Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
권호 표지

Enhancement of Virus-induced Gene Silencing in Tomato by Low Temperature and Low Humidity

  • Fu, Da-Qi (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Zhu, Ben-Zhong (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Zhu, Hong-Liang (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Zhang, Hong-Xing (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Xie, Yuan-Hong (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Jiang, Wei-Bo (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Zhao, Xiao-Dan (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University) ;
  • Luo, Yun-Bo (Laboratory of Fruit Biology, College of Food Science and Nutritional Engineering, China Agricultural University)
  • Received : 2005.09.06
  • Accepted : 2005.10.20
  • Published : 2006.02.28
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Abstract

Virus-induced gene silencing (VIGS) is an attractive reverse-genetics tool for studying gene function in plants. We showed that silencing of a phytoene desaturase (PDS) gene is maintained throughout TRV-PDS-inoculated tomato plants as well as in their flowers and fruit and is enhanced by low temperature ($15^{\circ}C$) and low humidity (30%). RT-PCR analysis of the PDS gene revealed a dramatic reduction in the level of PDS mRNA in leaves, flowers and fruits. Silencing of PDS results in the accumulation of phytoene, the desaturase substrate. In addition, the content of chlorophyll a, chlorophyll b and total chlorophyll in the leaves of PDS-silenced plants was reduced by more than 90%. We also silenced the LeEIN2 gene by infecting seedlings, and this suppressed fruit ripenning. We conclude that this VIGS approach should facilitate large-scale functional analysis of genes involved in the development and ripening of tomato.

Keywords

Acknowledgement

Supported by : National Nature Science Foundation of China

References

  1. Alkharouf, N. W. and Matthews, B. F. (2004) SGMD: the soybean genomics and microarray database. Nucleic Acids Res. 1, D398-D400
  2. Angell, S. M. and Baulcombe, D. C. (1999) Potato virus X amplicon- mediated silencing of nuclear genes. Plant J. 20, 357-362 https://doi.org/10.1046/j.1365-313X.1999.t01-1-00597.x
  3. Baulcombe, D. C. (1999a) Fast forward genetics based on virus induced gene silencing. Curr. Opin. Plant Biol. 2, 109−113 https://doi.org/10.1016/S1369-5266(99)80022-3
  4. Baulcombe, D. C. (2004b) RNA silencing in plants. Nature 431, 356-363 https://doi.org/10.1038/nature02874
  5. Benedito, V. A., Visser, P. B., Angenent, G. C., and Krens, F. A. (2004) The potential of virus-induced gene silencing for speeding up functional characterization of plant genes. Genet. Mol. Res. 3, 323−341
  6. Bramley, P. M. (2002) Regulation of carotenoid formation during tomato fruit ripening and development. J. Exp. Bot. 53, 2107−2113 https://doi.org/10.1093/jxb/erf059
  7. Brigneti, G., Martin-Hernandez, A. M., Jin, H., Chen, J., Baulcombe, D. C., et al. (2004) Virus-induced gene silencing in Solanum species. Plant J. 39, 264-272 https://doi.org/10.1111/j.1365-313X.2004.02122.x
  8. Burch-Smith, T. M., Anderson, J. C., Martin, G. B., and Dinesh- Kumar, S. P. (2004) Applications and advantages of virusinduced gene silencing for gene function studies in plants. Plant J. 39, 734-746 https://doi.org/10.1111/j.1365-313X.2004.02158.x
  9. Chen, J. C., Jiang, C. Z., Gookin, T. E., Hunter, D. A., Clark, D. G., et al. (2004) Chalcone synthase as a reporter in virusinduced gene silencing studies of flower senescence. Plant Mol. Biol. 55, 521−530 https://doi.org/10.1007/s11103-004-0590-7
  10. Chung, E., Seong, E., Kim, Y. C., Chung, E. J., Oh, S. K., et al. (2004) A method of high frequency virus-induced gene silencing in chili pepper (Capsicum annuum L. cv. Bukang). Mol. Cells 17, 377−380
  11. Constantin, G. D., Krath, B. N., Macfarlane, S. A., Nicolaisen, M., Elisabeth, J. I., et al. (2004) Virus-induced gene silencing as a tool for functional genomics in a legume species. Plant J. 40, 622−631 https://doi.org/10.1111/j.1365-313X.2004.02233.x
  12. Davies, J. N. and Hobson, G. E. (1981) The constituents of tomato fruit-the influence of environment, nutrition, and genotype. Crit. Rev. Food Sci. Nutr. 15, 205-580 https://doi.org/10.1080/10408398109527317
  13. Dinesh-Kumar, S. P., Anandalakshmi, R., Marathe, R., Schiff, M., and Liu, Y. (2003) Virus-induced gene silencing. Methods Mol. Biol. 236, 287-294
  14. Faivre-Rampant, O., Gilroy, E. M., Hrubikova, K., Hein, I., Millam, S., et al. (2004) Potato virus X-induced gene silencing in leaves and tubers of potato. Plant Physiol. 134, 1308-1316 https://doi.org/10.1104/pp.103.037507
  15. Fei, Z., Tang, X., Alba, R. M., White, J. A., Ronning, C. M., et al. (2004) Comprehensive EST analysis of tomato and comparative genomics of fruit ripening. Plant J. 40, 47-59 https://doi.org/10.1111/j.1365-313X.2004.02188.x
  16. Fofana, I. B., Sangare, A., Collier, R., Taylor, C., and Fauquet, C. M. (2004) A geminivirus-induced gene silencing system for gene function validation in cassava. Plant Mol. Biol. 56, 613-624 https://doi.org/10.1007/s11103-004-0161-y
  17. Fraser, P. D., Truesdale, M. R., Bird, C. R., Schuch, W., and Bramley, P. M. (1994) Carotenoid biosynthesis during tomato fruit development. Plant Physiol. 105, 405–413
  18. Fraser, P. D., Pinto, M. E., Holloway, D. E., and Bramley, P. M. (2000) application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids. Plant J. 24, 551-558 https://doi.org/10.1046/j.1365-313x.2000.00896.x
  19. Fray, R. G. and Grierson, D. (1993) Molecular genetics of tomato fruit ripening. Trends Genet. 9, 438–443 https://doi.org/10.1016/0168-9525(93)90108-T
  20. Fu, D. Q., Zhu, B. Z., Zhao, X. D., and Luo, Y. B. (2005a) Advance of the Virus-induced gene silencing in plant. J. Chinese Biol. 1, 89-95
  21. Fu, D. Q., Zhu, B. Z., Zhu, H. L., Jiang, W. B., and Luo, Y. B. (2005b) Virus-induced gene silencing in tomato fruit. Plant J. 43, 299-308 https://doi.org/10.1111/j.1365-313X.2005.02441.x
  22. Giovannoni, J. J. (2001) Molecular biology of fruit maturation and ripening. Annu. Rev. Plant Physiol. Plant Mol. Biol. 52, 725−749 https://doi.org/10.1146/annurev.arplant.52.1.725
  23. Giovannoni, J. J. (2004) Genetic regulation of fruit development and ripening. Plant Cell 16 (Suppl), S170-S180 https://doi.org/10.1105/tpc.019158
  24. Gray, J., Picton, S., Shabbeer, J., Schuch, W., and Grierson, D. (1992) Molecular biology of fruit ripening and its manipulation with antisense genes. Plant Mol. Biol. 19, 69−87
  25. Gregory, T. W., Margaret, G. R., and Jone, G. S. (1988) A procedure for the small-scale isolation of plant suitable for RNA blot analysis. Anal. Biochem. 172, 279-283 https://doi.org/10.1016/0003-2697(88)90443-5
  26. Hajirezaei, M. R., Peisker, M., Tschiersch, H., Palatnik, J. F., Valle, E. M., et al. (2002) Small changes in the activity of chloroplastic $NADP^{+}$-dependent ferredoxin oxidoreductase lead to impaired plant growth and restrict photosynthetic activity of transgenic tobacco plants. Plant J. 29, 281-293 https://doi.org/10.1046/j.0960-7412.2001.01209.x
  27. Hedden, P. and Phillips, A. L. (2000) Manipulation of hormone biosynthetic genes in transgenic plants. Curr. Opin. Biotech. 11, 130−137 https://doi.org/10.1016/S0958-1669(00)00071-9
  28. Herrera-Estrella, L., Simpson, J., and Martinez-Trujillo, M. (2005) Transgenic plants: an historical perspective. Methods Mol. Biol. 286, 3-32
  29. Holzberg, S., Brosio, P., Gross, C., and Pogue, G. P. (2002) Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J. 30, 315-327 https://doi.org/10.1046/j.1365-313X.2002.01291.x
  30. Kumagai, M. H., Donson, J., della-Cioppa, G., Harvey, D., Hanley, K., et al. (1995) Cytoplasmic inhibition of carotenoid biosynthesis with virusderived RNA. Proc. Natl. Acad. Sci. USA 92, 1679–1683
  31. Lichtenthaler, H. K. (1987) Chlorophylls and carotenoides: pigments of photosynthetic biomembranes. Meth. Enzymol. 148, 350−382 https://doi.org/10.1016/0076-6879(87)48036-1
  32. Liu, Y., Schiff, M., and Dinesh-Kumar, S. P. (2002a) Virusinduced gene silencing in tomato. Plant J. 31, 777-786 https://doi.org/10.1046/j.1365-313X.2002.01394.x
  33. Liu, Y., Schiff, M., Marathe, R., and Dinesh-Kumar, S. P. (2002b) Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J. 30, 415-429 https://doi.org/10.1046/j.1365-313X.2002.01297.x
  34. Liu, Y., Nakayama, N., Schiff, M., Litt, A., Irish, V. F., et al. (2004) Virus induced gene silencing of a DEFICIENS ortholog in Nicotiana benthamiana. Plant Mol. Biol. 54, 701−711 https://doi.org/10.1023/B:PLAN.0000040899.53378.83
  35. Lu, R., Martin-Hernandez, A. M., Peart, J. R., Malcuit, I., and Baulcombe, D. C. (2003) Virus-induced gene silencing in plants. Methods 30, 296-303 https://doi.org/10.1016/S1046-2023(03)00037-9
  36. Mayer, M. P., Nievelstein, V., and Beyer, P. (1992) Purification and characterisation of a NADPH-dependent oxidoreductase from chloroplasts of NarcissusÐa redox mediator possibly involved in carotene desaturation. Plant Physiol. Biochem. 30, 389-398
  37. Moore, S., Vrebalov, J., Payton, P., and Giovannoni, J. (2002) Use of genomics tools to isolate key ripening genes and analyse fruit maturation in tomato. J. Exp. Bot. 53, 2023–2030 https://doi.org/10.1093/jxb/erf057
  38. Peart, J. R., Cook, G., Feys, B. J., Parker, J. E., and Baulcombe, D. C. (2002) An EDS1 orthologue is required for N-mediated resistance against tobacco mosaic virus. Plant J. 29, 569-579 https://doi.org/10.1046/j.1365-313X.2002.029005569.x
  39. Peele, C., Jordan, C. V., Muangsan, N., Turnage, M., Egelkrout, E., et al. (2001) Silencing of a meristematic gene using geminivirus- derived vectors. Plant J. 27, 357-366 https://doi.org/10.1046/j.1365-313x.2001.01080.x
  40. Ratcliff, F., Martin-Hernandez, A. M., and Baulcombe, D. C. (2001) Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J. 25, 237-245
  41. Richmond, T. and Somerville, S. (2000) Chasing the dream: plant EST microarrays. Curr. Opin. Plant Biol. 3, 108–106 https://doi.org/10.1016/S1369-5266(99)00049-7
  42. Robertson, D. (2004) VIGS vectors for gene silencing: many targets, many tools. Annu. Rev. Plant Biol. 55, 495-519 https://doi.org/10.1146/annurev.arplant.55.031903.141803
  43. Ronen, G., Cohen, M., Zamir, D., and Hirschberg, J. (1999) Regulation of carotenoid biosynthesis during tomato fruit development: expression of the gene for lycopene epsiloncyclase is down-regulated during ripening and is elevated in the mutant Delta. Plant J. 17, 341-351 https://doi.org/10.1046/j.1365-313X.1999.00381.x
  44. Ryu, C. M., Anand, A., Kang, L., and Mysore, K. S. (2004) Agrodrench: a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species. Plant J. 40, 322-331 https://doi.org/10.1111/j.1365-313X.2004.02211.x
  45. Stearns, J. C. and Glick, B. R. (2003) Transgenic plants with altered ethylenebiosynthesis or perception. Biotechnol. Adv. 21, 193-210 https://doi.org/10.1016/S0734-9750(03)00024-7
  46. Szittya, G., Silhavy, D., Molnar, A., Havelda, Z., Lovas, A., et al. (2003) Low temperature inhibits RNA silencing-mediated defence by the control of siRNA generation. EMBO J. 22, 633-640 https://doi.org/10.1093/emboj/cdg74
  47. Turnage, M. A., Muangsan, N., Peele, C. G., and Robertson, D. (2002) Geminivirus-based vectors for gene silencing in Arabidopsis. Plant J. 30, 107-114 https://doi.org/10.1046/j.1365-313X.2002.01261.x
  48. Valentine, T., Shaw, J., Blok, V. C., Phillips, M. S., Oparka, K. J., et al. (2004) Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector. Plant Physiol. 136, 3999-4009 https://doi.org/10.1104/pp.104.051466
  49. Zegzouti, H., Jones, B., Frasse, P., Marty, C., Maitre, B., et al. (1999) Ethylene-regulated gene expression in tomato fruit: characterization of novel ethylene-responsive and ripeningrelated genes isolated by differential display. Plant J. 18, 589-600 https://doi.org/10.1046/j.1365-313x.1999.00483.x