title: Recovery of transgenic plants by pollen-mediated transformation in Brassica juncea.
abstract: The aroA-M1 encoding the mutant of 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) was introduced into the Brassica juncea genome by sonication-assisted, pollen-mediated transformation. The plasmid DNA and collected pollen grains were mixed in 0.3 mol/L sucrose solution and treated with mild ultrasonication. The treated pollen was then pollinated onto the oilseed stigmas after the stamens were removed artificially. Putative transgenic plants were obtained by screening germinating seeds on a medium containing glyphosate. Southern blot analysis of glyphosate-resistant plants indicated that the aroA-M1 gene had been integrated into the oilseed genome. Western blot analysis further confirmed that the EPSPS coded by aroA-M1 gene was expressed in transgenic plants. The transgenic plants exhibited increased resistance to glyphosate compared to untransformed plants. Some of those transgenic plants had considerably high resistance to glyphosate. The genetic analysis of T1 progeny further confirmed that the inheritance of the introduced genes followed the Mendelian rules. The results indicated that foreign genes can be transferred by pollen-mediated transformation combined with mild ultrasonication.
authors: School of Life Science and Technology, Shanxi University, Taiyuan 030006, P.R. China. jingxuew@hotmail.com
authors: Jingxue
authors: J
authors: Wang
authors: Yonghu
authors: Y
authors: Li
authors: Chao
authors: C
authors: Liang
mesh_terms: D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors; D018525:Germination / Q000187:drug effects / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D053209:Herbicide Resistance / Q000235:genetics; D009149:Mustard Plant / Q000235:genetics*; D030821:Plants, Genetically Modified; D011058:Pollen / Q000235:genetics*; D054817:Pollination / Q000235:genetics; D013048:Specimen Handling; D014170:Transformation, Genetic; D000097797:Glyphosate
chemical_list: D005998:Glycine; D000097797:Glyphosate
references: Annu Rev Plant Physiol Plant Mol Biol. 1999 Jun;50:473-503
references: 15012217
references: Plant Cell Rep. 1995 Dec;15(3-4):159-63
references: 24185767
references: Plant Cell Rep. 2006 May;25(5):410-6
references: 16331457
references: J Plant Res. 2003 Dec;116(6):455-60
references: 12955571
references: Vet Hum Toxicol. 1992 Dec;34(6):531-43
references: 1287975
references: Plant Cell Rep. 1990 Aug;9(4):207-10
references: 24226704
references: Biosci Biotechnol Biochem. 2003 Jun;67(6):1405-9
references: 12843673
references: Proc Natl Acad Sci U S A. 1986 Apr;83(8):2571-5
references: 3458219
references: Arch Environ Contam Toxicol. 1999 Feb;36(2):193-9
references: 9888965
references: Plant Cell Rep. 1995 Feb;14(5):273-8
references: 24186759
references: Plant Cell Rep. 2005 Nov;24(9):513-22
references: 16172896
references: Theor Appl Genet. 1985 Mar;69(5-6):571-4
references: 24254015
references: Trends Plant Sci. 1999 Jun;4(6):226-231
references: 10366879
references: Proc Natl Acad Sci U S A. 1986 Feb;83(3):715-9
references: 16593654
references: Theor Appl Genet. 1995 Sep;91(4):627-31
references: 24169890
references: Biochim Biophys Acta. 2001 Nov 7;1568(1):1-6
references: 11731078
references: Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2767-72
references: 9501164
references: Plant Cell Rep. 2003 Feb;21(6):599-604
references: 12789436
references: Methods Cell Biol. 1995;50:375-82
references: 8531809