes and screening of somatic cell mutants, and is suitable for gene transformation mediated by agrobacterium. technical background [0002] Chinese tallow tree (Sapium Sebiserum) is a deciduous tree of the genus Sapium in the Euphorbiaceae family. It is the main tree species in the forests of the seven autumn red leaf viewing scenic spots in my country. The leaves of tallow tree are a black dye used to dye silk fabrics. The leaf extract is a kind of soil pesticide, which is used to kill pests such as cotton aphids, and can also prevent and treat rice fever. Chinese tallow tree is also a unique economic tree species for oil in my country. There is a layer of white wax on the surfac...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 李霞何小兰周月兰刘桂华
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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