Fungal Endophytes from Three Cultivars of Panax ginseng Meyer Cultivated in Korea

Sang Un Park,Hyoun-Sub Lim,Kee-Choon Park,Young-Hwan Park,Hanhong Bae 고려인삼학회 2012 Journal of Ginseng Research Vol.36 No.1

In order to investigate the diversity of endophytes, fungal endophytes in Panax ginseng Meyer cultivated in Korea were isolated and identifi ed using internal transcribed spacer (ITS) sequences of ribosomal DNA. Three cultivars of 3-year-old ginseng roots (Chunpoong, Yunpoong, and Gumpoong) were used to isolate fungal endophytes. Surface sterilized ginseng roots were placed on potato dextrose agar plates supplemented with ampicilin and streptomycin to inhibit bacterial growth. Overall, 38 fungal endophytes were isolated from 12 ginseng roots. According to the sequence analysis of the ITS1-5.8S-ITS2, 38 fungal isolates were classifi ed into 4 different fungal species, which were Phoma radicina, Fusarium oxysporum, Setophoma terrestris and Ascomycota sp. 2-RNK. The most dominant fungal endophyte was P. radicina in 3 cultivars. The percentage of dominant endophytes of P. radicina was 65.8%. The percentage of colonization frequency of P. radicina was 80%, 52.9%, and 75% in Chunpoong, Yunpoong, and Gumpoong, respectively. The second most dominant fungal endophyte was F. oxysporum. The diversity of the fungal endophytes was low and no ginseng cultivar specifi city among endophytes was detected in this study. The identifi ed endophytes can be potential fungi for the production of bioactive compounds and control against ginseng pathogens.

Fungal Endophytes from Three Cultivars of Panax ginseng Meyer Cultivated in Korea

Park, Sang-Un,Lim, Hyoun-Sub,Park, Kee-Choon,Park, Young-Hwan,Bae, Han-Hong The Korean Society of Ginseng 2012 Journal of Ginseng Research Vol.36 No.1

In order to investigate the diversity of endophytes, fungal endophytes in Panax ginseng Meyer cultivated in Korea were isolated and identified using internal transcribed spacer (ITS) sequences of ribosomal DNA. Three cultivars of 3-year-old ginseng roots (Chunpoong, Yunpoong, and Gumpoong) were used to isolate fungal endophytes. Surface sterilized ginseng roots were placed on potato dextrose agar plates supplemented with ampicilin and streptomycin to inhibit bacterial growth. Overall, 38 fungal endophytes were isolated from 12 ginseng roots. According to the sequence analysis of the ITS1-5.8S-ITS2, 38 fungal isolates were classified into 4 different fungal species, which were Phoma radicina, Fusarium oxysporum, Setophoma terrestris and Ascomycota sp. 2-RNK. The most dominant fungal endophyte was P. radicina in 3 cultivars. The percentage of dominant endophytes of P. radicina was 65.8%. The percentage of colonization frequency of P. radicina was 80%, 52.9%, and 75% in Chunpoong, Yunpoong, and Gumpoong, respectively. The second most dominant fungal endophyte was F. oxysporum. The diversity of the fungal endophytes was low and no ginseng cultivar specificity among endophytes was detected in this study. The identified endophytes can be potential fungi for the production of bioactive compounds and control against ginseng pathogens.

Endophytic Fungal strains Specifically Modified the Biochemical Status of Grape Cells

Li-Hua Huang,Ming-Zhi Yang,Xiu-Jin Ao,An-Yun Ren,Ming-Quan Yuan,Han-Bo Zhang 한국식물학회 2018 Journal of Plant Biology Vol.61 No.4

Previously, specific interactions in morphologywere observed between grape cells and endophytic fungalstrains during a dual culture experiment. However, thebiochemical impacts of these fungal endophytes on grapecells is also expected due to their potential application in grapequality management. After assessed multiple physiochemicaltraits to those grape cells which have co-cultured with differentendophytic fungal strains in this study, and found the presenceof fungal endophytes obviously triggered ROS stress responsesin grape cells, and the biochemical status in grape cells weredifferentially modified by different fungal strains. In thosetested endophytic fungal strains, RH37 (Epicoccum sp.),RH6 (Alternaria sp.), RH32 (Alternaria sp.) and RH34(Trichothecium sp.) conferred greater metabolic impacts ongrape cells. And soluble protein (SPr), total flavonoids (TF),total phenols (TPh) and malondialdehyde (MDA) on the otherhand, were sensitive biochemical parameters which can beinfluenced in greater ranges than other detected parameters. Most interestedly, fungal endophytes shaped metabolitespatterns in grape cells during the dual culture appeared fungalgenus/species/strain-specificity. The work confirmed thesignificance of fungal endophytes in grape metabolic regulationand elucidated the possibility to purposely manage grapequality using tool of fungal endophytes.

Diversity Distributions and the Anthocyanin Associations of Fungal Endophytes in Diferent Colored Grapevine Leaves

Jing‑Chao Chen,Man Yu,Fang Liu,Jin‑Zhuo Qu,Xiao‑Xia Pan,Han‑Bo Zhang,Ming‑Zhi Yang 한국식물학회 2020 Journal of Plant Biology Vol.63 No.2

Anthocyanins composed the major pigments which conferred sensory and chemical values to red grapes and wines. Multiple factors involved in grape anthocyanin biosynthesis have been greatly covered to date. However, the endophytes which closely associate with plants, their possible roles in plant coloration are still under clear. Our present research frstly investigated the diversity distributions of culturable fungal endophytes (CFE) in diferent colored grapevine leaves which harvested from same vineyard and cultivar, and then proposed method to analyze the possible efects of these fungal endophytes on anthocyanin concentrations of grape cells. In totally 532 endophytic fungal isolates, 19 OTUs belong to 13 genera were isolated from fve color diferent grapevine leaves. Obvious leaf color specifcity of CFE communities were observed distribution in diferent colored grapevine leaves in this experiment. Assessing the infuences of those isolated CFEs on anthocyanin concentrations of a teinturier grape pulp cells via dual culture system revealed that anthocyanin promotion fungal strains were more possibly isolated from red and purple colored grape leaves. And elite CFE strains such as DQ53 and DQ55, both belong to the fungal genus Epicoccum, which conferred signifcant promotion efects on grape cellular anthocyanin contents were selected out. And the transcriptions of anthocyanin biosynthesis association genes MYBA1, UFGT and F3′5′H in grape cells, were signifcantly infuenced by these endophytic fungi, furtherly implicated the involvements of these genes in fungal endophytes-mediated grape cell anthocyanin promotion process. Additionally, endophytic fungi triggered anthocyanin promotion in grape cells seem independently to that of the light mediated anthocyanin synthesis pathway. This work demonstrated that beside other environmental factors, endophytes may also involve in grape anthocyanin metabolisms. And the study provided methods and clues to explore fungal endophytes in colored grape leaves which possibly applied in grape pigmentation processing.

Identification and characterization of type III polyketide synthase genes from culturable endophytes of ethnomedicinal plants

Manoharan, Gomathi,Sairam, Thiagarajan,Thangamani, Rajesh,Ramakrishnan, Dhivya,K.Tiwari, Manish,Lee, Jung-Kul,Marimuthu, Jeya IPC Science and Technology Press 2019 Enzyme and microbial technology Vol.131 No.-

<P><B>Abstract</B></P> <P>Endophytic fungi provide benefits to host plants by producing a diverse class of secondary metabolites (natural products). Arrays of polyketide natural products are synthesized by specific classes of polyketide synthases (PKS I, II and III) in host organisms. In the present study, we attempt to screen and identify type III PKSs in culturable fungal endophytes isolated from the ethno medicinal plants including <I>Arbus precatorius</I>, <I>Bacopa monnieri,Citrus aurantifolia</I> and <I>Datura metel</I> to detect the genetic potential of endophytic fungi in producing bioactive compounds. A total of seventeen endophytic fungal strains belonging to eight genera were identified using fungal morphology and rDNA-ITS phylogenetic analyses. A CODEHOP-PCR based strategy was followed to design degenerate primers for the screening of type III PKS genes from fungal endophytes. We had successfully amplified partial PKS genes from eight endophytes. The amplified PKS sequences showed 60–99% identity to already characterized/putative PKS genes. From the partial sequence of FiPKS from <I>Fusarium incarnatum</I> BMER1, a full-length gene was amplified, cloned and characterized. FiPKScDNA was cloned and expressed in <I>E. coli</I> Lemo21 (DE3) and the purified protein was shown to produce pyrones and resorcinols using acyl-CoA thioesters as substrates. FiPKS showed the highest catalytic efficiency of 7.6 × 10<SUP>4</SUP> s<SUP>−1</SUP> M<SUP>-1</SUP> with stearoyl CoA as a starter unit. This study reports the identification and characterization of type III PKS from endophytes of medicinal plants by CODEHOP PCR.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A CODEHOP PCR based screening method was employed for type III polyketide synthase gene identification in fungal endophytes. </LI> <LI> By this approach, partial type III PKS genes from eight fungal endophytes were amplified and sequenced. </LI> <LI> FiPKS gene from Fusarium incarnatum BMER1, an endophyte of Bacopa monnieri was cloned and functionally characterized. </LI> <LI> FiPKS produced pyrones and resorcinols with the highest catalytic efficiency of 7.6 x 104 s-1 M-1towards stearoyl CoA. </LI> </UL> </P>

An insight into endophytic antimicrobial compounds: an updated analysis

Digra Shivani,Nonzom Skarma 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.4

Resistance in micro-organisms against antimicrobial compounds is an emerging phenomenon in the modern era as compared to the traditional world which brings new challenges to discover novel antimicrobial compounds from different available sources, such as, medicinal plants, various micro-organisms, like, bacteria, fungi, algae, actinomycetes, and endophytes. Endophytes reside inside the plants without exerting any harmful impact on the host plant along with providing ample of benefits. In addition, they are capable of producing diverse antimicrobial compounds similar to their host, allowing them to serve as useful micro-organism for a range of therapeutic purposes. In recent years, a large number of studies on the antimicrobial properties of endophytic fungi have been carried out globally. These antimicrobials have been used to treat various bacterial, fungal, and viral infections in humans. In this review, the potential of fungal endophytes to produce diverse antimicrobial compounds along with their various benefits to their host have been focused on. In addition, classification systems of endophytic fungi as well as the need for antimicrobial production with genetic involvement and some of the vital novel antimicrobial compounds of endophytic origin can further be utilized in the pharmaceutical industries for various formulations along with the role of nanoparticles as antimicrobial agents have been highlighted.

Effects of Water Stress on the Endophytic Fungal Communities of Pinus koraiensis Needles Infected by Cenangium ferruginosum

( Sun Keun Lee ),( Seung Kyu Lee ),( Han Hong Bae ),( Sang Tae Seo ),( Jong Kyu Lee ) 한국균학회 2014 Mycobiology Vol.42 No.4

To examine the effects of water stress and Cenangium ferruginosum (CF) on the fungal endophytic community of needles of Pinus koraiensis (PK), fungal endophytes isolated from the needles of 5-year-old PK seedlings were compared before and after exposure to water stress conditions and artificial inoculation with CF ascospores. Artificial CF inoculation was successfully confirmed using PCR with CF-specific primers (CfF and CfR). For comparison of the degree of water deficit in water-stressed and control groups of PK seedlings infected with CF, the water saturation deficit and water potential were measured. Lower water potential estimates were found in the water-stressed seedlings than in the control group. The fungal endophytes isolated from the second-year needles of non-water-stressed seedlings before and after CF inoculation revealed that primary saprobes were approximately 30% and 71.7%, respectively, and the remaining endophytes were rot fungi or pathogens. Sixty days after CF inoculation, diverse fungal endophytes in the first-year needles were isolated from the water-stressed seedlings. However, some fungal endophytes isolated from the non-water-stressed seedlings were also identified. Fungal endophytes in the second-year needles of the water-stressed and non-water-stressed seedlings were approximately 8% and 71.7% of saprobes, respectively, and the remaining endophytes were rot fungi or pathogens. On the basis of the results, we conclude that water deficit and CF can have an effect on fungal endophytic communities in the needles of PK seedlings.

Age-dependent Distribution of Fungal Endophytes in Panax ginseng Roots Cultivated in Korea

Young-Hwan Park,Young-Chang Kim,Sang Un Park,Hyoun-Sub Lim,Joon Bum Kim,Byoung-Kwan Cho,Hanhong Bae 고려인삼학회 2012 Journal of Ginseng Research Vol.36 No.3

Fungal endophytes were isolated from 1-,2-, 3- and 4- year-old ginseng roots(Panax ginseng Meyer) cultlivated in Korea. The isolated fungal endophytes were identified based on sequence analysis of the internal transcribed spacer and morphological characterization by microscopic observations. A total of 81 fungal endophytes were isolated from 24 ginseng roots. Fungal endophytes were classified into 9 different fungal species and 2 unknown species. Ginseng roots that were 1.-2,-3-, and 4- years old were colonized by 2, 6, 8 and 5 species of fungal endophytes, respectively. While Phoma radicina was the most frequent fungal endophyte in 2,-3- and 4- years old ginseng roots, Fusarium solani was the dominant endophyte in 1-year-old ginseng roots. The colonization frequencies (CF) varied with the host age. The CF were 12%, 40%, 31% and 40% for 1.-2,-3-, and 4- years old ginseng roots, respectively. We found a variety of fungal endophytes that were distributed depending on the age of ginseng plants.

Fungal Diversity and Plant Growth Promotion of Endophytic Fungi from Six Halophytes in Suncheon Bay

( Young Hyun You ),( Hyeokjun Yoon ),( Sang Mo Kang ),( Jae Ho Shin ),( Yeon Sik Choo ),( In Jung Lee ),( Jin Man Lee ),( Jong Guk Kim ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.11

Endophytic fungi were isolated from roots of six halophytes in Suncheon Bay. The endophytic fungi of 35 species isolated from halophytes were identified by internal transcribed spacer (ITS) containing the ITS1, 5.8s, and ITS2 regions. All fungal strains were analyzed to diversity at the genus level. Fungal culture filtrates (FCF) of endophytic fungi were treated to Waito-c rice (WR) seedling for plant growth-promoting verification. It was confirmed that fungal strain Sj-2-2 provided plant growth promotion (PGP) to WR seedling. Then, PGP of Suaeda japonica was confirmed by treating culture filtrate of Sj-2-2. As a result, it was verified that culture filtrate of Sj-2-2 had more advanced PGP than positive control when treated to S. japonica. The secondary metabolites involved in culture filtrate of Sj-2-2 were identified by HPLC and GC-MS SIM analysis. The presence of physiologically bioactive gibberellins (GAs) and other inactive GAs in culture filtrate of Sj-2-2 was detected. The molecular analysis of sequences of Sj-2-2 showed the similarity to Penicillium sp. of 99% homology. The PGP of Sj-2-2 as well as symbiosis between endophytic fungi and halophytes growing naturally in salt marsh was confirmed. Sj-2-2 was identified as a new fungal strain producing GAs by molecular analysis of sequences. Consequently, the Sj-2-2 fungal strain was named as Penicillium sp. Sj-2-2. In this study, the diversity of endophytic fungi isolated from roots of halophytes in salt marsh and the PGP of a new gibberellin-producing fungal strain were confirmed.

Fungal Community Analyses of Endophytic Fungi from Two Oak Species, Quercus mongolica and Quercus serrata, in Korea

( Manh Ha Nguyen ),( Keum Chul Shin ),( Jong Kyu Lee ) 한국균학회 2021 Mycobiology Vol.49 No.4

Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.