Biolistic co-transformation of rice using gold nanoparticles

Authors

  • S. E. Mortazavi Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Karaj, I. R. Iran
  • Z. Zohrabi Department of Plant Inbreeding, College of Agriculture, Zanjan University, Zanjan,, I. R. Iran
Abstract:

ABSTRACT- In order to produce transgenic rice lines lacking selectable marker gene, biolistic co-transformation technique using gold nanoparticles was adopted. In the first step, the efficiency of different sizes of gold particles was evaluated. The results showed that the efficiency of the nanoparticles in the transformation was comparable to that of the micro particles. Subsequently, two separate plasmids including p UBC harboring cry IA (c) gene and pTRA132 carrying hph gene as selectable marker gene were mixed and coated with gold nanoparticles. Embryo genic calli originated from mature rice seeds were targeted with plasmid-coated nan projectiles. Putative transgenic cells were selected from selection media after three subcultures in MS medium were supplemented with 50 mg hygromycine B l-1 in three weeks' intervals. The selected calli were then grown into putative transgenic plantlets. Molecular analysis performed on the regenerated lines showed that at least one copy of both genes was inserted into the genome of some transgenic lines. RT-PCR analysis also confirmed successful transcription of the cry IA (c) gene. Accordingly, it could be concluded that the biolistic co-transformation method using gold nanoparicles could be a successful method for gene delivery into rice varieties. Nevertheless, expression of cry IA (c) gene and loci of both genes should still be investigated in future studies.

Download for Free

Sign up for free to access the full text

Already have an account?login

similar resources

Biolistic transformation of a procaryote, Bacillus megaterium.

We present a simple and rapid method for introducing exogenous DNA into a bacterium, Bacillus megaterium, utilizing the recently developed biolistic process. A suspension of B. megaterium was spread onto the surface of nonselective medium. Plasmid pUB110 DNA, which contains a gene that confers kanamycin resistance, was precipitated onto tungsten particles. Using a biolistic propulsion system, t...

full text

Biolistic gun-mediated maize genetic transformation.

Biolistic gun-mediated transformation is one of the two most effective and popular methods for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype, Hi II. The protocol uses 0.6-microm gold particles as microcarriers and the herbicide resistance bar gene as a selective marker. Both immature zygotic embryos and immature e...

full text

Generation of transgenic C. elegans by biolistic transformation.

The number of laboratories using the free living nematode C. elegans is rapidly growing. The popularity of this biological model is attributed to a rapid generation time and short life span, easy and inexpensive maintenance, fully sequenced genome, and array of RNAi resources and mutant animals. Additionally, analysis of the C. elegans genome revealed a great similarity between worms and higher...

full text

GENETIC TRANSFORMATION AND HYBRIDIZATION Unintended consequence of plant transformation: biolistic transformation caused transpositional activation of an endogenous retrotransposon Tos17 in rice ssp. japonica cv. Matsumae

Genetic instability could be provoked as an unintended consequence of genetic engineering in plants. Here, we report that the rice endogenous long terminal repeat (LTR) retrotransposon Tos17 was transpositionally activated only in transgenic calli and their regenerated plants produced by biolistic transformation in rice (Oryza sativa L.) ssp. japonica cv. Matsumae. Moreover, the transpositional...

full text

Green synthesis of gold nanoparticles using plant extract: Mini-review

In this review, we examine the greenest nanoparticles of zero-valent metals, metal oxides and metal salts, with emphasis on recent developments routes. Products from nature or those derived from natural products, such as extracts of several plants or parts of plants, tea, coffee, banana, simple amino acids, as well as wine, table sugar and glucose, have been used as reductants and as capping ag...

full text

Cationic gold microparticles for biolistic delivery of nucleic acids.

Here we report preparation and properties of positively charged gold microparticles, and their use for biolistic DNA delivery. Micron-sized gold microparticles were modified by building self-assembling polyethylenimine monolayers on their surfaces, which enabled their electrostatic interaction with negatively charged molecules such as DNA. One milligram of the surface-modified microparticles wa...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 37  issue 1

pages  75- 82

publication date 2018-08-23

By following a journal you will be notified via email when a new issue of this journal is published.

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023