Agrobacterium-mediated genetic transformation method using Cinnamomum camphora embryo-cultured seedling stem segment as receptor

Abstract

The invention provides an Agrobacterium-mediated method genetic transformation using Cinnamomum camphora embryo-cultured seedling stem segment as a receptor. The method comprises the following steps: pre-culturing stem segment of Cinnamomum camphora embryo-cultured seedling clone EL6 tissue culture seedling in an adventitious bud induction medium S1; infecting the Cinnamomum camphora stem segment subjected to pre-culture as a transformation acceptor material in an Agrobacterium infection liquid; absorbing excessive liquid on the infected explants with sterile filter paper, transferring the explants into a co-culture medium for co-cultivation; transferring the stem segments into a selective medium for selective culture, so as to obtain a transgenic positive plant material. The method provided by the invention is convenient for acquiring materials, uses abundant materials, and is easy to obtain regeneration plants; the genetic transformation of the clone stem segments of Cinnamomum camphora embryo-cultured seedling has the characteristics of simple method, simplified steps and short period; the application of high-frequency regeneration Cinnamomum camphora clone plant material greatly improves the transgenic plant acquisition rate of Agrobacterium mediated genetic transformation, shortens the Cinnamomum camphora genetic transformation cycle, and shortens the period to obtain novel germplasm of transgenic Cinnamomum camphora.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a genetic transformation method mediated by Agrobacterium using the stem section of camphor embryo cultured seedlings as receptors. Background technique [0002] Cinnamomum camphora (Cinnamomum camphora L.), a plant of the family Lauraceae, is a representative tree species of subtropical evergreen broad-leaved forest. Its tree shape is beautiful, with luxuriant branches and leaves, green shade covering the sun, and majestic momentum. It is an excellent tree species commonly used in urban landscaping. The distribution in my country is generally bounded by the north of the Yangtze River, south to Guangdong, Guangxi and southwest, with Jiangxi, Zhejiang, Fujian and other southeast coastal provinces as the majority, so the cultivation range is limited by low temperature. In recent years, with the development of urban construction and the new requirement...

AI Technical Summary

Problems solved by technology

The superposition of two unfavorable factors makes it difficult to obtain transgenic regenerated plants by using the embryogenic callus of Cinnamomum camphora as explants for genetic transformation

[0005] Nanyang Normal University's patent application "Cinnamomum camphora Embryo Cultivated Seedling Stem Cut Section Breeding Method for High-frequency Plant Regeneration Clones" (Acceptance No.: 201410188756.3) proposes to use the Cinnamomum camphora cotyledon cultured seedling stem cut section as explants, through direct Organogenesis pathway, a method for establishing a high-frequency plant regeneration system, because it does not have specific genetic transformation steps, so it is impossible to achieve the improvement of camphor varieties

[0006] At present, there is no report about the transgenic plants of Cinnamomum camphora obtained through the organogenesis pathway at home and abroad.

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