Origin traceability of Cordyceps sinensis based on trace elements and stable isotope fingerprints.

Cordyceps sinensis is a kind of valuable Chinese herbal medicine, and its quality and price depend on the place of origin. Building a traceability system for Cordyceps sinensis products is an effective way to protect Cordyceps sinensis geographical indication products and consumers. In this study, concentrations of 45 trace elements and stable C, N, and Pb isotopes were used to distinguish Cordyceps sinensis samples from different habitats and different varieties (natural and artificial). The results showed that there were significant differences in the isotope compositions of N and Pb and trace elements contents in the Cordyceps sinensis samples from different sources. Stepwise discriminant analysis was used to select effective traceability indicators, and three discriminant models were successfully established. A combination of Co, Sr, Cu, Tl, and Zr indexes was selected to distinguish the naturally grown samples from the artificially cultivated ones, with an overall cross-validation correctness rate of 90.0 %; while a combination of As, Cu, Rb, Tl, W, and Zr indexes was adopted to distinguish the naturally grown samples from different regions, with a corresponding 100.0 % overall cross-validation correctness rate. To simultaneously distinguish samples between natural and artificial and between different regions, a combination of As, Cu, Rb, Tl, U, W, and δ15N indexes was employed, with an overall cross-validation correctness rate of 89.3 %.

Lanostane triterpenoids from artificially cultivated fruiting bodies of Ganoderma cf. mastoporum.

In the quest for bioactive compounds from Ganoderma, artificially cultivated fruiting bodies of Ganoderma cf. mastoporum, strain TBRC-BCC 47851 were chemically investigated. The study led to the isolation of three undescribed lanostane triterpenoids (1-3) together with twelve known compounds. The structures were elucidated on the basis of NMR spectroscopic and mass spectrometry data. The new compounds were inactive in the antimalarial and antitubercular activity assays.

Artificial Cultivation Changes Foliar Endophytic Fungal Community of the Ornamental Plant Lirianthe delavayi.

Many wild ornamental plant species have been introduced to improve the landscape of cities; however, until now, no study has been performed to explore the composition and function of foliar endophytes associated with cultivated rare plants in cities after their introduction. In this study, we collected the leaves of the healthy ornamental plant Lirianthe delavayi from wild and artificially cultivated habitats in Yunnan and compared their diversity, species composition, and functional predictions of their foliar endophytic fungal community based on high-throughput sequencing technology. In total, 3125 ASVs of fungi were obtained. The alpha diversity indices of wild L. delavayi populations are similar to those of cultivated samples; however, the species compositions of endophytic fungal ASVs were significantly varied in the two habitats. The dominant phylum is Ascomycota, accounting for more than 90% of foliar endophytes in both populations; relatively, artificial cultivation trends to increase the frequency of common phytopathogens of L. delavayi, such as Alternaria, Erysiphe. The relative abundance of 55 functional predictions is different between wild and cultivated L. delavayi leaves (p < 0.05); in particular, chromosome, purine metabolism, and peptidases are significantly increased in wild samples, while flagellar assembly, bacterial chemotaxis, and fatty acid metabolism are significantly enhanced in cultivated samples. Our results indicated that artificial cultivation can greatly change the foliar endophytic fungal community of L. delavayi, which is valuable for understanding the influence of the domestication process on the foliar fungal community associated with rare ornamental plants in urban environments.

Ultra-performance liquid chromatography-tandem mass spectrometry revealed the significantly different metabolic profiles of Auricularia cornea growing on weakly acidic and weakly alkaline substrates.

Auricularia cornea is a widely prized basidiomycetous mushroom with culinary and medicine value, which is cultivated artificially on a large scale in China. However, little attention has been paid to the differences in metabolic profiles under different pH growth conditions. In the present study, weakly acidic and weakly alkaline artificial substrates were developed and used for the cultivation of A. cornea, and the metabolic profiles of its fruiting bodies were determined using ultra performance liquid chromatography-tandem mass spectrometry. The results show that the weakly alkaline substrate environment promoted mycelial growth, increased body surface area, and improved yield and transformation efficiency, but attenuated metabolite accumulation by A. cornea. A total of 412 different metabolites were identified in negative and positive ion mode, of which 99 had significantly different amounts and covered 51 metabolic pathways. Principal component analysis and orthogonal patrial least squares discriminant analysis showed clear separation between two treatments, indicating different metabolic profiles. The different metabolites mainly included seven chemical categories, including amino acids and derivatives, nucleotides and derivatives, phenolic acids, organic acids, lipids, flavonoids and alkaloids. This study revealed the biological significance of these metabolites, which could be useful for further unexplored compounds and possible biological functions of A. cornea.

HS-SPME coupled with GC-MS for elucidating differences between the volatile components in wild and cultivated Atractylodes chinensis.

INTRODUCTION: Atractylodes chinensis is a Chinese herb that is used in traditional medicine; it contains volatile components that have enormous potential for pharmaceutical, food, and cosmetic applications. The destruction of wild resources demands significant improvement in the quality of artificial cultivation of Atractylodes chinensis. However, little is known about the compositional differences in the volatile substances derived from the wild and cultivated varieties of Atractylodes chinensis. OBJECTIVES: We aimed to evaluate the specific components of Atractylodes chinensis and analyse the similarities and differences between the volatile components and metabolic pathways in the wild and cultivated varieties. MATERIAL AND METHODS: Metabolomic analysis using gas chromatography-mass spectrometry (GC-MS) was employed following the extraction of volatile components from Atractylodes chinensis using headspace solid-phase microextraction (HS-SPME). RESULTS: A total of 167 volatile metabolites were extracted, and 137 substances were matched with NIST and Wiley databases. Among them, 76 compounds exhibited significant differences between the two sources; these mainly included terpenes, aromatics, and esters. KEGG enrichment analysis indicated that the differential metabolites were primarily involved in the biosynthesis of secondary metabolites, terpene biosynthesis, and limonene and pinene degradation; all these pathways have geranyl diphosphate (GDP) as the common link. CONCLUSION: The total content of volatile substances extracted from wild Atractylodes chinensis was 2.5 times higher than that from the cultured variety; however, each source had different dominant metabolites. This study underscores the necessity for protecting wild Atractylodes chinensis resources, while enhancing the quality of cultivated Atractylodes chinensis.

Medicinal, nutritional, and nutraceutical potential of Sparassis crispa s. lat.: a review.

Sparassis crispa is an edible mushroom exhibiting a wide range of medicinal properties. It is recognized for therapeutic value because of the high ß-glucan content in the basidiomes. The broad range of its reported curative effects include anti-tumour, anti-cancer, immune-enhancing, hematopoietic, anti-angiogenic, anti-inflammatory, anti-diabetic, wound-healing, antioxidant, anti-coagulant, and anti-hypertensive properties. However, most of the studies are conducted on immunomodulatory and anticancer activities. Besides this, it also exhibits anti-microbial properties due to the presence of sparassol. Technology is now available for the cultivation of S. crispa on coniferous sawdust. This review is an attempt to focus on its distribution, taxonomy, chemical composition, medicinal properties, potential applications, and artificial cultivation.

Analysis of Internal and External Microorganism Community of Wild Cicada Flowers and Identification of the Predominant Cordyceps cicadae Fungus.

The parasitoid fungus Cordyceps cicadae, whose fruiting bodies are known in China as "chan hua," literally "cicada flower," has been used as a traditional Chinese medicinal ingredient for centuries. However, systematic disclosure of the vital factors responsible for the formation of wild cicada flower is limited. Here, we determined the physicochemical properties of soil and simultaneously analyzed the diversities and the structures of microbial community inhabiting the coremia, sclerotia, and soil around wild cicada flowers through high-throughput sequencing. Our results indicated that cicada flower more preferentially occurred in acidic soil (pH 5.9) with abundant moisture content (MC), total nitrogen (TN), and organic matter (OM). The dominant fungal genera in soil mainly included Isaria, f__Clavariaceae_Unclassified, Umbelopsis, f__Chaetomiaceae_Unclassified, Mortierella, f__Sordariaceae_Unclassified, and Arcopilus. Among them, C. cicadae was the only fungus that was massively detected in both the coremia and sclerotia with abundance of 83.5 and 53.6%, respectively. Based on this, a C. cicadae strain named AH10-4 with excellent adenosine- and N 6-(2-hydroxyethyl)-adenosine (HEA)-producing capability was successfully isolated. However, to the aspect of bacteria, Burkholderia-Caballeronia-Paraburkholderia, Bacillus, Acidibacter, f__Xanthobacteraceae_Unclassified, and Candidatus_Solibacter were the dominant genera in soil. Pedobacter, f__Enterobacteriaceae_Unclassified, Pandoraea, Achromobacter, Stenotrophomonas, Burkholderia-Caballeronia-Paraburkholderia, and Chitinophaga were the dominant genera in the coremia and sclerotia. Notably, Burkholderia-Caballeronia-Paraburkholderia was the shared bacteria among them with high abundance of 3.1, 11.4, and 5.2% in the sclerotia, coremia, and soil, respectively. However, the possible role of these bacteria to the occurrence of cicada flower has been unclear to our knowledge. By analyzing the correlation between physicochemical properties and microbial community of soil, we found that MC, Fe, and Zn were significantly negatively correlated with soil Isaria and that Cu was significantly negatively correlated with most dominant soil bacterial genera. But Mg was significantly positively correlated with most dominant taxa. This study provides new insight into the formation mechanisms of cicada flower and may contribute to the large-scale cultivation of cicada flowers.

Different Cultivation Environments Affect the Yield, Bacterial Community and Metabolites of Cordyceps cicadae.

Cordyceps cicadae is an entomogenous fungus with important uses in traditional Chinese medicine. However, its wild resources have not met consumers' demand due to excessive harvesting practices. Artificial cultivation is therefore an important alternative, but research on cultivating C. cicadae in natural habitats has not been reported. In this study, we aimed to explore the viability of cultivating C. cicadae in a natural habitat, in the soil of Pinus massoniana forest. We assessed and compared the yield, metabolite contents and bacterial community composition of C. cicadae grown in the Antheraea pernyi pupae at different growth stages, and under different cultivation conditions, in the soil of a natural habitat and in sterile glass bottles. Our results showed that cultivating C. cicadae in a natural habitat is feasible, with up to 95% of pupae producing C. cicadae fruiting bodies. The content of nitrogen compounds (amino acids) in C. cicadae cultivated in a natural habitat was significantly higher than in glass bottles, while the yield and carbon compound (mannitol and polysaccharide) and nucleoside (cordycepin and adenosine) contents were lower. Different bacterial genera were enriched in C. cicadae at different growth stages and cultivation environments, and these bacterial genera were closely related to metabolites contents during growth. This study demonstrated the viability of a novel cultivation method of C. cicadae, which could be used as an alternative to wild stocks of this fungus. These findings provided new insights into the growth mechanism of C. cicadae and its interaction with soil microorganisms.

Artificial Cultivation Characteristics and Bioactive Effects of Novel Tropicoporus linteus (Syn. Phellinus linteus) Strains HN00K9 and HN6036 in Korea.

Phellinus strains were collected from different areas in Korea. Of them, the fast mycelial growing strains were artificially cultivated on the oak logs to produce fruiting body. The varieties, Phellinus linteus ASI26099 (Korea Sanghwang) and P. baumii PBJS (Jangsoo Sanghwang) were grown under the same conditions as controls. Their cultivating characteristics including mycelial colonization, pinhead formation, and fruiting body formation rate were investigated on the logs. Basidiocarps of Phellinus strains HN00K9, HN6036, and ASI26099 were concentrically zonate and shallowly sulcate, and dark chestnut showing typical characteristics of Tropicoporus linteus (synonyum: P. linteus, Inonotus linteus, polyporus linteus), which is distinguishably different to PBJS. HN00K9 showed the highest yield of fruiting body among the mushroom strains. The ß-glucan content in fruiting bodies of HN00K9 was 20% higher than those of other strains. Bioactive effects of polysaccharide samples from fruiting bodies of Phellinus strains, HN00K9, HN6036, ASI26099, and PBJS were assessed on cell viability and cytokine (IL-6 and TNF-α) inhibition and finally on anticancer to different human cancer cells.

The fungal community in non-rhizosphere soil of Panax ginseng are driven by different cultivation modes and increased cultivation periods.

Continuous cropping obstacles severely hindered the sustained development of the ginseng industry. Among the obstacles, an imbalance of soil microbiome community was considered one of the major culprits. The fungal community is an essential part of the soil microbiome community. Extensive characterization of the fungal community composition and variation during ginseng cultivation will help us understand the mechanism underlying continuous cropping obstacles. By using a high-throughput amplicon sequencing method, the non-rhizospheric fungal community of farmland cultivated ginseng of 2 years old (C2) and 5 years old (C5), understory wild ginseng of 15 years old (W15) and 35 years old (W35), fallow fields which have been abandoned for 10 (F10) years were characterized. Farmland cultivated ginseng and understory wild ginseng harbored distinct non-rhizospheric fungal communities, and extension of cultivation periods enlarged the fungal community difference between two cultivation modes. Extended cultivation periods significantly decreased the OTU richness and PD whole tree indices, and OTU number and cultivation periods were negatively correlated. Extension of cultivation periods led to an increased abundance of pathotrophs. Still, the increased abundance of pathotrophs may not be the leading cause of severe continuous cropping obstacles in farmland cultivated ginseng. Compared with understory wild ginseng, farmland cultivated ginseng had a lower abundance of symbiotrophs and a higher abundance of saprotrophs. This changed symbiotrophs/saprotrophs ratio may have some correlation with the severe continuous cropping obstacles that occurred in farmland cultivated ginseng. Fallowing on the fungal community of the non-rhizosphere soil was generally opposite of that of extension of ginseng cultivation periods. The impacts of farmland cultivation on the fungal community of the non-rhizosphere soil can last for decades, even if the following is practiced.

The changes of morphological and physiological characteristics in hemiparasitic Monochasma savatieri before and after attachment to the host plant.

BACKGROUND: Monochasma savatieri is an endangered hemiparasitic medicinal plant with a variety of antioxidant, antimicrobial and anti-inflammatory properties. Despite the urgent need to understand the parasitic biology of M. savatieri, parasite-host associations have long been neglected in studies of M. savatieri. METHODS: We conducted a pot cultivation experiment to analyze changes in the growth traits, physiological performance and anatomical structures of M. savatieri grown with the potential host Gardenia jasminoides E., before and after the establishment of the parasite-host association. RESULTS: Prior to the establishment of the parasite-host association, the presence of the host had no significant effect on the maximum root length, leaf indexes or total dry weight of M. savatieri seedlings, but had significant positive effect on seedling height, number of roots or number of haustoria. When it was continuously grown without a host, M. savatieri growth was rather slow. The establishment of the parasite-host association enhanced the growth of M. savatieri, and higher levels of photosynthetic pigments, increased antioxidant enzyme activity and lower malondialdehyde accumulation were observed in M. savatieri with an established parasite-host association. Furthermore, an analysis of the anatomical structures of M. savatieri showed that the establishment of the parasite-host association enabled better development of the seedling vegetative organs than that in seedlings without parasite-host associations. CONCLUSIONS: Our study demonstrates the physiological and anatomical changes that occurred in M. savatieri after connection with a host and suggests that the enhanced growth and development of M. savatieri were highly dependent on the parasite-host association.

Effects of one haustorium-inducing quinone DMBQ on growth and development of root hemiparasitic plant Monochasma savatieri / Efeitos de quinona indutor de haustório (DMBQ) no crescimento e desenvolvimento de plantas hemiparasitárias de raiz Monochasma savatieri

ABSTRACT: Monochasma savatieri Franch. ex Maxim is a perennial, parasitic herb used in traditional Chinese medicine and its wild resources have decreased sharply in recent years due to destructively harvesting and habitat destruction. Haustorium formation is a key event of parasites, but the concentrations of haustorium-inducing factors vary with species and cultivation conditions. In this study, we investigated the effects of the 2,6-dimethoxy-p-benzoquinone (DMBQ) concentration and cultivation density on the growth traits, haustorium formation and biomass of M. savatieri in the absence of a host plant. The results showed that both the DMBQ concentration and cultivation density regulated growth traits, haustorium formation and biomass in M. savatieri. The number of haustoria was significantly positively correlated with seedling height, maximum root length, the number of root tips and total dry weight. Membership function analysis revealed an overall greater increase in growth traits, haustorium formation and biomass when M. savatieri was treated with 10 μmol·L-1DMBQ and grew solitarily. These results offer an understanding of growth in M. savatieri influenced by the DMBQ concentration and cultivation density, which may aid in the establishment of a comprehensive cultivation system for M. savatieri or similar plants.

RESUMO: Monochasma savatieri Franch. O ex Maxim é uma erva parasitária aperene usada na medicina tradicional chinesa suas fontes diminuíram acentuadamente nos últimos anos devido à colheita destrutiva e à destruição de habitats e condições de sobrevivência no campo. Neste estudo, investigamos os efeitos da concentração de 2,6-dimetoxi-p-benzoquinona (DMBQ) e densidade de cultivo sobre as características de crescimento, formação de haustório e biomassa de M. savatieri na ausência de uma planta hospedeira. Os resultados mostraram que a concentração de DMBQ e a densidade de cultivo regularam as características de crescimento, a formação de haustório e a biomassa em M. savatieri. O número de haustórios foi significativamente correlacionado positivamente com a altura das plântulas, comprimento máximo das raízes, número de pontas das raízes e peso seco total. Revelou também um aumento geral nas características de crescimento, formação de haustório e biomassa quando M. savatieri foi tratado com 10 μmol • L-1DMBQ e cresceu solitariamente. Esses resultados oferecem uma compreensão do crescimento de M. savatieri influenciado pela concentração de DMBQ e densidade de cultivo, o que pode ajudar no estabelecimento de um sistema abrangente de cultivo para plantas similares de M. savatierior.

Artificial Cultivation of the Chinese Cordyceps From Injected Ghost Moth Larvae.

The Chinese cordyceps, regarded as the 'Himalayan Viagra', is highly valued for its medicinal benefits. The decline of its yield due to over-exploitation and increased market demand have stimulated efforts to artificially cultivate Chinese cordyceps for over half a century. However, successful cultivation of Chinese cordyceps through caterpillar infection by the fungus Ophiocordyceps sinensis (Berk.) and the induction of the fruiting body from each mummified cadaver remains difficult for its complex life cycle. Herein, we report the developmental dynamics of hyphal bodies in hemolymph of injected Thitarodes xiaojinensis (Tu, Ma & Zhang) larvae and the success in artificial cultivation of sexual fruiting bodies from the mummified cadavers in the low-altitude area. We find that not only the numbers of hyphal bodies but also the conversion of hyphal bodies into hyphae played important roles in the mummification of the injected larvae. This cultivation will be beneficial for sustainable utilization of natural resources and provides the possibility for further research on the mechanism of the interaction between pathogenic fungus and host insect.

A breakthrough in the artificial cultivation of Chinese cordyceps on a large-scale and its impact on science, the economy, and industry.

Chinese cordyceps, an entity of the Chinese caterpillar fungus (Ophiocordyceps sinensis, syn. Cordyceps sinensis) that parasitizes ghost moth larvae, is one of the best known traditional Chinese medicines and is found exclusively on the Tibetan Plateau with limited natural resources. Although the fungus O. sinensis can grow on artificial substrates and the ghost moth has been successfully reared, the large-scale artificial cultivation of Chinese cordyceps has only recently been accomplished after several decades of efforts and attempts. In this article, research progress related to this breakthrough from living habitats, the life history of the fungus, its host insect, fungal isolation and culture, host larvae rearing, infection cycle of the fungus to the host, primordium induction, and fruiting body development have been reviewed. An understanding of the basic biology of O. sinensis, its host insect and the simulation of the Tibetan alpine environment resulted in the success of artificial cultivation on a large scale. Practical workshop production has reached annual yields of 2.5, 5, and 10 tons in 2014, 2015, and 2016, respectively. There was no difference in the chemical components detected between the cultivated and natural Chinese cordyceps. However, the artificial cultivation system can be controlled to avoid heavy metal contamination and results in high-quality products. Although omics studies, including genomic, transcriptomic, proteomic, and metabolomic studies, have helped to understand the biology of the fungus, the success of the artificial cultivation of the Chinese cordyceps is clearly a milestone and provides the possibility for research on the in-depth mechanisms of the interaction between the fungus and host insects and their adaptation to the harsh habitats. This cultivation will not only result in a large industry to alleviate the pressure of human demand but also protect the limited natural resources for sustainable utilization.

Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis.

To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield, it is important to identify molecular fingerprint of dominant O. sinensis. In the present study, we collected 3 batches of industrially cultivated O. sinensis product with higher yield than the others and compared their internal transcribed spacer (ITS) sequences with the wild and the reported. The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O. sinensis collected. The ITS sequences of O. sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship. The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples. The results showed that all wild and cultivated samples were identified as O. sinensis and all sequences were divided into seven phylogenetic groups in the tree. Those groups were precisely distributed on the map and the process of their system evolution was clearly presented. The three cultivated samples were clustered into two dominant groups, showing the correlation between the industrially cultivated samples and the dominant wild samples, which can provide references for its optimized breeding in the future.

Artificial cultivation of true morels: current state, issues and perspectives.

Morels (Morchella, Ascomycota), which are some of the most highly prized edible and medicinal mushrooms, are of great economic and scientific value. Morel cultivation has been a research focus worldwide for more than 100 years, and the outdoor cultivation of morels has succeeded and expanded to a large scale in China in recent years. In this study, we review the progress in recent research regarding the life cycle and reproductive systems in the genus Morchella and the current state of outdoor cultivation. Sclerotia formation and conidia production are two important phases during the life cycle. The morel species cultivated commercially in America is M. rufobrunnea based on molecular phylogenetic analysis. The species currently cultivated in China are black morels, including M. importuna, M. sextalata and M. eximia. The field cultivation of morels expanded in the majority of the provinces in China with a yield of fresh morels of 0-7620 kg per ha. The key techniques include spawn production, land preparation and spawning, the addition of exogenous nutrition, fruiting management and harvesting. The application of exogenous nutrition is the most important breakthrough in the field of morel cultivation, but the mechanism remains unclear. It was estimated that the total amount of field cultivated fresh morels was ∼500 t in 2015-2016. We also discuss the potential issues remaining in the current literature and suggest directions for future studies.

Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis / 中国天然药物

To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield, it is important to identify molecular fingerprint of dominant O. sinensis. In the present study, we collected 3 batches of industrially cultivated O. sinensis product with higher yield than the others and compared their internal transcribed spacer (ITS) sequences with the wild and the reported. The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O. sinensis collected. The ITS sequences of O. sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship. The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples. The results showed that all wild and cultivated samples were identified as O. sinensis and all sequences were divided into seven phylogenetic groups in the tree. Those groups were precisely distributed on the map and the process of their system evolution was clearly presented. The three cultivated samples were clustered into two dominant groups, showing the correlation between the industrially cultivated samples and the dominant wild samples, which can provide references for its optimized breeding in the future.

Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis / 中国天然药物

To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield, it is important to identify molecular fingerprint of dominant O. sinensis. In the present study, we collected 3 batches of industrially cultivated O. sinensis product with higher yield than the others and compared their internal transcribed spacer (ITS) sequences with the wild and the reported. The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O. sinensis collected. The ITS sequences of O. sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship. The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples. The results showed that all wild and cultivated samples were identified as O. sinensis and all sequences were divided into seven phylogenetic groups in the tree. Those groups were precisely distributed on the map and the process of their system evolution was clearly presented. The three cultivated samples were clustered into two dominant groups, showing the correlation between the industrially cultivated samples and the dominant wild samples, which can provide references for its optimized breeding in the future.

[Ecological suitability assessment for Notopterygium plants transplanting along altitudinal gradients].

A comprehensive field research had been focused on growing status, underground biomass and active constituents of Notopterygium incisum and N. franchetii to evaluate the ecological suitability and appropriate cultivation zones by growing the two species seedlings along different elevation gradient. The results showed that compared to the survival rate and underground biomass, the beneficial altitude region to N. incisum was ranged from 2 600 m to 4 100 m, while N. franchetii required a lower altitude which ranges from 1 700 m to 3 600 m. For the active constituent contents, the values were higher in the range of 2 600 to 3 600 m for N. incisum, but for N. franchetii, the range was form 1 700 to 3 600 m. This result provides instructional guidance and scientific basis for artificial cultivation of N. incisum and N. franchetii.

Plant Resource Availability of Medicinal Fritillaria Species in Traditional Producing Regions in Qinghai-Tibet Plateau.

The genus Fritillaria (Liliaceae) comprises of ~140 species of bulbous perennials, which are distributed in the temperate zone of the Northern Hemisphere. Fritillaria species have attracted much attention because of their commercial value, partly as ornamental plants but principally as a source of material for use in traditional medicine. The use of Fritillaria extracts is well established in some countries in Eastern Europe (e.g., Turkey), and Asia (e.g., China, Japan). In traditional Chinese medicine, the medicinal Fritillaria species is called Bulbus Fritillariae Cirrhosae (BFC), which has been used as a traditional medicine for thousands of years. However, to the best of our knowledge, there are no reports on resource investigation of plants of BFC in the last ten years. In this study, we chose 32 traditional producing regions in Qinghai-Tibet Plateau to perform an investigation on resource availability of BFC. In five sites we did not find any plants of BFC. Results show that the average number of the plants of BFC per quadrat in 26 sites was less than 7, and the average resource density was <22 mg/m2. Habitat types and plant morphology of BFC plants were recorded. Our investigation shows that the area for artificial cultivation of BFC is larger than 400 hm2 and productivity was higher than 180 t. In addition, the total alkaloid contents of samples from cultivated bases and plantations are higher than that from wild fields. This study suggests that the wild populations of BFC are still at the risk of depletion. Artificial cultivation of BFC might be an important way to resolve the current contradiction between resource protection and resource utilization. In addition, identifying the closest European relatives of the Fritillaria species used in traditional medicine may resolve this contradiction.