Production of doubled haploid plants from anther cultures of borage (<Emphasis Type="Italic">Borago officinalis</Emphasis> L.) by the application of chemical and physical stress

Cibotium barometz is an endangered tree fern, used both as ornamental plant and traditional Chinese medicinal plant. In this study, an effective in vitro propagation protocol was obtained through formation of green globular bodies (GGBs) from in vitro juvenile sporophytes. The effect of plant growth regulators (PGRs) on GGB induction and multiplication, as well as mineral salt concentration and active charcoal (AC) on plantlet regeneration from GGBs was evaluated. Thidiazuron (TDZ; 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea) had a significant effect on GGB induction and multiplication (P?<?0.001), while a-naphthaleneacetic acid (NAA) did not (P?>?0.05). GGB induction rate was above 80?% on 1/2 Murashige and Skoog (MS) media supplemented with TDZ (1.0 mg L^??1) and NAA (0.1, 0.3 or 0.5 mg L^??1). The same media were also optimal for GGB multiplication. GGBs cultured on 1/4 MS media supplemented with 0.1 or 0.2?% (w/v) AC showed a high rate of GGB development into plantlets above 90?%. 1/2 MS media supplemented with 0.1 or 0.2?% AC were the most effective for plantlet growth. Regenerated plantlets were successfully acclimatized (80?%) in greenhouse conditions. Morphological and histological analysis revealed that C. barometz GGBs was a yellow-green globular structure composed of the single GGB with meristems and hair-like structures, and new single GGBs were initiated from the epidermal cells of meristem zone.     

A refined method for ovule culture in sugar beet (Beta vulgaris L.)

Induction of gynogenesis through ovule culture is a valuable tool to produce haploid and doubled haploid plants in sugar beet (Beta vulgaris L.). However, there is still large room for refining the method. In this study we investigated the gynogenic response of cultured ovules of three sugar beet genotypes, the effect of the application to inflorescences of different pretreatments with mannitol at 4ºC and with 5-azacytidine and 2,4-D, and the effect of the use of different basal culture media and sucrose concentrations. The response was evaluated in terms of percentages of induction of gynogenesis, embryogenesis and callogenesis, as well as of regenerated plants. We showed that a pretreatment with 0.5 M mannitol at 4 °C for 4 days, and with 50 µM 5-AzaC for 1 h, notably improved the percentage of embryogenesis and plant regeneration. Besides, the use of MS basal medium and 60 g/L sucrose was also found beneficial. This study provides new ways to improve the efficiency of haploid induction and plant regeneration through ovule culture in sugar beet, and is potentially applicable to ovule culture in other crops.