拟南芥嫁接实验

Tonni Grube  Andersen; Dacheng Liang; Barbara  Ann Halkier; Rosemary White

doi:10.21769/BioProtoc.1164

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Peer-reviewed

Grafting Arabidopsis

拟南芥嫁接实验

TA Tonni Grube Andersen

DL Dacheng Liang

BH Barbara Ann Halkier

RW Rosemary White email

发布: 2014年07月05日第4卷第13期 DOI: 10.21769/BioProtoc.1164 浏览次数: 16656

评审: Ru Zhang

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参见作者原研究论文

The authors used this protocol in:

Cover of The Plant Cell, featuring study using the protocol.

Aug 2013

实验方案合集

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Abstract

In Arabidopsis thaliana, hypocotyl micrografting has been used to investigate transport of flowering signals, mobile silencing signals and other peptides, proteins and secondary compounds. The effects of transported signals on target tissues require that a good vascular connection is re-established across the graft junction between the cut hypocotyls (stumps) of the root (rootstock) and shoot (scion) tissues. We outline here a method that requires only that the cut stumps be placed in close proximity, so that they touch, followed by 3-5 days of undisturbed recovery time during which the grafts are allowed to dry out somewhat. This method is quick, easy to monitor and has up to 90% success rate.

Keywords: Grafting (嫁接)

Materials and Reagents

Young Arabidopsis seedlings (grown for 3-6 days on ½ MS containing agar plates)
Sterile distilled water
96 % ethanol

Equipment

Sterile 9 cm petri dishes
Sterile standard Whatman No. 1 filter paper circles Grade 1:11 µm with diameter about 0.5 cm smaller than the sterile petri dish (autoclaved wrapped in foil beforehand) (8.5 cm diameter) (Whatman)
Sterile cellulose nitrate filters (Whatman, catalog number: NC 45 ST)
Sterile forceps
Sterile 1 ml pipette tips
1 ml pipettor
Micropore tape (3 M, Micropore^TM)
Sapphire knife holder (World Precision Instruments, catalog number: 500317 ) or diamond knife plus blade (Electron Microscopy Sciences, Diamond Dissecting Knife, Type M-M, catalog number: 72025 )
Sapphire blade (World Precision Instruments, catalog number: 504072 ) or diamond blade (Electron Microscopy Sciences, Blade Resharpening, Type M-M, catalog number: 72025-R ; Electron Microscopy Sciences, Replacement Blade, Type M-M, catalog number: 72025-L )
Flow cabinet mounted dissecting microscope with a minimum of 5x magnification
Note: Any brand we have tried worked.
Parafilm
Hot glass bead sterilizer (any brand) unless using 96 % ethanol for sterilization