Cold hardiness evaluation is important for screening woody species in cold areas. We compared cold hardiness by estimating the 50% lethal temperature (LT50) using electrolyte leakage test (ELLT50) and triphenyltetrazolium chloride test (TTCLT50) for 26 woody species in the Bashang region of China. One-year-old shoots were collected in January and exposed to five subfreezing temperatures in a programmable temperature and humidity chamber. LT50 was estimated by fitting relative electrolyte leakage and percentage of dead tissue against test temperature. For all tested species, triphenyltetrazolium chloride (TTC) staining of the pith was weak and the cambium TTCLT50 was lower than the extreme minimum temperature (−37 °C) recorded in the region. The cambium TTCLT50 and the sd were lower than that for the phloem and xylem. The phloem TTCLT50 was lower than the xylem TTCLT50, and the two sds were similar. The ELLT50 showed no significant correlation with any TTCLT50. For most species, the ELLT50 was higher than the cambium and phloem TTCLT50 and was not significant different with the xylem TTCLT50. The ELLT50 showed higher sd than any tissue TTCLT50. Based on results obtained in this study, when choosing cold hardiness of single stem tissue as an indicator for screening woody species, the xylem should be considered first, followed by the phloem; the cambium and pith were unsuitable. The cold hardiness estimated by ELLT50 was more suitable as indicator for screening woody species than that of stem tissue in winter estimated by TTCLT50.
Dendrobium officinale Kimura et Migo is a famous traditional Chinese medicinal plant. It produces various phytochemicals, particularly polysaccharides, which have nutraceutical and pharmaceutical values. To increase its biomass production and polysaccharide content, our breeding program has generated a series of polyploid cultivars through colchicine treatment of protocorm-like bodies (PLBs). The present study compared two tetraploid cultivars, 201-1-T1 and 201-1-T2, with their diploid parental cultivar, 201-1, in an established in vitro culture system. Tetraploid ‘201-1-T1’ and ‘201-1-T2’ had shorter leaves and shorter and thicker stems and roots, and they produced higher biomass compared with the diploid cultivar. The length and width of stomata significantly increased, but stomatal density decreased in tetraploid cultivars. The PLB induction rates from the stem node explants of the tetraploid cultivars were significantly higher than those of diploid. However, the PLB proliferation of tetraploids was lower than that of the diploid. The mean number of plantlets regenerated from tetraploid PLBs was also lower than that of the diploid after 4 months of culture. Polysaccharide contents in stems, leaves, and roots of 6-month-old tetraploid plantlets were significantly higher than those of diploids. The polysaccharide content in the stem of ‘201-1-T1’ was 12.70%, which was a 2-fold increase compared with the diploid cultivar. Our results showed that chromosome doubling could be a viable way of improving D. officinale in biomass and polysaccharide production.
The procedure for Heuchera villosa ‘Caramel’ propagation was investigated, which involves shoot regeneration, rooting of regenerated shoots, and acclimation of regenerated plantlets. Petioles, as explants, were cultured on MS medium supplemented with 1-naphthylacetic acid (NAA), benzylaminopurine (BA), thidiazuron (TDZ) and callus formed on all media. Shoots were observed to proliferate from callus on media with BA and NAA, whereas no shoots regenerated on media with TDZ and NAA. On media containing 0.5 or 1.0 mg·L−1 BA in combination with NAA, the regenerated shoots showed severe hyperhydricity, whereas on media containing 0.1 mg·L−1 BA in combination with NAA, the regenerated shoots grew normally. The highest shoot induction rate, 90.6%, was obtained on media containing 0.1 mg·L−1 BA and 0.01 mg·L−1 NAA. The effects of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and NAA on rooting of H. villosa ‘Caramel’ was explored. The highest rooting rate (95%) was obtained on 1/2 MS medium containing 0.2 mg·L−1 NAA. In the subsequent acclimation experiments, about 85% of rooted plantlets survived and grew normally.