Thesis (Ph. D.)--University of Hong Kong, 1998. / Includes bibliographical references (leaves 203-225).

Thesis (M.S.)--University of Kentucky, 2007. / Title from document title page (viewed on June 14, 2007). Document formatted into pages; contains: vii, 57 p. : ill. Includes abstract and vita. Includes bibliographical references (p. 48-56).

Magister Scientiae - MSc (Biotechnology) / Endophytic bacteria are known to have an endosymbiotic relationship with plants and provide them with many beneficial properties. These bacteria stimulate plant hormones, provide protection from pathogens and increase nutrient availability in the environment. In this study some of these potential growth factors were tested. Endophytic bacteria have the potential to be of great value for the increase of crop production. They offer a variety of processes that aid in plant growth promotion in an ecofriendly manner. The use of endophytic bacteria provides a cheaper and cleaner approach compared to industrial made fertilizers. They also have potential uses in bioremediation to clean the environment polluted by industrial processes. Endophytes were isolated and showed significant growth improvement. Each isolate displayed different morphologies. Isolates were tested for classical growth promotion mechanisms such as the ability to solubilize phosphate, Indole-3-acetic acid and siderophore production. Inductively Coupled Plasma Optical Emission Spectrometry was performed to measure the effect of the isolates on the plants nutrient profile. The isolates were then tested again while the plants were under heavy metal stress to determine if they were still capable of growth promotion. The plants were then assayed for cell death using Evans blue and biomass was measured to determine the effect of vanadium stress. Inductively Coupled Plasma Optical Emission Spectrometry was performed again to assess the change in nutrient profile while under vanadium stress. / 2021-08-31

Made available in DSpace on 2015-04-20T12:31:50Z (GMT). No. of bitstreams: 1 Eliandra de Freitas Sia.pdf: 1557550 bytes, checksum: e4d21c384795b53b83656a535e1bf31f (MD5) Previous issue date: 2012-09-04 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The filamentous fungi are an important group of microorganisms, being not only one of the most numerous among the microorganisms due to their potential in biotechnological processes such as the production of antibiotics, enzymes, vitamins and others. The fungi are also plant, animals and human pathogens, acting sometimes as toxins producers. The fungal growth is a routine practice at microbiological laboratories and the Potato Dextrose Agar (PDA) medium is the most used medium due to be a rich source of starch. However, in some regions of Brazil, the potato production has a quite high cost. For this reason the aiming of the present work was the search for new alternative sources of starch. Thus, initially several tubercles: cassava (Manihot esculenta), sweet potato (Ipomoea batatas), carrot (Daucus carota), ginger (Zingiber officinale) and yam (Dioscorea alata) were tested as a starch source to fungal growth. The cassava (Manihot esculenta) showed the best results to fungal growth. This species is a tropical plant with a great commercial importance to Brazil, Africa and other parts of the world. In brazilian north and northeast regions, the cassava is hugely cultivated by a low cost. After the first screening, it was optimized the utilization of Cassava Dextrose Agar (CDA) media. Many fungal species of biotecnological importance (Beauveria bassiana, Aspergillus niger, Penicillium sp., Fusarium sp., Phanerochaete chrysosporium) were evaluated to growth, development sporulation and hyphal formation. The BDA media was used as a control. The present results demonstrated a high efficiency of MDA to fungal growth. However, in MDA was observed a reduction of fungal sporulation comparing with BDA. Finally, both media BDA and MDA, were used to the endophytic fungal isolation from: guarana (Paullinia cupana) and olives (Olea europea). The fungal identification demonstrated that isolates varied according to host plant. The fungal diversity was higher in guaraná being higher using the MDA in comparison with the BDA media. To both media, it was obtained a low number of isolated fungi from O. europea. The present results clearly demonstrated that the cassava is an feasible important starch source, being a potential alternative media, mainly in tropical countries / Os fungos filamentosos são um importante grupo de micro-organismos, não apenas por serem os mais numerosos dentre todos os micro-organismos, mas devido ao seu potencial em processos biotecnológicos como a produção de antibióticos e enzimas, fontes de vitaminas, alimentação e outros. Os fungos também são conhecidos como patógenos de plantas, animais e humanos, e produtores de toxinas. Cultivar fungos é uma rotina em laboratórios de microbiologia e um dos meios muito utilizado é o Batata Dextrose Ágar (BDA), que utiliza a batata como uma fonte rica em amido. Em algumas regiões do país, como a Região Norte, a batata tem um elevado custo e por este motivo a busca por novas fontes de amido é uma alternativa. Assim, outros vegetais devem ser pesquisados visando a substituição da batata. No presente trabalho foram inicialmente utilizados mandioca (Manihot esculenta), batata-doce (Ipomoea batatas), cenoura (Daucus carota), gengibre (Zingiber officinale) e cará (Dioscorea alata), sendo demonstrado que a mandioca (Manihot esculenta) é uma alternativa relevante para o cultivo de fungos. Esta espécie vegetal é uma planta tropical de grande importância comercial no Brasil, na África e em outras regiões do mundo, sendo que para a região Norte a mandioca é muito cultivada e também de baixo custo. Assim, após a triagem inicial com outros tubérculos, foi realizada a avaliação e otimização do meio de cultura de Mandioca Dextrose Ágar (MDA). Para o ensaio do meio com mandioca foram utilizadas diversas espécies de fungos com importância biotecnológica (Beauveria bassiana, Aspergillus niger, Penicillium sp., Fusarium sp., Phanerochaete chrysosporium). Foi avaliada a taxa de crescimento, a esporulação e a formação de hifas no meio de cultura BDA (utilizado como controle) e MDA. Os resultados comprovaram a eficiência do MDA para o crescimento dos fungos analisados, destacando-se a taxa de crescimento radial em meio sólido e peso seco do micélio em meio líquido. Entretanto, em MDA há uma menor esporulação dos fungos quando comparado ao BDA. Finalmente, foi realizado, utilizando meio BDA e MDA, o isolamento de fungos endofíticos de duas plantas: guaraná (Paullinia cupana) e oliveira (Olea europea), nos quais o meio de cultura MDA também se mostrou eficiente. A identificação de fungos endofíticos revelou que eles variaram conforme a planta hospedeira. A diversidade de fungos endofíticos foi maior em guaraná no meio MDA que em BDA, e o número de isolados foi menor em oliveira nos dois meios utilizados. Assim, do ponto de vista econômico e social, o substrato de mandioca pode ser considerado uma alternativa de alta potencialidade na obtenção de meios de cultura para laboratórios de micologia, especialmente em países tropicais

En raison de la nature symbiotique des micro-organismes endophytes, cette étude visait à étudier l'activité antibactérienne, antifongique et cytotoxique chez les métabolites secondaires des extraits endophytes foliaires de plantes de l'Amazonie brésilienne et du Cerrado. Dans ce travail de thèse 147 micro-organismes cultivables ont été isolés (130 champignons, 3 bactéries e 14 champignons non-identifiés ou inconnus) à partir de 28 plantes (4 espèces collectées au Brésil et 24 en Guyane Française). Tous les micro-organismes furent identifiés par analyse moléculaire des régions spécifiques de l'ADNr en utilisant des techniques de séquençage génomique. Des champignons endophytes de l’ordre des Xylariales furent ceux de plus importante fréquence d’isolation dans cette étude, représentés par 25 isolats. Des extraits bruts à l’AcOEt furent produits à partir de cultures de chaque micro-organisme isolé. Une proportion relative signifiante (23,1%) des extraits démontra une activité sur Candida albicans ATCC 10213, pendant que 4% furent actifs sur Staphylococcus aureus ATCC 29213. Le potentiel cytotoxique des extraits fut évalué pour des lignées cellulaires humaines KB (carcinome cervical utérin), MDA-MB-435 (mélanome), et MRC-5 (fibroblastes de poumon normal), résultant en proportion signifiante avec activité d’inhibition de la prolifération cellulaire (24,4%, 23,1% e 16,3%, respectivement).Dix-huit métabolites secondaires furent isolés et identifiés à partir du fractionnement des extraits bruts de huit endophytes. Dix-sept de ces substances furent déjà décrites précédemment dans la littérature: l’acide piliformique (24) et la griséofulvine (25) isolés à partir de Xylaria cubensis SNB-GCI02; La phomopirone (26), la pyrenocine (27), l’alterpérilenol (28) et la novae-zelandine (29), isolés de Lewia infectoria SNB-GTC2402; 5-métilmelleina (31) et la dihidrosporothrioride (32) isolées à partir de Xylaria sp. SNB-GTC2501; La mycoleptodiscine (34) et la mycoleptodiscine B (35) isolées à partir de Mycoleptodiscus sp. SNB-GTC2304; le mycoepoxidiène (36) et la altiloxina A (37) isolés à partir de Diaporthe pseudomangiferae SNB-GSS10; L’acremonisol (40), le semicochliodinol (41) et le cochliodinol (42) isolés à partir de Chaetomium globosum SNB-GTC2114; la colletofragarone A2 (43) isolée à partir de Colletotrichum sp. SNB-GTC0201; la flavoglaucine (44) isolée à partir de Eurotium rubrum BBS01. Parmi ces substances, la flavoglaucine (44) isolée à partir de E. rubrum BBS01 démontra une activité comparable au contrôle fluconazol en C. albicans (CIM de 4 µg.mL-1). La substance (44) présenta IC50 >10 µM sur cellules normales MRC-5, ce qui en fait un candidat pour des études ultérieures. Dans ce travail, fut identifié pour la première fois l’activité de la colletofragarone A2 (43) isolée à partir de Colletotrichum sp. SNB-GTC0201. La substance inédite nommée pyrrocidine C (30) fut isolée à partir de L. infectoria SNB-GTC2402 et identifiée par des analyses spectroscopiques (Casella et al., 2013). La pyrrocidine C (30) fut active en S. aureus ATCC 10213 (CIM de 2 µg.mL-1) et ne fut pas considérées cytotoxique pour les cellules normales MRC-5 (IC50 >10 µM), démontrant une sélectivité dans l’action antimicrobienne. Ces résultats démontrent la grande diversité des endophytes fongiques chez les plantes de l'Amazonie brésilienne et du Cerrado et la grande chimiodiversité associée aux métabolites secondaires de ces micro-organismes. Des champignons endophytes tropicaux, comme ceux présentés dans cette étude, peuvent apparaître comme une nouvelle source de substances antimicrobiennes et cytotoxiques. / Because of the symbiotic nature of endophytes, this survey aims to investigate the probality of discovering antibacterial, antifungal and cytotoxic activities in secondary metabolites of leaf endophytes isolated from plants of Amazon and Cerrado biomes. In this study, 147 cultivable microorganisms were isolated (130 fungi, 3 bacteria and 14 unidentified or unknown microbes) from 28 plant species (4 species collected in Brazil and 24 in French Guyana). All endophytes were identified by molecular analyses of specific rDNA regions, with genomic sequencing techniques. Fungal endophytes belonging to Xylariales order were the most frequently isolated in this study, represented by 25 isolates. Crude AcOEt extracts were produced from cultures of each isolated endophyte. A significant relative proportion (23,1%) of extracts showed activity in Candida albicans ATCC 10213, while 4% were active in Staphylococcus aureus ATCC 29213. The cytotoxic potencial of the extracts was evaluated for human cell lines KB (uterin cervical carcinome), MDA-MB-435 (melanoma) and MRC-5 (normal lung fibroblasts), and a significant proportion of them showed cellular proliferation inhibition (24,4%, 23,1% e 16,3%, respectively).Eighteen secondary metabolites were isolated by the fractionation of eight endophytic extracts. Seventeen of these substances had already been previously described in the literature: piliformic acid (24) and griseofulvin (25) isolated from Xylaria cubensis SNB-GCI02; phomopiron A (26), pyrenocin A (27), alterperilenol (28) and novae-zelandin A (29) isolated from Lewia infectoria SNB-GTC2402; 5-metilmellein (31) and dihidrosporothriorid (32) isolated from Xylaria sp. SNB-GTC2501; mycoleptodiscin A (34) and mycoleptodiscin B (35) isolated from Mycoleptodiscus sp. SNB-GTC2304; mycoepoxidien (36) and altiloxin A (37) isolated from Diaporthe pseudomangiferae SNB-GSS10; acremonisol A (40), semicochliodinol A (41) and cochliodinol (42) isolated from Chaetomium globosum SNB-GTC2114; colletofragaron A2 (43) isolated from Colletotrichum sp. SNB-GTC0201; flavoglaucin (44) isolated from Eurotium rubrum BBS01. Among these substances, flavoglaucin (44), isolated from E. rubrum BBS01, showed comparable antifungal activity with the positive control fluconazol in C. albicans (MIC of 4 µg.mL-1). Flavoglaucin (44) also showed IC50 >10 µM in normal MRC-5 cells, becoming a good candidate for further studies. In this work, the cytotoxic activity of colletofragaron A2 (43), isolated from Colletotrichum sp. SNB-GTC0201 was described for the first time. The unpublished substance named pyrrocidin C (30) isolated from L. infectoria SNB-GTC2402 was identified by spectroscopic analyses (Casella et al., 2013). The pyrrocidin C (30) was active in S. aureus ATCC 10213 (MIC of 2 µg.mL-1), and was not considered cytotoxic for normal MRC-5 cells (IC50 >10 µM), showing selectivity in antimicrobial activity. These results demonstrate the great endophytic fungal diversity in plants of Amazon and Cerrado biomes, along with the chemodiversity associated to the secondary metabolites of these endophytes. Tropical fungal endophytes, like those seen in this work, may emerge as a new source of antimicrobial and cytotoxic substances. / Devido à natureza simbiótica dos micro-organismos endofíticos, este estudo teve como objetivo investigar a atividade antibacteriana, antifúngica e citotóxica em metabólitos secundários de extratos de fungos endofíticos foliares de plantas do bioma Amazônia e Cerrado. Neste trabalho de tese foram isolados 147 micro-organismos cultiváveis (130 fungos, 3 bactérias e 14 fungos não-identificados ou desconhecidos) a partir de 28 plantas (4 espécies coletadas no Brasil e 24 na Guiana Francesa). Todos os micro-organismos foram identificados por análise molecular de regiões específicas de DNAr, com uso de técnicas de sequenciamento genômico. Fungos endofíticos da ordem Xylariales foram os de maior frequência de isolamento neste estudo, representados por 25 isolados. Extratos brutos em AcOEt foram produzidos a partir de culturas de cada micro-organismo isolado. Uma proporção relativa significante (23,1%) dos extratos demonstrou atividade em Candida albicans ATCC 10213, enquanto 4% foram ativos em Staphylococcus aureus ATCC 29213. O potencial citotóxico dos extratos foi avaliado para as linhagens celulares humanas KB (carcinoma cervical uterino), MDA-MB-435 (melanoma), e MRC-5 (fibroblastos de pulmão normal), resultando em proporção significante com atividade de inibição da proliferação celular (24,4%, 23,1% e 16,3%, respectivamente). Dezoito metabólitos secundários foram isolados a partir do fracionamento de oito extratos brutos endofíticos. Dezessete destas substâncias já tinham sido descritas anteriormente na literatura: ácido pilifórmico (24) e griseofulvina (25) isoladas de Xylaria cubensis SNB-GCI02; phomopirona A (26), pyrenocina A (27), alterperilenol (28) e novae-zelandina A (29) isoladas de Lewia infectoria SNB-GTC2402; 5-metilmelleina (31) e Dihidrosporothriolida (32) isoladas de Xylaria sp. SNB-GTC2501; mycoleptodiscina A (34) e mycoleptodiscina B (35) isoladas de Mycoleptodiscus sp. SNB-GTC2304; mycoepoxidieno (36) e altiloxina A (37) isoladas de Diaporthe pseudomangiferae SNB-GSS10; acremonisol A (40), semicochliodinol A (41) e cochliodinol (42) isoladas de Chaetomium globosum SNB-GTC2114; colletofragarona A2 (43) isolada de Colletotrichum sp. SNB-GTC0201; flavoglaucina (44) isolada de Eurotium rubrum BBS01. Dentre estas substâncias, a flavoglaucina (44) isolada de E. rubrum BBS01, demonstrou atividade comparável ao controle fluconazol em C. albicans (CIM de 4 µg.mL-1). Esta substância (44) apresentou IC50 >10 µM em células normais MRC-5, tornando-se candidata para estudos posteriores. Neste trabalho foi identificado pela primeira vez a atividade citotóxica da colletofragarona A2 (43), isolada de Colletotrichum sp. SNB-GTC0201. A substância inédita nomeada pyrrocidina C (30) foi isolada a partir de L. infectoria SNB-GTC2402 e identificada através de análises espectroscópicas (Casella et al., 2013). A pyrrocidina C (30) foi ativa em S. aureus ATCC 10213 (CIM de 2 µg.mL-1), e não foi considerada citotóxica para as células normais MRC-5 (IC50 >10 µM), demonstrando seletividade na ação antimicrobiana. Estes resultados demonstram a grande diversidade fúngica endofítica em plantas do bioma Amazônia e Cerrado, e a quimiodiversidade associada aos metabólitos secundários destes micro-organismos. Fungos endofíticos tropicais, como os vistos neste trabalho, podem emergir como uma nova fonte de substâncias antimicrobianas e citotóxicas.

Abstract Mycorrhizal symbiosis is generally advantageous for plants in nutrient-poor soils. Arcto-alpine areas are relatively nutrient-poor, but abundantly inhabited by non-mycorrhizal species. Possibly, mycorrhizal symbiosis is not favoured due to the harsh climatic conditions and the short growing season, which constrain the photosynthetic gain and growth of the arcto-alpine plants. This hypothesis was theoretically evaluated by assuming that optimal mycorrhizal colonization maximizes the net carbon gain of the host plant. In addition, the prevalence of arbuscular mycorrhizal (AM) and dark-septate endophytic (DSE) fungi along an altitudinal gradient was studied in the field, and their effects on the plant performance were tested in the laboratory. In the model, the photosynthetic nutrient use efficiency (PNUE) had a key role in determining whether mycorrhizal strategy would be optimal for the plant net carbon gain. The model generated several colonization patterns depending on possible changes in PNUE and soil nutrient concentrations along altitudinal gradients. Field studies indicated that species-level colonizations do not yield a consistent pattern along the altitude except for fine endophyte, which increased along an altitudinal gradient. In a high-alpine field site root fungal colonizations were rare. Seasonal shifts in colonizations in low-alpine conditions were not found. DSE fungi were common root-associates in the field. In the laboratory, AM had a positive impact on the performance of Gnaphalium norvegicum at 15°C, but not at 8°C. DSE-inoculation did not colonize the roots, but it had a positive impact on seedling performance, which may be due to the saprophytic activity of the fungus in the substrate. Additionally, mycorrhizal inoculum was found to decrease the performance of a non-mycorrhizal plant in a competition experiment. Species-level mycorrhizal colonization patterns may differ from community-level pattern along altitudinal gradients and the relative abundance of different fungal symbionts may change along with the altitude. The performance of mycorrhizal plants in high-alpine conditions may be decreased due to several factors e.g. low temperature constraints on plant and fungal physiology and allocation, soil disturbances and low availability of inoculum. Climatic constraints for plant photosynthesis may thus affect the mycorrhizal colonization patterns in arcto-alpine conditions, but are not necessarily the primary cause for lower performance of mycorrhizal plants at higher altitudes.

The Botryosphaeriaceae is a family of fungi that includes many species, which are well-known as pathogens, saprophytes and endophytes of plants and especially of trees. As a result of their pathogenic nature and potential threat to plantations and agricultural crops, much research has been devoted to their identification. The main focus of studies that make up this thesis has been on the fungal complex referred to as Diplodia pinea sensu lato. These fungi are members of the Botryosphaeriaceae and studies have specifically concentrated on their taxonomy, host associations and mycovirus infections associated with them. Diplodia pinea sensu lato represents a species complex of highly similar morphological types that mainly infect Pinus spp., world-wide. The species complex includes what have in the past been known as the A, B and C morphological types of D. pinea. Multiple gene genealogies based on sequences of partial protein-coding genes and microsatellite markers were used to resolve the species complex into two genera, D. pinea and D. scrobiculata (= B morphotype). Diplodia-like isolates from Australia, Greece and Cyprus were characterized using both morphological and molecular characteristics. Morphologically, these isolates all have dark, thick-walled conidia (Diplodia-like) but phylogenetically, they could belong to three distinct genera of the Botryosphaeriaceae namely Diplodia, Lasiodiplodia and Dothiorella. Results of this study led to the description of Dothiorella casuarini from Casuarina spp. in Australia and they highlight the fact that similar morphological characteristics and disease etiology does not necessarily provide a true reflection of the evolutionary history of a pathogen. Phylogenetic studies on species of the Botryosphaeriaceae with Diplodia-like anamorphs revealed intriguing host association patterns. The availability of sequence data for many species of the Botryosphaeriaceae made it possible to extend the phylogeny to include six of the ten lineages as previously described for the Botryosphaeriaceae. Angiosperms appeared to be the most common, and possibly ancestral, host group of the Botryosphaeriaceae, with the exception of Macrophomina, Guignardia, Saccharata and “Botryosphaeria” quercuum. Infection of gymnosperms most likely occurred more recently, only in specific groups (Diplodia and Lasiodiplodia) via host shifts. Three distinct viruses have now been characterized from isolates of D. pinea sensu lato. Two of these were previously characterized and are known as Sphaeropsis sapinea RNA virus 1 and 2 (SsRV1 and SsRV2). The third dsRNA element more commonly found in association with D. scrobiculata was characterized in this dissertation and named Diplodia scrobiculata RNA virus 1 (DsRV1). It has a genome of 5018 bp with a unique genome organization characterized by two open reading frames (ORFs). One ORF codes for a putative polypeptide similar to proteins of the vacuolar protein-sorting (VPS) machinery and the other one for a RNA dependent RNA polymerase (RdRp). The hypothetical protein probably has a role in transport or protection of this unencapsulated virus into membranous vesicles. Phylogenetically, DsRV1 groups closest to a dsRNA element from Phlebiopsis gigantea (PgV2) and they both group separately from other families in which fungal viruses have been classified. The frequency and distribution of DsRV1, SsRV1 and SsRV2 were determined in a collection of D. pinea and D. scrobiculata isolates using Real-time PCR. Infections with SsRV1 and SsRV2 occurred in both D. pinea and D. scrobiculata, while DsRV1 was mainly found in D. scrobiculata. DsRV1 was also found to always occur in combination with SsRV1 and/or SsRV2. Therefore, DsRV1 probably selected against a coat protein as the result of a fitness trade-off. Although earlier studies indicated that these viruses have no effect on the phenotype or virulence of D. pinea and D. scrobiculata isolates, the presence of specific viruses in their host populations serve as a useful marker in studying movement of fungal pathogens. The ultimate aim of studies making up this dissertation was to expand the base of knowledge regarding species in the D. pinea species complex. This was justified by the fact that D. pinea is one of the most important tree pathogens in South Africa and that an expanded knowledge might contribute to reducing diseases caused by it. Clearly understanding the identity of the fungus must clearly underpin many elements of a management strategy and this was one of the aims of the suite of studies conducted. Furthermore, I attempted to augment the knowledge base regarding dsRNA elements in D. pinea sensu lato. These studies were of a basic nature and relatively far removed from the practical application level. Nonetheless, it is my hope that they have pushed ahead knowledge barriers and that in some way they will contribute to reducing the impact of Diplodia-associated diseases in the future. / Thesis (PhD)--University of Pretoria, 2011. / Microbiology and Plant Pathology / Unrestricted

>Magister Scientiae - MSc / Salinity is one of the major limiting factors to crop production, which consequently contributes to the risk of reduced food security. Among other factors, food security depends on availability of sufficient and nutritious food for humans. Livestock such as cattle and sheep are fed with various plant-based feeds; with Medicago sativa (commonly known as alfalfa or lucerne) being a very important forage/feed crop, so much that it is regarded as the queen of forage crops. However, alfalfa is severely affected by high soil salinity and thus its growth and yield are drastically reduced in soils with high NaCl content. Among the various alfalfa genotypes/varieties examined in this study, Agsalfa was identified as salt tolerant because it performed better under salt treatment compared to Magna601.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21149号 / 農博第2275号 / 新制||農||1059(附属図書館) / 学位論文||H30||N5123(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 田中 千尋, 教授 本田 与一, 准教授 刑部 正博 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

Mutualistic relationships between endophytic fungi and grasses have shown to improve the hardiness of the host. This relationship is common in grasses, including North American native grasses that are important in both forage and grassland ecosystems. Elymus species, such as Canada wildrye (CWR), commonly host the endophytic fungi, Epichloё, while southeastern wildrye (SEWR) may not. In this study, seed of CWR and seed and leaves from local SEWR were assessed for endophyte infection. Infection status was confirmed via histological analysis of seed squashes and leaf peels. Presence/absence of endophyte was determined by scanning for mycelia within seed and between leaf epidermal cells. Following assessment, all SEWR germplasm were endophyte-free (E-), while CWR was endophyte-infected (E+). Endophyte- infected CWR seed were used to isolate and culture the symbiotic endophyte. Isolated endophyte was used to infect E- SEWR. Infection status of SEWR was determined using leaf peels and PCR. This research will help determine if artificial endophyte infection can be performed in SEWR.