Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana

S. F. Ee, M. B. Khairunnisa, Zeti Azura Mohamed Hussein, S. Noor Azmi, Zamri Zainal

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The floral-dip transformation method for Arabidopsis thaliana has long been established. Following transformation, an important step is involving the selection for transgenic plants through antibiotics or herbicides. Hygromycin has been widely applied for transgenic plants selection. However, the concentrations used were varied between laboratories mostly in a range of 10 mg/L up to 200 mg/L. In the present study, the hygromycin sensitivity test was performed on wild-type seeds to obtain the most effective hygromycin concentration for selection of the transgenic. A standard curve on average mean of hypocotyls lengths against hygromycin concentrations was constructed. Based on hypocotyls length and leaf colour, the most effective concentration of hygromycin was determined in the range of 20 to 30 mg/L. For screening the transformed seeds, hygromycin concentration at 25 mg/L was used. Non-transgenic plants demonstrated a clear decrease in hypocotyls lengths and no root elongation, as compared to transgenic plants. The identified transgenic seedlings were further verified through polymerase chain reaction (PCR), giving fragment with an expected size of 365 bp.

Original languageEnglish
Pages (from-to)119-123
Number of pages5
JournalMalaysian Applied Biology
Volume43
Issue number1
Publication statusPublished - 2014

Fingerprint

Agrobacterium
Arabidopsis
hypocotyls
transgenic plants
Arabidopsis thaliana
genetically modified organisms
Hypocotyl
Genetically Modified Plants
seeds
root growth
Seeds
herbicides
polymerase chain reaction
antibiotics
screening
seedlings
color
Herbicides
Seedlings
hygromycin A

Keywords

  • Agrobacterium-mediated
  • Arabidopsis thaliana
  • Floral-dip transformation
  • Hygromycin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Effective hygromycin concentration for selection of Agrobacterium-mediated transgenic Arabidopsis thaliana. / Ee, S. F.; Khairunnisa, M. B.; Mohamed Hussein, Zeti Azura; Noor Azmi, S.; Zainal, Zamri.

In: Malaysian Applied Biology, Vol. 43, No. 1, 2014, p. 119-123.

Research output: Contribution to journalArticle

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