Laser-based micromanipulation for separation and identification of individual Frankia vesicles.
In studies of symbiotic efficiency it is of great importance to identify and separate individual Frankia strains from a nodule. Therefore, a new laser-based micromanipulation technique has been developed in which individual vesicles from root nodules of two Frankia-Alnus symbioses have been successfully cut loose and separated from clusters of vesicles in sterile conditions under light microscopy using a laser scalpel and optical tweezers. Vesicles from the Alnus incana-Frankia AvCI1 symbiosis were successfully isolated and grown in culture using this technique. The DNA from both Frankia sources was amplified by polymerase chain reaction (PCR). The work shows that a combination of laser-based manipulation techniques and PCR can be used for the separation and study of individual vesicles. This novel laser-based micromanipulation technique opens up various new possibilities, for instance, to study whether several Frankia strains can grow simultaneously in the same root nodule.
Leitz
Guenther
G
Department of Experimental Physics, Umeå University, 901 87 Umeå, Sweden.
Lundberg
Carina
C
Fällman
Erik
E
Axner
Ove
O
Sellstedt
Anita
A
D022162:Cytoplasmic Vesicles / Q000502:physiology*; D004269:DNA, Bacterial / Q000032:analysis; D040161:Frankia / Q000235:genetics / Q000648:ultrastructure*; D005798:Genes, Bacterial; D007834:Lasers; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods*; D009586:Nitrogen Fixation / Q000235:genetics; D018517:Plant Roots / Q000382:microbiology; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial