ABSTRACT
This study was designed to establish a multiplex allele-specific polymerase chain reaction method for simultaneous identification of Dendrobium huoshanense, D. officinale and D. devonianum, which may resolve identification problems of caulis dendrobii. Internal transcribed spacer sequences and trnL-trnF sequences of the Dendrobium species were aligned by BioEdit software, then specific SNPs of the three species were analyzed for designing allele-specific primers and the multiplex allele-specific PCR reaction system was established. The different origin of Dendrobium huoshanense, D. officinale and D. devonianum was amplified and identified by the sizes of respective band. The results showed that 584 bp, 397 bp and 211 bp bands could be amplified by D. devonianum, Dendrobium officinale and Dendrobium huoshanense respectively, when the annealing temperature was 61 ℃ and the number of cycles was 35. The limit of detection (LOD) of D. devonianum and D. huoshanense were both 1.2 ng, while D. officinale was low than 0.24 ng. The detection limit of adulterates in D. devonianum, D. devonianum and D. huoshanense mixture sample was 1%, 1% and 5% respectively. This result suggests that the method of multiplex allele-specific PCR is useful to identify D. huoshanense, D. officinale and D. devonianum is accurate and specific.
ABSTRACT
To obtain seedling growth-promoting fungi is a key step in restoration-friendly cultivation of medicinal Dendrobium species, since there are a large number of functionally-unknown endophytic fungi in the roots of Dendrobium plants.In this study, six functionally-unknown endophytic fungal strains were isolated from roots of D.devonianum using single peleton isolation technology, and used in inoculation experiments to test their effectiveness for seedling growth in D.devonianum.After 90 days of inoculation, comparing with the control treatment, FDdS-1, FDdS-2 and FDdS-4 showed strong pathogenic or fatal effects on seedlings; while, FDdS-12, FDdS-9 and FDdS-5 had different effects on seedling growth.FDdS-5 had significant promoting effects on height, fresh and dry weight, stem diameter and root numbers, while FDdS-9 only had significant promoting effect on seedling height, and FDdS-12 had a negative effect on seedling growth.According to the anatomical features of the inoculated roots, FDdS-5 fungi could infect the velamina of seedlings and the existence of symbiosis pelotons in the cortex cells, suggesting that FDdS-5 is a mycorrhiza fungi of D.devonianum.FDdS-5 and FDdS-9 were identified as Sebacina vermifera and Sebacina sp.by molecular technologies.By using FDdS-5 in the restoration-friendly cultivation of D.devonianum, it could effectively promote seedling growth and shorten the seedling growth periods.The results will aid in reintroduction and cultivation of D.devonianum.
ABSTRACT
OBJECTIVE: To evaluate the effect on mice immunity activity of Dendrobium devonianum polysaccharides and Dendrobium candidum polysaccharides. METHODS: Cultured splenic lymphocytes of mice in vitro to observe the influence of Dendrobium devonianum polysaccharides and Dendrobium candidum polysaccharides separately cooperated with ConA on proliferation of splenic lymphocyte and content of IFN-γ and IL-2. In animal experiments, respectively offered Dendrobium devonianum polysaccharides and Dendrobium candidum polysaccharides to mouse in different dosage, after 14d, copied the model of toe swelling induced by SRBC to observe the effect of Dendrobium polysaccharides on toe swelling and viscera index. RESULTS: Medium dosage groups of two Dendrobium polysaccharides both can stimulate the proliferation of splenic lymphocyte observably; medium and high dosage groups both can promote the secretion of IFN-γ and IL-2 in splenic lymphocyte: In addition, high dosage groups of two Dendrobium polysaccharides both can increase toe swelling and spleen index of mouse obviously, but every dosage groups have no effect on thymus index of mouse. CONCLUSION: To some extent, Dendrobium devonianum polysaccharides and Dendrobium candidum polysaccharides can improve the immunity activity of mouse.
ABSTRACT
OBJECTIVE: To study the chemical constituents of Dendrobium devonianum. METHODS: The compounds were isolated and purified by means of column chromatography with silica gel, Sephadex LH-20 and prep- HPLC. Their structures were identified on the basis of physicochemical parameters and spectroscopic data comparison with standard literature data. RESULTS: Twenty-five compounds were isolated from the 95% ethanol extract of D. devonianum, and identified as 3-hydroxy-4', 5-dimethoxybibenzyl(1), tristin(2), 3, 4', 5-trihydroxybibenzyl(3), N-trans-coumaroyltyramine(4), N-trans- caffeoyltyramine(5), p-hydroxyphenylpropionyltyramine(6), N-cis-coumaroyltyramine(7), 2, 5-dihydroxy-4-methoxy-9, 10-dihydrophenanthrene(8), 2, 4, 7-trihydroxy-9, 10-dihydrophenanthrene(9), 4', 5, 7-trihydroxy-flavonol(10), dihydroconifery acetate(11), ethyl 4-hydroxycinnamate(12), p-hydroxycinnamic acid(13), p-hydroxyphenylpropionic acid(14), 4-hydroxybenzoic acid(15), protocatechuic acid(16), erigeside II(17), citrusin C(18), dihydrosyringin(19), leonuriside A(20), adenosine(21), β-sitosterol(22), daucosterol(23), (+)-syringgaresinol-O-β-D-glucopyranoside(24) and sucrose(25). CONCLUSION: Compounds 1-21 and 24-25 are obtained from this plant for the first time. Compound 1 is isolated from the Dendrobium genus for the first time. Compounds 5, 7, 11, 12 and 18 are obtained from Orchidaceae for the first time.