In vitro embryo culture of Epimedium wushanense / 中国中药杂志

<p><b>OBJECTIVE</b>To study the in vitro embryo culture of Epimedium wushanense and provide scientific basis for large scale production of tissue culture.</p><p><b>METHOD</b>Cullus and buds were induced from embryo of E. wushanense on a MS medium supplemented with different 2,4-D,6-BA, NAA, IBA.</p><p><b>RESULT</b>The optimal compositions of medium that induced callus and buds from embryo were the MS medium supplemented with 2,4-D 2 mg x L(-1), IBA 2 mg x L(-1) and NAA 0.5 mg x L(-1) and the MS medium supplemented with IBA 2 mg x L(-1) and 6-BA 0.5 mg x L(-1), respectively. The optimum medium for callus differentiation was MS + 6-BA 1 mg x L(-1) + NAA 0.5 mg x L(-1) + IBA 1 mg x L(-1), and MS +6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) for shoots proliferation.</p><p><b>CONCLUSION</b>Using embryo as explants, the method of induction and culture of E. wushanense was established by the callus and buds, and the embryo of E. wushanense can be quickly propagated.</p>

Infection progress of arbuscular mycorrhizae on tissue-cultured plantlets of Pinellia ternata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the Arbuscular mycorrhizal (AM) formation progress and infection characteristics between tissue culture plantlets of Pinellia ternata and Glomus mosseae.</p><p><b>METHOD</b>The tissue culture plantlets of P. ternata were inoculated with G. mosseae, the formation of AM were sampled and observed with microscopy by staining.</p><p><b>RESULT AND CONCLUSION</b>The hyphae of G. mosseae began to penetrate the root epidermis after 10 days of inoculation. Lots of intracellular hyphae formed in cortex cells at the 15th day. Arbuscules started to form and there were some hyphae on the root at the 20th day. At the 25th day, many arbuscules formed and most as Arum type. Some arbuscles started to disintegrate at the 30th day, and a few of vesicles occurred. Lots of spores formed after 35 days. At the 40th day, some vesicles began to decline. The hand section showed that the intercellular hyphae gradually formed in intercellular space, and the hyphae branched in cortex cells and occupied most cell lumen finally. It is expounded that P. ternata and G. mosseae could recognize each other quickly and form a symbiont system.</p>

Dormancy characteristics and breaking method of seeds from Epimedium wushanense / 中国中药杂志

<p><b>OBJECTIVE</b>To research the mechanism of dormancy and find out the breaking method for the seeds of Epimedium wushanense.</p><p><b>METHOD</b>The water permeability of seed coat was tested by weighing seeds. The germination inhibitor of the seeds were determined with biotic measurement. The development of embryos, germination rate and germination potential were determined after stratification.</p><p><b>RESULT</b>The water permeability of seed coat was 41.86% after 5 h. The extracts of seeds had strong inhibition effects to the length growth of cabbage seedlings. The growth and development of embryos under the cold stratification (5 degrees C) were better than that under the other conditions. The embryo rate extended from 15.39% to 86.21% after 90 d. Germination rate and germination potential after stratification under 5 degrees C were significantly higher than that under other temperatures.</p><p><b>CONCLUSION</b>The results showed that there was no obstacle of water permeability on the test of E. wushanense, after-ripening of embryogenesis and the germination inhibitor of the seed were the main reason for the seed dormancy. The cold stratification would be an effective way for breaking of the dormancy, which could significantly promote the seed embryogenesis and increase germination rate comparing to other methods.</p>

Study on influence of arbuscular mycorrhizal fungi Pinellia ternata yield and chemical composition / 中国中药杂志

<p><b>OBJECTIVE</b>To study the influence of arbuscular mycorrhizal fungi to Pinellia ternata leaf index, propagation coefficient, tuber yield and chemical composition.</p><p><b>METHOD</b>The Pinellia ternata leaf length, width, plant height and leafstalk diameter were measured during growing time till P. ternata loded. The leaf index was calculated and the tuber yield were measured after harvest. The contents of guanosine was determined by HPLC method, and the alkaloid contents were determined by spectrophotometric method.</p><p><b>RESULT AND CONCLUSION</b>Inoculated AM fungi could increase P. ternata propagation coefficient and the yield of tuber. Inoculated AM fungi could increase P. ternata leaf index, made leaf tends to be more like bamboo leaf type. Inoculated AM fungi could delayed P. ternata lodging time; G. mosseae + G. intraradices treatment (early inolulation time) proved better influence on increasing fresh weight and dry weight of P. ternata tuber and the propagation coefficient. Inoculated AM fungi could increase guanosine and alkaloid contents of Pinellia ternata tuber, and the contents of guanosine and alkaloid in inoculated treatments were higher than those in cultivated P. ternata from Hezhang; G. mosseae + G. intraradices treatment (later inolulation time) proved good effect on increasing the contents of guanosine and alkaloid.</p>

Arbuscular mycorrhiza of cultivated and wild Pinellia ternata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the arbuscular mycorrhiza and arbuscular mycorrhizal fungi associated with cultivated and wild Pinellia ternata in Guizhou province.</p><p><b>METHOD</b>Wild and cultivated P. ternata roots were observed through staining and microscopic examination, the arbuscular mycorrhizal fungi spores were isolated through wet thieving according to Gerdemann & Nicolson (1963), the spores were identified following the description of Schenck & Pérez (1988), and some previous publications.</p><p><b>RESULT</b>The typical arbuscular mycorrhiza (AM) structure was showed according to a research of wild and cultivated P. ternata. In the survey of AM fungi species in the rhizosphere of wild and cultivated P. ternata, 3 genera and 21 species were found, 3 genera and 7 species were identified. 5 species of them belong to Glomus, 1 species belongs to Scutellospora, 1 species belongs to Gigaspora, including Glomus mosseae, G. intraradices, G. melanosporum, G. deserticola, G. aggregatum, Scutellospora castanea, Gigaspora albida, and one of them was a new record, i.e., Scutellospora castanea which was the dominant species in Bijie.</p><p><b>CONCLUSION</b>The diversity of AM fungi between wild and cultivated Pinellia ternata was showed on this survey, the fungi associated with wild ones are different form the cultivated ones, such as Gigaspora albida only occurs in cultivated ones, Glomus melanosporum only occurs in wild ones, while Glomus mosseae and Glomus intraradices occur in both wild and cultivated ones, and there were specialization species in Bijie, all these can provide new though for solving degradation problem of cultivated Pinellia ternata.</p>

Extraintestinal lesions caused by rotavirus:An experimental study on suckling mice / 第三军医大学学报

Objective To study the extraintestinal lesions induced by rotavirus(RV)infection and explore the pathogenesis.Methods Simian rotavirus SA11 was cultured in MA-104 cells.After inoculation,the pathological changes in brain,lung,heart,liver,pancreas and kidney tissues were observed,the RV antigens detected,and the apoptotic cells observed.Besides,we stained the filamentous actin(F-actin)with Phalloidine-TRITC,and then quantified the F-actin amount.Results Several pathological changes,inclusive of myocardial interstitial edema,granular degeneration in cardiacytes,hepatic congestion,and hepatocellular vacuolar degeneration,were found,but not found in the brain,lung,and pancreas tissues.Meanwhile,several ultrastructural changes,inclusive of dissolved myocardial F-actin,extended smooth endoplasmic reticulum,swollen mitochondria,and widened perinuclear space,were found.No difference was found in the quantity of myocardial F-actin.Apoptosis was found in liver cells,but not in myocardial cells.RV RNA was detected in tissues such as brain,lung,heart,liver and pancreas tissues.Conclusion All the results suggest that RV may spread from the intestine to various extraintestinal organs and hence induce injury.Filamentous actin depolymerization,cytoskeleton damage and apoptosis induced by RV infection are the important mechanism not only in intestinal damages,but also in extraintestinal lesions.

On pathogenesis of rotavirus diarrhea in suckling mouse / 第三军医大学学报

Objective To study the pathogenesis of rotavirus(RV) diarrhea.Methods Simian rotavirus(SA-11) was grown in cultured MA-104 cell.The viral titers of the culture supernatant were determined by plaque forming assay.KM mice aged 7 days were inoculated with the viral supernatant via feeding tube(gavage).Histological and ultramicrostructure changes of the small intestines were observed under light microscope and electron microscope.The values of crypt depth and villi height were measured with software(image pro plus 5.1,IPP5.1).The distribution of the RV antigen in small intestine and the filamentous actin of the small intestine chorioepithelium were observed with immunohistochemical techniques.The apoptosis of the small intestine epithelium cells was observed with an in situ apoptosis detection kit.Results There were mild hyperemia,dropsy and extensive vacuolar degeneration of small intestine villi under light microscope.Plenty of lipid droplet-like structure at the top of the villi,microvilli malalignment or defluxion and enterocyte defluxion could be seen by electron microscope,but no obvious structure changes at the cell junctions were seen.The RV antigen mainly distributed at the top of the villi.The quantity of small intestine filamentous actin decreased and enterocyte apoptosis increased after RV infection.Conclusion RV mainly infects the mature villous epithelium.The presentation of RV diarrhea relates to the lesion of cytoskeleton,the microvilli lesion of the small intestine,enterocyte apoptosis and defluxion,villi atrophy,etc.,but may have no relationship with the structural changes of cell junctions in the small intestine epithelium.