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RNAi-mediated male sterility of tobacco by silencing TA29

Molecular Biotechnology volume 36, pages 159–165 (2007)Cite this article

Abstract

The superior performance of F1 hybrids has a significant impact on agricultural productivity. For commercial application, the availability of an efficient system for obtaining male-sterile lines of crops is an essential prerequisite. Here we have investigated the use of RNA interference (RNAi) technology to silence a male-specific gene in the model host tobacco. TA29 is expressed exclusively in anthers at the time of microspore development. About 10 out of 13 tobacco lines transformed with a hairpin RNAi construct containing TA29 sequences were male sterile. Transgenic plants were phenotypically indistinguishable from non-transgenic plants. At the anthesis stage, pollen grains from transgenic, male-sterile plants were aborted and lysed in comparison to the round and fully developed pollen in non-transgenic plants. Microscopic analysis of anthers showed selective degradation of tapetum in transgenic plants with no microspore development. One week after self-pollination, the ovules of non-transgenic plants were double the size of those in transgenic plants, due to successful self-fertilization. Male sterile transgenic plants set seed normally, when cross-pollinated with pollen from non-transgenic plants, confirming no adverse effect on the female parts of the flower. These results show that silencing of male-specific genes by RNAi is potentially a useful tool for generating male-sterile lines for producing hybrid seed.

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References

  1. Peacock, J. (1990). Ways to pollen sterility. Nature, 347, 714–715.

    Article  Google Scholar 

  2. Dewey, R. E., Timothy, D. H., & Levings, D. C. S. (1997). A mitochondrial protein associated with cytoplasmic male sterility in the T cytoplasm of maize. Proceedings of the National Academy of Sciences of the United States of America, 84, 5374–5378.

    Article  Google Scholar 

  3. Mariani, C., De-Beuckeleer, M., Truettner, J., Leemans, J., & Goldberg, R. B. (1990). Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature, 347, 737–741.

    Article  CAS  Google Scholar 

  4. Shivanna, K. R., Cresti, M., & Ciampolini, F. (1997). Pollen development and pollen-pistil interaction. In K. R. Shivanna & V. K. Sawhney (Eds.), Pollen biotechnology for crop production and improvement (pp. 15–39). Cambridge: Cambridge University Press.

    Google Scholar 

  5. van der Meer, I. M., Stam, M. E., van Tunen, A. J., Mol, J. N., & Stuitje, A. R. (1992). Antisense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell, 4, 253–262.

    PubMed  Article  Google Scholar 

  6. Mansoor, S., Amin, I., Hussain, M., Zafar, Y., & Briddon, R. W. (2006). Engineering of novel traits in plants through RNA interference. Trends in Plant Science, 11, 559–565.

    PubMed  Article  CAS  Google Scholar 

  7. Smith, N. A., Singh, S. P., Wang, M.-B., Stoutjesdijk, P., Green, A., & Waterhouse, P. M. (2000). Total silencing by intron-spliced hairpin RNAs. Nature, 407, 319–320.

    PubMed  Article  CAS  Google Scholar 

  8. Doyle, J. J., & Doyle, J. L. (1989). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19, 11–15.

    Google Scholar 

  9. Horsch, R., Fry, J., Hoffman, N., Neidermeyer, J., Rogers, S. G., & Fraley, R. T. (1988). Leaf disc transformation. In S. Gelvin & R. Schilperoort (Eds.), Plant molecular biology manual (pp. 23–50). Dordrecht: Kluwer.

    Google Scholar 

  10. Koltunow, A. M., Truettner, J., Cox, K. H., Wallroth, M., & Goldberg, R. B. (1990). Different temporal and spatial gene expression patterns occur during anther development. Plant Cell, 2, 1201–1224.

    PubMed  Article  CAS  Google Scholar 

  11. Chang, M. T., & Neuffer, M. G. (1989). A simple method for staining nuclei of mature and germinated maize pollen. Stain Technology, 64, 181–184.

    PubMed  CAS  Google Scholar 

  12. Karnovsky, M. J. (1965). A formaldehyde–glutaraldehyde fixative of light osmolality for use in electron microscopy. The Journal of Cell Biology, 27, 137A–138A.

    Google Scholar 

  13. O’Brien, T. P., Feder, N., & McCully, M. E. (1964). Polychromatic staining of plant cell walls by toluidine blue. Protoplasma, 59, 368–373.

    Article  CAS  Google Scholar 

  14. Seurinck, J, Truettner, J., & Goldberg, R. B. (1990). The nucleotide sequence of an anther specific gene. Nucleic Acids Research, 18, 3403.

    PubMed  Article  CAS  Google Scholar 

  15. Goldberg, R., Beals, T., & Sanders, P. (1993). Anther development: Basic principles and practical applications. Plant Cell, 5, 1217–1229.

    PubMed  Article  CAS  Google Scholar 

  16. Kriete, G., Niehaus, K., Perlick, A. M., Puhler, A., & Broer, I. (1996). Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compounds N-acetyl-l-phosphinothricin. The Plant Journal, 9, 809–818.

    PubMed  Article  CAS  Google Scholar 

  17. Theis, R., & Robbelen, G. (1990). Anther and microspore development in different male sterile lines of oilseed rape (Brassica napus L.). Angewandte Botanik, 64, 419–434.

    Google Scholar 

  18. Gourret, J. P., Delourme, R., & Renard, M. (1992). Expression of ogu cytoplasmic male sterility in cybrids of Brassica napus. Theoretical and Applied Genetics, 83, 549–556.

    Article  Google Scholar 

  19. Bartkowiak-Broda, I., Rousselle, P., & Renard, M. (1979). Investigations of two kinds of cytoplasmic male sterility in rapeseed (Brassica napus L.). Genetica Polonica, 20, 487–497.

    Google Scholar 

  20. Pellan-Delourme, R., & Renard, M. (1987). Identification of maintainer genes in Brassica napus L. for the male-sterility-inducing cytoplasm of Diplotaxis muralis L. Plant Breeding, 99, 89–97.

    Article  Google Scholar 

  21. Goldberg, R. B. (1988). Plants: Novel developmental processes. Science, 240, 1460–1467.

    PubMed  Article  CAS  Google Scholar 

  22. Esau, K. (1977). Anatomy of seed plants, 2nd edn. New York: Wiley.

    Google Scholar 

  23. Knox, R. B. (1979). Pollen and allergy. In: Studies in biology, no. 107, London: Edward Arnold.

  24. Cigan, A. M., Unger-Wallace, E., & Haug-Collet, K. (2005) Transcriptional gene silencing as a tool for uncovering gene function in maize. The Plant Journal, 43, 929–940.

    PubMed  Article  CAS  Google Scholar 

  25. Greenland, A., Bell, P., Hart, C., Jepson, I., & Nevshemal, T. (1998). Reversible male sterility: A novel system for production of hybrid corn. Symposia of the Society for Experimental Biology, 51, 141–147.

    PubMed  CAS  Google Scholar 

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Acknowledgements

This project was funded by the Pakistan Agricultural Research Council, Islamabad under the “Agriculture Linkage Program”. We are thankful to Dr. Rubina Tabassum for use of the light microscope, Mr. Javed Iqbal for assistance with microscopy and Dr. Richard Jorgenson for providing pFGC5941. RWB acknowledges the support of the Higher Education Commission (Pakistan) under the “Foreign Faculty Hiring Scheme”.

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Affiliations

  1. National Institute for Biotechnology and Genetic Engineering, Jhang Road, Faisalabad, Pakistan

    Muhammad Shah Nawaz-ul-Rehman, Shahid Mansoor, Yusuf Zafar & Rob W. Briddon

  2. Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad, Pakistan

    Asif Ali Khan

Authors
  1. Muhammad Shah Nawaz-ul-Rehman
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  2. Shahid Mansoor
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  3. Asif Ali Khan
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  4. Yusuf Zafar
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  5. Rob W. Briddon
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Correspondence to Rob W. Briddon.

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Nawaz-ul-Rehman, M.S., Mansoor, S., Khan, A.A. et al. RNAi-mediated male sterility of tobacco by silencing TA29 . Mol Biotechnol 36, 159–165 (2007). https://doi.org/10.1007/s12033-007-0025-1

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  • DOI: https://doi.org/10.1007/s12033-007-0025-1

Keywords

  • Anther
  • Hybrid
  • Male-sterility
  • Silencing
  • Tapetum