Progress 10/01/14 to 01/29/19
Outputs
Target Audience:The Principal Investigator Professor Sharman D. O'Neill collaborated with important stakeholders in California: 1) those in the California floriculture and horticulture industries, especially major commercial orchid growers, and 2) those in the California-based vanilla industry. For the first group, Professor O'Neill engaged with nursery/greenhouse operations in the Bay Area, south into the Salinas area, and north into Sebastopol. Professor O'Neill communicated with a number of California-based grower and horticulture businesses involved particularly in potted orchid flowering plant production (e.g., Matsui Nursery, Salinas, CA) and marketing of flowers. For the second group, Professor O'Neill worked with California-based companies involved in the vanilla industry including the sale of fruit (From The Field Trading Company, Inc.; Saffron Imports, Inc.) or the sale of fruit extract (Cook Flavoring and Manufacturing Company). She also engaged with international and USA corporations involved in crop improvement and food and beverage manufacturing (e.g., General Mills Inc., Minneapolis, Minnesota). All these groups were supportive of Professor O'Neill's research on orchid reproduction and orchid fruit flavor and aroma for crop improvement. During the final years of the project, Dr. O'Neill engaged in a public-private partnership involving research sponsorship by the California-based flavor manufacturing business, Cook Flavoring Company and Lochhead Manufacturing Company, Paso Robles, CA. She also engaged with manufacturers and suppliers of greenhouse hydroponics equipment for her experimental use for growing orchids hydroponically. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples of California-based outreach were to the San Francisco Conservatory of Flowers, the San Francisco Exploratorium, the San Francisco Orchid Society, the Orchid Conservation Alliance, and Humboldt State University, Arcata, CA. In addition, PI O'Neill engaged in formal collaboration on Phalaenopsis and Vanilla orchid research with scientists at the USDA-ARS, Pacific Basin Agricultural Research Center (Hawaii) and Hawaiian orchid producers and farmers. She also continued to seek collaboration with the Tropical Agricultural Research Station at the USDA-ARS Mayaguez, Puerto Rico, and had planned to visit the USDA-ARS Mayaguez, but the trip was postponed due to current natural disasters in that country making travel impossible. Finally, Professor O'Neill served as the Director of the Vanilla Genetic Diversity Center at UC Davis, and as and lead organizer of the Vanilla Genomics Project called "The Vanilla Sustainability Project," and as such, she communicated with many scientists, farmers, business and industry leaders, and stakeholders worldwide. She was actively involved in Vanilla germplasm conservation and participated in activities of the California-based Orchid Conservation Alliance (Encinitas, CA). During the final years of the project, Professor O'Neill continued her professional service as Editor for Plant Biology (five subfields from Applied to Basic Research in Plant Biology), as a major part of Elsevier's Major Reference Works in the Life Sciences. This activity reached a large international target audience of scientists and students through its continually updated online resource in plant biology. In summary, Professor O'Neill continued to share her basic science and applied knowledge expertise with California orchid producers, scientists, agronomists, business and industry leaders, especially in the area of molecular biology and genetics of commercially important orchids, especially Phalaenopsis and Vanilla. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This CRIS project provided training to one third-year Horticulture and Agronomy Graduate Group graduate student, Ms. Amy Bump, for whom Dr. O'Neill served as graduate faculty mentor and Major Professor. Ms. Bump has finished her thesis. Other training of undergraduate students, University Honors students, and other volunteer interns was previously reported. How have the results been disseminated to communities of interest?The results have been disseminated to communities of interest, especially in the form of public outreach on behalf of the University of California, Davis and through publications and presentations. This is described above. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples include invitations to speak at public education institutions and at California State Universities. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The overall goal of this project was to develop fundamental knowledge about commercially important orchids. Specifically, this project investigated important reproductive processes using the unique biological system of the orchids, with Phalaenopsis and Vanilla as the primary biological and genomic models, respectively. The overall focus is on the unique program in orchids for: 1) flower development with a focus on the gynostemium, and male and female gametophytes; 2) pollination and post-pollination development; 3) floral aroma; 4) floral longevity and perianth senescence in response to pollination, emasculation, or hormones; 5) post-pollination ovary and ovule development; 6) fertilization and early seed (embryo and endosperm) development; and 7) fruit development, maturation and ripening. The long-term goal was to obtain a detailed understanding of molecular genetic events that control these reproductive processes in important commercial orchid cultivars. Knowledge was gained about floral longevity and senescence, floral scent product, fruit quality and aroma, and stress responses (abiotic and biotic). Development of improved varieties of premier-quality Phalaenopsis orchids that have fragrant as well as long-lived flowers, are disease resistant, and more resilient to climate stress, would be a major output achievement. During the final year of the project, many projects were brought to fruition in the form of publications, presentations, and new data sets for bioinformatics analysis. In one case, we completed a detailed quantitative analysis of the effect of pollination and pollination-associated events or factors, such as emasculation, auxin, ethylene, and ACC on the expression of ethylene biosynthetic genes in Phalaenopsis orchids that leads to flower senescence or development. This investigation provides the first report of localization by in situ hybridization of ACC synthase in the gynoecium of pollinated orchid flowers over an early and detailed time course and in response to ethylene treatment. In addition, it addresses important questions regarding the induction of ethylene by pollination through an in-depth examination of ACC oxidase gene expression at the earliest times after pollination in stigma tissue. The results are novel and made important contributions to the body of knowledge about plant hormone regulation of development as well as cell-specific expression of two important ethylene biosynthetic genes. In addition, the use of quantitative methods to measure gene expression using real-time quantitative PCR provides a test and validation of earlier reports in the literature on gene expression in flower tissue measured by qualitative methods, such as RNA blot hybridization. Finally, this research provides new insight into the possible mode of regulation of seed plant ACC synthases by examining the question of post-transcriptional regulation of orchid ACC synthases. A novel and unique to orchids consensus motif is reported for orchid ACC synthases that may be involved in their unique mode of post-transcriptional regulation similar to Type 3 ACC synthase genes/proteins in Arabidopsis and tomato. Another example included a project in which we conducted a detailed analysis of 8 vanillin pathway-associated compounds in unripe green vanilla fruit and the effects of ethylene and abiotic stress (high temperature shock or stress) on the levels of the compounds. Eight compounds examined were present in different relative amounts in six-month-old green unripe fruit of Vanilla planifolia, and most were present in both their non-glycosylated and glycosylated forms. Ethylene clearly induced the biosynthesis of certain intermediates in the vanillin biosynthetic pathway, as proven by the use of the ethylene action inhibitor, 1-MCP. Ethylene was involved in increasing the amounts of key pathway compounds, whereas heat was the important factor for other compounds. Some of these increases were supported in enzyme-treated samples by statistical analysis. Principal axis factor analysis was also conducted. This is the first report of the use of 1-MCP to examine the role of ethylene in vanilla fruit ripening. A third investigation was completed on two endangered orchid species, Vanilla grandifolia, and Vanilla pompona, useful for fruit production and useful as highly suitable parents in interspecific hybridizations. Both species have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We characterized flowering and fruit development as never before reported and provided important basic knowledge about Vanilla orchids. This information is novel because there are few to no publications about these two species and few complete developmental studies of vanilla reproduction. This research line provides fundamental information about reproduction to guide breeding efforts with Vanilla planifolia, the orchid of commerce, through interspecific hybridization. During the fourth year, this research was presented at the World Orchid Conference in Guayaquil, Ecuador (November, 2017) and at the annual meeting of the American Society of Plant Physiologists in Quebec, Canada (July 2018). In another CRIS project study of Phalaenopsis was brought to completion providing new insight into the mechanism of pollination of orchids. Professor O'Neill proposed a four-stage process involving pollen-derived signals and the novel differentiation of fertile stigmatic cells that are required for fertility and pollination success. In another project, Professor O'Neill and her graduate student, Amy Bump, conducted a project aimed at establishing and evaluating via real-time quantitative Polymerase Chain Reaction (RT-qPCR) analysis and statistical analysis a set of Reference Genes to use in our projects aimed at evaluating gene expression in the transcriptomes. Reference Genes are critical to standardize and evaluate gene expression across tissues and developmental stages, with good Reference Genes being expressed stably at low levels across all time points and in all tissues. We we established a number of reliable Reference Genes and disqualified other candidates as unstable and not suitable. The second aim of the project was to use the Reference Genes to assess the role of the vanillin synthase gene (VpVAN) during fruit development and ripening. These results will be submitted for publication in a peer-reviewed journal shortly and will credit NIFA support. The poster presentation described preliminaries of this research, which won the only award of "First Prize for a Scientific Poster" at the World Orchid Conference, Ecuador (November, 2018). Ms. Amy Bump, Graduate Student, completed her Masters' thesis describing the Reference Gene project and the VpVan project. The Reference Genes were used as controls for analysis of gene expression of a potential candidate vanillin biosynthetic gene in the phenylpropanoid pathway and in other developmental pathways of interest to Professor O'Neill. This work was carried out in collaboration with staff of the UC Davis Real-time PCR Research & Diagnostic Core Facility, Dept. of Medicine & Epidemiology, School of Veterinary Medicine, University of California, Davis and with the Bioinformatics Core of the UCD Genome Center.
Publications
- Type:
Books
Status:
Awaiting Publication
Year Published:
2019
Citation:
A. Bump, C. Wademan, S. Barnum, E. Hodzic, and S. D. O'Neill. 2019.
Evaluation of reference genes and a potential vanillin synthase gene during fruit development in Vanilla planifolia. In Press.
|
Progress 10/01/17 to 09/30/18
Outputs
Target Audience:The Principal Investigator Professor Sharman O'Neill collaborates with important stakeholders in the California floriculture and horticulture industries, especially major commercial orchid growers. Professor O'Neill continues to engage with other nursery/greenhouse operations in the Bay Area, south into the Salinas area, and north into Sebastopol. Professor O'Neill communicates with a number of California-based grower and horticulture businesses involved in orchid potted flowering plant production (e.g., Matsui Nursery, Salinas, CA) and marketing of flowers, and with other California-based companies involved in the vanilla industry including the sale of fruit (From The Field Trading Company, Inc.; Saffron Imports, Inc.; Cook Flavoring and Manufacturing Company) and other international and USA corporations involved in crop improvement and food and beverage manufacturing (General Mills Inc.). All these groups are interested in Professor O'Neill's research on orchid reproduction and orchid fruit flavor and aroma. During the fourth year of the project, Dr. O'Neill continued the public-private partnership involving research sponsorship with a California-based flavor manufacturing business, Cook Flavoring Company and Lochhead Manufacturing Company, Paso Robles, CA. She has also engaged with manufacturers and suppliers of greenhouse hydroponics equipment for her experimental use for growing orchids for research. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples include outreach to San Francisco Orchid Society (March 2018) and Humboldt State University, Arcata, CA. The PI has ongoing formal collaboration on Phalaenopsis and Vanilla orchid research with scientists at the USDA-ARS, Pacific Basin Agricultural Research Center (Hawaii) and Hawaiian orchid producers and farmers. She also continued to seek collaboration with the Tropical Agricultural Research Station at the USDA-ARS Mayaguez, Puerto Rico, and had planned to visit the USDA-ARS Mayaguez, but the trip was postponed due to current natural disasters in that country making travel impossible. Finally, Professor O'Neill is the Director of the International Vanilla Users Group and the Vanilla Genomics Project called "The Vanilla Sustainability Project" and as such has communicated with many scientists, farmers, business leaders and other stakeholders worldwide. She is also actively involved in Vanilla germplasm conservation and participates in activities of the California-based group, the Orchid Conservation Alliance (Encinitas, CA). During the fourth year, Professor O'Neill continued her professional service as Editor for Plant Biology (five subfields from applied to basic research), as a major part of Elsevier's Major Reference Works in the Life Sciences. This activity reached a large international target audience of scientists and students through its continually updated online resource in plant biology. In summary, Professor O'Neill continued to share her basic science and applied knowledge expertise with California orchid producers, scientists, agronomists, and industry leaders, especially in the area of molecular biology and genetics of commercially important orchids, especially Phalaenopsis and Vanilla. ? Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This CRIS project provided training to one second-year Horticulture and Agronomy Graduate Group graduate student, Ms. Amy Bump, for whom Dr. O'Neill serves as graduate faculty mentor and Major Professor, and 6 UC Davis undergraduate students, including one Honors Program student. The six Undergraduate Research Interns listed below conducted independent research projects with Professor O'Neill and her lab team in either the lab or greenhouse for which they received academic credit in PLB 192 or MCB 099/199 courses during Year 4. For 2017-2018: Oliver Abundez-Dominguez Anne Ashmore Miyako Noguchi Abigail Stevens Erik Toivonen Carolina Tweedy Professor O'Neill also served as the Faculty Mentor and adviser for Ms. Anne Ashemore for her Senior Honors Thesis project administered through the University Honors Program as a kind of capstone course. Her Honor's Thesis project resulted in two different posters that were presented at the UC Undergraduate Research Biology Conference (Spring 2017) and at the World Orchid Conference (November 2017). Both of Ms. Ashemore's presentations credited the USDA-CRIS support to Professor O'Neill. Two new volunteers joined the O'Neill Lab to focus on the Vanilla Greenhouse: Fall 2018: Thomas Mason, B.A. Sadie Nikki Brooks, M.A. Finally, three graduate students received training and academic credit during the reporting period: Amy Bump Trent Spriochaeren Lanxuan Wang Another 6 undergraduate students received mentoring, training, and formal course credit in how to be an Undergraduate Teaching Assistant, an excellent experience for their future careers in STEM. How have the results been disseminated to communities of interest?The results have been disseminated to communities of interest, especially in the form of public outreach on behalf of the University of California, Davis and through publications and presentations. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples include invitations to speak at public education institutions and at California State Universities. In November 2017, Professor O'Neill disseminated her research to the general public by giving a high-profile talk about her on Vanilla planifolia. Professor O'Neill's talk was targeted to an educated non-scientist audience that are interested in science. San Francisco Exploratorium, Pier 15 Observatory, Exploratorium Museum, SF, CA. Invited talk in "Pairings: Cultivating a Taste for Science through Food" series to large audience (120 adult members) of the SF Exploratorium (including Board Members and some Donors). This event was advertised on Facebook to "hundreds of thousands of Exploratorium members." Abstract of Professor O'Neill's talk: Title: Vanilla: From Flower to Fertilization and Flavor. Sharman D. O'Neill, M.A., Ph.D. Professor Department of Plant Biology College of Biological Sciences University of California, Davis, Davis, CA 95616 U.S.A. E-mail: sdoneill@ucdavis.edu Vanilla planifolia, an orchid of Mesoamerican origin, is exceptional among the large family of orchid species in that it produces an extraordinary aromatic fruit, known as the "vanilla bean." Vanilla beans have the highest value of any legal crop due to their sustainable mode of production and process. Recently though the price of one kilogram of cured vanilla beans soared to ~$600.00 reflecting serious problems in the vanilla industry. A key issue is that of fruit quality. There is a conundrum that when the price of vanilla fruit is high, their quality is low. But what determines fruit quality? From scientific investigation, the developmental pathway from flower to fertilization to fruit determines fruit quality and involves complex biological processes of pollination and fertilization. The growth and development of a vanilla fruit requires a full 9 months. During this time, the coordinated development of new reproductive structures in the ovary largely determine fruit quality measures, such as aroma. Our research has focused on vanilla flowers and fruit using genomics and bioinformatics to understand the underlying molecular genetic basis of fruit quality. In order to understand these changes, it was essential to obtain detailed knowledge of reproduction in this species, beginning with flower pollination. This research will be presented in the broader context of The Vanilla Sustainability Project, an international collaboration for crop improvement in Vanilla planifolia, germplasm conservation, sustainability, and for biodiversity conservation in Madagascar, the center of world vanilla production. This presentation will also provide an overview of the history of vanilla cultivation in Mexico, the patrimonial birthplace of vanilla, and in Europe and islands of the Southwest Indian Ocean including Madagascar, Réunion Island, and the Comoros Islands. Flower diversity will be shown of different vanilla species, time permitting. Demonstrations will include vanilla beans from these regions and live vanilla plants. This talk was extremely well received by the public and staff of the Exploratorium. Feedback shared later with Professor O'Neill stated that the talk was "fantastic." The USDA-CRIS funding was acknowledged. What do you plan to do during the next reporting period to accomplish the goals?Ms. Amy Bump, Graduate Student, will complete her thesis describing the Reference Gene project and the VpVan project. The Reference Genes will be used as controls for analysis of gene expression of an important biosynthetic gene in the phenylpropanoid pathway and in other developmental pathways of interest to Professor O'Neill. This work is being done in collaboration with staff of the UC Davis Real-time PCR Research & Diagnostic Core Facility, Dept. of Medicine & Epidemiology, School of Veterinary Medicine, University of California, Davis and with the Bioinformatics Core of the UCD Genome Center. We will continue our analysis of genomics data for Vanill
Impacts
What was accomplished under these goals?
This project aims to develop fundamental knowledge that is currently lacking for commercially important orchids. Specifically, this project investigates important reproductive processes using the unique biological system of the orchids, with Phalaenopsis and Vanilla as the primary biological and genomic models, respectively. The overall focus is on the unique program in orchids for: 1) flower development with a focus on the gynostemium, and male and female gametophytes; 2) pollination and post-pollination development; 3) floral aroma; 4) floral longevity and perianth senescence in response to pollination, emasculation, or hormones; 5) post-pollination ovary and ovule development; 6) fertilization and early seed (embryo and endosperm) development; and 7) fruit development, maturation and ripening. The long-term goal is to obtain a detailed understanding of molecular genetic events that control these reproductive processes in commercially important orchid cultivars. For example, we will gain knowledge about floral longevity and senescence, floral scent product, fruit quality and aroma, and stress responses (abiotic and biotic). Development of improved varieties of premier-quality Phalaenopsis orchids that have fragrant as well as long-lived flowers, are disease resistant, and more resilient to climate stress, would be a major output achievement. During the fourth year of the project, many projects were brought to fruition in the form of publications, presentations and new data sets under bioinformatics analysis. In one case, we completed a detailed quantitative analysis of the effect of pollination and pollination-associated events or factors, such as emasculation, auxin, ethylene, and ACC on the expression of ethylene biosynthetic genes in Phalaenopsis orchids that leads to flower senescence or development. This investigation provides the first report of localization by in situ hybridization of ACC synthase in the gynoecium of pollinated orchid flowers over an early and detailed time course and in response to ethylene treatment. In addition, it addresses important questions regarding the induction of ethylene by pollination through an in-depth examination of ACC oxidase gene expression at the earliest times after pollination in stigma tissue. The results are novel and make important contributions to the body of knowledge about plant hormone regulation of development as well as cell-specific expression of two important ethylene biosynthetic genes. In addition, the use of quantitative methods to measure gene expression using real-time quantitative PCR provides a test and validation of earlier reports in the literature on gene expression in flower tissue measured by qualitative methods, such as RNA blot hybridization. Finally, this research provides new insight into the possible mode of regulation of seed plant ACC synthases by examining the question of post-transcriptional regulation of orchid ACC synthases. A novel and unique to orchids consensus motif is reported for orchid ACC synthases that may be involved in their unique mode of post-transcriptional regulation similar to Type 3 ACC synthase genes/proteins in Arabidopsis and tomato. In a second project, we conducted a detailed analysis of 8 vanillin pathway-associated compounds in unripe green vanilla fruit and the effects of ethylene and abiotic stress (high temperature shock or stress) on the levels of the compounds. Eight compounds examined were present in different relative amounts in six-month-old green unripe fruit of Vanilla planifolia, and most were present in both their non-glycosylated and glycosylated forms. Ethylene clearly induced the biosynthesis of certain intermediates in the vanillin biosynthetic pathway, as proven by the use of the ethylene action inhibitor, 1-MCP. Ethylene was involved in increasing the amounts of key pathway compounds, whereas heat was the important factor for other compounds. Some of these increases were supported in enzyme-treated samples by statistical analysis. Principal axis factor analysis was also conducted. This is the first report of the use of 1-MCP to examine the role of ethylene in vanilla fruit ripening. A third investigation was completed on two endangered orchid species, Vanilla grandifolia, and Vanilla pompona, useful for fruit production and useful as highly suitable parents in interspecific hybridizations. Both species have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We characterized flowering and fruit development as never before reported and provided important basic knowledge about Vanilla orchids. This information is novel because there are few to no publications about these two species and few complete developmental studies of vanilla reproduction. This research line provides fundamental information about reproduction to guide breeding efforts with Vanilla planifolia, the orchid of commerce, through interspecific hybridization. During the fourth year, this research was presented at the World Orchid Conference in Guayaquil, Ecuador (November, 2017) and at the annual meeting of the American Society of Plant Physiologists in Quebec, Canada (July 2018). In another CRIS project study of Phalaenopsis was brought to completion and a manuscript was submitted and re-submitted following revisions. This project provides totally new insight into the mechanism of pollination of orchids. Professor O'Neill has proposed a four-stage process involving pollen-derived signals and the novel differentiation of fertile stigmatic cells that are required for fertility and pollination success. In another project, Professor O'Neill and her graduate student, Amy Bump, conducted a project aimed at establishing and evaluating via real-time quantitative Polymerase Chain Reaction (RT-qPCR) analysis and statistical analysis a set of Reference Genes to use in our projects aimed at evaluating gene expression in the transcriptomes described in Products: Entries 1 & 2. Reference Genes are critical to standardize and evaluate gene expression across tissues and developmental stages, with good Reference Genes being expressed stably at low levels across all time points and in all tissues. We were able to establish a number of reliable Reference Genes and disqualify other candidates as unstable and not suitable. The second aim of the project was to use the Reference Genes to assess the role of the vanillin synthase gene (VpVAN) during fruit development and ripening. These results will be submitted for publication in a peer-reviewed journal shortly and will credit NIFA support and be reported in the Annual Report for year 4 (2018-2019). The poster presentation (Products: Entry 2) describing preliminaries of this research won the only award of "First Prize for a Scientific Poster" at the World Orchid Conference, Ecuador (November, 2018).
Publications
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2019
Citation:
Citation: O'Neill, S. D. 2017. A novel secretory structure is involved in fertilization and required for fruit development in Vanilla planifolia. Published September 2017.
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2019
Citation:
Citation: A. Bump, C. Wademan, S. Barnum, E. Hodzic, and S. D. O'Neill. 2017.
Evaluation of reference genes and a potential vanillin synthase gene during fruit development in Vanilla planifolia. Published September 2017.
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:The Principal Investigator Professor Sharman O'Neill collaborates with several different types of groups within the target audience. Professor O'Neill has established relationships with important stakeholders in the California floriculture and horticulture industries, especially major commercial orchid growers. Professor O'Neill continues to engage with other nursery/greenhouse operations in the Bay Area, south into the Salinas area, and north into Sebastopol. Professor O'Neill communicates with a number of California-based grower and horticulture businesses involved in orchid potted flowering plant production (e.g., Matsui Nursery, Salinas, CA) and marketing of flowers, and with other California-based companies involved in the vanilla industry including the sale of fruit (From The Field Trading Company, Inc.; Saffron Imports, Inc.; Cook Flavoring and Manufacturing Company) and other international and USA corporations involved in crop improvement and food and beverage manufacturing (General Mills Inc., Synthrex, and International Flavour and Fragrance). All these groups are interested in Professor O'Neill's research on orchid reproduction and orchid fruit flavor and aroma. During the third year of the project, Dr. O'Neill established a new public-private partnership involving research sponsorship with a California-based flavor manufacturing business, Cook Flavoring Company and Lochhead Manufacturing Company, Paso Robles, CA. She has also engaged with manufacturers and suppliers of greenhouse hydroponics equipment for her experimental use for growing orchids for research. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples include invitations to speak at major public education institutions in San Francisco including the Conservatory of Flowers in Golden Gate Park, San Francisco, the San Francisco Exploratorium at Pier 15 Embarcadero, and at public California State Universities. Also, Professor O'Neill was invited to present her research on vanilla in a major Vanilla Symposium to a scientific audience and to a public audience of orchid growers, conservationists, and representatives of most major research institutions involved in orchid horticulture and conservation. Specifically, Professor O'Neill was an invited Symposium Speaker at the 22nd World Orchid Congress, Ecuador, and gave a scientific talk to an international audience of scientists, horticultural producers of orchids, and dedicated conservationists of orchids, with participants from many agricultural nations worldwide specializing in orchid floricultural production. She was invited to present her research at two additional international meeting in the near future 2018 (Mexico International Network on Vanilla) and 2019 (Taiwan 23rd World Orchid Conference) that will further credit this CRIS Project. The PI has ongoing formal collaboration on Phalaenopsis and Vanilla orchid research with scientists at the USDA-ARS, Pacific Basin Agricultural Research Center (Hawaii) and Hawaiian orchid producers and farmers. She also continued a collaboration with the Tropical Agricultural Research Station at the USDA-ARS Mayaguez, Puerto Rico, and had planned to visit the USDA-ARS Mayaguez, but the trip was postponed due to current natural disasters in that country making travel impossible. Finally, Professor O'Neill is the Director of the International Vanilla Users Group and the Vanilla Genomics Project called "The Vanilla Sustainability Project" and as such has communicated with many scientists, farmers, business leaders and other stakeholders worldwide. She is also actively involved in Vanilla germplasm conservation and participates in activities of the California-based group, the Orchid Conservation Alliance (Encinitas, CA) for whom she is planning to give a series of talks on Vanilla conservation in Southern California and Arizona during project year 4. During the third year, Professor O'Neill continued in her role as Editor for Plant Biology (five subfields from applied to basic research), as a major part of Elsevier's Major Reference Works in the Life Sciences. This activity reached a large international target audience of scientists and students through its continually updated online resource in plant biology. In summary, Professor O'Neill continued to share her basic science and applied knowledge expertise with California orchid producers, scientists, agronomists, and industry leaders, especially in the area of molecular biology and genetics of commercially important orchids, especially Phalaenopsis and Vanilla. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This CRIS project provided training to one second-year Horticulture and Agronomy Graduate Group graduate student, Ms. Amy Bump, for whom Dr. O'Neill serves as graduate faculty mentor and Major Professor, and 14 UC Davis undergraduate students, including one Honors Program student. Another Lab Research Affiliate has been Mr. Garrett Cunha, who continued his histological studies of orchid root anatomy. Another 6 undergraduate students received mentoring and training in how to be a Teaching Assistant, an excellent experience for their university experience. The 8 Undergraduate Research Interns listed below conducted independent research projects with Professor O'Neill and her lab team in either the lab or greenhouse for which they received academic credit in PLB 192 or MCB 099/199 courses. For 2016-2017: Annie Ashmore Amy Bump Alexander Cain Illiana Castenada Michael Mears Miyako Noguchi Heewon Shin Abigail Stevens. Professor O'Neill also served as the Faculty Mentor and adviser for Ms. Anne Ashemore for her Senior Honors Thesis project administered through the University Honors Program as a kind of capstone course. Her Honor's Thesis project resulted in two different posters that were presented at the UC Undergraduate Research Biology Conference (Spring 2017) and at the World Orchid Conference (Fall 2017). Both of Ms. Ashemore's presentations credited the USDA-CRIS support to Professor O'Neill. How have the results been disseminated to communities of interest?The results have been disseminated to communities of interest, especially in the form of public outreach on behalf of the University of California, Davis and through publications and presentations. Professor O'Neill's target audience also included the general public and members of the academic community. She presented her research on orchids in several high-profile public outreach presentations, each time representing the University of California, Davis, allowing her to share the science and research activities of the University with the general public and academic audiences. Some examples include invitations to speak at major public education institutions in San Francisco including the Conservatory of Flowers in Golden Gate Park, San Francisco, the San Francisco Exploratorium at Pier 15 Embarcadero, and at public California State Universities. One example is Professor O'Neill's public outreach to the San Francisco general public. She gave a talk about the research she is conducting on Vanilla planifolia at UC Davis. Professor O'Neill's talk was targeted to an educated non-scientist audience that are interested in science. San Francisco Exploratorium, Pier 15 Observatory, Exploratorium Museum, SF, CA. Invited talk in "Pairings: Cultivating a Taste for Science through Food" series to large audience (120 adult members) of the SF Exploratorium (including Board Members and some Donors). This event was advertised on Facebook to "hundreds of thousands of Exploratorium members." Abstract of Professor O'Neill's talk: Title: Vanilla: From Flower to Fertilization and Flavor. Sharman D. O'Neill, M.A., Ph.D. Professor Department of Plant Biology College of Biological Sciences University of California, Davis, Davis, CA 95616 U.S.A. E-mail: sdoneill@ucdavis.edu Vanilla planifolia, an orchid of Mesoamerican origin, is exceptional among the large family of orchid species in that it produces an extraordinary aromatic fruit, known as the "vanilla bean." Vanilla beans have the highest value of any legal crop due to their sustainable mode of production and process. Recently though the price of one kilogram of cured vanilla beans soared to ~$600.00 reflecting serious problems in the vanilla industry. A key issue is that of fruit quality. There is a conundrum that when the price of vanilla fruit is high, their quality is low. But what determines fruit quality? From scientific investigation, the developmental pathway from flower to fertilization to fruit determines fruit quality and involves complex biological processes of pollination and fertilization. The growth and development of a vanilla fruit requires a full 9 months. During this time, the coordinated development of new reproductive structures in the ovary largely determine fruit quality measures, such as aroma. Our research has focused on vanilla flowers and fruit using genomics and bioinformatics to understand the underlying molecular genetic basis of fruit quality. In order to understand these changes, it was essential to obtain detailed knowledge of reproduction in this species, beginning with flower pollination. This research will be presented in the broader context of The Vanilla Sustainability Project, an international collaboration for crop improvement in Vanilla planifolia, germplasm conservation, sustainability, and for biodiversity conservation in Madagascar, the center of world vanilla production. This presentation will also provide an overview of the history of vanilla cultivation in Mexico, the patrimonial birthplace of vanilla, and in Europe and islands of the Southwest Indian Ocean including Madagascar, Réunion Island, and the Comoros Islands. Flower diversity will be shown of different vanilla species, time permitting. Demonstrations will include vanilla beans from these regions and live vanilla plants. This talk was extremely well received by the public and staff of the Exploratorium. Feedback shared later with Professor O'Neill stated that the talk was "fantastic." The USDA-CRIS funding was acknowledged. At another major international conference, Professor O'Neill was invited to present her research on vanilla to a scientific audience as well as to a public audience of orchid growers, conservationists, and representatives of most major research institutions involved in orchid horticulture and conservation. Specifically, Professor O'Neill was an invited Vanilla Symposium Speaker at the 22nd World Orchid Congress, Ecuador, and gave a scientific talk to an international audience of scientists, horticultural producers of orchids, and dedicated conservationists of orchids, with participants from many agricultural nations worldwide specializing in orchid floricultural production. She was invited to present her research at two additional international meeting in the near future 2018 (Mexico International Network on Vanilla) and 2019 (Taiwan 23rd World Orchid Conference) that will further credit this CRIS Project. Feedback at the conference about the presentation was stated that Professor O'Neill had contributed "academic gravitas" with her research on Vanilla planifolia reproduction and that the WOC Organizing Committee recognized the importance of this research and that it was associated with the University of California, Davis, and Plant Biology. In this talk, the USDA-CRIS funding was acknowledged. What do you plan to do during the next reporting period to accomplish the goals?Ms. Amy Bump, Graduate Student, will complete her data analyses on the Reference Gene project and do the same with the VpVan project. The Reference Genes will be used as controls for analysis of gene expression of an important biosynthetic gene in the phenylpropanoid pathway and in other developmental pathways of interest to Professor O'Neill. This work is being done in collaboration with staff of the UC Davis Real-time PCR Research & Diagnostic Core Facility, Dept. of Medicine & Epidemiology, School of Veterinary Medicine, University of California, Davis. We will complete an additional study of the effects of auxin, ethylene, gibberellic acid and inhibitors on post-pollination development in Phalaenopsis flowers. We will continue research on the importance of the organ re-purposing and the critical role of the perianth remainder in orchids. We will continue our analysis of genomics data for Vanilla, with a comparison to Phalaenopsis genomics data.
Impacts
What was accomplished under these goals?
This project aims to develop fundamental knowledge that is currently lacking for commercially important orchids. Specifically, this project investigates important reproductive processes using the unique biological system of the orchids, with Phalaenopsis and Vanilla as the primary biological and genomic models, respectively. The overall focus is on the unique program in orchids for: 1) flower development with a focus on the gynostemium, and male and female gametophytes; 2) pollination and post-pollination development; 3) floral aroma; 4) floral longevity and perianth senescence in response to pollination, emasculation, or hormones; 5) post-pollination ovary and ovule development; 6) fertilization and early seed (embryo and endosperm) development; and 7) fruit development, maturation and ripening. The long-term goal is to obtain a detailed understanding of molecular genetic events that control these reproductive processes in commercially important orchid cultivars. For example, we will gain knowledge about floral longevity and senescence, floral scent product, fruit quality and aroma, and stress responses (abiotic and biotic). Development of improved varieties of premier-quality Phalaenopsis orchids that have fragrant as well as long-lived flowers, are disease resistant, and more resilient to climate stress, would be a major output achievement. During the third year of the project, many projects were brought to fruition in the form of publications and presentations. In one case, we completed a detailed quantitative analysis of the effect of pollination and pollination-associated events or factors, such as emasculation, auxin, ethylene, and ACC on the expression of ethylene biosynthetic genes in Phalaenopsis orchids that leads to flower senescence or development. This investigation provides the first report of localization by in situ hybridization of ACC synthase in the gynoecium of pollinated orchid flowers over an early and detailed time course and in response to ethylene treatment. In addition, it addresses important questions regarding the induction of ethylene by pollination through an in-depth examination of ACC oxidase gene expression at the earliest times after pollination in stigma tissue. The results are novel and make important contributions to the body of knowledge about plant hormone regulation of development as well as cell-specific expression of two important ethylene biosynthetic genes. In addition, the use of quantitative methods to measure gene expression using real-time quantitative PCR provides a test and validation of earlier reports in the literature on gene expression in flower tissue measured by qualitative methods, such as RNA blot hybridization. Finally, this research provides new insight into the possible mode of regulation of seed plant ACC synthases by examining the question of post-transcriptional regulation of orchid ACC synthases. A novel and unique to orchids consensus motif is reported for orchid ACC synthases that may be involved in their unique mode of post-transcriptional regulation similar to Type 3 ACC synthase genes/proteins in Arabidopsis and tomato. See Products: Entry 1. In a second project, we conducted a detailed analysis of 8 vanillin pathway-associated compounds in unripe green vanilla fruit and the effects of ethylene and abiotic stress (high temperature shock or stress) on the levels of the compounds. Eight compounds examined were present in different relative amounts in six-month-old green unripe fruit of Vanilla planifolia, and most were present in both their non-glycosylated and glycosylated forms. Ethylene clearly induced the biosynthesis of certain intermediates in the vanillin biosynthetic pathway, as proven by the use of the ethylene action inhibitor, 1-MCP. Ethylene was involved in increasing the amounts of key pathway compounds, whereas heat was the important factor for other compounds. Some of these increases were supported in enzyme-treated samples by statistical analysis. Principal axis factor analysis was also conducted. This is the first report of the use of 1-MCP to examine the role of ethylene in vanilla fruit ripening. See Products: Entry 3. A third investigation was completed on two rare orchid species, Vanilla calopogon, an IUCN Red List species, and Vanilla imperialis, a contrasting species, but both highly suitable parents in interspecific hybridizations. Both species have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We characterized flowering and fruit development as never before reported and provided important basic knowledge about Vanilla orchids. This information is novel because there are few to no publications about these two species and few complete developmental studies of vanilla reproduction. This research line provides fundamental information about reproduction to guide breeding efforts with Vanilla planifolia, the orchid of commerce, through interspecific hybridization. See Products: Entry 8. Another study involved ex situ conservation of rare Vanilla orchid species germplasm collection that have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We describe the development of a modified improved procedure for the in vitro propagation of American wild non-cultivated Vanilla species in our ex situ collection. This information is novel because there are no publications about the in vitro culture of these delicate and rare Vanilla species that are important as breeding partners with Vanilla planifolia, the orchid of commerce. It is also an important contribution because it provides information about ex situ conservation efforts for the genus Vanilla using newly developed in vitro methods. See Products: Entry 9. In another CRIS project study of Phalaenopsis was brought to completion and a manuscript was submitted and re-submitted following revisions (Products: Entry 10). This project provides totally new insight into the mechanism of pollination of orchids. Professor O'Neill has proposed a four-stage process involving pollen-derived signals and the novel differentiation of fertile stigmatic cells that are required for fertility and pollination success. In another project, Professor O'Neill and her graduate student, Amy Bump, conducted a project aimed at establishing and evaluating via real-time quantitative Polymerase Chain Reaction (RT-qPCR) analysis and statistical analysis a set of Reference Genes to use in our projects aimed at evaluating gene expression in the transcriptomes described in Products: Entries 1 & 2. Reference Genes are critical to standardize and evaluate gene expression across tissues and developmental stages, with good Reference Genes being expressed stably at low levels across all time points and in all tissues. We were able to establish a number of reliable Reference Genes and disqualify other candidates as unstable and not suitable. The second aim of the project was to use the Reference Genes to assess the role of the vanillin synthase gene (VpVAN) during fruit development and ripening. These results will be submitted for publication in a peer-reviewed journal shortly and will credit NIFA support and be reported in the Annual Report for year 4 (2018-2019). The poster presentation (Products: Entry 6) describing preliminaries of this research won the only award of "First Prize for a Scientific Poster" at the World Orchid Conference, Ecuador.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Citation: ONeill, S. D., A. Q. Bui, D. Potter, and X. S. Zhang. 2017.
Pollination of Orchid Flowers: Quantitative and Domain-Specific Analysis of Ethylene Biosynthetic and Hormone-Induced Gene Expression. International Journal of Plant Sciences 178: 188-210.
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Citation: O'Neill, S.D. 2016. Plant Biology. Feature article. Major Reference Works in the Life Sciences. Elsevier, Inc., Oxford, England. United Kingdom. Published October 31, 2016.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2018
Citation:
Citation: Little, S.M, S. T. de Freitas, F. Negre-Zakharov, A. E. Mitchell, and S. D. ONeill. Effect of postharvest abiotic stress on vanillin production in Vanilla planifolia fruit. Journal of Postharvest Biology and Biotechnology.
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2018
Citation:
Citation: O'Neill, S. D. 2018. A novel secretory structure is involved in fertilization and required for fruit development in Vanilla planifolia. In: A. Pridgeon, Ed.. Proceeding of the 22nd World Orchid Conference.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Citation: O'Neill, S. D. 2017. A novel secretory structure is involved in fertilization and required for fruit development in Vanilla planifolia. Published September 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Citation: A. Bump, C. Wademan, S. Barnum, E. Hodzic, and S. D. O'Neill. 2017.
Evaluation of reference genes and a potential vanillin synthase gene during fruit development in Vanilla planifolia. Published September 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Citation: A. Ashmore, A. Bump, and S. D. O'Neill. 2017.
Fruit morphology and developmental deviation within Vanilla pompona subspecies. Published September 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Citation: G. M. Abatchev and S. D. ONeill. 2017. Flowering and fruit development in two Old World species of Vanilla: Vanilla calopogon (Asia) and Vanilla imperialis (Africa). Published September 2017.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2018
Citation:
Citation: Abatchev, A. M., and S. D. O'Neill. Ex situ conservation of Vanilla odorata (America) and other endangered genus Vanilla (Orchidaceae) species using an improved nodal tissue culture propagation method. In Vitro Cellular & Developmental Biology- Plants
- Type:
Journal Articles
Status:
Under Review
Year Published:
2018
Citation:
Citation: O'Neill, S. D. Gynoecium remodeling and ovary development in Phalaenopsis orchid flowers (Orchidaceae) in response to pollen signals and auxin but without pollen tube signaling. American Journal of Botany (Under Review).
|
Progress 10/01/15 to 09/30/16
Outputs
Target Audience:The Principal Investigator Dr. O'Neill collaborates with several different groups within the target audience. She visits stakeholders in the floriculture industry, especially major California commercial orchid growers in Salinas, including Matsui Nursery. Dr. O'Neill will continue to engage other nursery/ greenhouse operations in Half Moon Bay, Salinas, and Sebastopol, as well as suppliers of greenhouse hydroponics equipment for use in growing research orchids. She will continue communication with a number of California-based businesses involved in orchid potted flowering plant production and marketing of flowers and fruits, and with other non-floricultural California-based companies (e.g., From The Field Trading Company, Inc., Saffron Imports, Inc.) and others (General Mills Inc., Syntex, and IFF interested in orchid flower and fruit aroma. Professor O'Neill is the Director of the International Vanilla Users Group and the Vanilla Genomics Project called "The Vanilla Sustainability Project" and as such has communicated with many scientists, farmers, business leaders and other stakeholders worldwide. During the second year of the project, Dr. O'Neill has newly established a funding support relationship with a California-based flavor extract business. During the reporting period in March 2016, Dr. O'Neill presented her research on vanilla sustainability to an international scientific congress of growers and producers of orchid flowers and fruit, with participants from many agricultural nations specializing in orchid floricultural and also wood agricultural production. The PI also has formal collaboration on Phalaenopsis and Vanilla orchid research with scientists at the USDA-ARS, Pacific Basin Agricultural Research Center (Hawaii) and Hawaiian orchid producers and farmers. She also has a collaboration with Dr. Brian Irish at the USDA-ARS Mayaguez, Puerto Rico, and had planned to visit the USDA-ARS Mayaguez in June of Year 2, but the trip was postponed. Instead, she plans to visit USDA-ARS Mayaguez in June 2017. Finally, Dr. O'Neill will continue to share her basic science and applied knowledge expertise with California orchid producers, scientists, agronomists, and industry leaders, especially in the area of molecular biology and genetics of commercially important orchids, especially Phalaenopsis and Vanilla. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This CRIS project provided training to one first-year Horticulture and Agronomy Graduate Group graduate student, Ms. Amy Bump, for whom Dr. O'Neill serves as graduate faculty mentor and Major Professor, and 12 UC Davis undergraduate students, including one Honors Program student. Another Lab Research Affiliate has been Mr. Garrett Cunha, who continued his histological studies of orchid root anatomy. The Undergraduate Research Interns also received academic credit as PLB 192 or MCB 099/199 courses. For 2015-2016: Gamid Abatchev Taraneh Abudadi Adrienne Abu-Assal Amy Bump Yelin Hu. For 2016: Annie Ashmore Amy Bump Alexander Cain Illiana Castenada Michael Mears Miyako Noguchi Heewon Shin How have the results been disseminated to communities of interest?The results have been disseminated to communities of interest, especially in the form of publications and presentations. Dr. O'Neill was an invited international speaker at the I Congreso Internacional de Vainilla, Sistemas Agroforestales y Productos No Maderables del Bosque. Santa Clara, San Carlos, Costa Rica. March 7-11, 2016, where she gave a presentation on "Vanilla planifolia Reproductive Development & Vanilla Germplasm Conservation" to a large international audience. What do you plan to do during the next reporting period to accomplish the goals?Ms. Amy Bump, Graduate Student, is continuing research to analyze and validate gene expression using the analytic diagnostic procedure of RT-qPCR. We have developed a set of Reference Genes and validated them for all tissue types. Now we are using the Reference Genes as a control for analysis of gene expression of an important biosynthetic gene in the phenylpropanoid pathway. This work is being done in collaboration with the UC Davis Real-time PCR Research & Diagnostic Core Facility working with Ms. Samantha Mapes, Laboratory Manager, and Dr. John Madigan, DVM, Director and Professor, Dept. of Medicine & Epidemiology, School of Veterinary Medicine, University of California, Davis. We will complete an additional study of the effects of auxin, ethylene, gibberellic acid and inhibitors on post-pollination development in Phalaenopsis flowers. We will continue research on the importance of the perianth remainder using Phalaenopsis. We will continue our analysis of genomics data for Vanilla, with a comparison to Phalaenopsis genomics data.
Impacts
What was accomplished under these goals?
This project aims to develop fundamental knowledge that is currently lacking for commercially important orchids. Specifically, this project investigates important reproductive processes using the unique biological system of the orchids, with Phalaenopsis and Vanilla as the primary biological and genomic models, respectively. The overall focus is on the unique program in orchids for: 1) flower development with a focus on the gynostemium, and male and female gametophytes; 2) pollination and post-pollination development; 3) floral aroma; 4) floral longevity and perianth senescence in response to pollination, emasculation, or hormones; 5) post-pollination ovary and ovule development; 6) fertilization and early seed (embryo and endosperm) development; and 7) fruit development, maturation and ripening. The long-term goal is to obtain a detailed understanding of molecular genetic events that control these reproductive processes in commercially important orchid cultivars. For example, we will gain knowledge about floral longevity and senescence, floral scent product, fruit quality and aroma, and stress responses (abiotic and biotic). Development of improved varieties of premier-quality Phalaenopsis orchids that have fragrant as well as long-lived flowers, are disease resistant, and more resilient to climate stress, would be a major output achievement. During the second year of the project, many projects were brought to fruition in the form of publications and presentations. In one case, we completed a detailed quantitative analysis of the effect of pollination and pollination-associated events or factors, such as emasculation, auxin, ethylene, and ACC on the expression of ethylene biosynthetic genes in Phalaenopsis orchids that leads to flower senescence or development. This investigation provides the first report of localization by in situ hybridization of ACC synthase in the gynoecium of pollinated orchid flowers over an early and detailed time course and in response to ethylene treatment. In addition, it addresses important questions regarding the induction of ethylene by pollination through an in-depth examination of ACC oxidase gene expression at the earliest times after pollination in stigma tissue. The results are novel and make important contributions to the body of knowledge about plant hormone regulation of development as well as cell-specific expression of two important ethylene biosynthetic genes. In addition, the use of quantitative methods to measure gene expression using real-time quantitative PCR provides a test and validation of earlier reports in the literature on gene expression in flower tissue measured by qualitative methods, such as RNA blot hybridization. Finally, this research provides new insight into the possible mode of regulation of seed plant ACC synthases by examining the question of post-transcriptional regulation of orchid ACC synthases. A novel and unique to orchids consensus motif is reported for orchid ACC synthases that may be involved in their unique mode of post-transcriptional regulation similar to Type 3 ACC synthase genes/proteins in Arabidopsis and tomato. In a second project, we conducted a detailed analysis of 8 vanillin pathway-associated compounds in unripe green vanilla fruit and the effects of ethylene and abiotic stress (high temperature shock or stress) on the levels of the compounds. Eight compounds examined were present in different relative amounts in six-month-old green unripe fruit of Vanilla planifolia, and most were present in both their non-glycosylated and glycosylated forms. Ethylene clearly induced the biosynthesis of certain intermediates in the vanillin biosynthetic pathway, as proven by the use of the ethylene action inhibitor, 1-MCP. Ethylene was involved in increasing the amounts of key pathway compounds, whereas heat was the important factor for other compounds. Some of these increases were supported in enzyme-treated samples by statistical analysis. Principal axis factor analysis was also conducted. This is the first report of the use of 1-MCP to examine the role of ethylene in vanilla fruit ripening. A third investigation was completed on two rare orchid species, Vanilla calopogon, an IUCN Red List species, and Vanilla imperialis, a contrasting species, but both highly suitable parents in interspecific hybridizations. Both species have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We characterized flowering and fruit development as never before reported and provided important basic knowledge about Vanilla orchids. This information is novel because there are few to no publications about these two species and few complete developmental studies of vanilla reproduction. This research line provides fundamental information about reproduction to guide breeding efforts with Vanilla planifolia, the orchid of commerce, through interspecific hybridization. professor O'Neill and her student, Gamid Abatchev, also developed a three-phase predictive model that will be useful as a guide for future interspecific hybridization research. Another study involved ex situ conservation of rare Vanilla orchid species germplasm collection that have been under conservation in the Vanilla Genetic Diversity Center at the University of California, Davis, USA. We describe the development of a modified improved procedure for the in vitro propagation of American wild non-cultivated Vanilla species in our ex situ collection. This information is novel because there are no publications about the in vitro culture of these delicate and rare Vanilla species that are important as breeding partners with Vanilla planifolia, the orchid of commerce. It is also an important contribution because it provides information about ex situ conservation efforts for the genus Vanilla using newly developed in vitro methods. Finally, another CRIS project study of Phalaenopsis was brought to completion and a manuscript was submitted (O'Neill). This project provides totally new insight into the mechanism of pollination of orchids. Professor O'Neill has proposed a four-stage process involving pollinia-derived signals and the novel differentiation of fertile stigmatic cells that are required for fertility and pollination success.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
O'Neill, S.D. 2016. Vanilla planifolia Reproductive Development & Vanilla Germplasm Conservation. I Congreso Internacional de Vainilla, Sistemas Agroforestales y Productos No Maderables del Bosque. Santa Clara, San Carlos, Costa Rica. March 7-11, 2016.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2017
Citation:
ONeill, S. D., A. Q. Bui, D. Potter, and Zhang, X. S. Pollination of orchid flowers: quantitative and domain-specific analysis of ethylene biosynthetic and hormone-induced gene expression. International Journal of Plant Sciences (In Press).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2017
Citation:
Little, S.M, S. T. de Freitas, F. Negre-Zakharov, A. E. Mitchell, and ONeill, S. D. (2016) Effect of postharvest abiotic stress on vanillin production in Vanilla planifolia fruit. Journal of Postharvest Biology and Biotechnology (Under Review).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2017
Citation:
O'Neill, S. D., and Abatchev, A. M. Vanilla calopogon (Asia) and Vanilla imperialis (Africa): identification, flowering, and comparison of fruit development in two Old World species of Vanilla subgenus Xanata section Tethya. Botanical Journal of the Linnean Society (Under Review).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2017
Citation:
Abatchev, A. M., and O'Neill, S. D.. Ex situ conservation of Vanilla odorata (America) and other endangered genus Vanilla (Orchidaceae) species using an improved nodal tissue culture propagation method. In Vitro Cellular & Developmental Biology- Plants (Under Review).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2017
Citation:
O'Neill, S. D. Gynoecium remodeling and ovary development in Phalaenopsis orchid flowers (Orchidaceae) in response to pollen signals and auxin but without pollen tube signaling. American Journal of Botany (Under Review).
- Type:
Other
Status:
Published
Year Published:
2016
Citation:
: O'Neill, S.D. 2016. Plant Biology. Feature article. In: Major Reference Module in Life Sciences. Major Reference Works in Plant Biology. Subject Editor Sharman O'Neill. Elsevier, Inc., Oxford, UK (Published online on October 31, 2016 - release date November 7, 2016 and viewable via ScienceDirect).
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2015
Citation:
Little, S.M. 2015. Effect of postharvest abiotic stress on vanillin production in Vanilla planifolia fruit. Master's Thesis. University of California, Davis. Published in ProQuest.
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:The Principal Investigator Dr. O'Neill collaborates with several different groups within the target audience. She visits stakeholders in the floriculture industry, especially major California commercial orchid growers in Salinas, including Matsui Nursery (collaborating producer). Dr. O'Neill will continue to engage other nursery/greenhouse operations in Half Moon Bay, Salinas, and Sebastopol, as well as suppliers of greenhouse hydroponics equipment for use in growing research orchids. She will continue communication with a number of California-based businesses involved in orchid potted flowering plant production and marketing of flowers and fruits, and with other non-floricultural California-based companies (e.g., From The Field Trading Company, Inc., Saffron Imports, Inc.) and others (General Mills Inc., Syntex, and IFF interested in orchid flower and fruit aroma. PI O'Neill is the Director of the International Vanilla Users Group and the Vanilla Genomics Project called "The Vanilla Sustainability Project" and as such has communicated with many scientists, farmers, business leaders and other stakeholders worldwide. She is also a collaborating primary "Investigator" on the ERANET VaBiome Project (European Union, France), which is a collaborating multinational project that utilizes data from the UC Davis-based genomics project on the orchid of commerce, Vanilla planifolia. Dr. O'Neill will give presentations to the Food and Beverage Manufacturing Industry and has present her research at national and international scientific community conferences. The PI also has formal collaboration on Phalaenopsis and Vanilla orchid research with scientists at the USDA-ARS, Pacific Basin Agricultural Research Center (Hawaii) and Hawaiian orchid producers and farmers. Finally, she will continue to share her expertise with California orchid producers, scientists, agronomists, and industry leaders, especially in the area of molecular biology and genetics of commercially important orchids, especially Phalaenopsis and Vanilla. Finally, Dr. O'Neill will continue to advise other UC Davis CAES faculty and their graduate students about orchid reproduction and genomic approaches to crop improvement. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project provided training to one new Horticulture and Agronomy Graduate Group graduate student, Ms. Amy Bump, (Fall Quarter 2105) for whom I serve as Major Professor. It also provided training to four Undergraduate Research Interns who also receive academic credit as PLB 192 or MCB 192 courses: Mr. Gamid Abatchev, Mr. Yelin Hu, Ms. Taraneh Abudadi, and Ms. Adrienne Abu-Assal. Another Lab Research Affiliate has been Mr. Garrett Cunha, who has continued his histology studies of orchid root anatomy. 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?Main Questions to be immediately addressed in Project Year 2: 1. Does pollination trigger perianth senescence or development in the wiling versus the converting lines? 2. How does ethylene impact pigments such as chlorophylls and anthocyanins during the shift? 3. What are the anatomical or structural changes that occur during this perianth development? Experiment 1: What is the role of ethylene? If the perianth is maintained and converts following pollination, then ethylene levels should be less than wilting flowers. Experiment 2: What happens to the anthocyanins? If pollination triggers the conversion, then the anthocyanin content will decrease due to low ethylene levels and vary depending on the pollen origin. Hybrids will have lower degradation of anthocyanins. Experiment 3: What happens to the chlorophyll and how does it vary? If pollination triggers the conversion, then the chlorophyll content will increase and vary depending on the pollen origin. Hybrids with wilting lines will have lower chlorophyll concentrations. Experiment 4: What are the anatomical changes that occur? If the perianth is being converted into a leaf-like organ, then the perianth should develop specific leaf cell organelles and cell types and organization, including chloroplasts, palisade parenchyma cells, spongy mesophyll cells, and stomatal complexes. We will also commence additional longer-term goals of the project in Year 2: Years 2-4: Identify and analyze the gene activity, hormonal signaling, and biosynthetic pathways that function in orchid flower and fruit development, perianth longevity, floral fragrance, and fruit aroma. As appropriate, genes involved in stress resistance will be analyzed (e.g., ethylene biosynthetic genes and related gene expression). Apply statistical analysis for the determination of differential gene expression. Years 2-5: Analyze gene families, signaling networks, pathways and their regulation. Years 2-5: Use knowledge obtained in these studies to initiate development of genetic engineering strategies for floricultural crop improvement (in collaboration with groups routinely conducting transformation and genetic engineering of floricultural orchids).
Impacts
What was accomplished under these goals?
This project seeks to gain a detailed understanding of the molecular basis of reproductive development in the Phalaenopsis orchid as a model for other orchids, with a particular focus on the flowers: floral aroma, perianth remainder, and possible ethylene regulation of floral organ identity change. For the first year of the project, there have been two primary objectives: Objective 1: Conduct studies that compare the post-pollination syndrome of the elite cultivar Phalaenopsis cv. V8 (non-fragrant hybrid; Matsui Nursery) with three Phalaenopsis "species" orchids (fragrant non-hybrid primary species) that exhibit unique post-pollination developmental responses that may be linked to aroma of both flower and fruit. Objective 2: Investigate the critical role of the perianth remainder in orchid flower senescence and fruit development. During the first year of the project, a new graduate student in the O'Neill Laboratory, Ms. Amy Bump, (Horticulture and Agronomy Graduate Group) has initiated her thesis project research on the Phalaenopsis project and has also conducted additional research to assist with The Vanilla Sustainability Project. Her thesis project is the primary work of this CAES project. During Year 1 she has focused on Objectives 1 and 2. Her research consists of physiological studies of post-pollination phenomena in Phalaenopsis and the role of the perianth remainder. For Project Objective 1, she has ongoing experiments to compare the post-pollination syndrome of the elite cultivar Phalaenopsis cv. V8 (non-fragrant hybrid; Matsui Nursery) with three or more Phalaenopsis "species" orchids (fragrant non-hybrid primary species) that exhibit unique post-pollination developmental responses that may be linked to aroma of both flower and fruit. All species orchids are related to the primary species Phalaenopis amablis that lack perianth color (like cv. V8) and does not exhibit a perianth floral organ identity change response. We have established a small collection of genetically-related lines of species orchids. Their breeding history provides us with a pedigree that enables us to phenotypically distinguish between those species that are aromatic and also show the perianth organ identity conversion (floral organ identity developmental change) from those that are not aromatic and that do not undergo floral organ identity change. We are confirming the phenotype of the various clonal lines. There is variation in the response to pollination in Phalaenopsis. Different species respond to pollination by wiling their perianths while others convert them into fleshy, green leaf-like structures. This split appears to be related to pigmentation and fragrance characteristics in particular species. Some species, such as the non-fragrant and non-pigmented P. amabillis, wilt their perianths; but other species, such as the fragrant and pigmented P. violaeca and P. amboinensis, can convert their perianths into useful leaf-like organs. Just why and how the perianth is converted is not well investigated. Past work explored chlorophyll changes in several wilting and converting species and their hybrids, but offered no anatomical or hormonal data; instead, focus was on chlorophyll changes, which indicated a relationship between pigment production and perianth maintenance. Some of the main questions we have been addressing include: 1. Does pollination trigger perianth senescence or development in the wilting versus converting lines? 2. What cellular changes occur and is ethylene involved in pigment changes, such as chlorophyll production and anthocyanin loss during the identity transition? 3.What are the anatomical and structural changes that occur during this perianth development? Also during Project Year 1, we advanced Project Objective 6. Ms. Bump is conducting a second line of research to analyze and validate gene expression using the analytic diagnostic procedure of RT-qPCR. This work is being done in collaboration with the UC Davis Real-time PCR Research & Diagnostic Core Facility working with Ms. Samantha Mapes, Laboratory Manager, and Dr. John Madigan, DVM, Director and Professor, Dept. of Medicine & Epidemiology, School of Veterinary Medicine, University of California, Davis.
Publications
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