Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Establishment of an Efficient Agrobacterium Transformation System for Eggplant and Study of a Potential Biotechnologically Useful Promoter

  • Claudiu Magioli (Universidiade Federal do Rio de Janeiro, Instituto de Biologia LGMV/DG, Brazil) ;
  • Ana Paula Machado da Rocha (Universidiade do Estado do Rio de Janeiro, Instituto de Biologia LAMBIT/DBCG, Brazil) ;
  • Pinheiro, Marcia-Margis (Universidiade Federal do Rio de Janeiro, Instituto de Biologia LGMV/DG, Brazil) ;
  • Martins, Gilberto-Sachetto (Universidiade Federal do Rio de Janeiro, Instituto de Biologia LGMV/DG, Brazil) ;
  • Elisabeth Mansur (Universidiade do Estado do Rio de Janeiro, Instituto de Biologia LAMBIT/DBCG, Brazil)
  • Published : 2000.01.01
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Abstract

An efficient and reliable Agrobacterium transformation procedure based on TDZ (thidiazuron)-induced organogenesis was established and applied to six Brazilian eggp1ant varieties. Optimum transgenic plants recovery was achieved upon the study of the following parameters affecting transformation efficiency, using F-100 variety as a model: i) explant source; ii) pre-culture period; iii) physical state of the pre-culture medium and iv) coculture conditions. The highest frequency of kanamycin-resistant calli derived from leaf explants (5%) was obtained without a pre-culture period and co-cultivation for 24 h in liquid medium followed by five days on solid RM (regeneration medium). For cotyledon explants, best results were achieved upon a pre-culture of 24 h in liquid RM and a co-cultivation period of 24 h in liquid RM followed by three days in solid RM, resulting in a transformation Sequency of 22.7%. Kanamycin-resistant organogenic calli were also obtained from cultivars Emb, Preta Comprida, Round nose Shaded, Campineira and Florida Market. The expression pattern of an epidermis-specific promoter was studied using transformants expressing a chimaeric construct comprised by the promoter Atgrp-5 transcriptionally fused to the coding region of the gus gene. The expression pattern was similar to that previously observed in tobacco and Arabidopsis thaliana, with preferential expression at the epidermis and the stem phloem. These results support the idea that the Atgrp-5 promoter can be used to drive defense genes in these tissues, which are sites of pathogen interaction and spread, in programs for the genetic improvement of eggplant.

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References

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