A stable and simple transformation of rice (Oryza sativa L. ssp indica) mature embryo using in planta method

Nasr-Ramzi, S; Sohani, M; Hasani, H; Asghari, J

A stable and simple transformation of rice (Oryza sativa L. ssp indica) mature embryo using in planta method

Document Type : Research Paper

Authors

University of Guilan

Abstract

Agrobacterium-mediated transformation technique is a powerful and essential tool for genetic transformation and transgenic rice plant production. In this study an in planta transformation method was used for rice plants transformation. Therefore, rice seeds were socked for two days and the mature rice embryos was inoculated by means of an Agrobacterium coated needle. An experiment with factorial design including two strains of Agrobacterium tumefaciens (EHA105 and LBA4404) harboring pCAMBIAl105.1R, three levels of Acetosyringone (0, 100 and 200 Mµ), three cultivars of rice (Hashemi, Hasani and Gharib) and two treatments of vacuum and no vacuum operation was carried out in a Completely Randomized Design with three replications. Integration of the transgene into the genome of putative transgenic rice plants were confirmed using resistance of leaf tissues to Hygromycin, the histiochemical GUS assay and PCR with at least three different genes. Accordingly, EHA105 strain and Hashemi cultivar in the presence of 100µM acetosyringone using vacuum in a vir genes induction medium had a significant transformation efficiency (37.46%). The success of transformation was further confirmed by analysis of T1 generation with 21% transgenic.

Keywords

20.1001.1.22520783.1391.2.2.5.7

Main Subjects

Genetic Engineering and Gene Transformation

References

REFERENCES

Alt-Moerbe J, Neddermann P, Von LJ, Weiler EM, Schroder J (1988) Temperature sensitive step in Ti plasmid vir region induction and correlation with cytokinin secretion by Agrobacterium. Mol. Gen. Genet. 213: 1-8.

An G, Lee S, Kim SH, Kim SR (2005) Molecular genetics using T-DNA in rice plant. Plant cell physiol. 46: 14-22.

Anand A, Uppalapati SR, Ryu CM, Allen SN, Kang L, Tang Y, Mysore KS (2008) Salicylic acid and systemic acquired resistance play a role in attenuating crown gall disease caused by Agrobacterium tumefaciens. Plant Physiol. 146: 703-715.

Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C. R. Acad Sci, Paris Life Sci. 316: 1194-1199.

Bratic A, Majic DB, Miljus JD, Jovanovic ZS, Maksimovic VR (2007) In planta transformation of buckwheat (Fagopyrum esculentum Moench). Arch. Biol. Sci. 59 (2): 135-138.

Chumakov MI, Rozhok NA, Vlikov VA, Tyrnov VS, Volokhina IV (2006) Agrobacterium-mediated in planta transformation of maize via pistil filaments. Russ J Genet. 42: 893-897.

Clarke JD (2009) Cetyle tri methyl Ammonium Bromide (CTAB) DNA Mini prep for Plant DNA Isolation. Cold Spring Herb Protoc. 10: 1101-5177.

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

Danilova SA (2007) The technologies for genetic transformation of cereals. J. Plant physiol.54: 569-581.

Ditt RF, Nester E, Comai L (2005) The plant cell defense and Agrobacterium tumefaciens. FEMS Microbiol. 247: 207-213.

Dye F, Berthelot K, Griffon B, Delay D, Delmotte FM (1997) Alkysyringamides, new inducers of Agrobacterium tumefaciensvir gene induction expression. Proc. Natl. Acad. Sci. USA. 83: 379-383.

Fierer N, Jackson RB (2006) The diversity and Biogeography of soil bacterial communities. Proc. Natl. Acad. Sci. USA. 103: 626-631.

Finnegan EJ, Peacock WJ, Dennis ES (2000) DNA methylation, a key regulator of plant development and other processes. Curr. Opin. Genet. Dev. 10: 217-223.

Gelvin SB (2006) Agrobacterium virulence gene induction.Mol Biol. 343: 77-84.

Gould J (1996) Transformation of the cereals using Agrobacterium. Mol Biol. 62: 491-501.

Hellens R, Mullineaux P, Klee H (2000) A guide to Agrobacterium binary Ti vectors. Trends Plant Sci. 5: 446-451.

Hiei Y, Komari T, Kubo T (1997) Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol Biol. 35: 205-218.

Hiei Y, Komari T (2006) Improved protocols for transformation of indica rice mediated by Agrobacterium tumefaciens .Plant Cell. Tiss. Org. 85: 271-283.

Hiei Y, Komari T (2008) Agrobacterium-mediated transformation of rice using immature embryo or calli induced from mature seed. Nature. 3: 824-834.

Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983)A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti plasmid. Nature. 303: 179-180.

Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobaterium helper plasmids for plasmids for gene transfer to plants. Transgenic Res. 2: 208-218.

Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol. 5: 387-450.

Jin W, Gong Z, Lu Y (2004) Studies on the technique of in planta Agrobacterium-mediated gene transfer by vacuum infiltration of Brassica Juncea.ActaAgronBoreali-Sinica. 19: 8-12.

Kumar KK, Maruthasalam M, Loganathan D, Rudhakar D, Balasubramanian P (2005) An improved Agrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars. Plant Mol. Biol. Rep. 23: 67-73.

Lersten NR (2004) Flowering Plant Embryology, Blackwell Publishing, p. 212.

Lin J, Zhou B, Yang Y, Mei J, Zhao X, Guo X, Huang X, Tang D, Liu X (2009) Piercing and vacuum in filtration of mature embryo: a simplified method for Agrobacterium-mediated transformation of indica rice. Plant Cell Rep. 28: 1065-1074.

Liu CN, Steck TR, Habeck LL, Meyer JA, Gelvin SB (1993) Multiple copies of vir G allow induction of Agrobacterium tumefaciens vir genes and T-DNA processing at alkaline pH. Mol. Plant-Microbe Interact. 6: 144-156.

Liu F, Cao MQ, Yao L (1998) In planta transformation of Pakchoi (Brassica campestris L.) by infiltration of adult plants with Agrobacterium. Acta Hortic. 467: 187-193.

Liu Z, Park BJ, Kanno A, Kameya T (2005) The novel use of a combination of sonication and vacuum infiltration in Agrobacterium-mediated transformation of kidney bean (Phaseolus vulgaris L.) with lea gene. Mol. Breed. 16: 189-197.

McCullen CA, Binns AN (2006) Agrobacterium tumefaciens and plant cell interactions activities required for interkingdom macromolecular transfer. Annu. Rev. Cell Dev. Biol. 22: 101-128.

Mohan JS (2001) Tissue culture-derived variation in crops improvement. J. Plant Breed. 118: 153-160.

Peng WT, Lee YW, Nester EW (1998) The phenolic recognition profiles of the Agrobacterium tumefaciens virA protein are broadened by high level of the sugar binding protein ChvE. J. Bacteriol. 180: 5632-5638.

Rashid H, Yokoi SL, Toriyama K, Hinanta K (1996) Transgenic plant production mediated by Agrobacterium in indica rice. Plant Cell Rep. 15: 727-730.

Rashid H, Afzal A, Khan MH, Chaaudhary Z, Malik SA (2010) Effect of bacterial culture density and acetosyringone concentration on Agrobacterium mediated transformation in wheat. J. Bot. 42(6): 4183-4189.

Supartana P, Shimizu T, Shiori H, Nogawa M, Nozue M, Kojima M (2005) Development of simple and efficient in planta transformation method for rice (Oryza sativa L.) Using Agrobacterium tumefaciens. Biosci and Bioeng. 100: 391-397.

Supartana P, Shimizu T, Nogawa M, Hidenari S, Nakajima T, Haramoto N, Nozue M, Kojima M (2006) Development of simple and efficient in planta transformation method for wheat (Triticum aestivum L.) using Agrobacterium tumefaciens. Biosci and Bioeng.102(2): 162-170.

Tzfira T, Jianxiong L, Lacroix B, Citovsky V (2004) Agrobacterium T-DNA integration: molecules and models. Trend in Genet. 8: 375-383.

Veena, JH, Doerge RW, Gelvin SB (2003) Transfer of T-DNA and Vir proteins to plant cells by Agrobacterium tumefaciens induces expression of host genes involved in mediating transformation and suppresses host defense gene expression. Plant J. 35: 219-226.

Veluthambi K, Aditya K, Sharma A (2003) The current status of plant transformation technologies. Curr Sci. 84: 368-380.

Wang MB, Waterhouse PM (1997) A rapid and simple method of assaying plants transformed with hygromycin or PPT resistance genes. Plant Mol. Biol. Rep. 15: 209-215.

Wang WC, Menon G, Hansen G (2003) Development of a novel Agrobacterium-mediated transformation method to recover transgenic Brassica napus plants. Plant Res. 22: 274-281.

Xu GS, Rao YQ, Chen Y, Zhang CY, Meng JL (2004) Genetic transformation of Brassica napus with in planta method. Acta Agron. Sin. 30: 1-5.

Yoshida S, Forno DA, Cock JH, Gomez KA (1971) Laboratory manual for physiological studies of rice. The International Rice Research Institute, Los Babos, Philippines.

Yuan ZC, Edlind MP, Liu P, Saenkham P, Banta LM, Wise AA, Ronzone E, Binns AN, Kerr K, Nester EW (2007). The plant signal salicylic acid shuts down expression of the vir regulation and activates quormone-quenching genes in Agrobacterium. Proc. Natl. Acad. Sci. USA. 104: 11790-11795.

Zhao Z, Sagulenko E, Ding Z, Christie PJ (2001) Activities of virE1 and virE1 secretion chaperone in export of the multifunctional virE2 effectors via an Agrobacterium type IV secretion pathway. J Bacteriol. 183: 3855-3865.