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Physiology and Molecular Biology of Plants

, Volume 24, Issue 3, pp 503–511 | Cite as

Agroinfiltration: a rapid and reliable method to select suitable rose cultivars for blue flower production

  • Masoume Zeinipour
  • Pejman AzadiEmail author
  • Ahmad Majd
  • Maryam Jafarkhani Kermani
  • Saeed Irian
  • Seyed Mohammad Hosseini
  • Masahiro Mii
Research Article
  • 243 Downloads

Abstract

Rose cultivars with blue flower color are among the most attractive breeding targets in floriculture. However, they are difficult to produce due to the low efficiency of transformation systems, interactive effects of hosts and vectors, and lengthy processes. In this study, agroinfiltration-mediated transient expression was investigated as a tool to assess the function of flower color genes and to determine appropriate host cultivars for stable transformation in Rosa hybrida. To induce delphinidin accumulation and consequently to produce blue hue, the petals of 30 rose cultivars were infiltrated with three different expression vectors namely pBIH-35S-CcF3′5′H, pBIH-35S-Del2 and pBIH-35S-Del8, harbouring different sets of flower color genes. The results obtained showed that the ectopic expression of the genes was only detected in three cultivars with dark pink petals (i.e. ‘Purple power’, ‘High & Mora’ and ‘Marina’) after 6–8 days. The high performance liquid chromatography analyses confirmed delphinidin accumulation in the infiltrated petals caused by transient expression of CcF3′5′H gene. Moreover, there were significant differences in the amounts of delphinidin among the three cultivars infiltrated with the three different expression vectors. More specifically, the highest delphinidin content was detected in the cultivar ‘Purple power’ (4.67 µg g−1 FW), infiltrated with the pBIH-35S-Del2 vector. The expression of CcF3′5′H gene in the infiltrated petals was also confirmed by real time PCR. In conclusion and based on the findings of the present study, the agroinfiltration could be regarded as a reliable method to identify suitable rose cultivars in blue rose flower production programs.

Keywords

Agroinfiltration Delphinidin Expression vectors Cultivar Rosa hybrida 

Notes

Acknowledgements

This work was supported by Agricultural Biotechnology Research Institute of Iran (ABRII). M Zeini Pour did the project as part of her PhD thesis.

Conflicts of interest

The authors declare that there are no conflicts of interest.

References

  1. Azadi P, Ntui VO, Chin DP, Nakamura I, Fujisawa M, Harada H, Misawa N, Mii M (2010) Metabolic engineering of Lilium×formolongi using multiple genes of the carotenoid biosynthesis pathway. Plant Biotech Rep 4:269–280CrossRefGoogle Scholar
  2. Borcke LV (2002) High efficiency transient expression system for plants. Patent Publication Number WO 01/038512Google Scholar
  3. Brugliera F, Tao GQ, Tems U, Kalc G, Mouradova E, Price K, Stevenson K, Nakamura N, Stacey I, Katsumoto Y, Tanaka Y, Mason JG (2013) Violet/Blue chrysanthemums metabolic engineering of the anthocyanin biosynthetic pathway results in novel petal colors. Plant Cell Physiol 54:1696–1710CrossRefPubMedGoogle Scholar
  4. Davies KM (2009) Modifying anthocyanin production in flowers. In: Gould K et al (eds) Anthocyanins. Springer, Berlin.  https://doi.org/10.1007/978-0-387-77335-3-3 Google Scholar
  5. Durst RW, Wrolstad RE (2001) Separation and characterization of anthocyanins by HPLC. Unit F1.3.1–13. In: Wrolstad RE (ed) Current protocols in food analytical chemistry. Wiley, New York, pp 49–83Google Scholar
  6. Forkmann G, Martens S (2001) Metabolic engineering and applications of flavonoids. Curr Opin Biotechnol 12:155–160CrossRefPubMedGoogle Scholar
  7. Gutterson N (1995) Anthocyanin biosynthetic genes and their application to flower colour modification through sense suppression. Hort Sci 30:964–966Google Scholar
  8. Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083CrossRefPubMedPubMedCentralGoogle Scholar
  9. Katsumoto Y, Fukuchi-Mizutani M, Fukui Y, Bruglier F, Holton TA, Karan M, Nakamura N, Yonekura-Sakakibara K, Togam J, Pigeaire A, Ta GQ, Nehra NS, Lu CY, Dyson BK, Tsuda S, Ashikari T, Kusumi T, Mason JG, Tanaka Y (2007) Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant Cell Physiol 48:1589–1600CrossRefPubMedGoogle Scholar
  10. Kuriakose B, Du Toit ES, Jordaan A (2012) Transient gene expression assays in rose tissues using a Bio-Rad Helios hand-held gene gun. S Afr J Bot 78:307–311CrossRefGoogle Scholar
  11. Morante-Carriel MJ, Marchart SS, Márquez MA, Esteso MMJ, Luque I, Martínez BR (2014) RNA isolation from loquat and other recalcitrant woody plants with high quality and yield. Anal Biochem.  https://doi.org/10.1016/j.ab.2014.02.010 PubMedGoogle Scholar
  12. Moriwaki M, Yamakawa T, Washino T, Kodama T, Igarashi Y (1999) Organspecific expression of â-glucuronidase activity driven by the Arabidopsis heat-shock promoter in heat stressed transgenic tobacco and Arabidopsis plants. Plant Mol Biol 28:73–82Google Scholar
  13. Mudalige RG, Kuehnle AR, Amore TD (2003) Pigment distribution and epidermal cell shape in Dendrobium Speciec and hybrids. Hort Sci 38:573–577Google Scholar
  14. Nakatsuka T, Abe Y, Kakizaki Y, Yamamura S, Nishihara M (2007) Production of red-flowered plants by genetic engineering of multiple flavonoid biosynthetic genes. Plant Cell Rep 26:1951–1959CrossRefPubMedGoogle Scholar
  15. Nishihara M, Nakatsuka T (2010) Genetic engineering of novel flower colors in floricultural plants: recent advances via transgenic approaches. In: Jain SM, Ochatt SJ (eds) Protocols for in vitro propagation of ornamental plants, vol 589. Methods in molecular biology. Humana Press, New York, pp 325–347.  https://doi.org/10.1007/978-1-60327-114-1-29 CrossRefGoogle Scholar
  16. Qi Y, Lou Q, Quan Y, Liu Y, Wang Y (2013) Flower-specific expression of the Phalaenopsis flavonoid 3′,5′-hydoxylase modifies flower color pigmentation in Petunia and Lilium. Plant Cell Tiss Organ Cult 115:263–273CrossRefGoogle Scholar
  17. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108CrossRefPubMedGoogle Scholar
  18. Shimada OY, Shimada YN, Ohbayashi R, Kiyokawa M, Kikuchi SY (2003) Selective accumulation of delphinidin derivatives in tobacco using putative flavonoid 3′,5′–hydroxylase cDNA from Campanula medium. Bio Sci Biotech Biochem 67:161–165CrossRefGoogle Scholar
  19. Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1:2019–2025CrossRefPubMedGoogle Scholar
  20. Tanaka Y (2006) Flower colour and cytochromes p450. Phytochem Rev 5:283–291CrossRefGoogle Scholar
  21. Tanaka Y, Tsuda S, Kusumi T (1998) Metabolic engineering to modify flower color. Plant Cell Physiol 39:1119–1126CrossRefGoogle Scholar
  22. Tanaka Y, Katsumuto Y, Brugliera F, Mason J (2005) Genetic engineering in floriculture. Plant Cell Tiss Organ Cult 80:1–24CrossRefGoogle Scholar
  23. Tanaka Y, Brugliera F, Chandler S (2009) Recent progress of flower colour modification by biotechnology. Int J Mol Sci 10:5350–5369CrossRefPubMedPubMedCentralGoogle Scholar
  24. Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium-mediated transient assays of gene expression in lettuce, tomato and Arabidopsis. Plant Biotech J 3:259–273CrossRefGoogle Scholar
  25. Yasmin A, Debener T (2010) Transient gene expression in rose petals via Agrobacterium infiltration. Plant Cell Tiss Organ Cult 102:45–250CrossRefGoogle Scholar
  26. Yuki S, Araki S, Suzuki T (2013) Flavonoid-3′, 5′-hydroxylase gene of Commelina communis. US Patent 8,440,879, 2013Google Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  • Masoume Zeinipour
    • 1
    • 2
  • Pejman Azadi
    • 1
    Email author
  • Ahmad Majd
    • 2
  • Maryam Jafarkhani Kermani
    • 1
  • Saeed Irian
    • 2
  • Seyed Mohammad Hosseini
    • 1
  • Masahiro Mii
    • 3
  1. 1.Department of Genetic Engineering, Agricultural Biotechnology Research Institute of Iran (ABRII)Agricultural Research, Education and Extension Organization (AREEO)KarajIran
  2. 2.Faculty of Biology ScienceKharazmi UniversityTehranIran
  3. 3.Laboratory of Plant Cell Technology, Graduate School of HorticultureChiba UniversityMatsudoJapan

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