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RTS, a rice anther-specific gene is required for male fertility and its promoter sequence directs tissue-specific gene expression in different plant species

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Abstract

A tapetum-specific gene, RTS, has been isolated by differential screening of a cDNA library from rice panicles. RTS is a unique gene in the rice genome. RNA blot analysis and in situ hybridization indicates that this gene is predominantly expressed in the anther’s tapetum during meiosis and disappears before anthesis. RTS has no introns and encodes a putative polypeptide of 94 amino acids with a hydrophobic N-terminal region. The nucleotide and deduced amino acid sequence of the gene do not show significant homology to any known sequences. However, a sequence in the promoter region, GAATTTGTTA, differs only by one or two nucleotides from one of the conserved motifs in the promoter region of two pollen-specific genes of tomato. Several other sequence motifs found in other anther-specific promoters were also identified in the promoter of the RTS gene. Transgenic and antisense RNA approaches revealed that RTS gene is required for male fertility in rice. The promoter region of RTS, when fused to the Bacillus amyloliquefaciens ribonuclease gene, barnase, or the antisense of the RTS gene, is able to drive tissue-specific expression of both genes in rice, creeping bentgrass (Agrostis stolonifera L.) and Arabidopsis, conferring male sterility to the transgenic plants. Light and near-infrared confocal microscopy of cross-sections through developing flowers of male-sterile transgenics shows that tissue-specific expression of barnase or the antisense RTS genes interrupts tapetal development, resulting in deformed non-viable pollen. These results demonstrate a critical role of the RTS gene in pollen development in rice and the versatile application of the RTS gene promoter in directing anther-specific gene expression in both monocotyledonous and dicotyledonous plants, pointing to a potential for exploiting this gene and its promoter for engineering male sterility for hybrid production of various plant species.

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Acknowledgements

This work was supported by grants from the Rockefeller foundation to Purdue University (T.K.H.) and in part by the USDA-SBIR Grants #2003-33610-13107, #2005-33610-15620 to HybriGene Inc. (H.L.) and by the USDA-BRAG grant #2005-39454-16511 to Clemson University (H.L.). Technical Contribution No. 5214 of the Clemson University Experiment Station.

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  1. Department of Genetics, Biochemistry and Life Science Studies, Clemson University, 100 Jordan Hall, Clemson, SC, 29634, USA

    Hong Luo & Qian Hu

  2. HybriGene, Inc., 530 Liberty Lane, West Kingston, RI, 02892, USA

    Hong Luo, Qian Hu, Kimberly Nelson-Vasilchik & Thomas K. Hodges

  3. National Institute of Crop Science, Suwon, 441-857, Korea

    Jang-Yong Lee

  4. Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN, 47907, USA

    Jang-Yong Lee & Thomas K. Hodges

  5. Department of Cell and Molecular Biology, University of Rhode Island, Kingston, RI, 02881, USA

    Timothy K. Eitas, Colin Lickwar, Albert P. Kausch & Joel M. Chandlee

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  1. Hong Luo
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Luo, H., Lee, JY., Hu, Q. et al. RTS, a rice anther-specific gene is required for male fertility and its promoter sequence directs tissue-specific gene expression in different plant species. Plant Mol Biol 62, 397–408 (2006). https://doi.org/10.1007/s11103-006-9031-0

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