Biological projectiles (phage, yeast, bacteria) for genetic transformation of plants

Bacteriophage lambda particles, yeast cells, and bacterial cells were tested as projectiles to deliver marker/reporter genes into plant cells via the biolistic process. When phage particles were complexed to tungsten or gold particles and used to bombard tobacco cells, fewer than 15 cell clusters pe...

Full description

Saved in:

Bibliographic Details
Published in:	In vitro cellular & developmental biology. Plant Vol. 35; no. 1; pp. 43 - 50
Main Authors:	Kikkert, J.R, Humiston, G.A, Roy, M.K, Sanford, J.C
Format:	Journal Article
Language:	English
Published:	Wallingford Society for In Vitro Biology 1999 Cambridge University Press
Subjects:	Agronomy. Soil science and plant productions Bacteria Bacteriophages beta-glucuronidase binding biochemical techniques Biolistics Biological and medical sciences Biotechnology Biotechnology/Genetic Transformation/Somatic Cell Genetics Chemical suspensions Corn DNA Escherichia coli evaluation Fundamental and applied biological sciences. Psychology gene expression Genetic engineering applications genetic markers genetic transformation genetic vectors Genetics and breeding of economic plants molecular weight mutants Nicotiana tabacum particle size Plant breeding: fundamental aspects and methodology Plant cells Plasmids Projectiles reporter genes Saccharomyces cerevisiae Tungsten Yeasts Zea mays Cell culture Monocotyledones Microprojectile bombardment Petunia hybrida Escherichia coli Siphoviridae Nicotiana tabacum Cereal crop Fungi Oryza sativa Gramineae Dicotyledones Tungsten Angiospermae Bacteria Ascomycetes Solanaceae Phage lambda Enterobacteriaceae Thallophyta Complexation Gold Genetic transformation Zea mays Method Virus Flower crop Genetic engineering Spermatophyta Experimental plant Saccharomyces cerevisiae Phage
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Bacteriophage lambda particles, yeast cells, and bacterial cells were tested as projectiles to deliver marker/reporter genes into plant cells via the biolistic process. When phage particles were complexed to tungsten or gold particles and used to bombard tobacco cells, fewer than 15 cell clusters per plate transiently expressed β-glucuronidase (GUS). Cells of wild-type Saccharomyces cerevisiae were too large to be effective projectiles, but use of a reduced-size mutant resulted in a small number of transformants. Escherichia coli cells complexed with tungsten were the most effective projectile for plant transformation. Various methods to prepare E. coli were tested to reduce particle size, improve binding of bacteria to metal particles, and/or minimize particle clumping. In maize, the number of transformants was highest when bacteria/tungsten particles were air-dried onto macrocarriers from an aqueous solution. When maize cells were bombarded with bacteria/tungsten projectiles, rates of transient gene expression (2000 per plate) and stable transformation (50 per plate) were only two- to threefold lower than when purified DNA was used. Transformation of tobacco with E. coli projectiles was improved when the bacteria were treated with a series of ethanol and ether washes, then dried into a powder. Nevertheless, tobacco transformation was still 24- (transient) and 200-fold (stable) less than when purified DNA was used. Biological projectiles can be effective for plant transformation and are advantageous because once a DNA construct is made and put into the appropriate microorganism, the need to isolate and purify DNA for the biolistic process is eliminated, which saves time and lessens DNA shear. Such projectiles may be especially well suited where high molecular weight DNA constructs are needed.
ISSN:	1054-5476 1475-2689
DOI:	10.1007/s11627-999-0008-y