Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 107, Issue 2, pp 325–332 | Cite as

Expression of CoQ10-producing ddsA transgene by efficient Agrobacterium-mediated transformation in Panicum meyerianum

  • Mi-Suk Seo
  • Sakiko Takahashi
  • Koh-ichi Kadowaki
  • Makoto Kawamukai
  • Manabu Takahara
  • Tadashi Takamizo
Original Paper

Abstract

Panicum meyerianum Nees is a wild relative of Panicum maximum Jacq. (guinea grass), which is an important warm-season forage grass and biomass crop. We investigated the conditions that maximized the transformation efficiency of P. meyerianum by Agrobacterium infection by monitoring the expression of the β-glucuronidase (GUS) gene. The highest activities of GUS in calli were achieved by the co-cultivation of plants with Agrobacterium at 28°C for 6 days. We transferred the ddsA gene, which encodes decaprenyl diphosphate synthase and is required for coenzyme Q10 (CoQ10) synthesis, into P. meyerianum by using our optimized co-cultivation procedure for transformation. We confirmed by PCR and DNA gel blot hybridization that all hygromycin-resistant plants retained stable insertion of the hpt and ddsA genes. We also demonstrated strong expression of S14:DdsA protein in the leaves of transgenic P. meyerianum. Furthermore, we showed that transgenic P. meyerianum produced CoQ10 at levels 11–20 times higher than that of non-transformants. By comparison, the CoQ9 level in transgenic plants was dramatically reduced. This is the first report of efficient Agrobacterium-mediated transfer of a foreign gene into the warm-season grass P. meyerianum.

Keywords

Agrobacterium-mediated transformation Panicum meyerianum Co-cultivation Coenzyme Q10 

Abbreviations

GUS

β-glucuronidase

ddsA

Decaprenyl diphosphate synthase gene

CoQ

Coenzyme Q

hpt

Hygromycin phosphotransferase gene

PCR

Polymerase chain reaction

References

  1. Blanc G, Baptiste C, Oliver G, Martin F, Montoro P (2006) Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Müll Arg. plants. Plant Cell Rep 24:724–733PubMedCrossRefGoogle Scholar
  2. Cao D, Hou W, Song S, Sun H, Wu C, Gao Y, Han T (2009) Assessment of conditions affecting Agrobacterium rhizogenes-mediated transformation of soybean. Plant Cell Tissue Organ Cult 96:45–52CrossRefGoogle Scholar
  3. Claudiu M, Ana P, Machado R, Marcia MP, Gilberto SM, Elisabeth M (2000) Establishment of an efficient Agrobacterium-mediated transformation system for eggplant and study of a potential biotechnologically useful promoter. J Plant Biotechnol 2:43–49Google Scholar
  4. Dillen W, Clercq JD, Kapila J, Zambre M, Montagu MV, Angenon G (1997) The effect of temperature on Agrobacterium tumefaciens-mediated gene transfer to plants. Plant J 12:1459–1463CrossRefGoogle Scholar
  5. Emani CJM, Garcia ELF, Pozo MJ, Uribe P, Kim DJ, Sunikumar G, Cook DR, Kenerley CM, Rathore KS (2003) Enhanced fungal resistance in transgenic cotton expressing an endochitinase gene from Trichoderma virens. Plant Biotechnol J 1:321–336PubMedCrossRefGoogle Scholar
  6. Espasandin FD, Collavino MM, Luna CV, Paz RC, Tarragó JR, Ruiz OA, Mroginski LA, Sansberro PA (2010) Agrobacterium tumefaciens-mediated transformation of Lotus tenuis and regeneration of transgenic lines. Plant Cell Tissue Organ Cult 102:181–189CrossRefGoogle Scholar
  7. Glowacka K, Ježowski S, Kaczmarek Z (2010) The effects of genotype, inflorescence developmental stage and induction medium on callus induction and plant regeneration in two Miscanthus species. Plant Cell Tissue Organ Cult 102:79–86CrossRefGoogle Scholar
  8. Goldman JJ, Hanna WW, Fleming GH, Ozias-Akins P (2004) Ploidy variation among herbicide-resistant bermudagrass plants of cv. TifEagle transformed with the bar gene. Plant Cell Rep 22:553–560PubMedCrossRefGoogle Scholar
  9. Gondo T, Tsuruta S, Akashi R, Kawanura O, Hoffmann F (2005) Green, herbicide-resistant plants by particle inflow gun-mediated gene transfer to diploid bahiagrass (Paspalum notatum). J Plant Physiol 162:1367–1375PubMedCrossRefGoogle Scholar
  10. Gondo T, Matsumoto J, Tsuruta S, Yoshida M, Kawakami A, Terami F, Ebina M, Yamada T, Akashi R (2009) Particle inflow gun-mediated transformation of multiple-shoot clumps in rhodes grass (Chloris gayana). J Plant Physiol 166:435–441PubMedCrossRefGoogle Scholar
  11. Haberl H, Geissler S (2000) Cascade utilization of biomass: strategies for a more efficient use of a scarce resource. Ecol Eng 16:S111–S121CrossRefGoogle Scholar
  12. Jonassen T, Larsen PL, Clarke CF (2001) A dietary source of coenzyme Q is essential for growth of long-lived Caenorhabditis elegans clk-1 mutants. Proc Natl Acad Sci USA 98:421–426PubMedCrossRefGoogle Scholar
  13. Kawamukai M (2009) Biosynthesis and bioproduction of coenzyme Q10 by yeasts and other organisms. Biotechnol Appl Biochem 53:217–226PubMedCrossRefGoogle Scholar
  14. Kohli A, Melendi PG, Abranches R, Capell T, Stoger E, Christou P (2006) The Quest to understand the basis and mechanisms that control expression of introduced transgenes in crop plants. Plant Signal Behav 1:185–195PubMedCrossRefGoogle Scholar
  15. Kosugi S, Ohashi Y, Nakajima K, Arai Y (1990) An improved assay for p-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous o-glucuronidase activity. Plant Sci 70:133–140CrossRefGoogle Scholar
  16. Li L, Li R, Fei S, Qu R (2005) Agrobacterium-mediated transformation of common bermudagrass (Cynodon dactylon). Plant Cell Tissue Organ Cult 83:223–229CrossRefGoogle Scholar
  17. Li X, Ahlman A, Yan X, Lindgren H, Zhu L (2010) Genetic transformation of the oilseed crop Crambe abyssinica. Plant Cell Tissue Organ Cult 100:149–156CrossRefGoogle Scholar
  18. Liu SJ, Wei ZM, Huang JQ (2008) The effect of co-cultivation and selection parameters on Agrobacterium-mediated transformation of Chinese soybean varieties. Plant Cell Rep 27:489–498PubMedCrossRefGoogle Scholar
  19. Matzke AJM, Matzke MA (1998) Position effects and epigenetic silencing of plant transgenes. Curr Opin Plant Biol 1:142–148PubMedCrossRefGoogle Scholar
  20. Matzke AJM, Neuhuber F, Park YD, Ambros PF, Matzke MA (1994) Homology-dependent gene silencing in transgenic plants: epistatic silencing loci contain multiple copies of methylated transgenes. Mol Gen Genet 244:219–229PubMedCrossRefGoogle Scholar
  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  22. Nakajima K, Mochizuki N (1983) Degrees of sexuality in sexual plants of guineagrass by the simplified embryo sac analysis. Jp J Breed 33:45–54Google Scholar
  23. Okada K, Kainou T, Tanaka K, Nakagawa T, Matsuda H, Kawamukai M (1998) Molecular cloning and mutational analysis of the ddsA gene encoding decaprenyl diphosphate synthase from Gluconobacter suboxydans. Eur J Biochem 255:52–59PubMedCrossRefGoogle Scholar
  24. Okada K, Ohara K, Yazaki K, Nozaki K, Uchida N, Kawamukai M, Nojiri H, Yamane H (2004) The AtPPT1 gene encoding 4-hydroxybenzoate polyprenyl diphosphate transferase in ubiquinone biosynthesis is required for embryo development in Arabidopsis thaliana. Plant Mol Biol 55:567–577PubMedCrossRefGoogle Scholar
  25. Panaia M, Bunn E, Turner SR, McComb J (2009) Incubation temperature critical to successful stimulation of in vitro zygotic embryo growth in four Australian native Cyperaceae species. Plant Cell Tissue Organ Cult 97:197–202CrossRefGoogle Scholar
  26. Salas MG, Park SH, Srivatanakul M, Smith RH (2001) Temperature influence on stable T-DNA integration in plant cells. Plant Cell Rep 20:701–705CrossRefGoogle Scholar
  27. Sanderson M, Reed R, McLaughlin S, Wullschleger S, Conger B, Parrish D, Wolf D, Taliaferro C, Hopkins A, Ocumpaugh W, Hussey M, Read J, Tischler C (1996) Switchgrass as a sustainable bioenergy source. Bioresour Technol 56:83–93CrossRefGoogle Scholar
  28. Seo MS, Takahara M, Ebina M, Takamizo T (2008) Evaluation of tissue culture response from mature seeds of Panicum spp. Grassl Sci 54:125–130CrossRefGoogle Scholar
  29. Seo MS, Takahara M, Takamizo T (2010) Optimization of culture condition for plant regeneration of Panicum spp. through somatic embryogenesis. Grassl Sci 56:1–7CrossRefGoogle Scholar
  30. Sharma M, Chajer AK, Chugh SJ, Kothari SL (2011) Factors influencing Agrobacterium tumefaciens-mediated genetic transformation of Eleusine coracana (L.) Gaertn. Plant Cell Tissue Organ Cult 105:93–104CrossRefGoogle Scholar
  31. Somleva M, Tomaszewski Z, Conger B (2002) Agrobacterium-mediated genetic transformation of switchgrass. Crop Sci 42:2080–2087CrossRefGoogle Scholar
  32. Takahashi S, Ogiyama Y, Kusano H, Shimada H, Kawamukai M, Kadowaki K (2006) Metabolic engineering of coenzyme Q by modification of isoprenoid side chain in plant. FEBS Lett 580:955–959PubMedCrossRefGoogle Scholar
  33. Takahashi S, Ohtani T, Iida S, Sunohara Y, Matsushita K, Maeda H, Tanetani Y, Kawai K, Kawamukai M, Kadowaki K (2009) Development of CoQ10-enriched rice from giant embryo lines. Breed Sci 59:321–326CrossRefGoogle Scholar
  34. Takahashi W, Oishi H, Ebina M, Komatsu T, Takamizo T (2010) Production of transgenic Italian ryegrass expressing the betaine aldehyde dehydrogenase gene of zoysiagrass. Breed Sci 60:279–285CrossRefGoogle Scholar
  35. Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H (2006) Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. Plant J 47:969–976PubMedCrossRefGoogle Scholar
  36. Toriyama K, Hinata K (1985) Cell suspension and protoplast culture in rice. Plant Sci 41:179–183CrossRefGoogle Scholar
  37. Toyama K, Bae CH, Kang JG, Lim YP, Adachi T, Riu KZ, Song PS, Lee HY (2003) Production of Herbicide-tolerant zoysiagrass by Agrobacterium-mediated transformation. Mol cells 16:19–27PubMedGoogle Scholar
  38. Vogel KP, Brejda JJ, Walters DT, Buxton DR (2002) Switchgrass biomass production in the Midwest USA: harvest and nitrogen management. Agron J 94:413–420CrossRefGoogle Scholar
  39. Wang ZY, Ge Y (2005) Agrobacterium-mediated high efficiency transformation of tall fescue (Festuca arundinacea). J Plant Physiol 162:103–113PubMedCrossRefGoogle Scholar
  40. Wu H, Sparks C, Amoah B, Jones HD (2003) Factors influencing successful Agrobacterium-mediated genetic transformation of wheat. Plant Cell Rep 21:659–668PubMedGoogle Scholar
  41. Yasmin A, Debener T (2010) Transient gene expression in rose petals via Agrobacterium infiltration. Plant Cell Tissue Organ Cult 102:245–250CrossRefGoogle Scholar
  42. Zhang K, Wang J, Hu X, Yang A, Zhang J (2010) Agrobacterium-mediated transformation of shoot apices of Kentucky bluegrass(Poa pratensis L.) and production of transgenic plants carrying a betA gene. Plant Cell Tissue Organ Cult 102:135–143CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Mi-Suk Seo
    • 1
    • 5
  • Sakiko Takahashi
    • 2
  • Koh-ichi Kadowaki
    • 3
  • Makoto Kawamukai
    • 4
  • Manabu Takahara
    • 1
  • Tadashi Takamizo
    • 1
  1. 1.Forage Crop DivisionNational Institute of Livestock and Grassland ScienceTochigiJapan
  2. 2.Transgenic Crop Research and Development CenterNational Institute of Agrobiological ScienceTsukuba, IbarakiJapan
  3. 3.National Institute of Agrobiological ScienceTsukuba, IbarakiJapan
  4. 4.Faculty of Life and Environmental ScienceShimane UniversityShimaneJapan
  5. 5.Genomics Division, Department of Agricultural Bio-resourcesNational Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)SuwonKorea

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