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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (01): 58-69.doi: 10.3724/SP.J.1006.2016.00058

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Expression Analysis of Auxin Efflux Carrier Gene CsPIN3 in Tea Plant (Camellia sinensis)

WANG Bo1,2,CAO Hong-Li1,2,HUANG Yu-Ting1,2,HU Yu-Rong1,2,QIAN Wen-Jun1,3,HAO Xin-Yuan1, WANG Lu1,YANG Ya-Jun1,*,WANG Xin-Chao1,*   

  1. 1 Tea Research Institute of Chinese Academy of Agricultural Sciences / National Center for Tea Improvement / Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, Hangzhou 310008, China; 2 Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 Northwest A&F University, Yangling 712100, China
  • Received:2015-06-08 Revised:2015-09-06 Online:2016-01-12 Published:2015-10-08
  • Contact: 杨亚军, E-mail: yjyang@mail.tricaas.com, Tel: 0571-86650266; 王新超, E-mail: xcw75@mail.tricaas.com, Tel: 0571-86653162 E-mail:wb32244@tricaas.com
  • Supported by:

    This research was supported by the National Natural Science Foundation of China (31370690), the Modern Agro-industry Technology Research System (CARS-23) and the Scientific Innovation Program of Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2014-TRICAAS),

Abstract:

On the basis of previous transcriptome study on tea plant cold acclimatization, we obtained a PIN homology gene named CsPIN3 and cloned its full-length cDNA sequence by reverse transcription-PCR (RT-PCR) combining with rapid amplification of cDNA ends (RACE). The full length cDNA of CsPIN3 was 2654 bp (GenBank accession No. KP896474) and contained a 1 926 bp open reading frame (ORF) encoding 641 amino acid residues. Bioinformatic analyses showed that CsPIN3 was not a secretory protein and had a molecular weight of 70.15 kD, atheoretical isoelectric point of 8.42. Subcellular localization prediction showed that CsPIN3 was a typical membrane protein mainly located in plasmalemma and then in endoplasmic reticulum. Moreover, Amino acid sequence analysis indicated that CsPIN3 protein contained hydrophobic regions in both ends and hydrophilic regions in the middle. Similar to PIN protein in rice, the hydrophobic regions of CsPIN3 consisted of several transmembrane helixes, among which five was in N motif and four in C motif. The hydrophilic regions of CsPIN3 had two unstable domains, several o-glycosylation sites, several phosphorylation sites like TPRXS (N/S) motif (a PID/PINOID phosphorylation site) and a well characterized conserved inner motif NPNXY regulating the endocytosis of PIN. Comparison of sequences similarity showed that the amino acid sequence coded by CsPIN3 had more than 80% similarity with reported PINs of Populus trichocarpa, Vitis vinifera, Citrus sinensis, Nicotiana tomentosiformis, Solanum lycopersicum, Solanum tuberosum, and Sesamum indicum. Phylogenetic tree analysis showed that CsPIN3 had the closest genetic relationship with Solanaceae and the highest identity with AtPIN3 of Arabidopsis thaliana PIN proteins. The CsPIN3 gene differentially expressed in different tea plant tissues, and transcript abundance in flower was much higher than that in leaf, stem and root. In addition, we analyzed the expression of CsPIN3 by qRT-PCR during the different phases of bud dormancy formation and break, and the results indicated that in cultivar Longjing 43, the expression level of CsPIN3 at growth stage was higher than that at dormant stage (initial dormant stage to expanding stage) and an obvious expression jump was detected at bud sprouting stage. These demonstrated that CsPIN3 could be associated with the regulation of tea plant bud dormancy formation and break.

Key words: Tea plant (Camellia sinensis), Dormancy, Auxin efflux carrier, Expression analysis

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