Karyotype analysis,genome organization,and stable genetic transformation of the root colonizing fungus Piriformospora indica |
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| Institution: | 1. Department of Systems Biology, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran;2. Department of Agronomy, Faculty of Agriculture, Malayer University, Malayer, Iran;3. Department of Entomology and Plant Pathology, University of Tennessee, Knoxville, TN, USA;4. Department of Crop Biotechnology and Breeding, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran;5. Department of Soil Science, Faculty of Agriculture, Isfahan University of Technology, Isfahan, Iran;6. Australian School of Advanced Medicine, Faculty of Human Sciences, Macquarie University, Sydney, NSW, Australia;7. Australian Proteome Analysis Facility (APAF), Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia;8. Agricultural and Plant Biochemistry and Proteomics Research Group, Dept. of Biochemistry and Molecular Biology, University of Córdoba, Córdoba, Spain;9. Department of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran |
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| Abstract: | Piriformospora indica (Basidiomycota, Sebacinales) is a root colonizing fungus which is able to increase biomass and yield of crop plants and to induce local and systemic resistance to fungal diseases and tolerance to abiotic stress. A prerequisite for the elucidation of the mode of action of this novel kind of symbiosis is knowledge of the genome organization as well as the development of tools to study and modify gene functions. Here we provide data on the karyotype and genetic transformation strategies. The fungus was shown to possess at least six chromosomes and a genome size of about 15.4–24 Mb. Sequences of the genes encoding the elongation factor 1-α (TEF) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used for genome size estimation through real-time PCR analysis. Chromosomal location investigated by Southern blot and expression analysis suggested that TEF and GAPDH are single-copy genes with strong and constitutive promoters. A genetic transformation system was established using a fragment of the TEF promoter region for construction of vectors carrying the selectable marker hygromycin B phosphotransferase. Results demonstrate that P. indica can be stably transformed by random genomic integration of foreign DNA and that it posses a relative small genome as compared to other members of the Basidiomycota. |
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