Progress 10/01/19 to 09/30/20
Outputs
Target Audience:The audiences that were targeted included farmers from underserved communities, Historically Black colleges and Universities, Oklahoma colleges and universities, general plant biology and agricultural scientists, daylilies growers and enthusiasts, and other biological scientists and students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project offered direct research skills and experiential learning opportunities to the three project students during the 2019-2020 reporting period. Three students including Zoe McGowan, Kameron Lindsay, and Maya Williams were directly involved in the project activities and benefited research skills and experiential learning. The project also included hands-on lab and field activities of students who participated in Crop Production and Plant Breeding and Genetics courses. Students of those courses also benefited a field trip to the Noble Research Institute, Ardmore, Oklahoma. How have the results been disseminated to communities of interest?In compliance with COVID-19 health requirement, the primarily targeted audience was the Langston University (LU) community and other minority communities of farmers and high school students in the vicinity of the university through field demonstrations and high school and 4-H student hands-on activities. The results were also disseminated through conference presentations and publications. What do you plan to do during the next reporting period to accomplish the goals?In addition to continuous seasonal biological characterizations of varietal activities, the next reporting period will cover partial tissue culture activities that emphasize developing an efficient protocol for in vitro plant regeneration using bract as the organ explant. It will also attempt to pool previous project discoveries and experiences to develop a guideline for a universal in vitro tissue culture protocol for shoot organogenesis. We might also provide preliminary update on in vitro mutation assessment and new genetic discoveries of PCD initial investigations.
Impacts
What was accomplished under these goals?
The project's third growing season was the year 2019-2020 during which we continued seasonal field biological profiling. Previous varietal characteristics such as plant height, blooming rate and period, flower shape and color, seasonal plant regrowth proportion, root type, crown width, planting density, environmental adaptation, and pest, disease and drought tolerance were confirmed. We initially planted 1-2 fans per hole, and by this third year the number of fans per hole has increased up to 24 in four varieties out of the total 250. However, with a few exceptions with a lower end most varieties averaged a dozen of fans per hole by this season, which still is not significant compared to the results from tissue culture approaches (tens per explant when including repetitive shoot organogenesis). Although most varieties were observed to be midseason bloomers, ten were early bloomers while nine were described as late bloomers. Three of late bloomers kept blooming through November. We also identified 12 varieties that formed proliferations, which was three more varieties than last year. We further ascertained that our genetic collection lacks evergreen varieties. Concerning lab activities, we successfully reproduced two of previous in vitro multiple shoot induction protocols, we developed applying stem and bud as explants, in new genotypes. Each protocol induced several folds greater than natural plant multiplication through divisions. This has moved us much closer to developing a universal protocol for daylily in vitro plant regeneration. Furthermore, we expanded preliminary investigations of potential genes responsible for programmed cellular death (PCD) in daylily flowers using a few genes that have been identified in daylilies and other relative plant species. The results raised more questions than answers. For examples, the three genes whose expression was confirmed during flowering were present only in buds and not blooms despite repeated samplings. This suggests that we may be dealing with different PCD genes that may be expressing during blooming that we need to look for. Efforts are underway to expand the screening targeting new genetic discoveries.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Kanyand Matand, Meordrick Shoemake, and Chenxin Li. 2020. High frequency in vitro regeneration of adventitious shoots in daylilies (Hemerocallis sp) stem tissue using thidiazuron. BMC Plant Biology. DOI: 10.1186/s12870-020-2243-7.
- Type:
Journal Articles
Status:
Submitted
Year Published:
2021
Citation:
Kanyand Matand, Meordrick Shoemake, and Chenxin Li. 2020. In vitro daylily (Hemerocallis spp.) bract multiple shoot induction. AJB (AJB/21.11.20/17281)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Matand K, Lindsay K*, McGowan Z*. 2019. Daylilies introductions and research progress at Langston University. 77th PAWC. Opelika, AL (December 8-10)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lindsay K*, McGowan Z*, Matand K. 2019. Molecular analysis for genes responsible for daylily flowers twenty-four-hour preprogrammed apoptosis. 77th PAWC. Opelika, AL (December 8-10).
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Meordrick Shoemake*, Mary Mitchell, Conrad Green. 2019. Daylily in vitro plant micropropagation. 77th PAWC. Opelika, AL (December 8-10).
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:The audiences that were targeted included farmers from underserved communities, Historically Black colleges and Universities, Oklahoma colleges and universities, general plant biology and agricultural scientists, daylilies growers and enthusiasts, and other biological scientists and students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project offered direct research skills and experiential learning opportunities to five students during the 2019 reporting period. Five students including Conrad Green, Mary Mitchell, Meordrick Shoemake, Zoey McGowan, and Kameron Lindsay were directly involved in the project activities and benefited from research skills and experiential learning; and two of the three students who competed in annual academic research activities won first and third place prizes. Each of the three students who completed their experiments participated in three conferences, including local, state and national meetings for the total of nine student presentations. The project director (mentor) presented the project activities at one state and three national meetings, for the grand total of 13 project meeting presentations during the year 2019. A peer-reviewed manuscript was submitted to BMC Plant Biology, and the revision is in progress. The project also included hands-on lab and field activities of students who participated in Crop Production and Plant Breeding and Genetics courses. The project director used the present activities to participate in a two-week summer training to explore ways to team up with scientists at the University of California (Davis, CA) and plan for best strategies to silence daylilies apoptosis genes using the method they have successfully applied in rice. How have the results been disseminated to communities of interest?The primary audiences of Langston University (LU) are African American and other minority communities, who are generally underserved. To reach out to those communities, we teamed up with LU-School of Agriculture and Applied Sciences (LU-SAAS)'s extension programs and tapped into their national networks of farmers and other shareholders and participated in two annual national conferences (Small Farmers and Plasticulture/Horticulture Conferences). The project director presented LU research activities on daylilies. During the discussions on- and offstage, we discovered that there were no daylilies farmers in the audiences. We also realized that few participants, who were familiar with daylilies, knew it only for the beauty of its flowers. We took the opportunities to educate the audiences, in group and one-on-one interactions, on broad uses of daylilies as a medicinal, food nutritional, anti-erosion, landscape, and fire-retardant crop, etc. Because of their great interest, we are currently developing strategies to reach out to broad communities for education about daylilies more effectively. The project director also reached out to the Central Oklahoma Daylilies Association and was invited to make a presentation on daylilies research at LU. Despite the group's specialized interest, participants were not aware of many uses of daylilies other than flower beauty and landscaping. From lessons we learned, LU-SAAS is currently exploring the possibility of leading a national conversation and efforts on reinventing shareholders' education on the true value and image of daylilies. The project director participated in the biannual activities of the Association of Research Directors, the largest academic research gathering of Historically Black Colleges and Universities in the Nation; he focused more on scientific advancement made in daylilies studies at LU. Dr. Kanyand Matand is currently reaching out to scientists at the Noble Research Institute (Ardmore, OK) and exploring the possibility of teaming up to hasten advancement of daylilies genomics studies. To enhance local communities' education about daylilies, student mentees participated in individual research projects and disseminate the results at annual academic research activities that were organized locally, regionally, and nationally. The initial LU genetic stock is located along a rural road that is frequented mostly by local residents. Since installing the experiment, we have experienced this year the most site visits by residents to learn about the crop and research activities, primarily because of the maximal crop blooming we reached this season. The most impressive and inspiring outreach experience we had was the interaction with an African American family of three generations (grandmother, daughter, and two high school granddaughters) living in the vicinity (about 10 miles) of the field experiment. In search of opportunities for better future of the granddaughters, the grandmother and daughter initially visited the project's field experiment for curiosity; then, they brought their two high school girls to work on the field the next weekend. Their main purpose was to expose the youngsters to a unique and beautiful research and learning environment that could inspire them to dream big and aspire going to college after high school. Since then, they have committed to coming to work in the field experiment during the weekends. After interactions with the daylilies project director, the two high schoolers are now planning to join agricultural science majors at LU and their aunt has applied for work in the daylilies project field experiment. What do you plan to do during the next reporting period to accomplish the goals?The next reporting period will focus on partial activities that will emphasize developing an efficient protocol for in vitro plant regeneration of daylilies using bract as the organ explant. It will also give preliminary insights into the potential involvement of other species' orthologs in triggering apoptosis in daylilies. We also plan to confirm the prominence of select biological characteristics of agronomic importance that will guide the selection of original progenitors for long-term plant breeding studies. In addition to continuous tissue culture and field experimental studies, the report could lay the foundation for broad genomics investigations of daylilies apoptosis genes.
Impacts
What was accomplished under these goals?
The year 2019 was the project's second growing season. During this period, we focused on developing an in vitro plant regeneration protocol that can be applied across varieties using flower bud as the main organ explant. We also continued carrying out field biological characterization of individual varieties to ensure successful introduction to the new environment conditions and confirm or discover new traits of agronomic importance that can be incorporated or targeted into long-term breeding or genomics research programs. Overall, this year's project activities were very productive. We developed a more effective protocol for mass-producing in vitro plants using the flower bud organ. Out of 25 varieties we screened for the potential to regenerate new plants in vitro, 15 formed shoots. Our method induced multiple shoots directly, without callus phase, and formed up to more than a dozen of new shoots in a single original bud (during the three-month study-period), which is more than four folds of new plants formed per hole in the field during the whole growing season. Sub-culturing of in vitro cultures occurred once monthly. Each subculture induced repetitive multiple shoots that cumulatively added up to several tens of new plants per explant over time, beyond the studies period. In the field experiment, we continued with biological characterization studies that were initiated during last year. This was essential to ensure successful integration or adaptation of new varieties into the new environment. In comparison to the last season, all 250 varieties were able to reach maturity by forming flowers and pods. This is the first time it has happened, which signals a great survival success. Although rudimentary, we were able to initiate the categorization of varieties based upon their anti-erosion potential we associated with leaves arrangement and stalk and crown presentation per hole. We initially planted 1-2 fans per hole, and by this second year the number has increased up to nine fans per hole, which is not significant compared to the results from tissue culture approaches. It should be noted that no cytokinins were applied to the plants in the field. Most varieties completed their flowering by the end of July or early August. However, nine varieties were still forming inflorescences and flowering beyond September. This is a general improvement compared to last year during which only four varieties' flowering perdured this far. Nine varieties formed proliferations, which was also an improvement of four more varieties compared to last year. However, it was not clear why those four varieties did not form proliferations in the previous year. We also confirmed that our genetic stock does not include evergreen varieties. We also investigated whether select genes that have been determined to be involved in the pre-programmed death of flowers in other species were accountable for the 24-hour preprogrammed death in daylilies' blooms. We were able to confirm the involvement of two apoptosis genes in one variety. Efforts are underway to expand the screening to other varieties.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
- Kanyand Matand, Meordrick Shoemake, and Chenxin Li. 2020. High frequency in vitro regeneration of adventitious shoots in daylilies (Hemerocallis sp) stem tissue using thidiazuron. BMC Plant Biology. DOI: 10.1186/s12870-020-2243-7.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
- Kanyand Matand. 2019. Initial biological profile of the newly established LU daylily (Hemerocallis sp) genetic stock and research progress. ARD 19th Biennial Research Symposium, Jacksonville, FL; March 30-April 3, 2019. Abstract #250, Pg. 155.
- Kanyand Matand. 2019. Daylilies interests and activities at Langston University. Annual Small Farmers Conference, Midwest City, OK; May 21-23.
- Kanyand Matand, Meordrick Shoemake. 2019. Daylilies Research Progress at Langston University. Central Oklahoma Hemerocallis Society Meeting, Oklahoma City, OK; March14.
- Kanyand Matand. 2019. Overview of Langston University daylily genetic stock and research outlook. Langston University Plasticulture/Horticulture Conference, Midwest City, OK; February 09.
- Conrad Green, Meordrick Shoemake, Mary Mitchell, Kanyand Matand. 2019. De novo regeneration of abundant in vitro shoots in daylily (Hemerocallis sp) bract tissue. ARD 19th Biennial Research Symposium, Jacksonville, FL; March 30-April 3. Abstract #511, Pg. 261.
- Conrad Green, Meordrick Shoemake, Mary Mitchell, Kanyand Matand. 2019. Induction of Multiple Adventitious In vitro Shoots in Daylily (Hemerocallis sp) Bract Tissue. Oklahoma State University Research Day, Stillwater, OK; February 19.
- Conrad Green. 2019. In vitro Induction of Multiple Adventitious Shoots in Daylily Bract Tissue. Langston University 19th Annual Research Day Symposium, Langston, OK; April 26. Abstract # 6, Pg. 9.
- Meordrick Shoemake, Mary Mitchell, Conrad Green, Kanyand Matand. 2019. Development of a protocol for mass shoot organogenesis in daylily (Hemerocallis sp) stem tissue. ARD 19th Biennial Research Symposium, Jacksonville, FL; March 30-April 3. Abstract # 512, Pg. 261.
- Meordrick Shoemake, Mary Mitchell, Conrad Green, Kanyand Matand. 2019. Daylily In vitro Mass Shoot Organogenesis in Stem Tissue. Oklahoma State University Research Day, Stillwater, OK; February 19.
- Meordrick Shoemake. 2019. Daylily In vitro Mass Shoot Organogenesis in Stem Tissue. Langston University 19th Annual Research Day Symposium, Langston, OK; April 26. Abstract # 13, Pg. 10.
- Mary Mitchell, Meordrick Shoemake, Conrad Green, Kanyand Matand. 2019. Effect of environmental conditions on daylily (Hemerocallis sp) total RNA preservation and quality for downstream applications. ARD 19th Biennial Research Symposium, Jacksonville, FL; March 30-April 3, 2019. Abstract # 513, Pg. 262.
- Mary Mitchell, Meordrick Shoemake, Conrad Green, Kanyand Matand. 2019. Assessing the Resilience of Daylily Total RNA Quality for Down Stream Applications Under Harsh Environmental Conditioning. Oklahoma State University Research Day, Stillwater, OK; February 19.
-Mary Mitchell. 2019. Assessing the Resilience of Daylily RNA Quality, Under Stressful Condition, for Down Stream Applications. Langston University 19th Annual Research Day Symposium, Langston, OK; April 26. Abstract # 13, Pg. 9.
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:The target audiences included agricultural scientists, daylilies growers and enthusiasts, and other biological scientists and students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the 2018 reporting period, three undergraduate students funded through this project were provided with research skills and experiences in plant biotechnology in the lab and crop production in the field experiment. Hands-on lab skills and research experience were also provided to students who took the Genetics course. ? How have the results been disseminated to communities of interest?During the first growing season, the results were disseminated to communities of interest primarily through interactions with farmers, daylilies enthusiasts, and high school and college students who visited the lab and/or field experiments. ? What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan to focus on developing a more effective plant regeneration protocol that applies flower bud as the main in vitro explant organ. We will also carry out continual field experiment activities. We plan to initiate the investigation of potential involvement of select current apoptosis genes that have been discovered in other species.
Impacts
What was accomplished under these goals?
In 2018, we completed the first growing season of the project during which the initial emphasis was to conduct routine field activities to ensure survival success of introduced genotypes. Sample activities included field preparation, planting, weed and pest control, and nutrients and water provisions, etc. As results, all 250 varieties we introduced survived. Further, we conducted stem micropropagation studies of 25 genotypes amongst which 19 responded positively to thidiazuron and induced cumulatively up to more than 150 new shoots per explant during the studies period. Although all varieties survived during the first growing season, the biological characterization studies showed that only 70 varieties out of 250 formed flowers amongst which only 4 varieties formed pods. We initially planted 1-2 fans of originally purchased plants per hole. By the end of the first growing season, the number of plants per hole ranged from 2 to 5. Except four varieties, most varieties that formed inflorescences completed flowering by the end of July. Exceptionally, one variety continued to flower until the first frost in the second half of November. This variety has been tagged as a potential source of longer flowering genes, for breeding program and genomics studies. The studies also showed that there were no evergreen varieties within the bulk of our genetic stock. A significant proportion of the varieties had bulbous roots and others had cylindrical roots. All flowers induced apoptosis signs within 24 hours of their blooming. However, we identified one variety that extended the bloom life by two hours. We also tagged this variety as a potential source of genes of longer bloom period. The studies also showed that only four varieties developed proliferations.
Publications
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