Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (11): 2271-.doi: 10.3864/j.issn.0578-1752.2014.11.020

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Expression Analysis of Two Novel PPR Genes in Gossypium hirsutum L.

 HE  Peng, HUANG  Peng, HUANG  Sheng, QIAN  Hui, YU  Jia-Ning   

  1. College of Life Sciences, Shaanxi Normal University, Xi’an 710062
  • Received:2013-12-16 Online:2014-06-06 Published:2014-03-31

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Abstract: 【Objective】 The aim of this study was to clone two novel PPR genes in cotton. Their sequence characteristics were investigated, and their expression in different tissues was analyzed and subcellular localization was identified. All these results could provide a support for further studying PPR genes function of cotton. 【Method】 Two PPR genes were cloned on the basis of ESTs from cotton using RT-PCR techniques. The bioinformatics method was used to analyze the putative amino acid sequence, semi- quantitative RT-PCR and real-time PCR were used to analyze the expression of target genes in tissues and different development periods of fiber. Transient expression system of cotton cotyledons was used to analyze subcellular localization. 【Result】 GhPPRH1 and GhPPRH2 belonged to PLS subfamily of PPR gene family. The full length ORF of both genes is 1 917 and 2 556 bp, encoding 638 and 851 amino acids, respectively. GhPPRH1 has 18 PPR motifs, including one E+ domain and DYW structure. GhPPRH2 has 17 PPR motifs with one E domain, E+ domain and DYW structure. Phylogenetic analysis of homologue PPR proteins of other plants and five cotton PPR proteins showed that GhPPRH1 has a close relationship with OsRF1b of rice. It implied that GhPPRH1 may involve in fertility restoration. Whereas GhPPRH2 might relate to RNA editing because of clustered together with ZmPPR5 of Zea mays. The expression pattern of GhPPRH1 and GhPPRH2 can be found in root, stem, leaf, petal and different development periods of fibers, and GhPPRH1 has relatively higher expression level in root, however high expression level of GhPPRH2 was observed in root, leaf and 15 DPA fibers. Subcellular localization showed that green fluorescence of GhPPRH1 was merged well with red fluorescence of mitochondria marker; green fluorescence of GhPPRH2 was merged very well with chloroplast autofluorescence, which indicated GhPPRH1 and GhPPRH2 might locate in mitochondria and chloroplast, respectively.【Conclusion】Two novel PPR genes were cloned and characterized from Gossypium hirsutum, which belonged to the typical PLS subclass family of PPR. The results of gene expression pattern and subcellular localization implied that two novel genes might participate in organelles RNA processing and modification.

Key words: cotton , PPR protein , gene cloning , expression analysis , subcellular localization

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