Progress 10/01/20 to 09/30/21
Outputs
Target Audience:Botanists, horticulturist, microbiologists, foresters, conservationists Guam Cooperative Extension personnel and the Forestry Division of the Guam Department of Agriculture, educators, environmentalists, and general community in the region and worldwide. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One undergraduate student was introduced to the study of orchid mycorrhizal taxonomy using traditional techniques of culturing fungal isolates for morphological characterization and more modern genetic techniques encompassing DNA extraction, PCR, and Sanger sequencing. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?We continue to collect data of the survey and DNA analysis of orchid mycorrhiza found in Guam.We will summarize data and prepare a manuscript for a publication.
Impacts
What was accomplished under these goals?
Objective 1.Determine the distribution of native epiphyte orchids and their host tree species in Guam. Field surveys were done on several epiphytic orchid species native to Guam, including:Taeniophyllum mariannense, Bulbophyllum guamense, Dendrobium guamense, Tuberolabium guamense, Luisia teretifolia,andCoelogyne guamense.Aside from previously surveyed populations of orchids atBolanos, Umatac; Mt. LamLam, Umatac; and Yigo Research Station, Yigo, the study this year was expanded to include smaller and more novel orchid populations around the island atMt. LamLam Overlook, Umatac; Sella Bay Overlook, Umatac; War in the Pacific Park, Agat; Northwest Field AAFB, Yigo; Windward Hills Golf Course, Yona; and Barrigada Heights, Barrigada.Taeniophyllum mariannense,a relatively common and leafless orchid species, was found at a much higher frequency in open canopy forests with more light. Other orchid species,Bulbophyllum guamense, Dendrobium guamense, Tuberolabium guamense, Luisia teretifolia,andCoelogyne guamense,were less common and found in select areas in southern Guam sites with limited access to light. A total of 121 orchid specimens were observed growing on 19 different host tree species as well as rocky substrate in different sites across Guam. Orchid host tree species of limestone and scrub forest primarily found in northern and southern Guam includedElaeocarpus joga, Hibiscus tiliaceus, Leucaena leucocephala, Morinda citrifolia, Pandanus tectorius, Premna serratifolia,andVitex parviflora. Host tree species of ravine forests mainly found in southern Guam includedAcacia auriculiformis,Calophyllum inophyllum,Casuarina equisetifolia, Cocos nucifera, Cycas micronesica, Glochidion marianum, Hibiscus tiliaceus, Leucaena leucocephala, Mangifera indica, Manilkara zapota, Spathodea campanulata,andVitex parviflora. Host tree species,Brachychiton acerifoliusandCalophyllum inophyllum,were found in coastal strand forest along the island's coastandAraucaria heterophyllaandTabebuia roseawere found in urban or residential areas in central and southern Guam. In the study of host tree diversity, the commonly found orchid,Taeniophyllum mariannene,was observed to have a greater number of different host tree species without any apparent preference for native or non-native tree species. Other orchid species were observed with a limited range of host tree species. Objective 2.Determine mycorrhizae associated with native orchids in Guam. Cytological analysis of root samples in all orchid species revealed that arbuscular mycorrhizal colonization occurred at the average rate of 32.7% (n=352). Mycorrhizal colonization occurred at similar rates inBulbophyllum guamense(34.4%,n=32),Dendrobium guamense(25.0%,n=32), andTaeniophyllum mariannense(31.9%,n=248). The least rate of colonization was observed inCoelogyne guamense(12.5%,n=8), and the higher withTuberolabium guamense(42.9%,n=28) and the highest withLuisia teretifolia(100%,n=4) with their smaller sample sizes. Mycorrhizal colonization was observed more at seedling stage or protocorms (83.3%,n=24) than mature orchid plants (29.0%,n=328). For morphological characterization ofmycorrhizal fungi,a very small portion of root samples ofTaeniophyllum mariannensewas cultured to isolate fungal colonies. However, it wasdifficult to isolate a single fungal species for morphological study due to overlapping structures of different fungal genus/species both within and outside theRhizoctoniacomplex.There was also a tendency offungal isolates in culturetoremainin anamorphic stage rather than developing reproductive myceliaatthe teleomorphic stagefor more definitive identification. We could not obtain information onmorphological characterizations of fungal colonies at this time. The DNA analysis of orchid mycorrhizal fungi was performed using PCR and Sanger sequencing.The DNA sequences of fungal samples were compared withthe NCBI BLAST Nucleotide Database for fungi identification.Representative samples showed BLASTN hits matching genera of theRhizoctoniacomplexand other fungal groups as possibly having mycorrhizal roles in orchid roots.The DNA sequences of fungal samples fromroots ofBulbophyllum guamensewere almost identical to those from roots of otherBulbophyllum sp.and closely matchedan uncultured mycorrhizal sample of the order Atractiellales from a root ofDendrobium chrysotoxumfrom China (KX587497.1) sharing an 85.89% identity.Fungalsamples from a root ofTaeniophyllum mariannenseshowed a top BLASTN hit closely matching the sequence of an uncultured fungal clone of the family Sebacinaceae (order Sebacinales) from a root ofEpidendrum firmum(JX998775.1)sharing 93.31% identity. Samples of a root ofTuberolabium guamenseshowed a top BLASTN hit closely matching the sequence of a vouchered mycorrhizal sample of the division Basidiomycota from a root ofTaeniophyllum aphyllumfrom Japan (AB449205.1)with 90.51% identity and matching with a fungal clone of the family Ceratobasidiaceae originally isolated from a root ofTaeniophyllum glandulosumfrom Japan (LC405936.1) with 89.78% identity. DNA sequence analysis ofthecultured mycorrhizalisolatesfrom roots ofTaeniophyllum mariannenseyielded mixed results with a large proportion of samples closely matching common endophytic orchid fungi whose mycorrhizal roles remain unclear. These orchid endophytes mainlybelonged tothe fungal genera,Annulohypoxylon,Aspergillus,Biscogniauxia,Clonostachys,Daldinia,Dothideomycetes,Fusarium, Hypoxylon, Pestalotiopsis, Tolypocladium,andXylaria,all showing a percent identity > 98%. Three fungal endophytes found in this study that are not common in orchids, but usually occurring in other plant species wereCampylocarpon,Rhytidhysteron, andSetophaeosphaeriawith a percent identity >97%. Objective 3.Disseminate the results of the research findings. Results of research findings are being summarized for a publication.
Publications
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Progress 10/12/18 to 09/30/21
Outputs
Target Audience:Botanists, Horticulturist, Microbiologists, Foresters, Conservationists Guam Cooperative Extension and the Forestry Division of the Guam Department of Agriculture, educators, environmentalists, and general community in the region and worldwide. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Undergraduate and graduate students were introduced to the study of orchid mycorrhizal taxonomy using techniques of fungal DNA extraction, PCR, and Sanger sequencing. How have the results been disseminated to communities of interest?The results of the project were disseminated by presentation of posters at two conferences during 2022, and earlier as MS thesis completed by a graduate student in 2019. Findings are currently being summarized to prepare a journal publication and a technical report to focus on importance of understanding host tree preferences and mycorrhizal associations of Guam's native epiphytic orchid species in conservation and reforestation efforts of the island ecosystem What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Objective 1. Determine the distribution of native epiphyte orchids and their host tree species in Guam. Field surveys were conducted on six epiphytic orchid species native to Guam, including: Taeniophyllum mariannense, Bulbophyllum guamense, Dendrobium guamense, Tuberolabium guamense, Luisia teretifolia, and Coelogyne guamensis and their host tree species. 147 orchid specimens were recorded growing on 22 different host tree species. T. mariannense, a relatively common and leafless orchid species, was found in open canopy forests and urban areas with more light. Other orchid species, B. guamense, D. guamense, T. guamense, L. teretifolia, and C. guamensis, were less common and found in select areas in southern Guam sites with limited access to light. Orchid host trees of the limestone and scrub forest in Guam included Elaeocarpus joga, Hibiscus tiliaceus, Leucaena leucocephala, Morinda citrifolia, Pandanus tectorius, Premna serratifolia, and Vitex parviflora. Host tree species of ravine forests mainly found in southern Guam included Acacia auriculiformis,Calophyllum inophyllum, Casuarina equisetifolia, Cocos nucifera, Cycas micronesica, Glochidion marianum, Hibiscus tiliaceus, Leucaena leucocephala, Mangifera indica, Manilkara zapota, Spathodea campanulata, and Vitex parviflora. Host tree species, Brachychiton acerifolius and Calophyllum inophyllum, were found in coastal strand forest along the island's coast and Araucaria heterophylla and Tabebuia rosea were found in urban or residential areas in central and southern Guam. The most common, leafless orchid T. mariannense was observed growing on 19 different host trees across all habitats with Calophyllum inophyllum, Hibiscus tiliaceus, and two of Guam's most invasive trees, Leucaena leucocephala, and Vitex parviflora. B. guamense, D. guamense, and C. guamensis were commonly observed growing on Cocos nucifera, Elaeocarpus joga, and Pandanus tectorius, common native tree species of undisturbed limestone forest and volcanic ravine forest. T. guamense was found growing exclusively on the invasive V. parviflora in volcanic ravine forest. L. teretifolia was found growing on V. parviflora in volcanic ravine forest and C. inophyllum in urban areas. In the study of host tree diversity, the commonly found orchid, Taeniophyllum mariannene, was observed to have a greater number of different host tree species without any apparent preference for native or non-native tree species. Other orchid species were observed with a limited range of host tree species. Objective 2. Determine mycorrhizae associated with native orchids in Guam. The DNA analysis of orchid mycorrhizal fungi was performed using PCR and Sanger sequencing. The DNA sequences of fungal samples were compared with the NCBI BLAST Nucleotide Database for fungi identification. From roots of six orchid species studied, three mycorrhizal fungi family were detected including Ceratobasidiaceae in the order Cantharellales, and two family, Sebacinaceae and Serendipitaceae in the order Sebacinales. There were fungal endophytes isolated from roots of some orchids belonged to class Basidiomycete and class Ascomycete, however, it is not certain if these fungal species were orchid mycorrhiza. Tuberolabium guamense was associated with Ceratobasidiaceae. Dendrobium guamense and Bulbophyllum guamense were associated with both Sebacinaceae and Serendipitaceae. In addition, from the roots of Bulbophyllum guamense, endophytes belonging to Basidiomycete and Ascomycete were isolated. Coelogyne guamensis was associated with Sebacinaceae and Luisia teretifolia with Serendipitaceae and an endophyte of Ascomycete. Taeniophyllum mariannense had shown association with orchid mycorrhizal fungi of Sebacinaceae and Ceratobasidiaceae and endophyte of Ascomycete. Although Tulasnellaceae was detected in some trials, we could not obtain a conclusive result. Objective 3. Disseminate the results of the research findings. Earlier work of the project was disseminated by a graduate student's MS thesis: Mario Martinez. 2019. Orchid Mycorrhizae in Taeniophyllum mariannense Schltr. in Guam. MS Thesis in Sustainable Agriculture, Food, Natural Resources, University of Guam. Results of research findings were presented as posters at the Botany 2022 conference in July 2022 at Anchorage, AK and at the conference of Society for Advancement of Chicanos/Hispanics & Native Americans in Science (SACNAS) in San Juan, PR in October 2022 Michael Fernandez and Mari Marutani. 2022. Host tree preferences and mycorrhizal associations of Guam's native epiphytic orchids. Poster presentation at the Botany 2022 Conference in Anchorage, AK from July 24 -27, 2022 Michael Fernandez, Mari Marutani and Mario Martinez. 2022. Host tree and mycorrhizal diversity of Guam's native epiphytic orchids. Poster presentation at SACNAS conference in San Juan, PR from October 27 - 29, 2022
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
2. Michael Fernandez, Mari Marutani and Mario Martinez. 2022. Host tree and mycorrhizal diversity of Guams native epiphytic orchids. Poster presentation at SACNAS conference in San Juan, PR from October 27 29, 2022
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
1. Michael Fernandez and Mari Marutani. 2022. Host tree preferences and mycorrhizal associations of Guams native epiphytic orchids. Poster presentation at the Botany 2022 Conference in Anchorage, Alaska from July 24 -27, 2022
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Progress 10/01/19 to 09/30/20
Outputs
Target Audience:Botanists, horticulturist, microbiologists, foresters, conservationists, Guam Cooperative Extension and the Forestry Division of the Guam Department of Agriculture, educators, environmentalists, and general community in the region and worldwide. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One undergraduate student was introduced to orchid mycorrhizal taxonomy based on genetic analysis. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?We continue to collect data of the survey and DNA analysis of orchid mycorrhiza found in Guam
Impacts
What was accomplished under these goals?
1. Determine the distribution of native epiphyte orchids and their host tree species in Guam. Twelve different host trees were recorded for 108 individual orchid specimens observed during this period of the study. In the northern part of Guam, tree species grown in open canopy limestone and scrub forests included Leucaena leucocephala, Vitex parviflora, Hibiscus tiliaceus, Premna serratifolia, Morinda citrifolia and Tabebuia rosea, while host trees in southern Guam at heavily shaded, high elevation ravine forests on volcanic substrate included Leucaena leucocephala, Elaeocarpus joga, Glochidion marianum, Cocos nucifera, Vitex parviflora, Hibiscus tiliaceus, Morinda citrifolia, Pandanus tectorius, Cycas micronesica, and Mangifera indica. A leafless orchid, Taeniophyllum mariannense, was commonly found growing on Leucaena leucocephala, Hibiscus tiliaceus, and Vitex parviflora. The main host trees of Dendrobium guamense were Cocos nucifera, Pandanus tectorius, and Elaeocarpus joga. Bulbophyllum guamense were mainly found on Cocos nucifera and Pandanus tectorius and the majority of Coelogyne guamense specimens were found growing on Pandanus tectorius. Tuberolabium guamense was exclusively found on Vitex parviflora in this study, and the single specimen of Luisia teretifolia was found on a host tree of Vitex parviflora. 2. Determine mycorrhizae associated with native orchids in Guam. Orchid mycorrhizal DNAs were isolated and selected primers were employed to determine taxonomic characters of mycorrhizae. Mycorrhizal DNAs isolated from Bulbophyllum guamense amplified with ITS1F/ITS2 showed a top BLAST hit with the sequence of an uncultured Atractiellales clone (Class: Atractiellomycetes, Order: Atractiellales) sharing 88% sequence identity (e value = 3.00e^-71). Taeniphyllum mariannense samples amplified with ITS1F/ITS2 showed a top BLAST hit with the sequence of an uncultured Sebacinales clone (Class: Agaricomycetes, Order: Sebacinales) sharing 93% sequence identity (e-value = 6.00e^-94). Tuberolabium guamense samples amplified with ITS1F/ITS2 showed a top BLAST hit with the sequence of vouchered mycorrhizae (Basidiomycota) genes sharing 91% sequence identity (e-value = 4.00e^-96). These samples also yielded subsequent BLAST results with an uncultured Thanatephorus isolate (Class: Agaricomycetes, Order: Cantharellales, Family: Ceratobasidiaceae, Genus: Rhizoctonia, Species: solani) and an uncultured Ceratobasidium clone (Class: Agaricomycetes, Order: Cantharellales, Family: Ceratobasidiaceae, Genus: Ceratobasidium) both sharing 86% sequence identity (e-value = 6.00e^-74 and e-value = 2.00e^-73, respectively). 3. Disseminate the results of the research findings Results of research findings are being summarized for a publication.
Publications
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Progress 10/12/18 to 09/30/19
Outputs
Target Audience:Botanists, horticulturists, microbiologists, foresters, conservationists, Guam Cooperative Extension and the Forestry Division of the Guam Department of Agriculture personnel, educators, environmentalists, and general community in the region and worldwide. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?We established collaboration work with Dr. Yuki Ogura-Tsujita and Keto Rammitsu of Saga University in Japan to exchange information on study and technology to advance orchid mycorrhizal research. How have the results been disseminated to communities of interest?Data collected by Mario Martinez were summarized and were disseminated and deposited to University of Guam Library as his thesis and as oral presentation in May 2019. What do you plan to do during the next reporting period to accomplish the goals?We continue to collect data of the survey and DNA analysis
Impacts
What was accomplished under these goals?
Determine the distribution of native epiphyte orchids and their host tree species in Guam. Fifteen host trees of a native epiphyte orchid, Taeniophyllum mariannense, were identified in Guam. In the limestone forest habitat, there were Aglaia mariannensis (Maliaceae), Averrhoa bilimbi (Oxalidaceae), Eugena reinwardtiana (Myrtaceae), Fagraea berteoana (Gentianaceae), Morinda citrifolia (Rubiaceae), Ochrosia mariannense (Apocynaceae), Ochrosia opposirifolia (Apocynaceae), Pandanus tectorius (Pandanaceae), Premma serratifolia (Lamiaceae) and Vitex parviflora (Lamiaceae), and in the southern ravine forest there were Areca catechu (Arecaceae), Cocos nucifera (Arecaceae), Hibiscus tiliaceus (Malvaceae), and Morinda citrifolia . T. mariannense was also found on ×Citrofortunella microcarpa (Rutaceae) in a farmland. 2. Determine mycorrhizae associated with native orchids in Guam. Mycorrhizae associated with Taeniophyllum mariannense in Guam were found to belong to the families Tulasnellaceae (Basidiomycota; Order Cantharellales), Ceratobasidicaeae (Basidiomycota, Order Cantharellales), Russuluaceae (Basidiomycota; Order Russulales) of Division according to the Sanger sequencing DNA analysis. Additional isolates were found to belong to Division Ascomycota and the other in Order Agaricales of Division Basidiomycota. 3.Disseminate the results of the research findings A MS degree thesis in Sustainable Agriculture, Food and Natural Resources was completed by Mario Martinez of with the title of "Orchid Mycorrhizae in Taeniophyllum mariannense Schltr. in Guam." and his oral report was presented as a seminar in May 2019.
Publications
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2019
Citation:
Mario Martinez. 2019. Orchid Mycorrhiza in Taeniophyllum mariannense Schltr. in Guam. MS Thesis in Sustainable Agriculture, Food and Natural Resources Progrem. University of Guam. 47 pp.
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