Structural characterization and evaluation the elicitors activity of polysaccharides from Chrysanthemum indicum.

This research evaluates the elicitors activity and structure characterization of four Chrysanthemum indicum polysaccharides (CIPs) which were isolated from C. indicum, obtained CIP1, CIP2, CIP3, CIP4. Results demonstrated that there was a distinct difference in inducibility and CIP3 was significantly stronger than other CIPs through bioactivity-tests. Taking CIP3 with total carbohydrate content 91.93 % as a representative, its structure was elucidated as a relative molecular weight of 8. 741 × 103 g/mol and mainly composed of xylose, galacturonic acid, galactose and glucuronic acid. Through GC, IR and NMR, CIP3 was determined to possess a backbone comprised of T-α-d-GalpA, 1,4-α-d-GlcpA, 1,2-α-d-Xylp, 1,3-α-l-Rhap, 1,2,4-α-l-Rhap and sidechains comprised of 1,3-ß-d-Galp, 1,6-α-d-Galp, T-α-Glcp, 1,3-ß-d-Glcp, 1,4-α-d-Glcp, 1,3,4-α-d-Manp, T-α-l-Fucp. Further results indicated that CIP3 with active sidechains could significantly increase the expression of defense genes in Atractylodes macrocephala Koidz (AM). It is believed that the sidechains of CIP3 were necessary to its elicitor activity via bioactivity tests.

Consequences of antagonistic interactions between endophytic fungus and bacterium on plant growth and defense responses in Atractylodes lancea.

Many studies have examined pair-wise interactions between plants and endophytes, while overlooking the interplays among multiple endosymbionts and their combined impacts on hosts. In this study, Atractylodes lancea plantlets were inoculated with endophytic fungus Acremonium strictum AL16, or endophytic bacterium Acinetobacter sp., or both, to investigate the impacts of the three-way symbiosis on the host. Our results showed that defense-related responses of the co-inoculated plantlets were delayed and weakened relative to plantlets with single inoculants, but no detrimental effects on phyto-physiology (growth, photosynthesis) were observed after combined inoculations. Quantitative PCR analysis verified a decrease in AL16 colonization density within plants after co-inoculation with the endobacteria. An in vitro assay was then performed to elucidate the suppressed plant defense responses and reduced fungal colonization by dual inoculation. The data showed that the presence of Acinetobacter sp. reduced AL16 colony diameter and spore germination rate without negatively affecting fungal morphology. Additionally, direct hyphal attachment of the bacterium to AL16 in vitro was visualized by scanning electronic microscopy. Therefore, we propose that a balanced and compatible symbiosis might require constraints conferred by the antagonistic endophyte Acinetobacter sp. on the fungus AL16 in the tripartite endophytic bacterium-fungus-plant system.

Taguchi approach for anti-heat stress prescription compatibility in mice spleen lymphocytes in vitro.

Heat stress (HS) may induce immunosuppression as well as inhibit the proliferation of lymphocytes. This study evaluated the effects on immune function of our prescription on splenic lymphocytes under HS as well as its compatibility. The effects of four herbal extracts from Agastache rugosa, Atractylodes lancea, Cortex Phellodendri, and Gypsum Fibrosum on heat treated splenic lymphocytes were investigated and the compatibility of the prescription was also explored by using the Taguchi method. This study revealed changes in proliferation by traditional Chinese medicines of splenic lymphocytes after HS. Proliferation in the HS group was significantly lower than the control group. Under HS, the effects of higher concentrations of Agastache rugosa (100 and 200 µg/mL), Atractylodes lancea (100 and 200 µg/mL), Cortex Phellodendri (50 and 100 µg/mL) and Gypsum Fibrosum (100 and 200 µg/mL) caused a significant increase on ConA/LPS-induced proliferation of lymphocytes than lower concentrations. We, therefore, conclude that the prescription of traditional Chinese medicines may recover splenic lymphocytes from the immunosuppression induced by HS. The Taguchi design, which allows rapid and high efficiency for the selection of the best conditions for our prescription on HS-treated splenic lymphocytes, demonstrated that Agastache rugosa (200 µg/mL), Atractylodes lancea (200 µg/mL), Cortex Phellodendri (100 µg/mL) and Gypsum Fibrosum (100 µg/mL) were the optimal conditions for the prescription. The validation experiment confirmed that our composition in optimum extraction conditions enhanced effects on ConA or LPS-stimulated lymphocytes under HS. The results showed that the Taguchi optimization approach is a suitable method for optimization of the composition of prescription.