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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (03): 376-388.doi: 10.3724/SP.J.1006.2016.00376

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

Cloning and Expression Analysis of a Neutral/alkaline Invertase Gene (CsINV10) in Tea Plant (Camellia sinensis L. O. Kuntze)

QIAN Wen-Jun1,2,YUE Chuan2,CAO Hong-Li2,HAO Xin-Yuan2,WANG Lu2,WANG Yu-Chun1,2,HUANG Yu-Ting2,WANG Bo2,WANG Xin-Chao2,XIAO Bin1,*,YANG Ya-Jun1,2,*   

  1. 1 College of Horticulture, Northwest Agriculture and Forestry University, Yangling 712100, China; 2 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
  • Received:2015-08-12 Revised:2015-11-20 Online:2016-03-12 Published:2015-12-18
  • Contact: 杨亚军, E-mail: yjyang@mail.tricaas.com, Tel: 0571-86653162; 肖斌, E-mail: xiaobin2093@sohu.com E-mail:jajyqian9066@mail.tricaas.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31170650), the Earmarked Fund for China Agriculture Research System (CARS-23), the Natural Science Foundation of Zhejiang Province (LY14C160001), the Major Project for New Agricultural Varieties Breeding of Zhejiang Province (2012C2905-3) and the Chinese Academy of Agricultural Sciences through an Innovation Project for Agricultural Sciences and Technology (CAAS-ASTIP-2014-TRICAAS).

Abstract:

Based on the comprehensive RNA-Seq analysis of tea plant during cold acclimation stage, we picked out six ESTs sequences with a high similarity to neutral/alkaline invertase gene to get a splicing. As a result, a full-length of 2101 bp nucleotide sequence was obtained from tea plant after validated by using RT-PCR technique. Bioinformatics analysis showed that the sequence containing 1923 bp ORF (Open Reading Fram) and encoding 640 amino acid residues with a putative molecular mass of 71.8 kD and theoretical isoelectric point of 5.69, was named as CsINV10 (GenBank accession number: KT359348). BlastX and phylogenetic analysis indicated that the protein encoded by CsINV10 shared the highest identity (80%) with LcNI in Litchi,and had a closest genetic relationship with MeNINV8 in Manihot esculenta Crantz. Also, CsINV10 as a neutral/alkaline invertase gene could be classified into G100 family. The protein had no N-terminal signal peptide and transmembrane domain, and was predicated to be a hydrophilic protein localized in chloroplast. The expression analysis of CsINV10 under different abiotic stress conditions showed that cold, PEG and salt stresses could gradually promote the expression of CsINV10 when treated for one day, however, it had a transient increase when treated by ABA after three hours. Consequently, we speculated that CsINV10 might be involved in the tea plant response to abiotic stresses. Moreover, we found that CsINV10 had a tissue-specific expression patterns in leaf and flower. This study will provide a theoretical basis for the functional analysis of invertase genes of tea plant in response to various stresses.

Key words: Tea plant (Camellia sinensis), Neutral/alkaline invertase gene, Expression analysis

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