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Full-length cDNA Cloning and Expression Analysis of RPT4 in Anther of Kenaf (Hibiscus cannabinus L.)

LIGang1;2;GUANHe-xin1;NIUYing3;ZHOUQiong1;ZhouRui-yang1   

  1. 1、College of Agriculture, Guangxi University, Nanning 530004;
    2、Guangxi Botanical Garden of Medicinal Plants, Nanning 530023;
    3、Guangxi Citrus Institute, Guilin 541001, Guangxi
  • Received:2010-08-20 Online:2011-04-02 Published:2010-11-05

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Abstract: 【Objective】 This study aims to clone the full-length cDNA of the corresponding gene of RPT4 that with differential expression between cytoplasmic male sterile (CMS) lines and their maintainers in kenaf (Hibiscus cannabinus L.) and to analyze its expression in anthers. 【Method】 With homology cloning approaches coupling with RACE techniques, the full-length cDNA of RPT4 was cloned from total RNA of the anthers of the maintainer line L23B in kenaf. Then the expressions of mRNA in the anthers before abortive stage, at abortive stage, and late abortive stage were analyzed by semi-quantitative RT-PCR. 【Result】 The longest 1197 bp ORF of RPT4 was detected from the cloned sequence with full-length of cDNA of 1 596 bp. The encoded peptide of 398 amino acids shares 91.93% homology with the one in Ricinus communis. It also contains conserved domain consisting of three motifs- Walker A, Walker B and arginine finger, which suggests that the domain has various important functions. RTP4 was confirmed to express in anthers of both CMS line L23A and its maintainer. The expressing level of RPT4 in the CMS line L23A was identical to that in the maintainer L23B at the tetrad stage. However, the expressing level of RPT4 in L23A was lower at the uninucleate (abortive) stage, and higher at the binucleate (late abortive) stage than that of L23B. 【Conclusion】 RPT4 is involved in the development of microspores in kenaf and may be related to CMS with complex molecular function.

Key words: kenaf (Hibiscus cannabinus L.) , cytoplasmic male sterility (CMS) , RPT4 , isobaric tags relative and absolute quantitation (iTRAQ)

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