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A xylem mutant (vse) was isolated from a Bambusa edulis (Odashima) Keng plantlet following vegetative micropropagation and subculture for 7 consecutive years and induced to proliferate in medium supplemented with 0.1 mg·L-1 (0.5 μm) thidiazuron (TDZ) and to develop roots in medium supplemented with 5 mg·L-1 (26.9 μm) α-naphthaleneacetic acid (NAA). Subsequent investigations comparing the growth habits of mutant plantlets with those of the wild type indicated that the growth of the former was retarded in a greenhouse. Several morphological abnormalities were observed in the vse mutant: it had thinner stems with fewer trichromes on the surface; the xylem vessels were smaller in diameter and contained crystal-like structures in the pith; the leaves were shorter and narrower with a sharp leaf blade angle; the roots were thinner and contained fewer xylem cells. The cation concentrations of both the mutant and wild type were similar in the in vitro analysis, except for those of iron and potassium, which were lower in mutant leaves in vivo. In 2-month-old mutant plants, iron chlorosis was observed on young leaves and a potassium deficiency was observed on older leaves. After 1 year of growth in the greenhouse, all of the wild-type plants had survived, but only 27% (16/60) of the mutant vse plants were alive.
The genus Dendrobium is important in traditional Chinese herbal medicine, and the precise identification of Dendrobium species is critical for the treatment and for pharmacological research. In the present study, a ribosomal DNA (rDNA) internal transcribed spacer (ITS) region-based analysis was used to ascertain the phylogenetic relationship among 20 Dendrobium species. The lengths of the ITS regions among the 20 species ranged from 636 to 653 bp, and the identities of the rDNA regions among the different species ranged from 75.7% to 99.1%. The results also showed that the ITS1 and ITS2 regions exhibit more variation than the 5.8S rDNA. A phylogenetic tree derived from the ITS sequence indicated that six medicinal Dendrobium species, of which five are common medicinal plants in the Taiwan market, were closely related and shared a common clade. Multiplex polymerase chain reaction (PCR) amplification was successfully performed to identify the six medicinal Dendrobium species, and amplification refractory mutation system (ARMS) PCR was used to distinguish D. tosaense specifically from the 19 other Dendrobium species. The established PCR-based (multiplex and ARMS) analyses can be used for the authentication of the raw materials of medicinal Dendrobium from other species.
To select resistant germplasm resources and understand the growth and physiological responses of kiwifruit (Actinidia sp.) to drought stress, five species, Actinidia macrosperma (Acma), Actinidia longicarpa (Aclo), Actinidia deliciosa (Acde), Actinidia hemsleyana (Ache), and Actinidia valvata (Acva), were assessed under tissue culture conditions. Rootless seedlings of five species were cultured in a medium containing polyethylene glycol [PEG (formula weight 8000)] to induce drought stress (0%, 5%, 10%, 15%, and 20%). After a 30-day culture, three growth indices [fresh weight (FW), plant height (PLH), and leaf number (LN)] and six physiological indices were determined, and the drought damage index (DDI) was determined. The DDIs of five species increased, and three growth indices decreased with increasing PEG concentrations. The following changes were observed under 20% PEG treatment conditions: superoxide dismutase (SOD) activities increased significantly in Acma, Aclo, and Ache specimens; peroxidase (POX) activities remained stable in Acde, Ache, and Acva specimens; and catalase (CAT) activities increased sharply in Acma and Acva. Furthermore, the results indicated that soluble sugar (SS) content increased slightly in Acma, Aclo, Acde, and Ache but it decreased in Acva specimens. Proline (PRO) content increased significantly in Acma and Acva, and malondialdehyde (MDA) contents tended to increase under drought stress in all five species. Principal component analysis (PCA) results indicated that the order of drought tolerance in the five genotypes examined in this study under tissue culture conditions was as follows: Acma > Acva > Acde > Aclo > Ache. Therefore, we concluded that Acma and Acva are more resilient germplasm resources that represent promising kiwifruit-breeding materials. Furthermore, tolerance to drought stress in these species should be further investigated under orchard conditions.