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Method for large-scale production of jatropha curcas plants by cell-tissue culture

A tissue culture, large-scale technology, applied in the fields of botany equipment and methods, plant cells, horticultural methods, etc., can solve the problem of high cost of cell culture methods, and achieve the effect of short cycle and low cost

Inactive Publication Date: 2010-01-13
玉溪市金树生物技术开发有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specific method is to transfer the plant cells from the test tube to the large-scale fermenter for microbial fermentation through the triangular flask, and give appropriate conditions for cultivation, so that the plant cells can multiply in large quantities like microorganisms in the fermenter, but the cost of cell culture is high.
Although the Chinese patent document with the application number 03135614.1 discloses a "cell-tissue culture method" in order to increase the proliferation rate and reduce the culture cost, its technical solution can only be used for Dendrobium officinale

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. The selection and cultivation of the explants of the tung tree: select the stem section of the tung tree, wash it with detergent, rinse it under running water for 10 minutes, transfer it to the clean workbench in the future, dry the water, and use 70% (volume than) alcohol sterilization for 30 seconds, then transfer to 0.1% (volume ratio) of HgCl 2 Sterilized in the solution for 5 minutes, rinsed with sterile distilled water for 5 times, cut the stem into 1cm long pieces, inoculated in the callus induction medium MS+6-BA2mg / L+NAAl mg / L+sucrose 30g / L, and used 0.8% (mass ratio) agar is solidified, the pH is adjusted to 5.8 before high-temperature sterilization, and the culture temperature is 25°C. Visible shoots were visible 20 days after inoculation of the explants.

[0016] 2. Suspension culture of A. japonicus cells: take the above-mentioned cultured buds, first add a little cell suspension medium to grind and inoculate in the research body. Use the triangular fl...

Embodiment 2

[0021] 1. The selection and cultivation of the explants of Tunga tungii Tung tree are the same as in Example 1.

[0022] 2. Suspension culture of A. japonicus cells: take the above-mentioned cultured buds, first add a little cell suspension medium to grind and inoculate in the research body. Use the triangular flask to hold the ground A. japonicus tissue and 2 times the weight of the cell suspension liquid medium, put it into a rotary shaker, and carry out the A. japonicus cell suspension culture under dark conditions at a temperature of 25°C for 2 days, and the shaker speed is 100r / min, cell suspension liquid medium is: MS+BA 0.5mg / L+NAA6mg / L+LH 1.5mg / L+sucrose 22g / L, pH value 5.8.

[0023] 3. Pour the cultured cell suspension into a Buchner funnel with filter paper, remove the culture medium, wash it several times with distilled water, then drain it, measure the cell growth, and set aside.

[0024] 4. Cumulus bud proliferation culture: Select A. tungii cells that have grown...

Embodiment 3

[0027] 1. The selection and cultivation of the explants of Tunga tungii Tung tree are the same as in Example 1.

[0028] 2. Suspension culture of A. japonicus cells: take the above-mentioned cultured buds, first add a little cell suspension medium to grind and inoculate in the research body. Use the triangular flask to hold the ground A. japonicus tissue and double the weight of the cell suspension liquid medium, put it in a rotary shaker, and carry out the suspension culture of the A. abaculata cells for 3 days at a temperature of 27°C in the dark, and the rotation speed of the shaker is 130r / min, the cell suspension liquid medium is: MS+BA 0.2mg / L+NAA 1mg / L+LH 0.5mg / L+sucrose 19g / L.

[0029] 3. Pour the cultured cell suspension into a Buchner funnel with filter paper, remove the culture medium, wash it several times with distilled water, then drain it, measure the cell growth, and set aside.

[0030] 4. Cumulus bud proliferation culture: Select A. tungii cells that have gr...

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PUM

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Abstract

The invention relates to a method for the large-scale production of jatropha curcas plants by cell-tissue culture, belonging to the culture technology of plant seedlings. The method comprises the following steps: (1) cell suspension culture: grinding and inoculating jatropha curcas tissue and adding a cell suspension culture substrate: 0.2-0.5 mg / L of MS+BA, 1-6 mg / L of NAA, 0.5-1.5 mg / L of LH and 19-22 g / L of sucrose; (2) cell growth measurement; (3) clumpy bud multiplication culture: selecting cells growing into a resting stage to access a clumpy bud multiplication culture substrate: 0.1-1 mg / L of MS+6-BA, 0.1-1.5 mg / L and 20-30 g / L of sucrose; (4) rooting culture: selecting well-growing clumpy buds to access a rooting culture substrate: 0.1-1 mg / L of MS+NAA; and (5) bottling culture. The invention organically combines the cell culture and the tissue culture of jatropha curcas and has the advantages of short period and low cost, provides a high-efficiency path for manually culturing the jatropha curcas, producing virus-free seedlings and culturing fine varieties and has a favorable prospect for developing and utilizing the jatropha curcas.

Description

technical field [0001] The invention belongs to the cultivation technology of plant seedlings, and in particular relates to a method for large-scale production of A. japonicus plants through cell culture and tissue culture. Background technique [0002] Jatropha curcas L., also known as Jatropha Curcas L., is distributed in tropical and subtropical regions, and is cultivated or semi-wild in my country. Tung tree is a shrub or small tree, 2-5m high, multi-branched, fast-growing, fertile soil, acidic soil or calcareous soil are suitable for growth, resistant to low temperature around 0 ℃ and light frost, strong wind resistance. In the past, it was mostly cultivated as fences and for viewing. The seed kernel contains about 50% oil, which can be used for lubricating oil, soap making, etc., and can induce vomiting and diarrhea, but it is poisonous and should not be eaten. In recent years, tung tree seed oil or modified tung tree seed oil has been used as biodiesel for various di...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 许继宏
Owner 玉溪市金树生物技术开发有限责任公司
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