Plant Cell Reports

, Volume 26, Issue 10, pp 1809–1819 | Cite as

Genotyping of somatic hybrids between Festuca arundinacea Schreb. and Triticum aestivum L.

  • Yunfei Cai
  • Fengning Xiang
  • Daying Zhi
  • Heng Liu
  • Guangmin Xia
Genetic Transformation and Hybridization

Abstract

In order to genotype hybrid genomes of distant asymmetric somatic hybrids, we synthesized hybrid calli and plants via PEG-mediated protoplast fusion between recipient tall fescue (Festuca. arundinacea Schreb.) and donor wheat (Triticum aestivum L.). Seventeen and 25 putative hybrid clones were produced from the fusion combinations I and II, each with the donor wheat protoplast treated by UV light for 30 s and 1 min, respectively. Isozyme and RAPD profiles confirmed that ten hybrid clones were obtained from combination I and 19 from combination II. Out of the 29 hybrids, 12 regenerated hybrid plants with tall fescue phenotype. Composition and methylation-variation of the nuclear and cytoplasmic genomes of some hybrids, either with or without regenerative ability, were compared by genomic in situ hybridization, restriction fragment length polymorphism, and DNA methylation-sensitive amplification polymorphism. Our results indicated that these selected hybrids all contained introgressed nuclear and cytoplasmic DNA as well as obvious methylation variations compared to both parents. However, there were no differences either in nuclear/cytoplasmic DNA or methylation degree between the regenerable and non-regenerable hybrid clones. We conclude that both regeneration complementation and genetic material balance are crucial for hybrid plant regeneration.

Keywords

Tall fescue (Festuca. arundinaceaCommon wheat (Triticum aestivumUV Asymmetric somatic hybridization Genotyping of hybrid nucleus/cytoplasm Methylation variation 

Notes

Acknowledgments

This work was supported by a grant form the National Natural Science Foundation of China (No.30370857) and National 863 High Technology Research and Development Project No. 2006AA100102.

References

  1. Bonnett HT, Kofer W, Hakansson G, Glimelius K (1991) Mitochondrial involvement in petal and stamen development studied by sexual and somatic hybridisation of Nicotiana species. Plant Sci 80:119–130CrossRefGoogle Scholar
  2. Cheng YJ, Guo WW, Deng XX (2003) Molecular characterization of cytoplasmic and nuclear genomes in phenotypically abnormal Valencia orange (Citrus sinensis) + Meiwa kumquat (Fortunella crassifolia) intergeneric somatic hybrids. Plant Cell Rep 21:445–451PubMedGoogle Scholar
  3. Cao MS, Chen WP, Liu DJ (1994) Cytogenetic studies of intergeneric hybrids F1 and amphiploid between Lolium multiflorum Lam. and Festuca arundinacea var. glaucescens Boiss. Sci Agr Sin 27:69–76Google Scholar
  4. Derks FHM, Hakkert JC, Derbeek WHJ (1992) Genome composition of asymmetric hybrids in relation to the phylogenetic distance between the parents nucleus-chloroplast interaction. Theor Appl Genet 84:930–940CrossRefGoogle Scholar
  5. Donaldson PA, Bevis EE, Pandeya RS (1994) Random chloroplast segregation and frequent mtDNA rearrangements in fertile somatic hybrids between Nicotiana tabacum L. and N. glutinosa L. Theor Appl Genet 87:900–908CrossRefGoogle Scholar
  6. Doyle JJ, Doyle JI (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  7. Forsberg J, Dixelius C, Lagercrantz U (1998) UV dose-dependent DNA elimination in asymmetric somatic hybrids between Brassica napus and Arabidopsis thaliana. Plant Sci 131:65–76CrossRefGoogle Scholar
  8. Fu CH, Guo WW, Liu JH, Deng XX (2003) Regeneration of Citrus sinensis (+) Clausena lansium intergeneric triploid and tetraploid somatic hybrids and their identification by molecular markers. In Vitro Cell Dev Biol Plant 39:360–364CrossRefGoogle Scholar
  9. Glimelius K (1988) Potentials of protoplast fusion in plant breeding programmers. Plant Cell Tissue Org Cult 12:163–172CrossRefGoogle Scholar
  10. Glimelius K, Fahleshon J, Landgren M, Sjodin C, Sundberg E (1991) Gene transfer via somatic hybridization in plants. Trends Biotechnol 9:24–30CrossRefGoogle Scholar
  11. Gray MW, Burger G, Lang BF (1999) Mitochondrial evolution. Science 283:1476–1481PubMedCrossRefGoogle Scholar
  12. Guo WW, Cheng YJ, Deng XX (2002) Regeneration and molecular characterization of intergeneric somatic hybrids between Citrus reticulata and Poncirus trifoliata. Plant Cell Rep 20:829–834CrossRefGoogle Scholar
  13. Guo WW, Prasad D, Cheng YJ, Serrano P, Deng XX, Grosser JW (2004) Targeted cybridization in citrus: transfer of Satsuma cytoplasm to seedy cultivars for potential seedlessness. Plant Cell Rep 22:752–758PubMedCrossRefGoogle Scholar
  14. Ishii T, Mori N, Ogihara Y (2001) Evaluation of allelic diversity at chloroplast microsatellite loci among common wheat and its ancestral species. Theor Appl Genet 103:896–904CrossRefGoogle Scholar
  15. Jacobsen E, De Jong JH, Kamstra SA, Van Der Berg PMMM, Ramanna MS (1995) Genomic in situ hybridisation (GISH) and RFLP analysis for the identification of alien chromosomes in the backcross progeny of potato (1) tomato fusion hybrids. Heredity 74:250–257Google Scholar
  16. Kumar A, Cocking EC (1987) Protoplast fusion: a novel approach to organelle genetics in higher plants. Am J Bot 74:1289–1303CrossRefGoogle Scholar
  17. Kisaka H, Lee H, Kisaka M, Kanno A, Kang K, Kameya T (1994) Production and analysis of asymmetric hybrid plants between monocotyledon (Oryza sativa L.) and dicotyledon (Daucus carota L.). Theor Appl Genet 89:365–371CrossRefGoogle Scholar
  18. Li ZY, Xia GM, Chen HM (1992) Somatic embryogenesis and plant regeneration from protoplasts isolated from embryogenic cell suspension of wheat (Triticum aestivum L.). Plant Cell Tissue Org Cult 28:72–85Google Scholar
  19. Li LL, Xia GM, Chen HM (2001) Asymmetric hybridization between wheat and millet. Acta Physiol Sin 27:445–460Google Scholar
  20. Liu LM, Zhang JM, Bates S, Li JJ, Peehl DM, Rhim JS, Pfeifer GP (2005) A methylation profile of in vitro immortalized human cell lines. Int J Oncol 26:275–285PubMedGoogle Scholar
  21. Lotfy S, Luro F, Carreel F, Froelicher Y, Rist D, Ollitrault P (2003) Application of cleaved amplified polymorphic sequence method for analysis of cytoplasmic genome among Aurantioideae intergeneric somatic hybrids. J Am Soc Hortic Sci 128:225–230Google Scholar
  22. Meyer P, Niedenhof I, ten Lohuis M (1994) Evidence for cytosine methylation of non-symmetrical sequences in transgenic Petunia hybrida. EMBO J 13:2084–2088PubMedGoogle Scholar
  23. Moreira CD, Chase CD, Gmitter FG Jr, Grosser JW (2000) Inheritance of organelle genomes in citrus somatic cybrids. Mol Breed 6:401–405CrossRefGoogle Scholar
  24. Oberwalder B, Schilde-Rentschler L, Ruoß B, Wittemann S, Ninnemann H (1998) Asymmetric protoplast fusion between wild species and breeding lines of potato-effect of recipients genome stability. Theor Appl Genet 97:1347–1354CrossRefGoogle Scholar
  25. Polgar ZS, Preiszner J, Dudits D, Feher A (1993) Vigorous growth of fusion products allows highly efficient selection of interspecific potato somatic hybrids: molecular proofs. Plant Cell Rep 12:399–402CrossRefGoogle Scholar
  26. Rasmussen JO, Nepper JP, Kirk HG, Tolstrup K, Rasmussen OS (1998) Combination of resistance to potato late blight in foliage and tubers by interspecific dihaploid protoplast fusion. Euphytica 102:363–370CrossRefGoogle Scholar
  27. Rossi V, Motto M, Pelligrini L (1997) Analysis of the methylation pattern of the maize Opaque-2 (O2) promoter and in vitro binding studies indicate that the O2 B-Zip protein and other endosperm factors can bind to methylated target sequences. J Biol Chem 272:3758–13765CrossRefGoogle Scholar
  28. Shaked H, Kashkush K, Ozkan H, Feldman M, Levy AA (2001) Sequence elimination and cytosine methylation are rapid and reproducible responses of the genome to wide hybridization and allopolyploidy in wheat. Plant Cell 13:1749–1759PubMedCrossRefGoogle Scholar
  29. Sheahan MB, Rose RJ, McCurdy DW (2004) Organelle inheritance in plant cell division: the actin cytoskeleton is required for unbiased inheritance of chloroplasts, mitochondria and endoplasmic reticulum in dividing protoplasts. Plant J 37:379–390PubMedCrossRefGoogle Scholar
  30. Skarzhinskaya M, Landgren M, Glimelius K (1996) Production of intertribal somatic hybrids between Brassica napus L. and Lesquerella fendleri (Gray) Wats. Theor Appl Genet 93:1242–1250CrossRefGoogle Scholar
  31. Spangenberg G, Valles MP, Wang ZY, Montavon P, Nagel J, Potrykus I (1994) Asymmetric somatic hybridization between tall fescue (Festuca arundinacea Schreb.) and irradiated Italian ryegrass (Lolium multiflorum Lam.) protoplasts. Theor Appl Genet 88:509–519CrossRefGoogle Scholar
  32. Trick HN, Bates GW (1996) Bromodeoxyuridine combined with UV light and gamma irradiation promotes the production of asymmetric somatic hybrid calli. Plant Cell Rep 15:986–990CrossRefGoogle Scholar
  33. Ulian EC, Magill JM, Magll CW, Smith RH (1996) DNA methylation and expression of NPT II in transgenic petunias and progeny. Theor Appl Genet 92:976–981CrossRefGoogle Scholar
  34. Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Fijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414PubMedCrossRefGoogle Scholar
  35. Wolters AMA, Schoenmakers HCH, Van der Muulen-Muisers JJ (1991) Limited DNA elimination from the irradiated potato parent in fusion products of albino Lycopersicon esculentum and Solanum tuberosum. Theor Appl Genet 83:225–232CrossRefGoogle Scholar
  36. Xia GM, Chen HM (1996) Plant regeneration from intergeneric somatic hybridization between Triticum aestivum L. and Leymus chinensis (Trin.) Tzvel Plant Sci 120:197–203CrossRefGoogle Scholar
  37. Xia GM, Li ZY, Wang SL, Xiang FN, Liu JY, Chen PD, Liu DJ (1998) Asymmetric somatic hybridization between haploid wheat and UV irradiated Haynaldia villosa. Plant Sci 37:217–223Google Scholar
  38. Xia GM, Xiang FN, Zhou AF, Wang H, Chen HM (2003) Asymmetric somatic hybridization between wheat (Triticum aestivum L.) and Agropyron elongatum (Host) Nevishi. Thero Appl Genet 107:299–305CrossRefGoogle Scholar
  39. Xiang FN, Xia GM, Chen HM (2002) Asymmetric somatic hybridization between wheat (Triticum aestivum) and Avena sativa L (in Chinese) Sci China (Series C) 32:299–305 Google Scholar
  40. Xiang FN, Xia GM, Zhou AF, Chen HM (1999) Asymmetric somatic hybridization between wheat (Triticum aestivum L.) and Bromus inermis. Acta Bot Sin 41:458–462Google Scholar
  41. Xiang FN, Xia GM, Chen HM (2003) Effect of UV dosage on somatic hybridization between common wheat (Triticum aestivum L.) and Avena sativa L. Plant Sci 164:697–707CrossRefGoogle Scholar
  42. Xiang FN, Xia GM, Chen HM (2004) Regeneration of somatic hybrids in relation to the nuclear and cytoplasmic genomes of wheat and Setaria italica. Genome 47:680–688PubMedCrossRefGoogle Scholar
  43. Xiong LZ, Xu CG, Saghai MA, Zhang QF (1999) Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique. Mol Gen Genet 261:439–446PubMedCrossRefGoogle Scholar
  44. Xu CH, Xia GM, Zhi DY, Xiang FN, Chen HM (2003) Integration of maize nuclear and mitochondrial DNA into the wheat genome through somatic hybridization. Plant Sci 165:1001–1008CrossRefGoogle Scholar
  45. Xu ML, Li XQ, Korban SS (2004) DNA-methylation alterations and exchanges during in vitro cellular differentiation in rose (Rosa hybrida L.). Theor Appl Genet 109:899–910PubMedCrossRefGoogle Scholar
  46. Yamagishi H, Landgren M, Forsberg J, Glimelius K (2002) Production of asymmetric hybrids between Arabidopsis thaliana and Brassica napus utilizing an efficient protoplast culture system. Theor Appl Genet 104:959–964PubMedCrossRefGoogle Scholar
  47. Yu ZY, Tan XY, Xue ZY, Xia GM (2006) Study on salt-tolerance and protein composition of somatic hybrids between common wheat and Agropyron elongatum. Chin High Technol Lett 16:511–516Google Scholar
  48. Zhi DY, Han XG, Zhao JS, Xue ZY, Xia GM (2004) Tissue culture and plant regeneration of eight genotypes of tall fescue. J ShanDong Uni 39:109–114Google Scholar
  49. Zhou AF, Xiang FN, Xia GM, Chen HM, Huang Y, Zhai XM (1996) Asymmetric somatic hybridization between monoploid wheat and Haynaldia villosa Schur. J ShanDong Uni 34:224–229Google Scholar
  50. Zhou AF, Xia GM, Zhang XY, Chen HM, Hu H (2001) Analysis of chromosomal and organellar DNA of somatic hybrids between Triticum aestivum and Haynaldia villosa Schur. Mol Genet Genom 26:5387–5393Google Scholar
  51. Zhou AF, Xia GM, Chen XL, Chen HM (2002a) Production of somatic hybrid plants between two types of wheat protoplasts and the protoplasts of Haynaldia villosa. Acta Bot Sin 44(8):1004–1008Google Scholar
  52. Zhou AF, Xia GM, Chen XL, Chen HM (2002b) UV-fusion between protoplasts of common wheat and Haynaldia villosa. J Plant Physiol Mol Biol 28(4):305–310Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Yunfei Cai
    • 1
  • Fengning Xiang
    • 1
  • Daying Zhi
    • 1
  • Heng Liu
    • 1
  • Guangmin Xia
    • 1
  1. 1.School of Life SciencesShandong UniversityJinanPeople’s Republic of China

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