Single-stranded DNA used as an efficient new vehicle for transformation of plant protoplasts

Plant Molecular Biology
K W RodenburgP J Hooykaas

Abstract

In relation to the question which DNA form (single- or double-stranded) is transferred by Agrobacterium tumefaciens to plant cells, we studied the behaviour of single-stranded DNA, as compared to double-stranded DNA, when it is introduced into plant protoplasts by electroporation. To this end, we cloned a construct with a plant NPTII gene as well as a CAT gene in the M13 vectors tg130 and tg131. We found that both complementary single-stranded molecules gave rise to substantial CAT activity in plant protoplasts, suggesting that single-stranded DNA is converted into double-stranded DNA by the plant cell replication machinery. Unexpectedly, we found that single-stranded DNA leads to a 3-10-fold higher frequency of stable transformation (selection for kanamycin resistance) than double-stranded DNA. These results indicate that the use of single-stranded DNA might be considered in experiments in which optimal transformation frequencies are needed, e.g. with protoplasts from recalcitrant plant species.

References

Aug 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·S RauthR Kucherlapati
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·C GietlB Hohn
Apr 11, 1985·Nucleic Acids Research·J Stanley, R Townsend
Jul 4, 1973·Nature: New Biology·P MaligaL Márton
Dec 21, 1984·Nucleic Acids Research·W KramerH J Fritz
Feb 5, 1984·Journal of Molecular Biology·G P DottoN D Zinder
Sep 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·T M Ou-LeeR Wu
Apr 26, 1907·Science·E F Smith, C O Townsend
Jan 1, 1987·Plant Molecular Biology·H P SpainkB J Lugtenberg

❮ Previous
Next ❯

Citations

May 1, 1992·Plant Molecular Biology·P J Hooykaas, R A Schilperoort
Sep 1, 1989·Plant Molecular Biology·P J Hooykaas
Jul 1, 1996·World Journal of Microbiology & Biotechnology·K Weising, G Kahl
Aug 16, 1994·Proceedings of the National Academy of Sciences of the United States of America·B TinlandH Puchta
Nov 9, 2002·Molecular Plant-microbe Interactions : MPMI·Tomas CantoPeter Palukaitis
Oct 1, 1995·Genome Génome / Conseil National De Recherches Canada·Z LiR L Jarret
Jun 29, 2010·Journal of Nanobiotechnology·Arcan GüvenUlrich Schwaneberg
Jun 1, 1992·Plant Physiology·S L Van Wert, J A Saunders
Jan 1, 1992·Molecular & General Genetics : MGG·U HalfterL Willmitzer
Feb 1, 1991·Genes & Development·G GheysenM Van Montagu

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.