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Partial male sterility in transgenic tobacco carrying an antisense gene for alternative oxidase under the control of a tapetum-specific promoter

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Abstract

The alternative oxidase of plant mitochondria is the terminal oxidase of the cyanide-insensitive respiratory pathway and is encoded by a nuclear gene. A 1 kb genomic fragment including exon 3 of the alternative oxidase was amplified by PCR from the genome of Arabidopsis thaliana. This fragment was connected to a tapetum-specific promoter in the antisense orientation and then introduced into tobacco. The pollen viability in three transgenic plants ranged from 2% to 60%. The reduced pollen viability cosegregated with the transgene in a selfed progeny. Immunolocalization of alternative oxidase protein in the immature flower bud section indicated that expression of alternative oxidase protein in tapetum of the transgenic plant was much lower than that of the non-transformant. The histological observation and protein gel-blot analysis showed that the development of pollen grains in the transgenic plant did not progress after the degradation of the tapetum, and the amount of alternative oxidase in pollen grains of the transgenic plant became lower than that of the non-transformant. These results suggested that the alternative oxidase activity in the tapetum has a significant effect on the pollen development.

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References

  1. Conley CA, Hanson MR: Tissue-specific protein expression in plant mitochondria. Plant Cell 6: 85–91 (1994).

    Article  PubMed  CAS  Google Scholar 

  2. Connett MB, Hanson MR: Differential mitochondrial electron transport through the cyanide-sensitive and cyanideinsensitive pathways in isonuclear lines of cytoplasmic male sterile, male fertile, and restored Petunia. Plant Physiol 93: 1634–1640 (1990).

    Article  PubMed  CAS  Google Scholar 

  3. Cruz-Hernandez A, Gomez-Lim MA: Alternative oxidase from mango (Mangifera indica L.) is differentially regulated during fruit ripening. Planta 197: 569–576 (1995).

    PubMed  CAS  Google Scholar 

  4. Day DA, Arron GP, Laties GG: Nature and control of respiratory pathways in plants: the interaction of cyanide-resistant respiration with the cyanide-sensitive pathway. In: Davies D (ed) The Biochemistry of Plants: A Comprehensive Treatise, vol. 2. Metabolism and Respiration, pp. 197–241. Academic Presss, New York (1980).

    Google Scholar 

  5. Day DA, Whelan J, Millar AH, Siedow JN, Wiskich JT: Regulation of the alternative oxidase in plants and fungi. Aust J Plant Physiol 22: 491–509 (1995).

    Google Scholar 

  6. Echlin P: The role of the tapetum during microsporogenesis of angiosperms. In: Heslop-Harrison J (ed) Pollen: Development and Physiology, pp. 41–61. Butterworths, London (1971).

    Google Scholar 

  7. Elthon TE, McIntosh L: Identification of the alternative terminal oxidase of higher plant mitochondria. Proc Natl Acad Sci USA 84: 8399–8403 (1987).

    Article  PubMed  CAS  Google Scholar 

  8. Elthon TE, Nickels RL, McIntosh L: Monoclonal antibodies to the alternative oxidase of higher plant mitochondria. Plant Physiol 89: 1311–1317 (1989).

    PubMed  CAS  Google Scholar 

  9. Goldberg RB, Beals TP, Sanders PM: Anther development: basic principles and practical applications. Plant Cell 5: 1217–1229 (1993).

    Article  PubMed  CAS  Google Scholar 

  10. Heslop-Harrison J: Incompatibility and the pollen-stigma interaction. Annu Rev Plant Physiol 26: 403–425 (1975).

    Article  CAS  Google Scholar 

  11. Hiei Y, Ohta S, Komari T, Kumashiro T: Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6: 271–282 (1994).

    Article  PubMed  CAS  Google Scholar 

  12. Hiser C, Kapranov P, McIntosh L: Genetic modification of respiratory capacity in potato. Plant Physiol 110: 277–286 (1996).

    Article  PubMed  CAS  Google Scholar 

  13. Horsch RB, Fry J, Hoffmann N, Neidermeyer J, Rogers SG, Fraley RT: Leaf disc transformation. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, A5/1- A5/9. Kluwer Academic Publisherss, Dordrecht, Netherlands (1988).

    Google Scholar 

  14. Johns C, Nickels R, McIntosh L, Mackenzie S: The expression of alternative oxidase and alternative respiratory capacity in cytoplasmic male sterile common bean. Sex Plant Reprod 6: 257–265 (1993).

    Article  Google Scholar 

  15. Kishitani S, Konno N: Inhibitors of cyanide-insensitive respiratory pathway induce male sterility in broccoli (Brassica oleracea). Japan J Breed 40: 217–222 (1990).

    Google Scholar 

  16. Kumar AM, Soll D: Arabidopsis alternative oxidase sustains Escherichia coli respiration. Proc Natl Acad Sci USA 89: 10842–10846 (1992).

    Article  PubMed  CAS  Google Scholar 

  17. Li Q, Ritzel RG, McLean LLT, McIntosh L, Ko T, Bertrand H, Nargang FE: Cloning and analysis of the alternative oxidase gene of Neurospora crassa. Genetics 142: 129–140 (1996).

    PubMed  CAS  Google Scholar 

  18. Mascarenhas JP: Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41: 317–338 (1990).

    Article  Google Scholar 

  19. Matsuda N, Tsuchiya T, Kishitani S, Tanaka Y, Toriyama K: Partial male sterility in transgenic tobacco carrying antisense and sense PAL cDNA under the control of a tapetum-specific promoter. Plant Cell Physiol 37: 215–222 (1996).

    CAS  Google Scholar 

  20. McCormick S: Male gametophyte development. Plant Cell 5: 1265–1275 (1993).

    Article  PubMed  Google Scholar 

  21. McIntosh L: Molecular biology of the alternative oxidase. Plant Physiol 105: 781–786 (1994).

    Article  PubMed  CAS  Google Scholar 

  22. Minagawa N, Koga S, Nakano M, Sakajo S, Yoshimoto A: Possible involvement of superoxide anion in the induction of cyanide-resistant respiration in Hansenula anomala. FEBS Lett 235: 76–80 (1992).

    Google Scholar 

  23. Moore AL, Siedow JN: The regulation and nature of the cyanide-resistant alternative oxidase of plant mitochondria. Biochim Biophys Acta 1059: 121–140 (1991).

    PubMed  CAS  Google Scholar 

  24. Moore AL, Umbach AL, Siedow JN: Structure-function relationships of the alternative oxidase of plant mitochondria: a model of the active site. J Bioenerg Biomembr 27: 367–377 (1995).

    Article  PubMed  CAS  Google Scholar 

  25. Murray MG, Thompson WF: Rapid isolation of high molecular weight plant DNA. Nucl Acids Res 8: 4321–4325 (1980).

    PubMed  CAS  Google Scholar 

  26. Nelson OE: The waxy locus in maize. II. The location of the controlling element alleles. Genetics 60: 507–524 (1968).

    PubMed  Google Scholar 

  27. Purvis AC, Shewfelt RL: Does the alternative pathway ameliorate chilling injury in sensitive plant tissues? Physiol Plant 88: 712–718 (1993).

    Article  CAS  Google Scholar 

  28. Rhoads DM, McIntosh L: Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott). Proc Natl Acad Sci USA 88: 2122–2126 (1991).

    Article  PubMed  CAS  Google Scholar 

  29. Sakajo S, Minagawa N, Komiyama T, Yoshimoto A: Molecular cloning of cDNA for antimycin A-inducible mRNA and its role in cyanide-resistant respiration in Hansenula anomala. Biochim Biophys Acta 1090: 102–108 (1991).

    PubMed  CAS  Google Scholar 

  30. Salehuzzaman SNIM, Jacobsen E, Visser RGF: Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato. Plant Mol Biol 23: 947–962 (1993).

    Article  PubMed  CAS  Google Scholar 

  31. Scott R, Hodge R, Paul W, Draper J: The molecular biology of anther differentiation. Plant Sci 80: 167–191 (1991).

    Article  CAS  Google Scholar 

  32. Shimada H, Tada Y, Kawasaki T, Fujimura T: Antisense regulation of the rice waxy gene expression using a PCR-amplified fragment of the rice genome reduces the amylose content in grain starch. Theor Appl Genet 86: 665–672 (1993).

    Article  CAS  Google Scholar 

  33. Siedow JN, Umbach AL, Moore AL: The active site of the cyanide-resistant oxidase from plant mitochondria contains a binuclear iron center. FEBS Lett 362: 10–14 (1995).

    Article  PubMed  CAS  Google Scholar 

  34. Tsuchiya T, Toriyama K, Ejiri S, Hinata K: Molecular characterization of rice genes specifically expressed in the anther tapetum. Plant Mol Biol 26: 1737–1746 (1994).

    Article  PubMed  CAS  Google Scholar 

  35. Tsuchiya T, Toriyama K, Yoshikawa M, Ejiri S, Hinata K: Tapetum-specific expression of the gene for an endo-δ-1,3-glucanase causes male sterility in transgenic tobacco. Plant Cell Physiol 36: 487–494 (1995).

    PubMed  CAS  Google Scholar 

  36. Umbach AL, Siedow JN: Covalent and noncovalent dimers of the cyanide-resistant alternative oxidase protein in higher plant mitochondria and their relationship to enzyme activity. Plant Physiol 103: 845–854 (1993).

    PubMed  CAS  Google Scholar 

  37. Umbach AL, Wiskich JT, Siedow JN: Regulation of alternative oxidase kinetics by pyruvate and intermolecular disulfide bond redox status in soybean seedling mitochondria. FEBS Lett 348: 181–184 (1994).

    Article  PubMed  CAS  Google Scholar 

  38. Van der Meer IM, Stam ME, van Tunen AJ, Mol JNM, Stuitje AR: Antisense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. The Plant Cell 4: 253–262 (1992).

    Article  PubMed  CAS  Google Scholar 

  39. Vanlerberghe GC, Day DA, Wiskich JT, Vanlerberghe AE, McIntosh L: Alternative oxidase activity in tobacco leaf mitochondria: dependence on tricarboxylic acid cycle-mediated redox regulation and pyruvate activation. Plant Physiol 109: 353–361 (1995).

    PubMed  CAS  Google Scholar 

  40. Vanlerberghe GC, McIntosh L: Mitochondrial electron transport regulation of nuclear gene expression: studies with the alternative oxidase gene of tobacco. Plant Physiol 105: 867–874 (1994).

    Article  PubMed  CAS  Google Scholar 

  41. Vanlerberghe GC, McIntosh L: Alternative oxidase: from gene to function. Annu Rev Plant Physiol Plant Mol Biol 48: 703–734 (1997).

    Article  PubMed  CAS  Google Scholar 

  42. Vanlerberghe GC, Vanlerberghe AE, McIntosh L: Molecular genetic alternation of plant respiration: silencing and overexpression of alternative oxidase in transgenic tobacco. Plant Physiol 106: 1503–1510 (1994).

    PubMed  CAS  Google Scholar 

  43. Vanlerberghe GC, Vanlerberghe AE, McIntosh L: Molecular genetic evidence of the ability of alternative oxidase to support respiratory carbon metabolism. Plant Physiol 113: 657–661 (1997).

    PubMed  CAS  Google Scholar 

  44. Watanabe M, Shiozawa H, Isogai A, Suzuki A, Takeuchi T, Hinata K: Existence of S-glycoprotein-like proteins in anthers of self-incompatible species of Brassica. Plant Cell Physiol 32: 1039–1047 (1991).

    CAS  Google Scholar 

  45. Whelan J, Millar AH, Day DA: The alternative oxidase is encoded in a multigene family in soybean. Planta 198: 197–201 (1996).

    Article  PubMed  CAS  Google Scholar 

  46. Whelan J, McIntosh L, Day DA: Sequencing of the soybean alternative oxidase cDNA clone. Plant Physiol 103: 1481 (1993).

    Article  PubMed  CAS  Google Scholar 

  47. Whelan J, Smith MK, Meijer M, Yu J-W, Badger MR, Price GD, Day DA: Cloning of an additional cDNA for the alternative oxidase in tobacco. Plant Physiol 107: 1469–1470 (1995).

    Article  PubMed  CAS  Google Scholar 

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Kitashiba, H., Kitazawa, E., Kishitani, S. et al. Partial male sterility in transgenic tobacco carrying an antisense gene for alternative oxidase under the control of a tapetum-specific promoter. Molecular Breeding 5, 209–218 (1999). https://doi.org/10.1023/A:1009606601521

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