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Diversity, Chemical Constituents, and Biological Activities of Endophytic Fungi Isolated From Ligusticum chuanxiong Hort.

ABSTRACT: The objective of this study was to evaluate the diversity of endophytic fungi of different parts of Ligusticum chuanxiong Hort (CX) and further characterize their biological activities and identify chemical compounds produced by these endophytic fungi. A total of 21 endophytic fungi were isolated and identified from CX. Penicillium oxalicum, Simplicillium sp., and Colletotrichum sp. were identified as promising strains by the color reaction. Comparing different organic extracts of the three strains, it was observed that the ethyl acetate extract of Penicillium oxalicum and Simplicillium sp. and the n-butanol extract of Colletotrichum sp. showed significant antioxidant and antibacterial activities. The ethyl acetate extracts of Penicillium oxalicum had outstanding antioxidant and antibacterial effects, and its radical scavenging rates for ABTS and DPPH were 98.43 ± 0.006% and 90.11 ± 0.032%, respectively. At the same time, their IC₅₀ values were only 0.18 ± 0.02 mg/mL and 0.04 ± 0.003 mg/mL. The ethyl acetate extract of Penicillium oxalicum showed MIC value of only 0.5 mg/mL against Escherichia coli and Staphylococcus aureus. By liquid chromatography-mass spectrometry (LC-MS), we found that Penicillium oxalicum could produce many high-value polyphenols, such as hesperidin (36.06 μmol/g), ferulic acid (1.17 μmol/g), and alternariol (12.64 μmol/g), which can be a potential resource for the pharmaceutical industry. In conclusion, these results increase the diversity of CX endophytic fungi and the antioxidant and antibacterial activities of their secondary metabolites.

SUBMITTER: Tang Z

PROVIDER: S-EPMC8636053 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:"Breeding on mountains, cultivation in dam areas" is a unique propagation method for the vegetatively propagated plant Ligusticum chuanxiong, including two transplants between the mountain and the dam area. It is well known that the environment can influence the endophytic community structure of plants. However, the change of host endophytic flora caused by transplanting in different places and its influence on asexual reproduction are still poorly understood. We carried out three cycles of cultivation experiments on L. chuanxiong and collected stem nodes (LZ), immature rhizomes (PX), medicinal rhizomes (CX), and rhizosphere. High-throughput sequencing was performed to analyze the endophytic fungi in all samples. We observed that the diversity and richness of endophytic fungi in L. chuanxiong increased as a result of transplanting cultivation from dam areas to mountains. Local transplantation caused minor changes in the endophytic fungus structure of L. chuanxiong, while remote transplantation caused significant changes. Compared with LZ after breeding in the dam area, the LZ after breeding on mountains has more abundant Gibberella, Phoma, Pericona, Paraphoma, and Neocosmospora. The regular pattern of the relative abundance of endophytic fungi is consistent with that of the fungus in the soil, while there are also some cases that the relative abundance of endophytic fungi is the opposite of that of soil fungi. In addition, there is a significant correlation among certain kinds of endophytic fungi whether in the soil or the plants. We have isolated more gibberellin-producing and auxin-producing fungi in the LZ cultivated in the mountains than that in the LZ cultivated in the dam area. The results of pot experiments showed that the three fungi isolated from LZ cultivated in mountainous areas can promote the development of shoots, stem nodes, and internodes of LZ, and increase the activity of plant peroxidase, catalase, phenylalanine ammonia lyase, and other enzymes. We can conclude that transplantation leads to the recombination of the host endophytic fungus, the more significant the difference in the environment is, the greater the reorganization caused by transplanting. Reorganization is determined by the soil environment, hosts, and the interaction of microorganisms. Remote transplantation is a crucial opportunity to reshuffle the micro-ecological structure of the asexual reproduction of plants, and regulate the growth, development, and resistance of plants, and prevent germplasm degradation caused by asexual reproduction.

Project description:BackgroundLong-term asexual reproduction can easily lead to the degradation of plant germplasm, serious diseases and insect pests, reduction of production and even catastrophic crop failure. "Mountain Breeding and Dam Cultivation" is the main cultivation mode of Ligusticum chuanxiong Hort., which successfully avoided the germplasm degradation caused by long-term asexual reproduction. The recombination of endophytic fungi of L. chuanxiong caused by off-site transplantation was considered to be an important reason for its germplasm rejuvenation. However, whether bacteria have the same regularity is not yet known.MethodsIn this study, we carried out the experiment of cultivating propagation materials of L. chuanxiong in different regions and transplanting them to the same region. High-throughput sequencing was performed to analyze the bacterial communities in L. chuanxiong and its soil.ResultsThe results showed that after transplanting, the plant height, tiller number, fresh weight, etc. of L. chuanxiong in mountainous areas were significantly higher than those in dam areas. At the same time, significant changes had taken place in the endophytic bacteria in reproductive material stem nodes (Lingzi, abbreviated as LZ). The diversity and abundance of bacteria in dam area LZ (YL) are significantly higher than those in mountainous area LZ (ML). The relative abundance of bacteria such as Xanthobacteraceae, Micromonosporaceae, Beijerinkiaceae, Rhodanobacteria, in ML is significantly higher than YL, mainly classified in Proteobateria and Actinobacteriota. In addition, the abundance advantage of Actinobacteriota still exists in MY (underground mature rhizomes obtained by ML). Meanwhile, the bacterial community was different in different area of transplanting. The diversity of bacterial communities in dam soil (YLS) is significantly higher than that in mountain soil (MLS). MLS had more Acidobacteriota than YLS. Comparative analysis showed that 74.38% of bacteria in ML are found in MLS, and 87.91% of bacteria in YL are found in YLS.ConclusionsWe can conclude that the community structure of endophytic bacteria recombined after the transplantation of L. chuanxiong, which was related to the bacterial community in soils. Moreover, after transplanting in mountainous areas, LZ accumulated more potentially beneficial Actinobacteriota, which may be an important reason for promoting the rejuvenation of germplasm in L. chuanxiong. However, this hypothesis requires more specific experiments to verify. This study provided a new idea that off-site transplanting may be a new strategy to restore vegetative plant germplasm resources.

Project description:The dried tuber of Alisma orientale (Sam.) Juzep. (AOJ) is a traditional Chinese medicine with high medicinal value. The endophytic fungi of medicinal plants are a treasure house of natural compounds. However, there is a lack of research on the diversity and biological activity of endophytic fungi of AOJ. In this study, high-throughput sequencing technology was used to study the diversity of endophytic fungi in the roots and stems of AOJ, and endophytic fungi with a high output of phenols and flavonoids were screened by chromogenic reaction, and the antioxidant and antibacterial activities and chemical constituents of crude extracts of their fermentation broth were studied. A total of 3,426 amplicon sequence variants (ASVs) belonging to 9 phyla, 27 classes, 64 orders, 152 families, and 277 genera were identified from AOJ. There were significant differences in the endophytic fungal communities of AOJ roots and stems, as well as in the endophytic fungal communities of triangular AOJ and circular AOJ. In addition, 31 strains of endophytic fungi were isolated from AOJ, of which 6 strains had good antioxidant and antibacterial activities. The crude extract of YG-2 had the strongest free radical scavenging ability and bacteriostatic ability, and its IC50 DPPH, IC50 ABTS, and IC50⋅OH values were 0.009 ± 0.000 mg/mL, 0.023 ± 0.002 mg/mL, and 0.081 ± 0.006 mg/mL, respectively. The results of LC-MS showed that the main component of the crude extract of YG-2 was caffeic acid (10.12 μmol/g). Overall, the results of this study preliminarily elucidated the diversity and community composition of endophytic fungi of AOJ, indicating that AOJ endophytic fungi have abundant secondary metabolites and good antioxidant and antibacterial activities. This study provides an important reference for further research, development and utilization of AOJ endophytic fungi and a theoretical basis for the further development of the endophytic fungus YG-2 (Chaetomium globosum) as a source of antioxidants.

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Diversity, Chemical Constituents, and Biological Activities of Endophytic Fungi Isolated From Ligusticum chuanxiong Hort.

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