Biolistic Delivery of Foreign DNA or Genomic Transcripts of Plant Virus Full-length cDNA Clones into Monocotyledonous and Dicotyledonous Plant Tissues

Abstract

Using an improved particle gun driven by compressed nitrogen gas (20kg/cm² pressure), embryos of oat (Avena sativa) and wheat (Triticum aestivum) or primary leaves of cowpea (Vigna unguiculata) were bombarded with gold particles coated with the vector DNA, pBI121, or genomic transcript RNAs of sweet clover necrotic mosaic virus (SCNMV). GUS gene expression in the oat seedlings was detected in the apical meristems of the primary and emerging lateral roots, and in the root hairs, procambium, stele, coleoptiles, phloem elements, and leaf tissues. However, the retention of the foreign gene in the engineered plants was transient and the gene was lost in the absence of Kanamycin stress. With genomic transcipts of SCNMV full-length cDNA clones of 5ng per bombardment, replication of the delivered genomic viral transcripts was supported by intact primary leaves of cowpea.