Validity of Selection Breeding for High Ephedrine-Alkaloid Content in E. sinica under Different Environments in Japan.

Dried terrestrial stems of Ephedra sinica are called 'Ephedra herb,' whose pharmacological effects are due mainly to two major ingredients, (-)-ephedrine and (+)-pseudoephedrine (total alkaloids which are defined in Japanese Pharmacopoeia (TA)). Ephedra herb is an important crude drug in Japan. However, E. sinica is widely distributed in arid areas of northeastern China and Mongolia. Recently, E. sinica has started to be cultivated in Japan. This study aimed to assess the validity of selection breeding on TA content of E. sinica in several locations in Japan. In this experiment, we grew approximately 350 seedlings and divided them randomly into seven groups. Nearly fifty plants were cultivated at each of seven locations. In Ibaraki, Yamanashi, and Shizuoka, average TA content of whole samples satisfied the criteria for Ephedra herb defined in Japanese Pharmacopoeia (7.0 mg/g of dry weight (DW)). Plants with high and intermediate TA content at four locations were selected and transplanted to Ibaraki. There were significant differences in TA content between selected plants with high and intermediate TA content before and after transplanting (p < 0.05). TA content of high-TA plants was significantly higher than that of control plants cultivated continuously at Ibaraki (p < 0.05). These results suggest that the selection on content of ephedrine alkaloids in E. sinica under various locations in Japan is valid, and high- TA E. sinica plants can be selected at various locations.

[Endophytic bacterial diversity in Codonopsis pilosula, Ephedra sinica, and Lamiophlomis rotate: a study with LH-PCR].

To investigate the endophytic bacterial diversity in the three medicinal plant species Codonopsis pilosula, Ephedra sinica, and Lamiophlomis rotata in Ganzi of Sichuan, Southwest China, the total DNA of the three species were extracted by stringent surface sterilization, and studied with length heterogeneity-PCR (LH-PCR) method. For the same plant species, their root-, stem-, and leaf LH-PCR profiles were in a high level of similarity, with little differences in band richness. However, there existed great differences in the LH-PCR profiles among different plant species. C. pilosula had the biggest band richness, followed by E. sinica, and L. rotata. In the three plant species, the endophytic bacteria with an approximately 474 bp DNA length were dominant. The endophytic bacterial diversity of the plants was negatively correlated with rhizosphere soil available phosphorus content, but positively correlated with rhizosphere soil pH. Elevation and rhizosphere soil total nitrogen content were the important environmental factors affecting the distribution of enophytic bacteria in these plant species. The information of population diversity obtained from LH-PCR could more intuitively reflect the differences of bacterial diversity among different plant species, and thus, LH-PCR would be available to be used for analyzing the endophytic bacterial diversity in medicinal plants, providing information and guidance for the further isolation of microbial resources.

[Surveys on resources and varieties on Chinese markets of crude drug mahuang].

Many species of the genus Ephedra plants are used as Chinese traditional medicines, in Chinese which are known as "Mahuang" (Ephedrae Herba). In order to get a clear picture of resources distribution and varieties on Chinese markets of the crude drug Mahuang and provide scientific basis for their resource conservation and sustainable use, during recent years we conducted field investigations and market researches many times. The results showed that the most common species on the Chinese markets was E. sinica (33/38 commercial samples), followed by E. intermedia (5/38 commercial samples), which was also used in local clinics in Qing-hai, Gansu and Xinjiang province, no E. equisetina was found in the market. We noticed that the resources of both official and non-official plants of Mahuang, especially Zhong-Mahuang and Muzei-Mahuang, were seriously damaged in the past decade because of the ecological environment damage and over-digging. Zhong-Mahuang was distinguished in Ningxia and north Gansu, which was once one of the most distribution areas and contains more than 10,000 t Zhong-Mahuang. Muzei-Mahuang was distinguished in most places and distributed sparsely around Altay Mountains in northeast in Xinjiang. Thus, Cultivation of Mahuang, especially Zhong-Mahuang and Muzei-Mahuang should be greatly developed. At the same time, wild resources of Mahuang must be preserved strictly, i.e., proper method of cutting Ephedra plant could prevent the damage of the resource.

[Accumulative trends of ephedrine and pesudoephedrine in Ephedra sinica].

In Yi Kezhao Alliance, the content of ephedrine and pseudoephedrine in Ephedra sinica is in a ratio of four to six. The total content of ephedine and pseudoephedrine form cultivated Ephedra sinica is higher than that of wild one, and in direct ratio with the years of plant. For the content enhance of total alkaloids, decreasing irrigation and applying middlingly fertilizer is more effective measure.

[Character of emergence for the seeds of Ephedra sinica].

The experimental results on the germinating of Ephedra sinica seeds and sowing show: the seeds of E. sinica are not sensitive to illumination, but very sensitive to humidity; the effects of soil, the depth of sowing and irrigation on emergence are very notable, but not remarkable to covering which can be used to prevent measure.