青蒿素的瞬时转化

Dongming Ma; Hong Wang

doi:10.21769/BioProtoc.1476

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Peer-reviewed

Transient Transformation of Artemisia annua

青蒿素的瞬时转化

Dongming Ma

Hong Wang

发布: 2015年05月20日第5卷第10期 DOI: 10.21769/BioProtoc.1476 浏览次数: 9022

评审: Tie LiuAnonymous reviewer(s)

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Cover of Plant & Cell Physiology, featuring study using the protocol.

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Abstract

Transient transformation of Artemisia annua does not depend on chromosomal integration of heterologous DNA, and recombinant DNA can be introduced into plant cells via Agrobacterium aided by vacuum. The leaves of 7th and 8th internode from 4-week-old seedlings were chosen as explants, a vacuum system was applied to facilitate agrobacteria into plant cells, the co-cultivation was in the dark at 25 °C for 36-72 h, then GUS or GFP maker genes were used for testing the efficiency of the transformation. The method is used for quick transferring of genes into Artemisia annua (A. annua) by transient transformation.

Keywords: Artemisia annua (阿尔泰米夏黄花)

Agrobacterium tumefaciens (农杆菌介导法)

Transient transformation (瞬时转化)

Materials and Reagents

Artemisia annua leaves from 4-week-old aseptic seedlings
Agrobacterium tumefaciens (A. tumefaciens) strain EHA105 (or LBA4404)
Lysogeny broth (LB) medium (pH 7.0, containing 50 mg/L rifampicin and 50 mg/L kanamycin)
Murashige and Skoog medium (MS) liquid medium (Sigma-Aldrich, catalog number: M5524 )
100 μM acetosyringone (Sigma-Aldrich, catalog number: D134406 )
10 mM MgCl₂
0.005% Silwet L-77 (Agri-Turf Supplies, catalog number: VIS-01 )
Glycerol
Filter paper (Lab Depot, catalog number: TLDCFP1-032 )
Yeast extract broth (YEB) medium (see Recipes)