Transfer of foreign DNA intoPhyscomitrella patens protonemal tissue by using the gene gun

Sawahel, Wagdy A.; Önde, Sertaç; Knight, Celia D.; Cove, David J.

doi:10.1007/bf02668906

Cited by 33 publications

(17 citation statements)

References 5 publications

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“…The biolistic transformation method is not restricted to specific plants or tissue types and functions with the same efficiency in monocotyledonous and dicotyledonous plants (Sawahel et al, 1992). In P. patens, transient transformation was achieved by shooting DNA-coated gold or tungsten particles onto living protonemal tissue (Cho et al, 1999, Cove et al, 2009c.…”

Section: Biolistic Delivery Of Dnamentioning

confidence: 99%

The moss Physcomitrella patens: methods and tools from cultivation to targeted analysis of gene function

Strotbek¹,

Krinninger²,

Frank³

2013

Int. J. Dev. Biol.

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To comprehensively understand the major processes in plant biology, it is necessary to study a diverse set of species that represent the complexity of plants. This research will help to comprehend common conserved mechanisms and principles, as well as to elucidate those mechanisms that are specific to a particular plant clade. Thereby, we will gain knowledge about the invention and loss of mechanisms and their biological impact causing the distinct specifications throughout the plant kingdom. Since the establishment of transgenic plants, these studies concentrate on the elucidation of gene functions applying an increasing repertoire of molecular techniques. In the last two decades, the moss Physcomitrella patens joined the established set of plant models based on its evolutionary position bridging unicellular algae and vascular plants and a number of specific features alleviating gene function analysis. Here, we want to provide an overview of the specific features of P. patens making it an interesting model for many research fields in plant biology, to present the major achievements in P. patens genetic engineering, and to introduce common techniques to scientists who intend to use P. patens as a model in their research activities.

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Section: Biolistic Delivery Of Dnamentioning

confidence: 99%

The moss Physcomitrella patens: methods and tools from cultivation to targeted analysis of gene function

Strotbek¹,

Krinninger²,

Frank³

2013

Int. J. Dev. Biol.

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“…Since then, the protocol has been optimized (107,109) and used routinely for transforming Physcomitrella and for transient gene expression assays (4, 41, 42, 46a, 47, 48, 52, 66, 79, 97, 101, 110, 119, 130, 131). Transient gene expression was also reported following biolistic delivery of DNA-coated microbeads into protonemal tissue (58,106), but this method seems to be less efficient than PEG-mediated transformation as there has been only one report of a transgenic strain produced this way (58). Genetic transformation of Physcomitrella using Agrobacterium-based methods has also been attempted without success in several laboratories (28,94,109).…”

Section: Physcomitrella Patensmentioning

confidence: 99%

A NEWMOSSGENETICS: Targeted Mutagenesis inPhyscomitrella patens

Schaefer

2002

Annu. Rev. Plant Biol.

162

110

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s Abstract The potential of moss as a model system to study plant biology is associated with their relatively simple developmental pattern that nevertheless resembles the basic organization of the body plan of land plants, the direct access to cell-lineage analysis, their similar responses to plant growth factors and environmental stimuli as those observed in other land plants, and the dominance of the gametophyte in the life cycle that facilitates genetic approaches. Transformation studies in the moss Physcomitrella patens have revealed a totally unique feature for plants, i.e., that foreign DNA sequences integrate in the genome preferentially at targeted locations by homologous recombination, enabling for the first time in plants the application of the powerful molecular genetic approaches used routinely in bacteria, yeast, and since 1989, the mouse embryonic stem cells. This article reviews our current knowledge of Physcomitrella patens transformation and its unique suitability for functional genomic studies.

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“…To take full advantage of this system, we need an effective and easy method to deliver DNA into moss cells. The most common ways to transform this moss are particle bombardment 10 and PEGmediated DNA uptake 11 . Although particle bombardment can produce a high transformation efficiency 12 , gene guns are not readily available to many laboratories and the protocol is difficult to standardize.…”

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confidence: 99%

Efficient Polyethylene Glycol (PEG) Mediated Transformation of the Moss <em>Physcomitrella patens</em>

Liu

Vidali

2011

JoVE

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A simple and efficient method to transform Physcomitrella pantens protoplasts is described. This method is adapted from protocols for Physocmitrella protonemal protoplast and Arabidopsis mesophyll protoplast transformation 1 . Due to its capacity to undergo efficient mitotic homologous recombination, Physcomitrella patens has emerged as an important model system in recent years 2 . This capacity allows high frequencies of gene targeting [3][4][5][6][7][8][9] , which is not seen in other model plants such as Arabidopsis. To take full advantage of this system, we need an effective and easy method to deliver DNA into moss cells. The most common ways to transform this moss are particle bombardment 10 and PEGmediated DNA uptake 11. Although particle bombardment can produce a high transformation efficiency 12 , gene guns are not readily available to many laboratories and the protocol is difficult to standardize. On the other hand, PEG mediated transformation does not require specialized equipments, and can be performed in any laboratory with a sterile hood. Here, we show a simple and highly efficient method for transformation of moss protoplasts. This method can generate more than 120 transient transformants per microgram of DNA, which is an improvement from the most efficient protocol previously reported 13 . Because of its simplicity, efficiency, and reproducibility, this method can be applied to projects requiring large number of transformants as well as for routine transformation.

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Transfer of foreign DNA intoPhyscomitrella patens protonemal tissue by using the gene gun

Cited by 33 publications

References 5 publications

The moss Physcomitrella patens: methods and tools from cultivation to targeted analysis of gene function

The moss Physcomitrella patens: methods and tools from cultivation to targeted analysis of gene function

A NEWMOSSGENETICS: Targeted Mutagenesis inPhyscomitrella patens

Efficient Polyethylene Glycol (PEG) Mediated Transformation of the Moss <em>Physcomitrella patens</em>

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