title: Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance.
abstract: BACKGROUND
  The recovery of high performing transgenic lines in clonal crops is limited by the occurrence of somaclonal variation during the tissue culture phase of transformation. This is usually circumvented by developing large populations of transgenic lines, each derived from the first shoot to regenerate from each transformation event. This study investigates a new strategy of assessing multiple shoots independently regenerated from different transformed cell colonies of potato (Solanum tuberosum L.).
  
  
  RESULTS
  A modified cry9Aa2 gene, under the transcriptional control of the CaMV 35S promoter, was transformed into four potato cultivars using Agrobacterium-mediated gene transfer using a nptII gene conferring kanamycin resistance as a selectable marker gene. Following gene transfer, 291 transgenic lines were grown in greenhouse experiments to assess somaclonal variation and resistance to potato tuber moth (PTM), Phthorimaea operculella (Zeller). Independently regenerated lines were recovered from many transformed cell colonies and Southern analysis confirmed whether they were derived from the same transformed cell. Multiple lines regenerated from the same transformed cell exhibited a similar response to PTM, but frequently exhibited a markedly different spectrum of somaclonal variation.
  
  
  CONCLUSIONS
  A new strategy for the genetic improvement of clonal crops involves the regeneration and evaluation of multiple shoots from each transformation event to facilitate the recovery of phenotypically normal transgenic lines. Most importantly, regenerated lines exhibiting the phenotypic appearance most similar to the parental cultivar are not necessarily derived from the first shoot regenerated from a transformed cell colony, but can frequently be a later regeneration event.
authors: The New Zealand Institute for Plant & Food Research Ltd, Private Bag 4704, Christchurch 8140, New Zealand. mei.meiyalaghan@plantandfood.co.nz
authors: Sathiyamoorthy
authors: S
authors: Meiyalaghan
authors: Philippa J
authors: PJ
authors: Barrell
authors: Jeanne M E
authors: JM
authors: Jacobs
authors: Anthony J
authors: AJ
authors: Conner
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods*; D015139:Blotting, Southern; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D014644:Genetic Variation; D006460:Hemolysin Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D007306:Insecticides / Q000378:metabolism / Q000494:pharmacology; D007814:Larva / Q000187:drug effects* / Q000502:physiology; D009036:Moths / Q000187:drug effects / Q000502:physiology; D018520:Plant Shoots / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D058439:Plant Somatic Embryogenesis Techniques; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012038:Regeneration; D011198:Solanum tuberosum / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D005819:Genetic Markers; D006460:Hemolysin Proteins; D007306:Insecticides; C030956:insecticidal crystal protein, Bacillus Thuringiensis