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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (09): 1332-1341.doi: 10.3724/SP.J.1006.2016.01332

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

Cloning and Characterization of Light Harvesting Chlorophyll a/b-Binding Protein Coding Gene (ScLhca3) in Sugarcane

ZHAI Yu-Shan, DENG Yu-Qing, DONG Meng, XU Qian, CHENG Guang-Yuan, PENG Lei, LIN Yan-Quan*,XU Jing-Sheng*   

  1. Fujian Agriculture and Forestry University / Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fuzhou 350002,China
  • Received:2016-01-11 Revised:2016-05-09 Online:2016-09-12 Published:2016-05-30
  • Contact: 林彦铨, E-mail: zwlinyq@163.com; 徐景升, E-mail: xujingsheng@126.com, Tel: 13959174787 E-mail:yushanzhai@126.com
  • Supported by:

    This study was supported by the National High-tech Research and Development Program of China (2013AA102604), the National Natural Science Foundation of China (31171605, 31371688), and the China Agriculture Research System (CARS-20-1-1).

Abstract:

The Lhca gene family in green plants encodes several light-harvesting chlorophyll a/b-binding proteins that harvest and transfer light energy to the reaction center of photosystem I (PSI) in photosynthesis. The cDNA sequence of Lhca3 gene was firstly obtained from sugarcane leaf full-length cDNA library through sequencing and validated by homology comparison. It was designated ScLhca3 and submitted to the GenBank (accession number: KU215669). ScLhca3 contains an 804 bp open reading frame (ORF) and encodes a deduced protein of 267 amino acids, with a molecular weight and pI of 28.91 kD and 8.96, respectively. Bioinformatics analysis showed that ScLhca3 is a hydrophilic non-secretory protein with three transmembrane domains and a chlorophyll a/b binding domain. Sequence multi-alignment and phylogenetic analysis demonstrated that the ScLhca3 protein sequence shared a high identity with Lhca3 from other plants, with the specificity of species. The protokaryotic expression vector pGEX-6P-1-ScLhca3 was constructed and expressed in E. coli cells under the induction of IPTG. The subcellular localization experiment showed that the ScLhca3 fused with GFP, a reporter protein, was located on chloroplast. Real-time quantitative PCR analysis showed the expression of ScLhca3 had a clear tissue specificity, and was upregulated by CdCl2, ABA, and H2O2, but downregulated by darkness, NaCl, and PEG.

Key words: Sugarcane, Photosystem I, Light harvesting chlorophyll a/b-binding protein, Real-time quantitative PCR

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