Progress 10/01/09 to 10/01/10
Outputs
OUTPUTS: During this reporting period, scientifc results and increases in the fundamental knowledge of the reproductive biology of endangered/threatened orchids, propagation procedures and resultant impact on orchid conservation continued to be disseminated through oral presentations made by graduate students and M. Kane at professional scientific meetings as well as the publication of several refereed research papers. These included the oral presentations: 1)Using local plants for restorations and reintroductions: local adaption, breeding systems, and suggestions for successful habitat rehabilitation; 2)Preventing loss of viability during seed storage of Eulophia alta (Orchidaceae. PARTICIPANTS: PARTICIPANTS: University of Florida graduate research assistant Tim Johnson and Jonathan Jasinski, and research associate Dr. Phil Kauth. University employees: Nancy Philman(Biological Scientist) and Dr. Michael Kane (Professor). US Fish and Wildlife Service collaborator/partner Larry Richardson (Wildlife Biologist). TARGET AUDIENCES: The target audiences for this research include, restoration conservationists, landscape managers and commerical native plant producers. The information has been communicated through publication in international refereed journals and presnetations at international professional meetings. In June 2010, a series of presentations were made as part of a workshop: "Recent Research and Idea-generating Discussion on Ecotypes in Plant Conservation and Restoration" held in conjunction with the Florida Native Plant Society Annual Meeting in Tallahassee, Florida. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Financial support from the U.S. Fish & Wildlife Service and the University of Florida was used to complete the various orchid conservation research projects. Leaf samples of the endangered orchid Cyrtopodium punctatum from existing plants on the Florida Panther National Wildlife Refuge were collected and DNA extracted for genetic analysis using AFLP. AFLP- generated data were generated and will be analyzed in early 2011. Morphometric data from field populations were collected to differentiate between Calopogon tuberosus var. tuberosus and C. tuberosus var. simpsonii. DNA was extracted from leaf samples of plants of each variety to further study the genetic relationships. Techniques for field establishment of Calopogon tuberosus were developed. The influence and importance of ecotypic differentiation on orchid seed germination and subsequent seedling developmental responses to environmental factors have been further elucidated by examining the role of chilling in aleviation of dormancy in Calopogon corms produced on plants from different geopgraphical sources. These results further support of the significance of making restoration/reintroduction decisions based on consideration of the impact of geographical source of plant materials used for orchid conservation/reintroduction. In vitro derived seedlings of orchid seedlings were maintained under greenhouse conditions and will be used in orchid restablishment/reintroduction studies to be initiated in spring 2011. Survival and establishment of seedlings and corms of Calopogon tuberosus were evaluated following outplanting. The effects of temperature and illumination on germination and early seedling development of the pink orchid (Bletia purpurea)were examined and a manuscript was submitted for publication review. The pollenation biology of Bletia purpurea have been delineated. This information is key to understanding the sexual reproductive biology and resultant population genetic diversity needed to develop effective conservation plans. Several additional publications have been submitted and are under review. In May 2010 Dr. Michael Kane was presented with the U.S Fish & Wildlife Service Regional Director's Conservation Award.
Publications
- 1. Stewart, S.L. and M.E. Kane. 2010. Effects of carbohydrate source on the in vitro asymbiotic seed germination of the terrestrial orchid Habenaria macroceratitis. J. Plant Nutrition 33: 1-11.
- 2. Kauth, P.J., M.E.Kane and T.R. Johnson. 2010. Techniques for field establishment of Calopogon tuberosus. North American Native Orchid Journal 16: 12-24.
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Progress 10/01/04 to 09/30/10
Outputs
OUTPUTS: Last report should have been marked final. This is to close project. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Last report should have been marked final. This is to close project.
Publications
- No publications reported this period
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Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: During this reporting period, scientifc results and increases in the fundamental knowledge of the reproductive biology of endangered orchids, propagation procedures and resultant impact on orchid conservation have continued to be disseminated through oral presentations made by graduate students and M. Kane at professional scientific meetings as well as the publication of numerous refereed research papers. These included the oral presentations: 1)The Florida Orchid restoration partnership - Proactive conservation of South Florida's native orchids through collaborative research and management; US Fish & Wildlfie Service Research Update; 2)Progress on development of conservation and reintroduction research on native orchids at the Florida Panther National Wildlife Refuge (2005-2009); and 3)Native orchid propagation presented to Gainesville Orchid Society. PARTICIPANTS: University of Florida graduate research assistants: Tim Johnson, Dr. Phil Kauth, and Dr. Scott Stewart. University employees: Nancy Philman(Biological Scientist) and Dr. Michael Kane (Professor). US Fish and Wildlife Service collaborator/partner Larry Richardson (Wildlife Biologist). TARGET AUDIENCES: The major target audiences are landscape managers, individuals involved in plant conservation. In addition to the U.S. Department of Interior, closer collaborative research relationships have been established between the University of Florida and the Florida Native Plant Society. During 2009, numerous scientific oral presentations were given to communicate our findings to these target audiences. An information brochure, previously developed as a source of information, was also disseminated to increase public awareness. A Symposium entitled, Plant Conservation and Restoration Research in Florida, was organized and held at the Florida Native Plant Society in May 2009. In February 2009, Daniela Dutra received the IFAS Excellence in Masters Research Award for her thesis on the reproductive biology of Cyrtopodium punctatum. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period. PARTICIPANTS: Individuals: Dr. Michael Kane, Professor, serving as the principle investigator and project manager, provided oversight on the various grant funded research projects. Nancy Philman, Biological Scientist, provided significant technical support. Graduate students Phil Kauth and Tim Johnson conducted both laboratory and field research. Training or Professional Development: Dr. Phil Kauth completed his doctoral studies during fall 2009. TARGET AUDIENCES: Presentaions were given to orchid society members, US Fish & Wildlife review team members and staff, native plant growers and orchid hobbyists. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Financial support from the U.S. Fish & Wildlife Service, and University of Florida was used to complete the various orchid conservation research projects. During this period, a doctoral student, Phil Kauth, completed his degree on the in vitro ecology of Calopogon tuberosus. The influence and importance of ecotypic differentiation on orchid seed germination and subsequent seedling developmental responses to environmental factors have been clearly defined using Calopogon seedlings from different geopgraphical sources. This information will have significant implications with respect to decisions made on the impact of geographical source of plant materials used for orchid conservation/reintroduction. The complete seed cutlure protocol to efficiently propagate Cyrtopodium punctatum was published. The seedlings produced are being maintained under greenhouse conditions and will be used in orchid restablishment/reintroduction experiments to be initiated in spring 2010. Survival and establishment of seedlings and corms of Calopogon tuberosus were evaluated following outplanting. The pollenation biology of two orchid species Cyrtopodium punctatum and Eulophia alta, have been delineated. This information is key to understanding the sexual reproductive biology and resultant population genetic diversity which is needed to develop effective conservation plants. DNA extraction of Cyrtopodium punctatum has been completed and AFLP is being used to assess genetic diversity and population structure of the limited populations. The reproductive biology of Eulophia. Several additional publications are under review.
Publications
- Dutra, D., M.E. Kane, C. Reinhardt Adams and L. Richardson. 2009. Reproductive Biology of Cyrtopodium punctatum: Implications for Conservation of an Endangered Florida Native Orchid. Plant Species Biology 24: 92-103.
- Johnson, T.R., S.L. Stewart, P. Kauth, M.E. Kane and N. Philman. 2009. Confronting assumptions about spontaneous autogamy in south Florida populations of Eulophia alta (Orchidaceae): Assessing the effect of pollen treatment on seed formation, seed germination and seedling development. Botanical Jour. Linnean Soc. 161: 78-88.
- Kauth, P. and M.E. Kane. 2009. In vitro ecology of Calopogon tuberosus var. tuberosus (Orchidaceae) seedlings from distant populations: implications for assessing ecotypic differentiation. J. Torrey Bot. Soc. 136: 433-444.
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Progress 10/01/07 to 09/30/08
Outputs
OUTPUTS: During this reporting period, scientifc results and increases in the fundamental knowledge of the reproductive biology of endangered orchids, propagation procedures and resultant impact on orchid conservation have been disseminated through both oral and posters presentations made by graduate students at professional scientific meetings and numerous research and educational publications. These included the poster presentations: 1) In vitro seed germination and seedling development of Calopogon tuberosus var. tuberosus from different geographic sources. 19th World Orchid Conference, Miami, Florida; 2)The Florida Orchid restoration partnership - Proactive conservation of South Florida's native orchids through collaborative research and management. 19th World Orchid Conference, Miami, FL; 3)Breeding system determination of Cyrtopodium punctatum: Implications for conservation of an endangered Florida native orchid. 19th World Orchid Conference, Miami, FL; and 4)A classroom exercise in hand pollination and in vitro asymbiotic orchid seed germination. World Congress on In Vitro Biology, Tucson, AZ. PARTICIPANTS: PARTICIPANTS: University of Florida graduate research assistants: Daniela Dutra, Tim Johnson, Phil Kauth, and Dr. Scott Stewart. University employees: Nancy Philman(Biological Scientist) and Dr. Michael Kane (Professor). US Fish and Wildlife Service collaborator/partner Larry Richardson (Wildlife Biologist). TARGET AUDIENCES: The major target audiences are landscape managers, individuals involved in plant conservation. In addition to the U.S. Department of Interior, closer collaborative research relationships have been established between the University of Florida and the Florida Native Plant Society. During 2007-2008, numerous scientific oral and poster presentations were given communicate our finds to these target audiences. A previously constructed three-panel display which outlines the University of Florida and U.S. Fish & Wildlife Florida native orchid research partnership was used as part of an interactive live orchid conservation display at the 19th World Orchid Conference in Miami, FL. Out of 1100 entries, the display received the third place education catgeory. An information brochure, developed as a source of information, was also disseminated to increase public awareness. A Symposium entitled: In Vitro Ecology was organized and held at the Society for In Vitro Biology in Tucson, AZ in June 2008. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
2007/10 TO 2008/09 Financial support from the U.S. Fish & Wildlife Service, American Orchid Society and University of Florida was used to complete the various orchid conservation research projects. During this period, a masters student completed her degree on the reproductive biology of Cyrtopodium punctatum. Significant progress was made in determining the procedures required to efficiently propagate Cyrtopodium punctatum, and Bleta purpurea and using asymbiotic seed culture. The seedlings produced have been established under greenhouse conditions and will be used in orchid restablishment/reintroduction experiments to be completed. The influence and importance of ecotypic differentiation on orchid seed germination and subsequent seedling developmental responses to environmental factors has been more clearly defined using Calopogon seedlings from different geopgraphical sources. This information will have significant implications with respect to decisions made on the impact of geographical source of plant materials used for orchid conservation/reintroduction.
Publications
- Dutra, D., T.J. Johnson, P.J. Kauth, S.L. Stewart, M.E. Kane and L. Richardson. 2008. Asymbiotic seed germination, in vitro seedling development and greenhouse acclimatization of the threatened terrestrial orchid Bletia purpurea. Plant Cell Tissue Organ Culture 94: 11-21.
- Kauth, P., M.E. Kane, W. Vendrame and C. Adams. 2008. Photoperiod and media affect asymbiotic germination and development of Calopogon tuberosus var. tuberosus seeds from different populations. Annals of Botany 102: 783-793.
- Dutra, D., M.E. Kane and L. Richardson. 2008. Asymbiotic seed germination and in vitro seedling development of Cyrtopodium punctatum: A propagation protocol for an endangered Florida native orchid. Plant Cell Tissue Organ Culture DOI 10.1007/s11240-008-9480-z.
- Johnson, T.R., M.E. Kane and H. Perez. 2008. Seed physiology of Bletia purpurea (pine pink; Ochidaceae)-fluctuating low temperature and dark slow development and inhibit germination. In Vitro Cellular & Developmental Biology 44: S37.
- Dutra, D., M.E. Kane and L. Richardson. 2008 Asymbiotic seed germination and in vitro seedling development of Cyrtopodium punctatum: A propagation protocol for an endangered Florida native orchid. In Vitro Cellular & Developmental Biology 44: S37. Kauth, P.J., M.E. Kane and W. Vendrame. 2008. Using in vitro techniques as tools to differentiate ecotypes of Calopogon tuberosus, A North American native orchid. In Vitro Cellular & Developmental Biology 44: S19.
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Progress 10/01/06 to 09/30/07
Outputs
OUTPUTS: Scientifc results and increases in the fundamental knowledge of the reproductive biology of endangered orchids, propagation procedures and resultant impact on orchid conservation have been disseminated through both oral and posters presentations made by graduate students at professional scientific meetings and publications. These included the poster presentations: 1) Asymbiotic seed germination of the threatened orchid Bletia purpurea in Florida. 2007 Society for In Vitro Biology Meeting, Indianapolis, IN; 2) Asymbiotic and symbiotic seed germination of Eulophia alta (Orchidaceae) - Preliminary evidence for symbiotic culture efficiency. 2007 Society for In Vitro Biology Meeting, Indianapolis, IN; 3) Pollination biology of Cyrtopodium punctatum, an endangered native Florida orchid. Florida Native Plant Society Annual Meeting, Gainesville, FL. Numerous oral presentations were made at regional and natioanl meetings including: 1) Comparative Photoperiod effects on in vitro seed
germination and seedling development in Calopogon tuberosus from differing geographic sources. 2007 Society for In Vitro Biology Meeting, Indianapolis, ID; 2)Comparative effects of photoperiod and culture media on asymbiotic seed germination and seedling development in Vanda hybrids. 2007 Society for In Vitro Biology Meeting, Indianapolis, ID.; 3)Asymbiotic seed germination as a tool for differentiating ecotypes of Calopogon tuberosus var. tuberosus, a Florida native orchid. Florida Native Plant Society Annual Meeting, Gainesville, FL; 4)Integrated conservation of Florida's Orchidaceae: A joint report from the University of Florida and U.S. Fish and Wildlife Service. Florida Native Plant Society Annual Meeting, Gainesville, FL. Hands-on teacher's plant tissue culture workshop on orhcid seed culture was held at the Society for In Vitro Biology Meeting, Indianapolis, IN., June 2007; 5) Orchid conservation in the Americas - lessions learned in Florida, 2007 International Orchid
Conservation Congress, San Jose, Costa Rica.
PARTICIPANTS: University of Florida graduate research assistants: Daniela Dutra, Tim Johnson, Phil Kauth, and Scott Stewart. University employees: Nancy Philman(Biological Scientist) and Dr. Michael Kane (Professor). US Fish and Wildlife Service collaborator/partner Larry Richardson (Wildlife Biologist).
TARGET AUDIENCES: During 2007, many scientific oral and poster presentations were given. A three-panel display was designed and constructed which outlines the University of Florida and U.S. Fish & Wildlife Florida native orchid research partnership that has been developed. A similar information brochure has been developed as a source of information dissemination and increased public awareness. The display and brochures are set up at professional meetings.
Impacts
Financial support from the Florida Department of Forestry, the U.S. Fish & Wildlife Service and University of Florida was used to complete the various research projects. During this period, a masters and doctoral student completed their degrees. Significant progress was made in determining the procedures required to efficiently propagate Eulophia alta, Cyrtopodium punctatum, and Bleta purpurea and Vanda hybrids using asymbiotic seed culture. The ability to produce plantlets now will enable completion of in situ orchid restablishment/reintroduction experiments to be completed. The influence and importance of ecotypic differentiation on orchid seed germination and subsequent seedling responses to cultural environments has begun to be defined. Studies completed on the ecology, demography, mycology, pollination biology and population genetics of Habenaria macroceratitis, Spiranthes floridana and S. brevilabris and S. cernua, aid in development of integrated conservation
plans for these species. These integrated conservation systems could be applicable outside of the United States.
Publications
- Dutra, D., T.J. Johnson, P.J. Kauth, S.L. Stewart, M.E. Kane and L. Richardson. 2008. Asymbiotic seed germination, in vitro seedling development and greenhouse acclimatization of the threatened terrestrial orchid Bletia purpurea. Plant Cell Tissue Organ Culture (submitted).
- Stewart, S.L. and M.E. Kane. 2007. Symbiotic seed germination and evidence for mycobiont specificity between two closely-related Florida terrestrial orchids, Spiranthes brevilabris and Spiranthes floridana (Orchidaceae). In Vitro Cellular Developmental Biology - Plant 43:178-186.
- Stewart SL & Kane ME 2007.Orchid conservation in the Americas:lessons learned in Florida. Lankesteriana 7:382-387.
- Johnson, T.R., S.L. Stewart, D. Dutra, M.E. Kane, and L. Richardson. 2007. Asymbiotic and symbiotic germination of Eulophia alta (Orchidaceae: Preliminary evidence for the symbiotic culture advantage. Plant Cell Tissue Organ Culture 90:313-323.
- Johnson, T.R. and M.E. Kane. 2007. Asymbiotic germination of ornamental Vanda: in vitro germination and development of three hybrids. Plant Cell Tissue Organ Culture 91:251-261.
- S.L. Stewart, D. Dutra, P.J. Kauth, T.R. Johnson, ,N. Philman, M. E. Kane. 2008. A classroom exercise in hand pollination and in vitro asymbiotic orchid seed germination. Plant Cell Tissue organ Culture (submitted).
- Stewart, S.L., D. Dutra, T. Johnson, P.J. Kauth, N. Philman, M.E. Kane and L. Richardson. Native Orchid conservation on the Florida Panther National Wildlife Refuge. Cooperative Research Projects Report on Eulophia alta, Cyrtopodium punctatum, Spiranthes ordorata, and Bletia purpurea, March 2007, 71 pp.
- Kauth , P.J, T.R. Johnson, S.L. Stewart, and M. E. Kane. 2008. A classroom exercise in hand pollination and in vitro asymbiotic orchid seed germination. Plant Cell Tissue organ Culture (submitted).
- Kauth, P.J., D. Dutra, T.R. Johnson, S.L. Stewart, M.E. Kane and W.A. Vendrame. 2008. Techniques and applications of in vitro orchid seed germination. In: J.A. Teixeira da Silva. (ed.) Floriculture, Ornamental and Plant Biotechnology: Advances and Topical Issues, Global Science Books, United Kingdom (in press).
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Progress 10/01/05 to 09/30/06
Outputs
Spiranthes floridana - Monitoring of this endangered species continued at the Starke, FL population site. A second population has been identified in Duval County, Fl. A breeding system has been proposed for this species based on field observations and population monitoring data. Ten fungal mycobionts have been isolated and all have been identified. The population genetics of this species are currently being explored using amplified fragment length polymorphism (AFLP). Early symbiotic seedling growth and development culture of Spiranthes brevilabris have been completed. Habenaria distans - A protocol for the ex vitro establishment of in vitro seedlings was tested. The population genetics of this species are currently being explored using AFLP. Habenaria macroceratitis - Fourth-year field data collection has continued in several populations around Brooksville, Florida. A breeding system has been determined. Sixteen mycobionts have been isolated and identified. The
population genetics of this species are being explored using the AFLP technique. In vitro culture requirements for seed culture of Calopogon tuberosus ecotypes is being investigated. A major obstacle to native orchid production is the difficulty in seed germination. Media and light effects on seed germination of Bletia purpurea, Spiranthes odorata, Cyrtopodium punctatum, and Eulophia alta are being studied. Pollenation biology of Eulophia alta and Cyrtopodium punctatum are being studied.
Impacts
Florida Rare and Endangered Orchids- Knowledge of the population and reproductive biology, genetic diversity, critical symbiotic asssociations and seed culture protocol development are critical to development of sound recovery plans. Development of seed culture protocls have allowed for production, conservation and potential restoration of native orchid populations in Florida.
Publications
- Stewart, S.L. and M.E. Kane. 2006. Asymbiotic seed germination and in vitro seedling development of Habenaria macrocertatitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue Organ Culture 86: 147-158.
- Stewart, S.L. and M.E. Kane. 2006. Symbiotic seed germination of Habenaria macrocertatitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue Organ Culture 86: 159-167.
- Kauth, P.J., W.A. Vendrame, and M.E. Kane. 2005 In vitro germination and seedling development of Calopogon tuberosus. In Vitro Cellular & Developmental Biology 41:27A.
- Stewart, S.L. and M.E. Kane. 2005 In vitro seed germination and seedling development of Habenaria macroceratitis, an endangered terrestrial orchid. In Vitro Cellular & Developmental Biology 41:27A.
- Stewart, S.L. and M.E. Kane. 2006. In vitro specificity exhibited by fungal mycobionts of Spiranthes floridana (Orchidaceae) within the congener Spiranthes brevilabris. In Vitro Cellular & Developmental Biology 42:37A.
- Kauth, P., W. Vendrame, and M.E. Kane. 2006. Micropropagation of Calopogon tuberosus, a native terrestrial orchid. In Vitro Cellular & Developmental Biology 42:23A.
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Progress 10/01/04 to 09/30/05
Outputs
Spiranthes floridana - Monitoring of this endangered species continued at the Starke, FL population site. A breeding system has been proposed for this species based on field observations and population monitoring data. Several fungal mycobionts have been isolated and tentatively identified. The population genetics of this species are currently being explored using amplified fragment length polymorphism (AFLP). Spiranthes brevilabris - As a congener to S. floridana, this species represents one avenue to explore the seed germination requirements and early seedling growth and development of the rarer S. floridana. A symbiotic seed germination survey has been conducted using seed from the more common congener and fungi from both species. Habenaria distans - A protocol for the ex vitro establishment of in vitro seedlings has been developed and is currently being tested. The population genetics of this species are currently being explored using AFLP. Habenaria
macroceratitis - Third-year field data collection has continued in several populations around Brooksville, Florida. A breeding system has been proposed for this species, based on studies of insect pollinator, pollination mechanism, and reproductive fitness. Several fungal mycobionts have been isolated from flowering, vegetative and seedling plants. These mycobionts are currently being identified and characterized. The population genetics of this species are being explored using the AFLP technique. Seed Culture of Calopogon tuberosa -A need exists for the introduction of new commercially valuable terrestrial orchids. A major obstacle to native orchid production is the difficulty in seed germination. Media and light effects on seed germination of Calopogon tuberosus, a native orchid with horticultural potential, were studied. Culture media included Knudson C, Malmgren modified terrestrial orchid, and PhytoTechnology orchid seed sowing (P723). Light treatments included 8 wks continual
darkness, 8 wks 16-h photoperiod, 2 wks dark followed by 6 wks 16-h photoperiod, 4 wks dark followed by 4 wks 16-h photoperiod, and 6 wks dark followed by 2 wks 16-h photoperiod. Percent seed germination was highest on KC after 8 wks culture; however, seedling development was enhanced on P723 in the 8-wk 16-h photoperiod. This suggests that while KC and darkness promoted seed germination, P723 and light were more beneficial for seedling development. Seedlings of C. tuberosus readily acclimated to greenhouse conditions. Preliminary results show that in vitro derived corms exhibit the capacity to generate new corms and shoots when divided and cultured on a medium with an auxin or cytokinin. Although asymbiotic seed germination of C. tuberosus provides an efficient means to produce large seedlings for commercial purposes, symbiotic seed germination should be explored for conservation and ecological purposes.
Impacts
Florida Rare and Endangered Orchids- Knowledge of the population and reproductive biology, genetic diversity, critical symbiotic asssociations and seed culture protocol development are critical to development of sound recovery plans. Calopogon tuberosus seed culture -The optimization of seed culture for native orchids provides an efficient method to produce larger quantities of vigorous plants and faster flowering times. In a commercial setting a seed culture protocol that produces a higher quantity as well as larger seedlings in a short period of time is valuable. This study also demonstrated the benefits KC for seed germination as well as light and P723 for seedling development of C. tuberosus. Culturing seeds first on KC and subsequent transfer to P723 can enhance seed germination and seedling development. The reproductive capacity of C. tuberosus to form corms in vitro provides an opportunity for propagule development of a commercially viable production system,
which facilitates storage.
Publications
- Stewart, S.L. and M.E. Kane. 2006. Symbiotic seed germination of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue and Organ Culture (in preparation).
- Kauth, P.J., W.A. Vendrame, and M.E. Kane. 2005. In vitro seed culture and seedling development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture (in press)
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