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The microecology of endophytic fungi in special habitats, such as the interior of different tissues from a medicinal plant, and its effects on the formation of metabolites with different biological activities are of great importance. However, the factors affecting fungal community formation are unclear. This study is the first to utilize "mini-community" remodeling to understand the above phenomena. First, high-throughput sequencing technology was applied to explore the community composition and diversity of endophytic fungi in the above-ground tissues (Ea) and below-ground tissues (Eb) of Ephedra sinica. Second, fungi were obtained through culture-dependent technology and used for "mini-community" remodeling in vitro. Then, the effects of environmental factors, partner fungi, and plant tissue fluid (internal environment) on endophytic fungal community formation were discussed. Results showed that environmental factors played a decisive role in the selection of endophytic fungi, that is, in Ea and Eb, 93.8% and 25.3% of endophytic fungi were halophilic, respectively, and 10.6% and 60.2% fungi were sensitive to high temperature (33 °C), respectively. Meanwhile, pH had little effect on fungal communities. The internal environment of the plant host further promoted the formation of endophytic fungal communities., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2023 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Ephedra sinica is one of the most famous Chinese medicinal plants. The insufficient supply of wild resources has led to the increased use of cultivated products. However, the related medicinal quality differs significantly. Although the influence of external environment on the quality of E. sinica has been studied, the impact of endophytic microbes on it remains vague. This study characterized differential metabolites and microbial community compositions in wild and cultivated E. sinica by combining metabolomics with microbiomics, and explored the effect of endophytes on the formation of differential metabolites further. The results showed that the difference in quality between wild and cultivated E. sinica was mainly in the productions of alkaloids, flavonoids, and terpenoids. The associated endophytes had special compositional characteristics. For instance, the distribution and abundance of dominant endophytes varied between wild and cultivated E. sinica. Several endophytes had significant or highly significant correlations with the formations of ephedrine, pseudoephedrine, D-cathinone, methcathinone, coumarin, kaempferol, rhamnetin, or phenylacetic acid. This study will deepen our understanding of the plant-endophyte interactions and provide a strategy for the quality control of E. sinica products., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Ephedra sinica, a well-known Chinese medicinal plant, is characterized as having the opposite medicinal effect among its root and stem. However, there is a lack of understanding to differentiate the active components present in the root and stem of E. sinica, as well as the molecular mechanisms underlying the formation of the differential compounds, which has significantly hampered the further development and utilization of E. sinica resource. In this study, forty-five differential metabolic markers are affiliated to alkaloids, flavonoids, terpenoids, and organic acids between root and stem of E. sinica, and sixty genes of key enzymes are involved in their biosynthesis distributed in metabolic pathway branches such as phenylalanine metabolism, flavonoid biosynthesis and phenylpropane biosynthesis, based on combination non-targeted metabolome with transcriptome technologies. The finding revealed that the expression activity changes of these enzyme genes had a direct impact on the distinction of differential metabolic markers in the root and stem of E. sinica. This study will help to understand the molecular mechanism of the differentiation and biosynthesis of the primary active metabolites in the root and stem of E. sinica, providing a theoretical foundation for its quality control and promotion in cultivation., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Fungal endophytic communities and metabolite profiles in the aerial and underground parts of plants differ exerting complex influences on each other through mechanisms that largely remain unknown. Ephedra sinica is a model for studying the interactions between endophytic fungi in different plant tissues that exhibit contrasting pharmacological activities. In this study, the endophytic fungal community and metabolites in the stems (Ea) and roots (Eb) of E. sinica were systematically investigated using metagenomic and metabolomic approaches, and their relationships were further analyzed. Results showed that OTU48 and OTU30589 were endophytic fungi shared by Ea and Eb, respectively. The genera of Phyllosticta fungi were specifically and abundantly present in Ea, whereas Talaromyces, Aporospora, and Aspergillus fungi were specific and abundant in Eb. A total of 17 significant differential metabolites were observed between Ea and Eb. Ephedrine and methylephedrine were mainly present in Ea, whereas ephedrannin A and mahuannin A were present in Eb. Nine endophytic fungi significantly (P < 0.05 or 0.01) correlated with several differential metabolites. The results of this study further substantiate the differential formation of secondary metabolites in specific plant tissues. [ABSTRACT FROM AUTHOR]

Ephedra sinica is widely used in the treatment of diseases due to its rich and diverse pharmacological effects. The quality of E. sinica is various in different years. However, the effect of endophytes on the accumulation of metabolites of E. sinica in different years is still obscure. In this study, the relationship between the ecological succession of bacterial endophytes and metabolite accumulation of E. sinica grown in different years were generally investigated based on 1H NMR and 16 s rDNA sequencing. The results showed that 38 differential marker metabolites existed in E. sinica grown in different years. The distribution and abundance of bacterial endophytes changed during the growth of cultivated E. sinica , and there was remarkable correlation between the dominant endophytes and differential metabolites. This study provides a critical reference in evaluating and monitoring the quality of E. sinica in the process of cultivation, and establish a solid foundation for the use of species interactions to improve the quality of E. sinica in different years. [Display omitted] • The first attempt with metabonomics on Ephedra sinica during the cultivation of E. sinica. • A total of 38 differential compounds including primary and secondary metabolites were obtained. • The succession characteristics of bacterial endophytes of E. sinica in different years was provided. • The phylum Proteobacteria plays an important role in the growth of E. sinica. • The correlation between differential constituents and dominant endophytes was analyzed. [ABSTRACT FROM AUTHOR]