Cloning of OfTCPs Gene from Sweet Osmanthus (Osmanthus fragrans) and Expression Analysis During Floral Bud Differentiation
ZHOU Dan, MIAO Yun-Feng, DONG Bin, YANG Li-Yuan, ZHAO Hong-Bo*
School of Landscape Architecture/Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants/Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants, Zhejiang A&F University, Hangzhou 311300, China
Abstract：Sweet osmanthus (Osmanthus fragrans) is traditional famous flowers in China, and its floral bud differentiation process is affected by ambient temperature. Plant specific transcription factors TCP (teosinte branched 1/cycloidea/proliferating cell) involved in regulation of plant cell, organ and tissue formation or other physiological processes, and plays a significant role during floral bud development in plants. In this study, 12 OfTCPs cDNA sequence were cloned based on the transcriptional data of O. fragrans 'Yanhonggui'. Among them, 7 OfTCPs had complete ORF lengths ranging from 807 to 1 365 bp. The length of the proteins encoded by TCP ranged from 268 to 454 aa with molecular weights from 28.24 to 49.49 kD and the isoelectric points were from 6.57 to 9.08. The protein domain analysis showed that OfTCP contained basic helix-loop-helix (bHLH) domain; The phylogenetic tree analysis showed that twelve OfTCPs could be divide into the ClassⅠand ClassⅡ, and the same subgroup proteins contained similar conservative element. The qRT-PCR results showed that three genes OfTCP5, OfTCP9 and OfTCP12 might be the key genes to promote flower bud differentiation in response to low temperature response after the different temperature treatment (19 and 25 ℃), however, the response of OfTCP9 to low temperature was relatively slow and play an important role in the late stage of flower bud differentiation. Meanwhile, OfTCP5 and OfTCP12 had no self-activating activity and toxicity. This study lay the foundation for further research on the molecular mechanism of the TCP transcription factor in response to environmental temperature changes in the regulation of flower bud differentiation of Osmanthus fragrans.
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