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Root-associated endophytic fungi modulate endogenous auxin and cytokinin levels to improve plant biomass and root morphology of trifoliate orange

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Horticultural Plant Journal 2023年第3期9卷 463-472页

作者： Ruicheng Liu Liu Yang Yingning Zou Qiangsheng Wu College of Horticulture and Gardening Yangtze UniversityJingzhouHubei434025China College of Arts and Sciences Yangtze UniversityJingzhouHubei434020China

Root-associated endophytic fungi like Serendipita indica and arbuscular mycorrhizal(AM) fungi can improve plant growth and root construction, but the potential mechanism is unclear. In this study, Funneliformis mosseae(an AM fungus) and S. indica, singly or in combination were inoculated into trifoliate orange(Poncirus trifoliata) seedlings, to assess changes in biomass and root morphological traits, coupled with auxins and cytokinins concentrations in leaves and roots and the expression of auxin synthesis and transporter protein genes. After 20 weeks of inoculation with these fungi, shoot and root biomass, root total length, taproot length, average diameter, surface area, volume, and the number of lateral roots in 1st-, 2nd-, and 3rd-order were improved, and S. indica showed a relatively greater effect than F. mosseae and dual inoculation. endophytic fungal inoculation also significantly increased the concentration of indoleacetic acid, indole butyric acid, trans-zeatin,dihydrozeatin, and isopentenyl adenine in leaves and roots, whilst F. mosseae and S. indica exhibited relatively greater effects on leaves and roots, respectively. Correlation analysis revealed that both biomass and root morphological traits(except root projected area) were significantly positively associated with endogenous auxins and cytokinins. In addition, the inoculated plants recorded comparatively higher expression levels of indoleacetic acid synthesis genes(PtTAA1, PtTAR2, PtYUC3, PtYUC4, PtYUC6, and PtYUC8) and indoleacetic acid transporter protein genes(PtAUX1, PtLAX1, PtLAX2, PtLAX3, PtPIN1, PtPIN3, PtPIN4, PtABCB1, and PtABCB19) than the non-inoculated plants, among which F. mosseae and S. indica showed better effects in leaves and roots, respectively. These results suggest that root-associated endophytic fungi improved plant growth and root architecture, which were associated with changes in endogenous auxins and cytokinins.

关键词： Citrus endophytic fungi Hormone Mycorrhiza IAA

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endophytic fungi harbored in Cannabis sativa L.:diversity and potential as biocontrol agents against host plant-specific phytopathogens

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Fungal Diversity 2013年第3期60卷 137-151页

作者： Parijat Kusari Souvik Kusari Michael Spiteller Oliver Kayser Department of Biochemical and Chemical Engineering Chair of Technical BiochemistryTU DortmundEmil-Figge-Str.6644227 DortmundGermany Institute of Environmental Research(INFU)of the Faculty of Chemistry Chair of Environmental Chemistry and Analytical ChemistryTU DortmundOtto-Hahn-Str.644221 DortmundGermany

The objective of the present work was isolation,phylogenetic characterization,and assessment of biocontrol potential of endophytic fungi harbored in various tissues(leaves,twigs,and apical and lateral buds)of the medicinal plant,Cannabis sativa L.A total of 30 different fungal endophytes were isolated from all the plant tissues which were authenticated by molecular identification based on rDNA ITS sequence analysis(ITS1,5.8S and ITS2 regions).The Menhinick’s index revealed that the buds were immensely rich in fungal species,and Camargo’s index showed the highest tissue-specific fungal dominance for the twigs.The most dominant species was Penicillium copticola that could be isolated from the twigs,leaves,and apical and lateral buds.A detailed calculation of Fisher’s log series index,Shannon diversity index,Simpson’s index,Simpson’s diversity index,and Margalef’s richness revealed moderate overall biodiversity of C.sativa endophytes distributed among its tissues.The fungal endophytes were challenged by two host phytopathogens,Botrytis cinerea and Trichothecium roseum,devising a dual culture antagonistic assay on five different media.We observed 11 distinct types of pathogen inhibition encompassing a variable degree of antagonism(%)on changing the media.This revealed the potential chemodiversity of the isolated fungal endophytes not only as promising resources of biocontrol agents against the known and emerging phytopathogens of Cannabis plants,but also as sustainable resources of biologically active and defensive secondary metabolites.

关键词： Cannabis sativa endophytic fungi Fungal diversity Antagonism Botrytis cinerea Trichothecium roseum

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Levels of rhizome endophytic fungi fluctuate in Paris polyphylla var.yunnanensis as plants age

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Plant Diversity 2017年第1期39卷 60-64页

作者： Tao Liu Alex Greenslade ShengchaoYang Yunnan Research Center on Good Agricultural Practice for Dominant Chinese Medicinal Materials Yunnan Agricultural University Kunming 650201China Biological Chemistry and Crop Protection Rothamsted Research Harpenden UK

Paris polyphylla var. yunnanensis is an important medicinal plant with abundant saponins that are widely used in the pharmaceuticals industry. It is unclear why the levels of active ingredients increase as these plants age. We speculated that the concentrations of those components in the rhizomes are mediated by fungal endophytes. To test this hypothesis, we took both culture-dependent and-independent(metagenomics) approaches to analyze the communities of endophytic fungi that inhabit those rhizomes in plants of different age classes(four, six, and eight years old). In all, 147 isolates representing 18 fungal taxa were obtained from 270 segments(90 per age class). Based on morphological and genetic characteristics, Fusarium oxysporum(46.55% frequency of occurrence) was the predominant endophyte,followed by Leptodontidium sp.(8.66%) and Trichoderma viride(6.81%). Colonization of endophytic fungi was maximized in the eight-year-old rhizomes(33.33%) when compared with four-year-old(21.21%) and six-year-old(15.15%) rhizomes. Certain fungal species were present only at particular ages. For example,Alternaria sp., Cylindrocarpon sp., Chaetomium sp., Paraphaeosphaeria sporulosa, Pyrenochaeta sp., Penicillium swiecickii, T. viride, and Truncatella angustata were found only in the oldest plants. Analysis of(metagenomics) community DNA extracted from different-aged samples revealed that, at the class level,the majority of fungi had the highest sequence similarity to members of Sordariomycetes, followed by Eurotiomycetes and Saccharomycetes. These results were mostly in accord with those we obtained using culture methods. Fungal diversity and richness also changed over time. Our investigation is the first to show that the diversity of fungi in rhizomes of P. polyphylla var. yunnanensis is altered as plants age, and our findings provide a foundation for future examinations of useful compounds.

关键词： Medicinal plants endophytic fungi Paris polyphylla var.yunnanensis

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Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第5期12卷 385-392页

作者： Jin-long CUI Shun-xing GUO Pei-gen XIAO Institute of Medicinal Plant Development Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100193 China

The purpose of this study was to isolate and characterize endophytic fungi from the stem tissue which can produce fragrant ingredients in Aquilaria sinensis(also called agarwood) to determine their antitumor and antimicrobial activities.Twenty-eight fungal endophytes were isolated from agarwood by strict sterile sample preparation and were classified into 14 genera and 4 taxonomic classes(Sordariomycetes,Dothideomycetes,Saccharomycetes,and Zy-gomycetes) based on molecular identification.Of the 28 isolates,13(46.4%) showed antimicrobial activity against at least one of the test strains by the agar well diffusion method,and 23 isolates(82.1%) displayed antitumor activity against at least one of five cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay.The diameters of inhibition zones of YNAS07,YNAS14,HNAS04,HNAS05,HNAS08,and HNAS11 were equal to or higher than 14.0 mm against Staphylococcus aureus,Escherichia coli,Bacillus subtilis,B.subtilis,Aspergillus fumigatus,and B.subtilis,respectively.The inhibition rates of YNAS06,YNAS08,and HNAS06 were not less than 60% to 293-T,293-T,and SKVO3 cells,respectively.These results suggest that the endophytic fungi associated with agarwood will provide us with not only useful micro-ecological information,but also potential antimicrobial and anti-tumor agents.

关键词： Agar diffusion method Agarwood Antimicrobial bioactivity Antitumor bioactivity endophytic fungi

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Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.)

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Asian Pacific Journal of Tropical Medicine 2017年第6期10卷 635-640页

作者： Minarni I Made Artika Heddy Julistiono Nurliani Bermawie Eny Ida Riyanti Hasim Akhmad Endang Zainal Hasan Department of Biochemistry Faculty of Mathematics and SciencesBogor Agricultural UniversityDarmaga CampusBogor 16680Indonesia Research Center for Biology The Indonesian Institute of Sciences(LIPI)Cibinong-Bogor Indonesia Research Institute of Medicinal and Aromatic Plants Agricultural Research AgencyBogor-Indonesia Center for Biotechnology and Genetic Resources of Agriculture Agricultural Research AgencyBogor-Indonesia

Objective: To analyze anticancer activity of an ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.). Methods: Anticancer activity of fungal extracts was determined by observing its toxicity against MCF-7(Michigan Cancer Foundation-7) cells in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay method. At an extract concentration of 100 μg/m L, 4 isolates out of 12 showed high activity against the cancer cell growth. The four isolates were then selected for further IC50 determination, by measuring the inhibition of cancer cell proliferation at extract concentration of 25 μg/m L, 50 μg/m L, 100 μg/m L, 200 μg/m L and 400 μg/m L. Results: Results showed that isolate Sir-G5 had the highest anticancer activity with an IC50 of 19.20 μg/m L. The best isolates were screened again using a normal cell(Chang cells) to determine its toxicity against normal cells. Results indicated that the extracts do not affect the proliferation of normal cells. Molecular identification showed that the fungal isolate Sir-G5 has a close relationship with Phomopsis sp. Conclusions: The endophytic fungi isolated from soursop leaf has the potential to be used as a source of anticancer agents.

关键词： Annona muricata L Anticancer MCF-7 endophytic fungi Soursop leaf

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Identification and Bioactivities of Secondary Metabolites Derived from endophytic fungi Isolated from Ethnomedicinal Plants of Tujia in Hubei Province:A Review

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Natural Products and Bioprospecting 2021年第2期11卷 185-205页

作者： Ke Ye Hong-Lian Ai Ji-Kai Liu School of Pharmaceutical Science South-Central University for NationalitiesWuhan 430074HubeiChina

Tujia is a national minority,inhabiting in the mountainous Wuling area in China.Since 1978,Tujia medicine has been studied,summarized and developed,leading to numerous achievements by Chinese researchers,such as the publishing of approximately 30 monographs of Tujia medicine.These publications are focused on summarizing and improving the theory of Tujia medicine and developing clinical therapies from this system of medicine.The shortage of natural medicinal plants used in Tujia medicine has created the need to discover new resources to replace them and protect endangered natural plant species.endophytic fungi are one of the conservation options,are considered a source of new bioactive natural products,and are a renewable and inexhaustible source of new drugs and agrochemicals.This review summarizes 260 compounds from endophytic fungi that have been previously isolated from the medicinal plants of Tujia.These compounds include steroids,terpenoids,meroterpenoids,polyketides,alkaloids,peptides,aliphatic compounds,aromatic compounds,and heterocyclic compounds.

关键词： Tujia medicine endophytic fungi Secondary metabolites Bioactivities

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A pilot study of antioxidant potential of endophytic fungi from some Sudanese medicinal plants

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Asian Pacific Journal of Tropical Medicine 2015年第9期8卷 689-692页

作者： Afra Khiralla Ietidal Mohamed Justinne Thomas Beno t Mignard Rosella Spina Sakina Yagi Dominique Laurain-Mattar Universit ede Lorraine SRSMC UMR 7565 BP 70239F-54506 Vandoeuvre-les-Nancy France CNRS SRSMC UMR 7565 BP 70239F-54506 Vandoeuvre-les-Nancy France Botany Department Faculty of Science University of KhartoumSudan Plant Advanced Technologies PAT c/o ENSAIA – INPLFrance

Objective: To evaluate the total phenolic content and total antioxidant capacity of ethyl acetate extracts of 21 endophytic fungi isolated from five Sudanese medicinal plants: Calotropis procera, Catharanthus roseus, Euphorbia prostrate, Vernonia amygdalina and Trigonella foenumgraecum. Methods: Crude extracts of endophytic fungi and their host plants were tested by classical Folin-Ciocalteu colorimetric method to determine the total phenolic content, also total antioxidant capacity was estimated using 1,1-diphenyl-2-picrylhydrazyl free radical scavenging in vitro method. Results: Among the endophytes, endophytic fungus Aspergillus sp. from Trigonella foenum-graecum seeds demonstrated the highest both total phenolic content in term of gallic acid equivalent(GAE) [(89.9±7.1 mg GAE/g)] and antioxidant activity for 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay [IC50:(18.0±0.1 μg/m L)]. A high positive linear correlation(R2=0.999 1) was found between total antioxidant capacity and total phenolic content of endophytic fungi isolated from Vernonia amygdalina. Conclusions: The present study revealed that some endophytic fungi from the five Sudanese medicinal plants could be a potential source of novel natural antioxidant compounds.

关键词： Sudanese medicinal plants endophytic fungi Antioxidant activity Total phenolic content

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Purification of four strains of endophytic fungi from Astragalus and their optimized liquid fermentations

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Journal of Forestry Research 2014年第3期25卷 701-706页

作者： Wei Ma Xiubo Liu Jiao Jiao Leiming Zhang Weichao Ren Ling Ma Xiangjun Kong Ning Zhang Xiwu Zhang College of Pharmacy Heilongjiang University of Chinese Medicine State Key Laboratory of Tree Genetics and Breeding Northeast Forestry University College of Forestry Northeast Forestry University The First Affiliated Hospital of Qiqihar Medical College

This study was designed to isolate endophytic fungi from A. mongholicus （growing in northeast China） to determine whether they can produce bioactive metabolites. Four strains of endophytic fungi （strains 16, 17, 23 and 75） were successfully isolated from A. mongholicus using the surface disinfection method. According to ITS-rDNA sequences analysis, strains 16 and 75 were identified as Fusarium oxysporum, and strains 17 and 23 were identified as Bionectria ochroleuca. We applied the Box-Behnken design （BBD） to optimize the liquid fermentation conditions and obtain the maximum cell dry weight （CDW） yield. Opti-mal parameters were obtained under the following experimental condi-tions：temperature of 28＆#176;C, potato dextrose agar （PDA） liquid medium of 80 mL and rotation speed of 150 rpm. The four isolated endophytic fungi did not produce astragalosides I-IV, flavonoids or polysaccharides. Iso-lation of additional species of endophytic fungi from A. mongholicus and determination of their capacity to produce biologically active substances are subjects in need of further research.

关键词： Astragalus endophytic fungi isolation purification liquid fermentation

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Factors shaping community structure of endophytic fungi–evidence from the Pinus-Viscum-system

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Fungal Diversity 2013年第3期60卷 55-69页

作者： Derek Peršoh Systematische Botanik und Mykologie Ludwigs-Maximilians Universität MünchenMenzinger Straße 6780638 MünchenGermany Abteilung Mykologie Universität BayreuthUniversitätsstraße 30–NW I95440 BayreuthGermany

endophytic fungal communities have been shown to be highly diverse in almost every host plant species analyzed so far.However,the factors shaping their compositions are largely unknown.To elucidate the impact of various factors,10 independent replicates of DNA extracts from each of 17 surface-sterilized leaf and stem samples were analyzed by pyrosequencing of fungal ITS1 rRNA gene amplicons.Altogether,154 fungal OTUs(operational taxonomic units),represented by 953,385 sequences,were found in at least 2 samples from Viscum album ssp.austriacum and/or its host Pinus sylvestris.Deviating from earlier,cultivation-based assessments,the communities were dominated by OTUs related to the genus Mortierella and OTUs not assignable to a certain fungal phylum.However,Ascomycota were still the most diverse group in terms of OTU richness and already hypothesized organ and host preferences of certain endophytic Xylariaceae isolated from the Pinus-Viscum-system could be confirmed.Host species and organ type were also the major factors shaping the detected fungal communities.The two plant species clearly differed according to the endophytic fungal communities,but only stems and needles of Pinus were inhabited by significantly different fungal assemblages.Interestingly,only the 1 and 3 year old stem sections differed according to the endophytic fungal community,while differently aged leaves of both plants were indistinguishable in this regard.Size of the organs had no impact on fungal communities in Pinus,but shorter internodes and smaller leaves showed at least a tendency to differ from the corresponding larger organs in Viscum.Fungal communities also differed slightly between the two sampling sites,lying 200 km apart,and between the three sampling campaigns.Because the samples were drawn within 15 days,this finding indicates that seasonal shifts clearly outweigh aging effects in host plant with perennial leaves.The results therefore provide strong evidence against a linear development of the endophytic fungal communities in Pinu

关键词： endophytic fungi Pyrosequencing Host preference Stems Leaves Organ age Pinus sylvestris Viscum album Genetic abundance Name assignment

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Distribution and diversity of endophytic fungi in Gentiana rigescens and cytotoxic activities

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Chinese Herbal Medicines 2020年第3期12卷 297-302页

作者： Li-li Xu Xiao-feng Ling Shu-juan Zhao Ru-feng Wang Zheng-tao Wang Institute of Science Technology and HumanitiesShanghai University of Traditional Chinese MedicineShanghai 201203China MOE Key Laboratory for Standardization of Chinese Medicines SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese MedicinesInstitute of Chinese Materia MedicaShanghai University of Traditional Chinese MedicineShanghai 201203China

Objective:In the present study,Gentiana rigescens was screened for fungi communities to clarify their diversity and community assemblage in hosts.Meanwhile,the identification and activity assays of the strains were also conducted.Methods:By culture-dependent(endophytic fungi isolations from plant sections)and culture-independent(metagenomic library and cloning from plant sections)techniques,fungi communities were studied.The metagenomic library was generated using direct DNA isolation of whole plants,plant radixes,plant stems,plant leaves,plant flowers and soils around the plant.Meanwhile,endophytes were isolated from all parts ofG.rigescens plants.After fermentation of the fungi isolations,all the isolates were evaluated for their cytotoxicity against four kinds of human cancer cell lines(HCT116,BEL7404,A549,MDA-MB-231).Results:Eventually,200 strains were isolated and 103 strains were further identified through the internal transcribed spacer(ITS,ITS1 and ITS2 regions)sequence by using the universal primers ITS5 and ITS4.A total of 59,106 fungal sequences corresponding to 374 putative operational taxonomic units(OTU)were identified by 454 pyrosequencing.Through 454 pyrosequencing,the main fungal genera were Sebacina,Botrytis,Mycosphaerella,Boletus and Gibberella,and the major fungal genera which were directly isolated were Aspergillus,Fusarium,Penicillium and Alternaria.Activity assays showed strains 5-26(Aspergillus sp.)and 6-2(Fusarium avenaceum)had the outstanding cytotoxicity to all the tested cell lines with IC5o valuesfungi isolation methods.Activity assays indicated that endophytes were a promising natural source of potential anticancer agents.

关键词： cytotoxic activities endophytic fungi Gentiana rigescens Franch. ITS metagenomic library

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