Onion, Leek and Garlic Transformation by Co-Cultivation with Agrobacterium

  • C. C. Eady
Part of the Molecular Methods of Plant Analysis book series (MOLMETHPLANT, volume 23)


Allium species are difficult to transform. Because of the difficulties, the genus is amongst the last commercially important vegetable genus for which gene transformation protocols are being developed. The protocols outlined in this chapter are still in their infancy and in the case of leek and garlic, they are the only successful reports of Agrobacterium-mediated transformation for those particular species (at the time of going to press). For onions, only two protocols have been published. Of these, only the one described below (Sects. 4.2.1 and 4.2.2) has been routinely used to produce transgenic onions containing several different traits. Therefore, the reader should be aware that while every effort has been made to accurately report the technology that is currently being used, there is every likelihood that these protocols can be improved upon or that new protocols may arise that will supersede what is reported here. This chapter deals with Agrobacterium-mediated transformation and does not cover direct gene transfer methods, such as biolistics and cell fusion-mediated transformation or interspecific gene integrations. For information on these gene transfer techniques in Allium species, the reader is referred to Eady (1995, 2002a,b), Buitveld (1998) and Kik (2002). This chapter does not cover the history of Allium transformation, gene delivery, gene regulation, and cell culture research, or the potential applications and risks of the technology to alliums. These have been covered in previous reviews (Eady 1995, 2002a,b). This chapter outlines the current state of the technology and how it is being applied. Rather inevitably, in this day and age, it is possible that some current applications have been omitted as some research may be subject to nondisclosure due to its commercial sensitivity. There may also be other cases for which published results are not yet available.


Vortex Carbohydrate Assimilation Microbial Degradation Pyruvate 


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  1. Bansal RK, Broadhurst PG (1992) An evaluation of Allium germplasm for resistance to white rot caused by Sclerotium cerpivorum Berk. NZ J Crop Hortic Sci 20: 362–365CrossRefGoogle Scholar
  2. Barrell P (2001) Expression of synthetic magainin genes in potato. PhD Thesis, Lincoln University Christchurch, New ZealandGoogle Scholar
  3. Block E (1992) The organosulfur chemistry of the genus Allium - implications for the organic chemistry of sulfur. Agnew Chem Int Ed Engl 31: 1135–1178CrossRefGoogle Scholar
  4. Buiteveld J (1998) Regeneration and interspecific somatic hybridization in Allium for transfer of cytoplasmic male sterility to leek. Wageningen Agricultural University 127. Wageningen Agricultural University, WageningenGoogle Scholar
  5. Clark SA (1993) Molecular cloning of a cDNA encoding alliinase from onion (Allium cepa L.). PhD dissertation, University of Canterbury, Christchurch, New ZealandGoogle Scholar
  6. Darbyshire B, Steer BT (1990) Carbohydrate biochemistry. In: Rabinowitch HD, Brewster JL (ed) Onions and allied crops, vol II. Agronomy, biotic interactions, pathology and crop protection. CRC Press, Boca Raton, pp 1–17Google Scholar
  7. Eady CC (1995) Towards the transformation of onions (Allium cepa) (review). NZ J Crop Hortic Sci 23: 239–250CrossRefGoogle Scholar
  8. Eady CC (2001) How green are your onions? Grower 56 (11): 23–24Google Scholar
  9. Eady CC (2002a) Genetic transformation of onions. In: Rabinowitch HD, Currah L (ed) Allium crop science: recent advances. CABI Publishing/CAB International, Wallingford, UK, pp 199–144Google Scholar
  10. Eady CC (2002b) The transformation of onions and related alliums. In: Khachatourians GG, McHughen A, Scorza R, NipW-K, Hui YH (eds) Transgenic plants and crops. Dekker, New York, pp 655–671Google Scholar
  11. Eady CC, Lister CE (1998) A comparison of four selective agents for use with Allium cepa L. immature embryos and immature embryo-derived cultures. Plant Cell Rep 18: 117–121CrossRefGoogle Scholar
  12. Eady CC, Lister CE, Suo YY, Schaper D (1996) Transient expression of uida constructs in in vitro onion (Allium cepa L.) cultures following particle bombardment and Agrobacterium-mediated DNA delivery. Plant Cell Rep 15: 958–962Google Scholar
  13. Eady CC, Butler RC, Suo Y (1998) Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Allium cepa L.). Plant Cell Rep 18: 111–116CrossRefGoogle Scholar
  14. Eady CC, Pither-Joyce M, Farrant J, Shaw M, Reader J (2000a) In vivo suppression of alliinase in onion (Allium cepa L.). Sixth international congress of plant molecular biology, Quebec, Canada, 18–24 June 2000, abstract S03–33. International society for plant molecular biology, Université LavalGoogle Scholar
  15. Eady CC, Weld RJ, Lister CE (2000b) Agrobacterium tumefaciens-mediated transformation and regeneration of onion (Allium cepa L.). Plant Cell Reports 19 (in press)Google Scholar
  16. Eady CC, Davis S, Farrant J, Reader J, Kenel F (2002a) Agrobacterium tumefaciens-mediated transformation and regeneration of herbicide resistant onion (Allium cepa L.) plants. Ann Appl Bot (in press)Google Scholar
  17. Eady CC, Reader J, Davis S, Dale T (2002b) Inheritance and expression of introduced DNA in transgenic onion plants (Allium cepa L.). Ann Appl Bot (in press)Google Scholar
  18. Eady CC, Davis SD, Catanach A, Kenel F (2002c) Transgenic allium fingerprinting for quality control and breeding. 10th International Association for Plant Tissue Culture and Biotechnology, 23–28 June, Orlando, Florida (Abstract)Google Scholar
  19. Galmarini CR, Goldman IL, Havey MJ (2001) Genetic analyses of correlated solids, flavor, and health-enhancing traits in onion (Allium cepa L.). Mol Genet Genom 265 (3): 543–551CrossRefGoogle Scholar
  20. Gilpin BJ, Leung DW, Lancaster JE (1995) Nucleotide sequence of a nuclear clone of ‘alliinase (accession no 148614) from onion (PGR 95–125). Plant Physiol 110: 336Google Scholar
  21. Hasegawa H, Sato M, Suzuki M (2002) Efficient plant regeneration from protoplasts isolated from long-term, shoot primordia-derived calluses of garlic (Allium sativum). J Plant Physiol (in press)Google Scholar
  22. Hell R (1997) Molecular physiology of plant sulfur metabolism. Planta 202 (2): 138–148PubMedCrossRefGoogle Scholar
  23. Hood EE, Helmer GL, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bacteriol 168: 1291–1301PubMedGoogle Scholar
  24. Hunger SA, McLean KL, Eady CC, Stewart A (2002) Seedling infection assay for resistance to Sclerotium cepivorum in onions (Allium cepa) and other allium species. Proceedings of the 55thGoogle Scholar
  25. New Zealand Plant Protection Society Conference. www.hortnet.co.nz/publications/NZPPS Keusgen M (2002) Health and alliums. In: Rabinowitch HD, Currah L (eds) Allium crop science: recent advances. CABI Publishing/CAB International, Wallingford, UK, pp 357–378Google Scholar
  26. Kik C (2002) Exploitation of wild relatives for the breeding of cultivated Allium species. In: Rabinowitch HD, Currah L (eds) Allium crop science: recent advances. CABI Publishing/CAB International, Wallingford, UK, pp 81–100CrossRefGoogle Scholar
  27. King J, Coley-Smith J (1969) Production of volatile alkyl sulphides by microbial degradation of synthetic alliin and alliin-like compounds, in relation to germination of sclerotia of Sclerotium cepivorum Berk. Annu Appl Biol 64: 303–314CrossRefGoogle Scholar
  28. Kondo T, Hasegawa H, Suzuki M (2000) Transformation and regeneration of garlic (Allium sativum L.) by Agrobacterium-mediated gene transfer. Plant Cell Rep 19: 989–993CrossRefGoogle Scholar
  29. Lancaster J, Shaw M, Pither-Joyce M, McCallum J, McManus M (2000) A novel alliinase from onion (Allium cepa L.) roots: biochemical characterization and cDNA cloning. Plant Physiol 122: 1269–1279PubMedCrossRefGoogle Scholar
  30. Leffel SM, Mabon SA, Stewart CN (1997) Applications of the green fluorescent protein in plants. Biotechniques 23: 912PubMedGoogle Scholar
  31. Leustek T, Saito K (1999) Sulfate transport and assimilation in plants. Plant Physiol 120 (3): 637–643PubMedCrossRefGoogle Scholar
  32. Liu Y-G, Whittier RF (1995) Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from pl and YAC clones for chromosome walking. Genomics 25: 674–681PubMedCrossRefGoogle Scholar
  33. McCallum JA, Leite D, Pither-Joyce M, Havey MJ (2001) Expressed sequence markers for genetic analysis of bulb onion (Allium cepa. L. ). Theor Appl Genet 103: 979–991Google Scholar
  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497CrossRefGoogle Scholar
  35. Novak FF (1990) Botany, physiology, and genetics. In: Rabinowitch HD, Brewster JL (eds) Onions and allied crops, vol I. Botany, physiology and genetics. CRC Press, Boca Raton, pp 233–250Google Scholar
  36. Randle WM, Bussard ML (1993) Streamlining onion pungency analysis. Hortscience 28: 60Google Scholar
  37. Randle WM, Lancaster JE (2002) Sulphur compounds in alliums in relation to flavour quality. In: Rabinowitch HD, Currah L (eds) Allium crop science: recent advances. CABI Publishing/CAB International, Wallingford, UK, pp 329–356CrossRefGoogle Scholar
  38. Seabrook J (1994) In vitro propagation and bulb formation in garlic. Can J Plant Sci 74: 155–158CrossRefGoogle Scholar
  39. Smith NA, Singh SP, Wang MB, Stoutjesdijk PA, Green AG, Waterhouse PM (2000) Gene expression–total silencing by intron-spliced hairpin RNAs. Nature 407 (6802): 319–320PubMedCrossRefGoogle Scholar
  40. Soni S, Ellis P (1990) Insect pests. In: Rabinowitch HD, Brewster JL (eds) Onions and allied crops, vol II. Agronomy, biotic interactions, pathology and crop protection. CRC Press, Boca Raton, pp 213–272Google Scholar
  41. van Heusden AW, Shigyo M, Tashiro Y, Vrielink-van Ginkel R, Kik C (2000) AFLP linkage group assignment to the chromosomes of Allium cepa L-via monosomic addition lines. Theor Appl Genet 100 (3–4): 480–486CrossRefGoogle Scholar
  42. Zheng SJ, Henken B, Wietsma W, Sofiari E, Jacobsen E, Krens FA, Kik C (2000) Development of bio-assays and screening for resistance to beet armyworm (Spodoptera Exigua Hubner) in Allium cepa L. and its wild relatives. Euphytica 114 (1): 77–85CrossRefGoogle Scholar
  43. Zheng SJ, Khrustaleva L, Henken B, Sofiari E, Jacobsen E, Kik C, Krens FA (2001) Agrobacterium tumefaciens-mediated transformation of Allium cepa L.: the production of transgenie onions and shallots. Mol Breed 7(2):101–115Google Scholar

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© Springer-Verlag Berlin Heidelberg 2003

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  • C. C. Eady

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