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• Establishment of ASimple and Efficient Transformation Protocol for Hairy Root Induction of Wild Bitter Melon (Momordica charantia L.var. abbreviata Ser.)

  Nguyen Thi Thu Trang, Vu Thi Dao, Bui Minh Tri

  Date Received: 28-09-2016 / Date Accepted: 13-02-2017

   Abstract PDF (VIETNAMESE)

  Wild bitter melon (Momordica charantia L. var. abbreviata Ser.) is known as a valuable vegetable and herb. It contains several important bioactive substances, including glycoside, saponin, alkaloide, triterpen and steroide. This study was carried out to establish a simple and efficient transformation protocol for inducting hairy roots of wild bitter melon by using strain TR7 of Agrobacterium rhizogenes which carries pPE113 binary vector harbouring e-gfp gene. Major parameters were optimized including, the age of plant and position for injection. The results indicated that highest hairy root induction frequencies was about 69.66-73.33% when seven-day-old seedlings were used to infect with the bacterium using hypocotyl. After regeneration and selection, thirty putative root clones of 4 specific phenotypes, i.e. white hairs, fast growth and branching on hormone-free MS medium, were obtained. The average GFP expression rate in 4 specific phenotypic groups were about 46.7%. PCR analysis using specific primers for rolA gene and virD gene confirmed the presence of the rolA gene and absence of virD gene in the genome of four hairy root lines. It was suggested that this simple transformation protocol can be applied for hairy root induction of other varieties as well.

• Construction and Development of Yeast-based Biosensor Applied for Detection of Genotoxic Chemical Compounds

  Bui Van Ngoc, Nguyen Huu Duc, Nguyen Duc Thang, Nguyen Cong Hieu

  Date Received: 06-05-2013 / Date Accepted: 26-08-2013 / Date Published: 13-06-2025

   Abstract PDF (VIETNAMESE)

  Biosensor was developed by design and construction of a reporter plasmid containing GFP gen under regulation of RNR2 promoter. This reporter plasmid was transformed in yeast cells which were then treated with different concentrations of MMS and menadione. Due to induction of genotoxic compounds, such as MMS, RNR2 promoter regulates transcription and translation of GFP leading to fluorescence signal measured at 485nm (excitation wave length) and 535nm (emission wave length). Fluorescence instensity was directly proportional to MMS concentrations when yeast cells treated with less than 1mM in the environment, but decreasing when exposed to higher MMS concentrations as a result of inhibition of cellular respiration and ATP synthesis. In contrast, promoter RNR2 was not induced by menadione resulting in no fluorescence signal. Thus, this kind of biosensor is able to not only determine and detect genotoxic chemical compounds, which are harmful to human and environment, but also identify new and uncharacterized compounds which possess the same chemical properties as MMS, and could be used in new drug discovery and drug screening.

  DOI: https://doi.org/10.31817/tckhnnvn.2013.11.6.