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An efficient method for in vitro regeneration of leafy spurge (Euphorbia esula L.)

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Abstract

Leafy spurge (Euphorbia esula L.) is a perennial, invasive weed used as a model to study invasive plant behavior, because molecular tools (such as a deep expressed sequence tag database and deoxyribonucleic acid microarrays) have been developed for this species. However, the lack of effective in vitro regeneration and genetic transformation systems has hampered molecular approaches to study leafy spurge. In this study, we describe an efficient in vitro regeneration system. Three highly regenerative lines were selected by screening the in vitro regeneration capabilities of stem explants of 162 seedlings. The effects of various culture conditions on in vitro regeneration were then evaluated based on explant competence to form calluses and shoots. High rates of shoot regeneration can be obtained using a growth medium containing 1× woody plant basal medium and 1× Murashige and Skoog (MS) basal salts, 1× MS vitamins, 1.11 μM 6-benzylaminopurine, 1.97 μM indole-3-butyric acid, and 3% sucrose, pH 5.6–5.8. After 30 d culture, multiple shoots formed either directly from the stem or indirectly from the callus. This method is a requisite for the development of genetic transformation systems for leafy spurge and may be used to develop in vitro regeneration techniques for other species in the Euphorbiaceae.

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References

  • Anderson J. V.; Horvath D. P.; Chao W. S.; Foley M. E.; Hernandez A. G.; Thimmapuram J. et al. Characterization of an EST database for the perennial weed leafy spurge: an important resource for weed biology research. Weed Sci. 55: 193–203; 2007.

    Article  CAS  Google Scholar 

  • Arokiaraj P.; Wan Abdul Rahaman W. Y. Agrobacterium mediated transformation of Hevea cells derived from in vitro and in vivo seedling cultures. J. Nat. Rubber Res. Malaysia 6: 55–61; 1991.

    CAS  Google Scholar 

  • Banno H.; Ikeda Y.; Niu Q. W.; Chua N. H. Overexpression of Arabidopsis ESR1 induces initiation of shoot regeneration. Plant Cell 13: 2609–2618; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Becerra D. C.; Forero A. P.; Góngora G. A. Age and physiological condition of donor plants affect in vitro morphogenesis in leaf explants of Passiflora edulis f. flavicarpa. Plant Cell, Tissue Organ Cult. 19: 87–90; 2004.

    Article  Google Scholar 

  • Bowes G. G.; Thomas A. G. Longevity of leafy spurge seeds in the soil following various control programs. J. Range Manag. 32: 137–140; 1978.

    Article  Google Scholar 

  • CAB International, Euphorbia esula. In: Crop protection compendium. CAB International, Wallingford, UK; 2004 [original text by Chao, W. and J. V. Anderson; CD-ROM].

  • Chao W. S.; Horvath D. P.; Anderson J. V.; Foley M. P. Potential model weeds to study genomics, ecology, and physiology in the 21st century. Weed Sci. 53: 929–937; 2005.

    Article  CAS  Google Scholar 

  • Davis D. G.; Olson P. A.; Stolzenberg R. L. Organogenesis in cell cultures of leafy spurge (Euphorbiaceae) accessions from Europe and North America. Plant Cell Rep. 7: 253–256; 1988.

    Article  Google Scholar 

  • de Langhe E.; Debergh P.; Van Rijk R. In vitro culture as a method for vegetative propagation of Euphorbia pulchurrima. Z. Pflanzenphysiol. 7: 271–274; 1974.

    Google Scholar 

  • Dewir Y. H.; Chakrabarty D.; Hahn E. J.; Paek K. Y. Reversion of inflorescence development in Euphorbia millii and its application to large-scale micropropagation in an air-lift bioreactor. J. Hortic. Sci. Biotechnol. 80: 581–587; 2005.

    CAS  Google Scholar 

  • Drew R. A. In vitro culture of adult and juvenile bud explants of Passiflora species. Plant Cell, Tissue Organ Cult. 26: 23–27; 1991.

    Article  Google Scholar 

  • Fortes A. M.; Pais M. S. Organogenesis from internode-derived nodules of Humulus lupulus var. Nugget (Cannabinaceae): histological studies and changes in the starch content. Am. J. Bot. 87: 971–979; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Fukuda H.; Komamine A. Establishment of an experimental system for the tracheary element differentiation from single cells isolated from the mesophyll of Zinnia elegans. Plant Physiol. 65: 57–60; 1980.

    Article  PubMed  CAS  Google Scholar 

  • Gelvin S. B. Agrobacterium and plant genes involved in T-DNA transfer and integration. Annu. Rev. Plant Physiol. Plant Mol. Biol. 51: 223–256; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Groll J. Effect of medium salt concentration on differentiation and maturation of somatic embryos of Cassava (Manihot esculenta Crantz). Ann. Bot. 89: 645–648; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Gulati A. W.; Jaiwal P. K. Culture conditions affecting plant regeneration from cotyledons of Vigna radiata (L.). Plant Cell, Tissue Organ Cult. 23: 1–7; 1990.

    Article  CAS  Google Scholar 

  • Henry Y.; Vain P.; Buyser J. D. Genetic analysis of in vitro plant tissue culture responses and regeneration capacities. Euphytica 79: 45–58; 1994.

    Article  Google Scholar 

  • Jasrai Y. T.; Thaker K. N.; D'Souza M. C. In vitro propagation of Euphorbia pulcherrima Willd. through somatic embryogenesis. Plant Cell, Tissue Organ Cult. 13: 31–36; 2003.

    Google Scholar 

  • Johansen D. A. Plant microtechnique. McGraw-Hill, New York; 1940.

    Google Scholar 

  • Jia Y.; Anderson J. V.; Horvath D. P.; Gu Y. Q.; Lym R. G.; Chao W. S. Subtractive cDNA libraries identify differentially expressed genes in dormant and growing buds of leafy spurge (Euphorbia esula). Plant Mol. Biol. 61: 329–344; 2006.

    Article  PubMed  CAS  Google Scholar 

  • Karp A. Somaclonal variation as a tool for crop improvement. Euphytica 85: 295–302; 1955.

    Article  Google Scholar 

  • Khanna H. K.; Raina S. K. Genotype x culture media interaction effects on regeneration response of three indica rice cultivars. Plant Cell, Tissue Organ Cult. 52: 145–153; 1998.

    Article  CAS  Google Scholar 

  • Lee C. W.; Yeches J.; Thomas J. C. Tissue culture propagation of Euphorbia lathyris and Asclepias erosa. Hortic. Sci. 17: 533; 1982.

    Google Scholar 

  • Lee T. T.; Starratt A. N. Growth substance requirements and major lipid constituents of tissue cultures of Euphorbia esula and E. cyparissias. Can. J. Bot. 50: 723–726; 1972.

    Article  CAS  Google Scholar 

  • Li H. Q.; Sautter C.; Potrykus I.; Puonti-Kaerlas J. Genetic transformation of cassava (Manihot esculenta Crantz). Nat. Biotechnol. 14: 736–740; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Murashige T.; Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497; 1962.

    Article  CAS  Google Scholar 

  • Nataraja K. Morphogenesis in embryonal callus of Euphorbia pulchurrima in vitro. Curr. Sci 44: 136–137; 1975.

    CAS  Google Scholar 

  • Pickens K. A.; Cheng Z. M.; Trigiano R. N. Axillary bud proliferation and organogenesis of Euphorbia pulchurrima winter rose. In Vitro Cell. Dev. Biol.—Plant 41: 770–774; 2005.

    Article  CAS  Google Scholar 

  • Raemakers C. J. J. M.; Sofiari E.; Taylor N.; Jacobsen E.; Visser R. G. F. Production of transgenic cassava (Manihot esculenta Crantz) plants by particle bombardment using luciferase activity as selection marker. Mol. Breed. 2: 339–349; 1996.

    Article  CAS  Google Scholar 

  • Raju M. V. S. Morphology and anatomy of leafy spurge. In: Watson A. K. (ed) Leafy spurge. Weed Science Society of America, Champaign, IL, pp 26–41; 1985.

    Google Scholar 

  • Ripley K. P.; Preece J. E. Micropropagation of Euphorbia lathyris L. Plant Cell, Tissue Organ Cult. 5: 213–218; 1986.

    Article  CAS  Google Scholar 

  • Schöpke C.; Taylor N.; Carcamo R.; Konan N. K.; Marmey P.; Henshaw G. G. et al. Regeneration of transgenic cassava plants (Manihot esculenta Crantz) from microbombarded embryogenic suspension cultures. Nature Biotechnol. 14: 731–735; 1996.

    Article  Google Scholar 

  • Skoog F.; Miller C. O. Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp. Soc. Exp. Biol. 54: 118–130; 1957.

    PubMed  CAS  Google Scholar 

  • Souissi T.; Kremer R. J. A rapid microplate callus bioassay for assessment of rhizobacteria for biocontrol of leafy spurge (Euhporbia esula L.). Biocontrol Sci. Technol. 8: 83–92; 1998.

    Article  Google Scholar 

  • Souissi T.; Kremer R. J.; White J. A. Interaction of rhizobacteria with leafy spurge (Euphoriba esula L.) callus tissue cells. Plant Cell, Tissue Organ Cult. 47: 279–287; 1997.

    Article  Google Scholar 

  • Sujatha M.; Sailaja M. Stable genetic transformation of castor (Ricinus communis L.) via Agrobacterium tumefaciens-mediated gene transfer using embryo axes from mature seeds. Plant Cell Rep. 23: 803–810; 2005.

    Article  PubMed  CAS  Google Scholar 

  • Taguchi-Shiobara F.; Lin S. Y.; Tanno K.; Komatsuda T.; Yano M.; Sasaki T. et al. Mapping quantitative trait loci associated with regeneration ability of seed callus in rice, Oryza sativa L. Theor. Appl. Genet. 95: 828–833; 1997.

    Article  CAS  Google Scholar 

  • Tideman J.; Hawker J. S. In vitro propagation of latex-producing plants. Ann. Bot. 49: 273–279; 1982.

    Google Scholar 

  • Vik N. I.; Hvoslef-Eide A. K.; Gjerde H.; Bakke K. Stable transformation of poinsettia via electrophoresis. Acta Hortic. 560: 125–128; 2001.

    Google Scholar 

  • Weber S.; Horn R.; Friedt W. High regeneration potential in vitro of sunflower (Helianthus annuus L.) lines derived from interspecific hybridization. Euphytica 116: 271–280; 2000.

    Article  Google Scholar 

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Authors and Affiliations

  1. Department of Plant Sciences, North Dakota State University, Fargo, ND, 58105, USA

    Bin Xu & Wenhao Dai

  2. Biosciences Research Laboratory, USDA—ARS, 1605 Albrecht Blvd., Fargo, ND, 58105-5674, USA

    Wun S. Chao

  3. Plant Science Research, USDA—Agricultural Research Service, 1605 Albrecht Blvd., Fargo, ND, 58105-5674, USA

    Wun S. Chao

Authors
  1. Bin Xu
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  2. Wenhao Dai
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  3. Wun S. Chao
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Correspondence to Wun S. Chao.

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Editor: Gregory C. Phillips

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Xu, B., Dai, W. & Chao, W.S. An efficient method for in vitro regeneration of leafy spurge (Euphorbia esula L.). In Vitro Cell.Dev.Biol.-Plant 44, 548–556 (2008). https://doi.org/10.1007/s11627-008-9139-9

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  • DOI: https://doi.org/10.1007/s11627-008-9139-9

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