Isolation of protoplasts and culture and regeneration into plants in Alstroemeria

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Abstract

An efficient system for the regeneration of plants from protoplasts was developed in Alstroemeria. Friable embryogenic callus (FEC) proved to be the best source for protoplast isolation and culture when compared with leaf tissue and compact embryogenic callus. Protoplast isolation was most efficient when FEC was cultured under vacuum for 5 min in an enzyme solution consisting of 4% cellulase, 0.5% Driselase and 0.2% Macerozyme, followed by culture for 12¿16 h in the dark at 24°C. Cell wall formation and colony formation were better in a liquid medium than on a semi-solid agarose medium. Micro-calluses were formed after 4 wk of culture. Ninety percent of the micro-calluses developed into FEC after 12 wk of culture on proliferation medium. FEC cultures produced somatic embryos on a regeneration medium and half of these somatic embryos developed shoots. Protoplast-derived plants showed more somaclonal variation than vegetatively propagated control plants
Original languageEnglish
Pages (from-to)505-510
JournalIn Vitro Cellular and Developmental Biology. Plant
Volume41
Issue number4
DOIs
Publication statusPublished - 2005

Keywords

  • somaclonal variation
  • cell-cultures
  • growth
  • callus
  • rice

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