title: Efficient soybean regeneration and Agrobacterium-mediated transformation using a whole cotyledonary node as an explant.
abstract: An optimized regeneration and Agrobacterium-mediated transformation protocol based on whole cotyledonary node explants was developed in soybean (Glycine max) cultivar Zhong Huang 13. Adding 6-benzylaminopurine (BAP) in a germinating medium could significantly increase regeneration efficiency; the optimal BAP concentration for shoot formation was 0.5 mg/L. The concentrations of plant growth regulators in a shoot induction medium were optimized by the orthogonal test [L9 (3(3))]. The best combination for shoot regeneration was a medium of Murashige & Skoog salts with B5 vitamins (MSB) supplemented with 3.5 mg/L BAP, 0.2 mg/L indole-3-butyric acid (IBA), and 0.2 mg/L kinetin (KT). Under this favorable condition, one node could regenerate 28-30 shoots. Soybean whole cotyledonary nodes were transformed by inoculation with A. tumefaciens strain EHA105 harboring a vector pBI121 containing a β-glucuronidase gene (gus). GUS assay, polymerase chain reaction, and Southern blot analysis indicated that the gus gene was transformed into soybean plants with 23.1% transformation efficiency. Transgenic plants could be obtained within 5-6 weeks, which was about 4 weeks less than that of a traditional single cotyledonary node method.
authors: Key Laboratory of Plant Genetics and Molecular Breeding, Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors: Fuli
authors: F
authors: Zhang
authors: Can
authors: C
authors: Chen
authors: Honglian
authors: H
authors: Ge
authors: Jinmei
authors: J
authors: Liu
authors: Yunling
authors: Y
authors: Luo
authors: Kun
authors: K
authors: Liu
authors: Long
authors: L
authors: Chen
authors: Kedong
authors: K
authors: Xu
authors: Yi
authors: Y
authors: Zhang
authors: Guangxuan
authors: G
authors: Tan
authors: Chengwei
authors: C
authors: Li
mesh_terms: D060054:Agrobacterium / Q000235:genetics*; D001593:Benzyl Compounds; D018548:Cotyledon / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D005966:Glucuronidase; D007701:Kinetin; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011687:Purines; D013025:Glycine max / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014170:Transformation, Genetic
chemical_list: D001593:Benzyl Compounds; D018744:DNA, Plant; D011687:Purines; D005966:Glucuronidase; C480551:benzylaminopurine; D007701:Kinetin
keywords: Agrobacterium-mediated transformation; Glycine max; whole cotyledonary node