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Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos

A bioreactor and plant regeneration technology, applied in the field of agricultural biology, can solve the problems of cultivating somatic embryos of Panax notoginseng that have not been reported, and achieve the effect of saving rooting process and efficient reproduction

Inactive Publication Date: 2013-02-06
NORTHEAST FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many studies on the somatic embryogenesis of Panax notoginseng and its plant regeneration, there is no report on the use of bioreactors to cultivate a large number of somatic embryos of Panax notoginseng

Method used

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  • Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos
  • Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos
  • Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] Example 1 Inducing Panax notoginseng somatic embryogenesis and development

[0021] 1) Induce the embryogenic callus of Panax notoginseng and proliferate

[0022] Firstly, the seeds of Panax notoginseng are germinated, and after the adventitious roots are induced with germinated aseptic seedlings, the adventitious roots are cut into root segments of about 1 cm, and inoculated with 1.0 mg / L 2, 4-D, 30 g / L sucrose and 3.0 G / L gelrite MS solid medium for dark culture. About 50% of adventitious roots induced callus after 5 weeks. Calli are produced from the incisions at both ends of adventitious roots, and are loose and milky white. These calli were subcultured in the same medium every 3 weeks and proliferated rapidly.

[0023] 2) Induction of somatic embryogenesis and development

[0024] The embryogenic callus induced to proliferate was inoculated into differentiation medium containing different concentrations of 2,4-D. After 4 weeks, under the condition of 0.5mg / L2,...

example 2

[0029] Example 2 Efficient cultivation of somatic embryos using small bioreactors

[0030] Transfer the induced and proliferated embryogenic callus to MS solid medium supplemented with 0-1.0mg / L 2,4-D, 30g / L sucrose and 3.0g / L gelrite for dark culture to induce somatic cells Embryogenesis. Inoculate 6-7 embryogenic calli with a diameter of 5.0 mm in each glass triangular flask, and inoculate 7-8 flasks for each treatment. After the spherical embryos are formed, 0.2-0.6g (approximately 300-1000) spherical embryos are inoculated into a triangular flask (100ml) equipped with 30mL of 1 / 2MS liquid medium without 2,4-D and cultivated for 3 weeks, 0.2 g The somatic embryos with the initial inoculation amount grew and developed best, and most of them developed to torpedo embryos.

[0031] According to the ratio of the initial inoculation amount to the amount of medium, 10.0g (approximately 1.65×10 4 1) spherical embryos are transferred to a 3L bioreactor for cultivation, reflecting...

example 3

[0035] Example 3 Somatic Embryo Germination and Soil Transplantation

[0036] (1) Germination of somatic embryos

[0037] The cotyledon somatic embryos obtained by the above liquid or solid culture are transferred to 1 / 2MS solid medium without any plant growth regulator and cultured for 2 weeks, and adult cell embryos will germinate, but their tops are difficult to elongate. We used a range of concentrations of GA 3 Treatment of cotyledon somatic embryos. Select normal cotyledon somatic embryos of about 2.0cm, and use GA of 0-4.0mg / L 3 After processing for 2 weeks, statistical results showed (Table 3), GA 3 The concentration significantly affected the growth of somatic embryo seedlings, 2.0mg / L GA 3 The growth promotion effect on somatic embryonic seedlings is the best, and the top growth and root growth are the best.

[0038] Table 3 Different concentrations of GA 3 Effects on somatic embryo germination

[0039]

[0040] Note: Duncan's multiple comparisons were used...

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Abstract

The present invention provides an efficient method for rapid asexual propagation of notoginseng regenerated plants-by using a bioreactor to cultivate notoginseng somatic embryos in large scale. After proliferation of notoginseng embryogenic callus obtained by inducing adventitious roots, under the condition of MS containing 0.5mg / L 2,4-D, 100% of the embryogenic callus produce spherical somatic embryos with a largest number of embryos. 10.0g (about 1.65x10<4>) spherical somatic embryos are cultured in a 3L-bioreactor which contains 1.5L 1 / 2MS liquid medium without any plant growth regulators, with an aeration rate of 0 .15 vvm. 4 weeks later, the spherical somatic embryos all develop into cotyledonary somatic embryos, and biological mass growth increases to approximately 8 times of the original mass. The cotyledonary somatic embryos obtained by the liquid or solid medium culture, germinate to be normal somatic embryo seedlings in 1 / 2MS medium containing 2.0mg / LGA3, then subculture in the 1 / 2MS medium without any plant growth regulators. Well-grown seedlings are transplanted into nutritious cups filled with artificial soil (peat moss and argillaceous rocks (3:1, v: v)), with relatively high acclimatization survival rate in greenhouse.

Description

technical field [0001] The invention relates to a method for cultivating somatic embryos in large quantities in a small-scale bioreactor, and for simple and rapid asexual propagation of Panax notoginseng regenerated plants. The invention belongs to the field of agricultural biotechnology. Background technique [0002] Panax notoginseng (Burk.) F.H.Chen, also known as Tianqi, Jinbuhuan, Panax notoginseng, etc., is a plant of the genus Panax of Araliaceae. Panax notoginseng is a traditional Chinese medicinal material in my country, and its roots are mainly used as medicine. Radix Notoginseng is warm in nature, sweet and bitter in taste, has the effects of stopping bleeding and analgesia, promoting blood circulation and removing blood stasis, nourishing and strengthening, eliminating fatigue and enhancing physical strength. Injuries and other diseases have various pharmacological effects and health care functions, and have broad development prospects. Since notoginseng belong...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 由香玲詹亚光李玉花谭啸代金玲陶雷尹静
Owner NORTHEAST FORESTRY UNIVERSITY
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