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Method for acquiring transgenic seedlings by converting adventitious buds of honeysuckle

A honeysuckle and transgenic technology, applied in the field of plant genetic engineering, to achieve the effects of high transformation frequency, simple operation and short transformation period

Inactive Publication Date: 2014-11-05
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] After searching, there is no report about improving the germplasm of honeysuckle by genetic engineering technology and the method of transforming adventitious buds of honeysuckle to obtain transgenic plants

Method used

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  • Method for acquiring transgenic seedlings by converting adventitious buds of honeysuckle
  • Method for acquiring transgenic seedlings by converting adventitious buds of honeysuckle
  • Method for acquiring transgenic seedlings by converting adventitious buds of honeysuckle

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Embodiment 1

[0050] The method for obtaining transgenic plants by transforming adventitious buds of Lonicerae japonica is specifically implemented as follows:

[0051] (1) Acquisition and cultivation of honeysuckle aseptic vaccine

[0052] Cut off the young shoots of the honeysuckle plant, wash them with tap water for 30 minutes, cut off the leaves, cut the stems into small sections of about 2 cm, sterilize with 70% ethanol for 1 minute, and then sterilize with 0.1% HgCl2 for 10 minutes after removing the ethanol. Rinse 4 times with sterile water. After absorbing the water with sterile filter paper, the stem segments were placed on a solid medium, and cultivated under light at 25°C. Wherein the solid medium formula is: MS+0.1-1.0mg / L NAA (naphthalene acetic acid)+30g / L sucrose+7g / L agar, pH5.8;

[0053] (2) pre-cultivation of honeysuckle stem segment

[0054] Cut off the leaves and roots of the aseptic seedlings of honeysuckle cultivated for 25 days, cut the stems into 1 cm sections, ea...

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Abstract

The invention discloses a method for acquiring transgenic seedlings by converting adventitious buds of honeysuckle. The method comprises the steps of: acquiring and culturing aseptic seedlings of honeysuckle, preculturing the honeysuckle pedicels, preparing plant expression vector-containing agrobacterium strains, activating the agrobacterium lysate and preparing agrobacterium infection fluid, infecting and co-culturing agrobacterium, removing agrobacterium and inducing the regeneration of adventitious buds, cutting and rooting the honeysuckle buds, screening antibiotics positive seedlings, transplanting the antibiotics positive seedlings, and extracting DNA (Deoxyribonucleic Acid) to obtain transgenic seedlings. The method has the characteristics of being fast in adventitious bud regeneration speed, short in conversion period, high in transgenic seedling acquisition rate, and simple to operate; the bad variation in callus culture can be avoided by taking the pedicels as explants to induce the adventitious buds; and the method is also suitable for adventitious bud generation systems of a plurality of honeysuckle varieties, thus hopefully being a reliable system for honeysuckle transgenic breeding.

Description

technical field [0001] The invention relates to a method for obtaining transgenic plants by transforming adventitious buds of honeysuckle, belonging to the field of plant genetic engineering. Background technique [0002] Honeysuckle (Lonicera japonica L.) is a perennial semi-evergreen twining woody vine of Lonicera genus Lonicera, known as a good medicine for clearing away heat and detoxification since ancient times. It has the functions of antibacterial and antiviral, antipyretic, increasing white blood cells, protecting liver and gallbladder, immune regulation, antitumor, lowering blood pressure, lowering blood fat, and scavenging free radicals. The main chemical components of honeysuckle include flavonoids, organic acids, triterpenoid saponins, volatile oil, etc. Among them, chlorogenic acid (Chlorogenic acid) is the main functional component, and its content is an important indicator for evaluating the quality of medicinal materials. Honeysuckle has a wide range of use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 向凤宁周冰谦
Owner SHANDONG UNIV
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