Tissue culture and seedling raising method for zinc cadmium hyperaccumulator plant Sedum plumbizincicola

A technology of Sedum sedum and tissue culture, which is applied in the field of plant tissue culture, to achieve the effects of fast propagation, widening the scope of material collection, and easy material collection

Inactive Publication Date: 2012-03-14
江苏萤火虫环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is suitable for the phytoremediation technology of polluted environment, but the molecular biological mechanism of its heavy metal tolerance and hyperaccumulation has not been studied yet. The maturity of molecular biology technology has opened up a new way to study the molecular mechanism of its heavy metal tolerance and hyperaccumulation. This method uses In its micropropagation and somatic mutant screening and molecular biology research
So far, tissue culture technology has not been reported on Sedum sedum. Therefore, the invention of the tissue culture method for the repair plant Sedum sedum has important significance for the study of the molecular mechanism of heavy metal tolerance and hyperaccumulation of Sedum sedum. It is of great significance to promote the popularization and application of phytoremediation technology in polluted environment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Rinse the stem section or the tip of the axillary buds of the repaired plant Sedum sedum from the zinc-cadmium mine with tap water, soak in 70% vt ethanol for 50 seconds, and then soak in 0.1% (g / mL) mercuric chloride for 10 minutes, After pouring out the mercuric acid and washing it with sterile water for 5 times, after obtaining sterile seedlings, take 0.3-0.5 cm stem segments or shoot tips with axillary buds on the ultra-clean workbench, and inoculate them into the autoclaved and cooled seedlings. 50mL of induction medium with a pH of 5.8 in a 200mL glass culture bottle, 100 bottles of induction medium (the composition is MS medium, which contains (0.05-0.15) mg / L 6-BA and (1.5-2.5) mg / L NAA) Each bottle was inoculated with 3 stem segments or shoot tips, and the culture bottle was placed in a temperature of 25±1°C, and the light intensity was maintained at 3000 lux (Lux) for 16 hours in an incubator for 16 hours for 25 days. For the shoot tips with clustered buds, se...

Embodiment 2

[0018] Rinse the stems of Sedum sedum with tap water, soak them in 70% vt ethanol for 50 seconds, then soak them in 0.1% (g / mL) mercuric chloride for 10 minutes, drain the mercuric chloride and use After washing with sterile water for 5 times, after obtaining the sterile seedlings, cut off the leaves and lateral buds from the stems on the ultra-clean workbench, cut them into small sections of 0.3-0.5 cm, and inoculate them into 200 mL of glass that has been autoclaved and cooled. On the 50mL induction medium with a pH of 5.5-5.8 in the culture bottle, the composition of the induction medium is MS medium, which contains (0.1-0.5) mg / L 6-BA and (1.5-2.0) mg / L 2,4 -D, put the culture bottle in an incubator with a temperature of 25±1°C and no light for 3 to 6 weeks to obtain callus, and transfer the callus to 50 mL of sterilized glass culture bottle for differentiation culture at pH 5.8 Among them, 50 bottles of differentiation medium are MS medium, which contains (0.1-0.5) mg / L 6...

Embodiment 3

[0020] Rinse the leaves of the repaired plant Sedum sedum from the zinc-cadmium mine with tap water, soak them in 70% vt ethanol for 50 seconds, then soak them in 0.1% (g / mL) mercuric chloride for 10 minutes, drain the mercuric liters, and rinse them with water-free After washing with bacterial water for 5 times, after obtaining the sterile seedlings, cut the leaves on the ultra-clean workbench, cut off the edge of the leaves, and cut them into 0.3-0.5cm 2Inoculated into 50mL induction medium with a pH of 5.5-5.8 in a 200mL glass culture bottle that had been autoclaved and cooled. The composition of the induction medium was MS medium containing a concentration of (2.5-3.5) mg / L NAA and (0.5~1.0)mg / L TDZ, put the culture bottle into the incubator with temperature of 25±1℃ and no light for 4~8 weeks to get the callus, transfer the callus to the sterilized 50 mL of differentiation medium with a pH of 5.8 in glass culture bottles, 50 bottles of which are MS medium containing (0.1-...

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Abstract

The invention provides a high-frequency plant regeneration technique for tissue culture of repairing plant Sedum plumbizincicola, which comprises the following steps: respectively taking the stem segment, the stem tip and the leaf of the repairing plant Sedum plumbizincicola for tissue culture and propagation, and respectively establishing high-frequency regeneration systems for the stem segment,the stem tip and the leaf. Since the stem segment, the stem tip and the leaf of the Sedum plumbizincicola are directly selected during propagation, the materials can be obtained easily, the scope of the materials for the propagation of the Sedum plumbizincicola is widened, the propagation method is rapid and high-efficiency, and conditions are provided for physiological biochemical researches andmolecular biological researches. Through respectively establishing the high-frequency plant regeneration systems for the stem segment, the stem tip or the leaf, the method is suitable for the screening and the micropropagation of somatic cell mutants and the transformation of the transgenic materials through agrobacterium-mediated transformation, and provides basic conditions for elaborating the heavy metal resistance of the Sedum plumbizincicola and the molecular biological mechanism of hyperaccumulators.

Description

technical field [0001] The invention relates to a high-frequency plant regeneration technology for in vitro culture of stem segments, stem tips or leaves of a repairing plant Sedum sedum, belonging to a plant tissue culture method. Background technique [0002] With the development of industry and the modernization of agricultural production, the problem of heavy metal pollution in the soil-plant-environment system is becoming more and more serious. Heavy metal pollution not only reduces the yield and quality of crops, leads to the degradation of soil function and quality, and affects the healthy and sustainable development of agricultural production, but also pollutes surface water and groundwater through runoff and leaching, deteriorates the hydrological environment, and may cause pollution through direct contact, food chains, etc. Ways to endanger human life and health. At present, the treatment of farmland soil polluted by heavy metals in our country should be paid atte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 吴龙华贲爱玲谭维娜王松凤骆永明
Owner 江苏萤火虫环境科技有限公司
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