title: Clovers (Trifolium spp.).
abstract: Clovers (Trifolium spp.) constitute one of the major forage legumes widely grown for its rich protein content and its major role in maintaining environmental sustainability by improving the soil fertility. Gene technology can assist plant improvement efforts in clovers (Trifolium spp.), aiming to improve forage quality, yield, and adaptation to biotic and abiotic stresses. An efficient and reproducible protocol for Agrobacterium-mediated transformation of a range of Trifolium species, using cotyledonary explants and different selectable marker genes, is described. The protocol is robust and allows for genotype and Agrobacterium strain-independent transformation of clovers. Stable meiotic transmission of transgenes has been demonstrated for selected transgenic clovers carrying single T-DNA inserts recovered from Agrobacterium-mediated transformation. This methodology can also be successfully used for "isogenic transformation" in clovers: the generation of otherwise identical plants with and without the transgene from the two cotyledons of a single seed. Stable transgenes may be used in further functional genomics, develop new traits and profile gene expression using reporters, and facilitate purification of tissue or single cells.
authors.affiliation: School of Applied Systems Biology, AgriBio, Centre for AgriBioscience, La Trobe University, Bundoora, VIC, 3083, Australia.
authors.forename: Samira
authors.initials: S
authors.lastname: Rahimi-Ashtiani
authors.forename: Sareena
authors.initials: S
authors.lastname: Sahab
authors.forename: Stephen
authors.initials: S
authors.lastname: Panter
authors.forename: John
authors.initials: J
authors.lastname: Mason
authors.forename: German
authors.initials: G
authors.lastname: Spangenberg
mesh_terms: D000383:Agriculture / Q000379:methods; D016960:Agrobacterium tumefaciens / Q000235:genetics; D004269:DNA, Bacterial; D005821:Genetic Techniques; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified; D012639:Seeds / Q000235:genetics; D012641:Selection, Genetic; D013242:Sterilization; D014169:Transformation, Bacterial; D029921:Trifolium / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA