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An establishing method of a rosemary somatic embryogenesis and regeneration system

A method of establishment, the technology of rosemary, applied in the field of plant cell engineering, to achieve the effects of easy genetic engineering operation, normal plant development and uniform traits

Inactive Publication Date: 2015-02-11
张家港芳香生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on rosemary tissue culture at home and abroad, and there is no report on the establishment of a regeneration system through the somatic embryogenesis pathway.

Method used

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  • An establishing method of a rosemary somatic embryogenesis and regeneration system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] In this test, rosemary seeds imported from the United States were used. The rosemary seeds were soaked in tap water at room temperature for 10 to 15 hours, then sterilized with 70% alcohol for 2 minutes, washed twice with sterile water, and then rinsed with 0.1% alcohol. Sterilize the surface with mercury chloride (HgCl2) for 10 minutes, rinse with sterile water 5 times, use tweezers and a scalpel to cut out the embryos and inoculate them in a medium containing 6-BA1.3mg / L, 2,4-D4.5mg / L, 1200mg / L L hydrolyzed milk protein on the modified B5 medium (Table 1). The medium contains 30g / L sucrose and 7.0g / L agar. Use 1mol / L NaOH or HCl to adjust the pH value to 5.8-6.2, sterilize with high-pressure steam for 20 minutes, cultivate at 18-22°C, light-dark cycle 16h / 8h, light intensity 2500-30001x, relative humidity 70%-80%, Cultured for 20 days and replaced with fresh medium. The callus began to appear in 5 to 7 days, and the callus grew rapidly after 12 days. In addition to...

Embodiment 2

[0018] The culture procedures were all carried out according to Example 1, the difference being that the first stage added 6-BA4.5mg / L, 2,4-D4.3mg / L and 700mg / L, and the second stage was subcultured for the first time. , 4-D0.8mg / L, 6-BA is 0.9mg / L, the second subculture is 2,4-D0.9mg / L, 6-BA is 0.4mg / L, the third subculture is 2,4 -D0.2mg / L, 6-BA 0.8mg / L, in the third stage, transfer the embryogenic callus obtained by subculture to the B5 medium without growth regulators and supplemented with 600mg / L hydrolyzed milk protein conduct.

[0019] Table 1: B5 minimal medium (Gamborg et al., 1968) and modified medium components

[0020]

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Abstract

The invention relates to a regeneration system establishing method by using a rosemary mature embryo as a material. The method includes following three culture stages: (1) callus culturing and inducing after the mature embryo is stripped; (2) callus subculturing; and (3) somatic embryogenesis in the callus and embryoid development into a plant. In the subculturing, a process of reducing the concentration of 2,4-dichlorophenoxyacetic acid step by step is adopted, and the somatic embryogenesis is 63%. The system can be adopted as a system for rosemary genetic transformation and rapid propagation.

Description

technical field [0001] The invention relates to a method for establishing a rosemary in vitro culture regeneration system. Specifically speaking, the mature embryos of rosemary seeds are used as explants to establish a regeneration system through somatic embryogenesis. It belongs to the field of plant cell engineering. Background technique [0002] Rosemary (Rosmarinus officinalis), also known as tansy, is an evergreen shrub belonging to the family Lamiaceae. It has a cool and sweet smell of camphor, and improves and strengthens memory. It mainly contains phenols, acids, ketones, terpenes and other compounds, and is currently recognized as one of the highly effective antioxidant plants. It can be used as a raw material for spices, seasonings, perfumes, and shampoos, and has broad application prospects in industries such as health drinks, oral liquids, and cosmetics. Rosemary is native to Europe and the Mediterranean region, and is now widely cultivated in France, Spain, ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 杨学军曾扬腾闫勇义
Owner 张家港芳香生物科技有限公司
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