title: Effect of Medium Supplements on Agrobacterium rhizogenes Mediated Hairy Root Induction from the Callus Tissues of Camellia sinensis var. sinensis.
abstract: Tea (Camellia sinensis L.) is recalcitrant to Agrobacterium-mediated genetic transformation largely due to the bactericidal effects of tea polyphenols and phenolics oxidation induced by necrosis of explant tissue over the process of transformation. In this study, different antioxidants/adsorbents were added as supplements to the co-cultivation and post co-cultivation media to overcome these problems for the transformation improvement. Tea-cotyledon-derived calli were used as explants and Agrobacterium rhizognes strain ATCC 15834 was used as a mediator. Results showed that Agrobacterium growth, virulence (vir) gene expression and browning of explant tissue were greatly influenced by different supplements. Murashige and Skoog (MS) basal salts medium supplemented with 30 g·L(-1) sucrose, 0.1 g·L(-1) l-glutamine and 5 g·L(-1) polyvinylpolypyrrolidone (PVPP) as co-cultivation and post co-cultivation media could maintain these parameters better that ultimately led to significant improvement of hairy root generation efficiency compared to that in the control (MS + 30 g·L(-1) sucrose). Additionally, the reporter genes β-glucuronidase (gusA) and cyan fluorescent protein (cfp) were also stably expressed in the transgenic hairy roots. Our study would be helpful in establishing a feasible approach for tea biological studies and genetic improvement of tea varieties.
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. ranabtri@yahoo.com.|Agronomy Division, Bangladesh Tea Research Institute, Srimangal-3210, Moulvibazar, Bangladesh. ranabtri@yahoo.com.
authors.forename: Mohammad M
authors.initials: MM
authors.lastname: Rana
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. hanzx1226@163.com.
authors.forename: Zhuo-Xiao
authors.initials: ZX
authors.lastname: Han
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. sdp20073882@163.com.
authors.forename: Da-Peng
authors.initials: DP
authors.lastname: Song
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. babcule1989@163.com.
authors.forename: Guo-Feng
authors.initials: GF
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. dxli@ahau.edu.cn.
authors.forename: Da-Xiang
authors.initials: DX
authors.lastname: Li
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. xcwan@ahau.edu.cn.
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Wan
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. mdukarthimicro@yahoo.com.
authors.forename: Alagarsamy
authors.initials: A
authors.lastname: Karthikeyan
authors.affiliation: State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China. weishu@ahau.edu.cn.
authors.forename: Shu
authors.initials: S
authors.lastname: Wei
mesh_terms: D060054:Agrobacterium / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D000975:Antioxidants / Q000494:pharmacology*; D028241:Camellia sinensis / Q000235:genetics*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant; D018517:Plant Roots / Q000187:drug effects / Q000254:growth & development*; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic
chemical_list: D000975:Antioxidants; D017077:Culture Media, Conditioned
keywords: Agrobacterium rhizogenes; antioxidants; browning; callus; hairy root; tea