May 29, 2004

Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses

Trehalose is a nonreducing disaccharide of glucose that functions as a compatible solute in the stabilization of biological structures under abiotic stress in bacteria, fungi, and invertebrates. With the notable exception of the desiccation-tolerant “resurrection plants,” trehalose is not thought to accumulate to detectable levels in most plants. We report here the regulated overexpression of Escherichia coli trehalose biosynthetic genes (otsA and otsB) as a fusion gene for manipulating abiotic stress tolerance in rice. The fusion gene has the advantages of necessitating only a single transformation event and a higher net catalytic efficiency for trehalose formation. The expression of the transgene was under the control of either tissue-specific or stress-dependent promoters. Compared with nontransgenic rice, several independent transgenic lines exhibited sustained plant growth, less photo-oxidative damage, and more favorable mineral balance under salt, drought, and low-temperature stress conditions. Depending on growth conditions, the transgenic rice plants accumulate trehalose at levels 3–10 times that of the nontransgenic controls. The observation that peak trehalose levels remain well below 1 mg/g fresh weight indicates that the primary effect of trehalose is not as a compatible solute. Rather, increased trehalose accumulation correlates with higher soluble carbohydrate levels and an elevated capacity for photosynthesis under both stress and nonstress conditions, consistent with a suggested role in modulating sugar sensing and carbohydrate metabolism. These findings demonstrate the feasibility of engineering rice for increased tolerance of abiotic stress and enhanced productivity through tissue-specific or stress-dependent overproduction of trehalose.

Source: Proc Natl Acad Sci U S A. (2002) vol 99, p. 15898–15903

April 24, 2004

Research

Doctoral Degree (2004 - Now):
* Part I:
1. Production of transgenic rice overexpressing AOS-LOX fusion gene (T0, T1, T2)

2. Genetic Analysis of transgenic rice
- Analysis of DNA (by PCR and/or Southern), RNA (by Northern), Protein (Western)
- Selection of homozygous lines of transgenic rice

3. Biochemical Analysis of transgenic rice
- Lipid analysis
- JA analysis
- Dihydro JA analysis

4. Stress response of overexpressed transgenic rice
- Ion leakage experiments in cold, salt stress
- Wound and HR (local, systemic) response
- Blast fungi resistance

5. Metabolomics of overexpressed transgenic rice
- Lipid peroxidation experiments
- Product analysis by HPLC, GC, GC-MS
- Metabolic changes by wound and other stresses


* Part II:
1. Characterization of Fusion Genes (AOS-LOX, AOS-AOC and AOC-AOS)


Master Degree (2000 - 2002):

1. Production of transgenic rice overexpressing allene oxide synthase gene

2. Kinetic properties of V311M mutant Bacillus subtilis protoporphyrinogene oxidase
- Protein expression
- Protein purification
- Enzyme kinetics

April 23, 2004

Publications

Updated on December 22, 2009
  1. Thavrak Huon, Sungkuk Jang, Kyoungwon Cho, Randeep Rakwal, Je Chang Woo, Ilchul Kim, Seung Wook Chi, Oksoo Han (2009) A Substrate Serves as a Hydrogen Atom Donor in the Enzyme-Initiated Catalytic Mechanism of Dual Positional Specific Maize Lipoxygenase-1. Bulletin of the Korean Chemical Society. 30: 719-723 [Abstract] [PDF Full Text]

  2. Sungkuk Jang, Thavrak Huon, Keumhwa Kim, Eunji Um, and Oksoo Han (2007) Regiochemical and stereochemical evidence for enzyme-initiated catalysis in dual positional specific maize lipoxygenase-1. Organic letters 16: 3113-3116 [Abstract] [PDF Full Text]

  3. Kyoungwon Cho, Sungkuk Jang, Thavrak Huon, Sangwook Park and Oksoo Han (2007) Biochemical Characterization of the Dual Positional Specific Maize Lipoxygenase and the Dependence of Lagging and Initial Burst Phenomenon on pH, Substrate, and Detergent during Pre-steady State Kinetics. Journal of Biochemistry and Molecular Biology 40: 100-106 [Abstract] [PDF Full Text]

  4. Eunjoo Jeong, Thavrak Huon, Yongin Kuk, Eun-Seon Kim, Hema Kumar Chandru, Myunggi Baik, Kyoungwhan Back, Ja-Ock Guh, and Oksoo Han (2003) A point mutation of valine-311 to methionine in Bacillus subtilis protoporphyrinogen oxidase does not greatly increase resistance to the diphenyl ether herbicide oxyfluorfen. Bioorganic Chemistry 31: 389-397 [Abstract] [PDF Full Text]

  5. Thavrak Huon (2002) Kinetic properties of V311M mutant Bacillus subtilis protoporphyrinogen oxidase. Thesis of Master degree, Department of Genetic Engineering, Chonnam National University. Gwangju, Republic of Korea

  6. Thavrak Huon (1997) The effect of organic and inorganic fertilization on the growth of Tilapia fingerling. Thesis of Bachelor degree, Faculty of Fisheries, Royal University of Agriculture. Phnom Penh, Cambodia