Factors affecting delivery and transient expression of gusA gene in Malaysian indica rice MR 219 callus via biolistic gun system

A. R. Zuraida, K. Rahiniza, Hafiza M R Nurul, Roowi Suri, Zamri Zainal, Subramaniam Sreeramanan

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The effect of the biolistic device parameters and callus factors affecting delivery and expression of gusA gene in indica rice MR219 was optimised. Four weeks old embryogenic callus were bombarded with gold microparticles coated with pCAMBIA1305.2 plasmid harbouring the gusA gene which encodes-glucuronidase. The physical and biological factors investigated were the helium pressure, distance from stopping screen to target tissues, vacuum pressure, gold microparticles size, spermidine and calcium on DNA precipitation, the number of bombardments, age of callus, osmotic pre-culture length and osmotic pretreatment medium. The embryogenic calli were subjected to histochemical GUS assay after three days bombardment. The GUS activity (recorded as blue spots) detected histochemically and fluorometrically were further analysed by microscopy method. Mostly, the variables used in this experiment showed significant difference on the delivery of DNA and transient expression of the gusA gene in embryogenic rice calli. Combinations of selected optimized parameters and an effective selection were developed which allowed high-efficiency of DNA delivery combined with minimum damage to target indica rice MR219 embryogenic callus tissue.

Original languageEnglish
Pages (from-to)8810-8818
Number of pages9
JournalAfrican Journal of Biotechnology
Volume9
Issue number51
Publication statusPublished - Dec 2010

Fingerprint

Biolistics
biolistics
Bony Callus
Firearms
callus
Gene Expression
rice
gene expression
Gold
gold
DNA
osmotic treatment
Pressure
Helium
helium
Spermidine
Glucuronidase
Biological Factors
spermidine
Vacuum

Keywords

  • Biolistic gun
  • GusA gene
  • Indica rice MR219
  • Transient expression

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Genetics
  • Molecular Biology
  • Agronomy and Crop Science

Cite this

Factors affecting delivery and transient expression of gusA gene in Malaysian indica rice MR 219 callus via biolistic gun system. / Zuraida, A. R.; Rahiniza, K.; Nurul, Hafiza M R; Suri, Roowi; Zainal, Zamri; Sreeramanan, Subramaniam.

In: African Journal of Biotechnology, Vol. 9, No. 51, 12.2010, p. 8810-8818.

Research output: Contribution to journalArticle

Zuraida, A. R. ; Rahiniza, K. ; Nurul, Hafiza M R ; Suri, Roowi ; Zainal, Zamri ; Sreeramanan, Subramaniam. / Factors affecting delivery and transient expression of gusA gene in Malaysian indica rice MR 219 callus via biolistic gun system. In: African Journal of Biotechnology. 2010 ; Vol. 9, No. 51. pp. 8810-8818.
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