Preliminary investigations of Agrobacteriu-mediated transformation in indica rice MR219 embryogenic callus using gusA gene

Zuraida Ab Rahman, Zulkifli Ahmad Seman, Naziah Basirun, Advina Lizah Julkifle, Zamri Zainal, Sreeramanan Subramaniam

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Preliminary steps in the genetic transformation of indica rice MR219 was investigated in the plant-Agrobacterium tumefaciens interaction. Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector pCAMBIA 1305.2 harboring the modified GUS gene driven by the CaMV 35S promoter was used. Various transformation parameters influences were optimized using embryogenic calli via β-glucuronidase (GUS) as a reporter marker. Various transformation parameters were optimized including bacterial concentration, age of embryogenic callus, pre-culture period, wounding technique, cocultivation period, immersion time and dry time before co-cultivation, acetosyringone (AS) concentration, pH of co-cultivation media and temperature of the co-cultivation period. The expression of the transient gusA gene in the plant genome was preliminary confirmed by histochemical GUS assay activity (as blue spots). The results from transient gusA gene expression of calli suggested that the Agrobacterium-mediated transfer system of T-DNA in indica rice MR219 was highly efficient. Therefore, the investigation of factors that influence T-DNA delivery is an important first step in the utilization of Agrobacterium in the transformation of indica rice MR219 calli.

Original languageEnglish
Pages (from-to)7805-7813
Number of pages9
JournalAfrican Journal of Biotechnology
Volume10
Issue number40
Publication statusPublished - 1 Aug 2011

Fingerprint

Bony Callus
Glucuronidase
callus
rice
Agrobacterium tumefaciens
Agrobacterium
Agrobacterium radiobacter
Genes
genes
Genetic Transformation
acetosyringone
Plant Genome
DNA
Immersion
genetic transformation
Coculture Techniques
promoter regions
Gene Expression
gene expression
Temperature

Keywords

  • Agrobacterium tumefaciens
  • GUS expression
  • Indica rice MR219

ASJC Scopus subject areas

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

Cite this

Preliminary investigations of Agrobacteriu-mediated transformation in indica rice MR219 embryogenic callus using gusA gene. / Rahman, Zuraida Ab; Seman, Zulkifli Ahmad; Basirun, Naziah; Julkifle, Advina Lizah; Zainal, Zamri; Subramaniam, Sreeramanan.

In: African Journal of Biotechnology, Vol. 10, No. 40, 01.08.2011, p. 7805-7813.

Research output: Contribution to journalArticle

Rahman, Zuraida Ab ; Seman, Zulkifli Ahmad ; Basirun, Naziah ; Julkifle, Advina Lizah ; Zainal, Zamri ; Subramaniam, Sreeramanan. / Preliminary investigations of Agrobacteriu-mediated transformation in indica rice MR219 embryogenic callus using gusA gene. In: African Journal of Biotechnology. 2011 ; Vol. 10, No. 40. pp. 7805-7813.
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