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Int. J. Mol. Sci. 2016, 17(7), 1132; doi:10.3390/ijms17071132

Effect of Medium Supplements on Agrobacterium rhizogenes Mediated Hairy Root Induction from the Callus Tissues of Camellia sinensis var. sinensis

1
State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang Blvd West, Hefei 230036, China
2
Agronomy Division, Bangladesh Tea Research Institute, Srimangal-3210, Moulvibazar, Bangladesh
*
Author to whom correspondence should be addressed.
Academic Editor: Jianhua Zhu
Received: 3 May 2016 / Revised: 29 June 2016 / Accepted: 4 July 2016 / Published: 15 July 2016
(This article belongs to the Section Molecular Botany)
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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. View Full-Text
Keywords: Agrobacterium rhizogenes; antioxidants; browning; callus; hairy root; tea Agrobacterium rhizogenes; antioxidants; browning; callus; hairy root; tea
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MDPI and ACS Style

Rana, M.M.; Han, Z.-X.; Song, D.-P.; Liu, G.-F.; Li, D.-X.; Wan, X.-C.; Karthikeyan, A.; Wei, S. Effect of Medium Supplements on Agrobacterium rhizogenes Mediated Hairy Root Induction from the Callus Tissues of Camellia sinensis var. sinensis. Int. J. Mol. Sci. 2016, 17, 1132.

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