Functional complementation analysis in potato via biolistic transformation with BAC large DNA fragments

Ercolano, Maria Raffaella; Ballvora, Agim; Paal, Jürgen; Steinbiss, Hans-Henning; Salamini, Francesco; Gebhardt, Christiane
January 2004
Molecular Breeding;Jan2004, Vol. 13 Issue 1, p15
Academic Journal
Gene isolation from plants by positional cloning frequently requires several rounds of transformation. To reduce the resources invested and to accelerate the process, we have used large DNA fragments in transformation experiments, followed by analysis of transgenic plants to assess functional complementation. Specifically, the transformation of potato with DNA from the 106 kb BAC plasmid BA87d17 is described. The large fragment was introduced into the potato genome by biolistic transformation, while attempting to clone the R1 gene conferring a race specific resistance to Phytophthora infestans. Thirty-one kanamycin resistant plants were regenerated of which thirteen showed the necrotic lesions typical for the hypersensitive response after infection with the incompatible P. infestans race 4, which carries the avirulence gene Avr1. The successful complementation supported the location of the R1 gene in the BAC insertion of the BA87d17 plasmid. Based on PCR and Southern gel blot analysis, both complete and incomplete integrations of the large construct into the recipient genome were demonstrated.


Related Articles

  • Mapping of Avirulence Genes in Phytophthora infestans With Amplified Fragment Length Polymorphism... van der Lee, Theo; Robold, Andrea; Testa, Antonino; van 't Klooster, John W.; Govers, Francine // Genetics;Mar2001, Vol. 157 Issue 3, p949 

    Investigates the genetic control of avirulence (Avr) in the diploid oomycete pathogen Phytophthora infestans. Number of resistance genes that match the Avr genes; Number of Avr genes found located on the most distal part of the linkage group; Reason for the use of high-density mapping.

  • Fighting potato blight.  // Plantsman: New Series;Dec2009, Vol. 8 Issue 4, p208 

    The article presents the findings of a study that the genome of Phytophthora infestans (P. infestans), a potato blight pathogen that is member of a oomycetes or water moulds, is large compared with P. sojae and P. ramorum due to its repetitive DNA.

  • Quantitative resistance to late blight from Solanum berthaultii cosegregates with R: insights in stability through isolates and environment. Rauscher, Gilda; Simko, Ivan; Mayton, Hilary; Bonierbale, Merideth; Smart, Christine D.; Grünwald, Niklaus J.; Greenland, Andrew; Fry, William E. // Theoretical & Applied Genetics;Nov2010, Vol. 121 Issue 8, p1553 

    Genetic resistance is a valuable tool in the fight against late blight of potatoes but little is known about the stability and specificity of quantitative resistance including the effect of defeated major resistance genes. In the present study we investigated the effect of different isolates of...

  • Genomics: Irish-famine pathogen decoded.  // Nature;05/30/2013, Vol. 497 Issue 7451, p538 

    The article discusses the genetic sequencing of microorganism of historical plant pathogen of the strain Phytophthora infestans that destroyed Irish potato crop.

  • Potato blight has the genome of death.  // New Scientist;9/12/2009, Vol. 203 Issue 2725, p16 

    The article focuses on a study on the genome of Phytophthora infestans (P. infestans), an oomycete or water mould that causes potato blight, conducted by researchers at the Sainsbury Laboratory in Norwich, England. The genome was sequenced by the research team and discovered it stores genes that...

  • Analyses of genome architecture and gene expression reveal novel candidate virulence factors in the secretome of Phytophthora infestans. Raffaele, Sylvain; Joe Win; Cano, Liliana M.; Kamoun, Sophien // BMC Genomics;2010, Vol. 11, p637 

    Background: Phytophthora infestans is the most devastating pathogen of potato and a model organism for the oomycetes. It exhibits high evolutionary potential and rapidly adapts to host plants. The P. infestans genome experienced a repeat-driven expansion relative to the genomes of Phytophthora...

  • Genetic relationships between the dominant genotypes of Phytophthora infestans in Hokkaido, Japan. Akino, Seishi; Kato, Masayasu; Gotoh, Kiyotaka; Naito, Shigeo; Ogoshi, Akira // Journal of General Plant Pathology;Jun2005, Vol. 71 Issue 3, p200 

    The genetic characteristics of the dominant genotypes of Phytophthora infestans in Japan (US-1, JP-1, Japanese A1-A, A1-B) were compared. Differences were evident in the peptidase genotype, amplified fragment length polymorphism, and RG57 DNA fingerprints. Almost all of the fingerprint bands for...

  • Species Tree Estimation for the Late Blight Pathogen, Phytophthora infestans, and Close Relatives. Blair, Jaime E.; Coffey, Michael D.; Martin, Frank N. // PLoS ONE;May2012, Vol. 7 Issue 5, p1 

    To better understand the evolutionary history of a group of organisms, an accurate estimate of the species phylogeny must be known. Traditionally, gene trees have served as a proxy for the species tree, although it was acknowledged early on that these trees represented different evolutionary...

  • Historic Late Blight Outbreaks Caused by a Widespread Dominant Lineage of Phytophthora infestans (Mont.) de Bary. Saville, Amanda C.; Martin, Michael D.; Ristaino, Jean B. // PLoS ONE;12/28/2016, Vol. 11 Issue 12, p1 

    Phytophthora infestans (Mont.) de Bary, the causal agent of potato late blight, was responsible for the Irish potato famine of the 1840s. Initial disease outbreaks occurred in the US in 1843, two years prior to European outbreaks. We examined the evolutionary relationships and source of the...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics