Abstract
Only few studies in the plant tissue culture literature have examined the impact of filter paper on in vitro plant organogenesis. In this study, using a model plant, hybrid Cymbidium Twilight Moon ‘Day Light’, the impact of a single or double layer of Advantec #2 or Whatman #1 filter paper on new protocorm-like body (neo-PLB) formation on Teixeira Cymbidium (TC) medium was examined for half-PLBs (transgenic and non-transgenic), PLB-derived transverse thin cell layers (tTCLs), and PLB synseeds. In addition, the response of half-PLBs or tTCLs to two antibiotics (kanamycin and cefotaxime, commonly used in plant genetic transformation studies) was investigated either directly on gelled medium or on filter paper-overlaid medium. Filter paper negatively affected most growth and developmental parameters of all the explants tested, both transgenic and non-transgenic. A double sheet of filter paper had a significantly (P ≤ 0.05) more negative impact than a single sheet, relative to the control values (i.e., no filter paper). Kanamycin inhibited neo-PLB formation on TC medium, the negative impact being greater on a single layer than on a double layer of filter paper, i.e., filter paper buffered the growth-inhibiting characteristics of kanamycin. Up to 100 mg/l, cefotaxime showed no apparent negative effects on neo-PLBs formation and growth, although hyperhydricity was observed when filter paper was not used.
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