Skip to main content
Log in

Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus

  • Genetic Transformation and Hybridization
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T1 plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Babu RC, Zhang J, Blum A, Ho DT-H, Wu R, Nguyen HT (2004) HVA1, a LEA gene from barley confers dehydration tolerance in transgenic rice (Oryza sativa L.) via cell membrane protection. Plant Sci 166:855–862

    Google Scholar 

  • Bray EA, Bailey-Serres J, Weretilnyk E (2000) Responses to abiotic stresses. In: Buchanan B, Gruissem W, Jones R (eds) Biochemistry & molecular biology of plants. American Society of Plant Physiologists, Rockville, MD, pp 1158–1176

    Google Scholar 

  • Chandler PM, Robertson M (1994) Gene expression regulated by abscisic acid and its relation to stress tolerance. Annu Rev Plant Physiol Plant Mol Biol 45:113–141

    Google Scholar 

  • Christou P (1997) Rice transformation: bombardment. Plant Mol Biol 35:197–203

    Google Scholar 

  • Curtis IS, Nam HG (2001) Transgenic radish (Raphanus sativusL. var. longipinnatus Bailey) by floral-dip method – plant development and surfactant are important in optimizing transformation efficiency. Transgenic Res 10:363–371

    Google Scholar 

  • Curtis IS, Nam HG, Yun JY, Seo K-H (2002) Expression of an antisense GIGANTEA (GI) gene fragment in transgenic radish causes delayed bolting and flowering. Transgenic Res 11:249–256

    Google Scholar 

  • Curtis IS (2003) The noble radish: past, present and future. Trends Plant Sci 8:305–307

    Google Scholar 

  • Dure L III (1993a) A repeating 11-mer amino acid motif and plant desiccation. Plant J 3:363–369

    Google Scholar 

  • Dure L III (1993b) Structural motifs in Lea proteins. In: Close TJ, Bray EA (eds) Plant responses to cellular dehydration during environmental stress. American Society of Plant Physiology, Rockville, MD, pp 91–103

    Google Scholar 

  • Effendi, Kisaka H, Kanno A, Kameya T (2000) Transformation of soybean by infecting embryonic calli with Agrobacterium tumefaciens and that of soybean and kidney bean by injecting the bacteria into germinating seeds. Plant Biotechnol 17:187–194

    Google Scholar 

  • Holsters M, de Waele D, Depicker A, Messens E, Van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187

    Google Scholar 

  • Hong B, Barg R, Ho T-HD (1992) Developmental and organ-specific expression of an ABA- and stress-induced protein in barley. Plant Mol Biol 18:663–674

    Google Scholar 

  • Honjoh KI, Oda Y, Takata R, Miyamoto T, Hatano S (1999) Introduction of the hiC6 gene, which encodes a homologue of a late embryogenesis abundant (LEA) protein, enhances freezing tolerance of yeast. J Plant Physiol 155:509–512

    Google Scholar 

  • Imai R, Chang L, Ohta A, Bray EA, Takagi M (1996) A LEA class gene of tomato confers salt and freezing tolerance when expressed in Saccharomyces cerevisiae. Gene 170:243–248

    Google Scholar 

  • Jeong WJ, Min SR, Liu JR (1995) Somatic embryogenesis and plant regeneration in tissue cultures of radish (Raphanus sativus L.). Plant Cell Rep 14:648–651

    Google Scholar 

  • Joersbo M, Brunstedt J (1992) Sonication: a new method for gene transfer to plants. Physiol Plant 85:230–234

    Google Scholar 

  • Matsubara S, Hegazi HH (1990) Plant regeneration from hypocotyl callus of radish. HortScience 25:1286–1288

    Google Scholar 

  • Pua E-C, Sim G-E, Chi G-L, Kong L-F (1996) Synergistic effect of ethylene inhibitors and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey) in vitro. Plant Cell Rep 15:685–690

    Google Scholar 

  • Rogers SO, Bendich AJ (1988) Extraction of DNA from plant tissues. In: Galvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp A6/1–A6/10

    Google Scholar 

  • Rohila JS, Jain RK, Wu R (2002) Genetic improvement of Basmati rice for salt and drought tolerance by regulated expression of a barley Hva1 cDNA. Plant Sci 163:525–532

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York, pp 1–808

    Google Scholar 

  • Santarém ER, Trick HN, Essig JS, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression. Plant Cell Rep 17:752–759

    Google Scholar 

  • Schrimer E, Lindquist S, Vierling E (1994) An Arabidoposis heat protein complements a thermotolerance defect in yeast. Plant Cell 6:1899–1909

    Google Scholar 

  • Sivamani E, Bahieldin A, Wraith JM, Al-Niemi T, Dyer WE, Ho THD, Qu R (2000) Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene. Plant Sci 155:1–9

    Google Scholar 

  • Skriver K, Mundy J (1990) Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2:503–512

    Google Scholar 

  • Swire-Clark GA, Marcotte WR Jr (1999) The wheat LEA protein Em functions as an osmoprotective molecule in Saccharomyces cerevisiae. Plant Mol Biol 39:117–128

    Google Scholar 

  • Takahata Y, Komatsu H, Kaizuma N (1996) Microspore culture of radish (Raphanus sativus L.): influence of genotype and culture conditions on embryogenesis. Plant Cell Rep 16:163–166

    Google Scholar 

  • Tingay S, McElroy D, Kalla R, Fieg S, Wang M, Thornton S, Bretell R (1997) Agrobacterium tumefaciens-mediated barley transformation. Plant J 11:1369–1376

    Google Scholar 

  • Trick NH, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean (Glycine max L. Merrill) embryogenic suspension culture tissue. Plant Cell Rep 17:482–488

    Google Scholar 

  • Trieu AT, Burleighet SH, et al (2000) Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium. Plant J 22:531–541

    Google Scholar 

  • Wakui K, Takahata Y (2002) Isolation and expression of Lea gene in desiccation-tolerant microspore-derived embryos in Brassica spp. Physiol Plant 116:223–230

    Google Scholar 

  • Xu D, Duan X, Wang B, Hong B, Ho T-HD, Wu R (1996) Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol 110:249–257

    Google Scholar 

  • Zhang LJ, Cheng LM, Xu N, Zhao NM, Li CG, Jing Y Jia SR (1991) Efficient transformation of tobacco by ultrasonication. Bio/Technology 9:996–997

    Google Scholar 

  • Zhang L, Ohta A, Takagi M, Imai R (2000) Expression of plant group 2 and group 3 lea genes in Saccharomyces cerevisiae revealed functional divergence among LEA proteins. J Biochem 127:611–616

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zaochang Liu.

Additional information

Communicated by H. Ebinuma

Rights and permissions

Reprints and permissions

About this article

Cite this article

Park, BJ., Liu, Z., Kanno, A. et al. Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus . Plant Cell Rep 24, 494–500 (2005). https://doi.org/10.1007/s00299-005-0973-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-005-0973-5

Keywords

Navigation