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| Simultaneous Over-expressions of AOD1 and HPS-PSI in Chloroplasts Creates a Novel Photosynthetic CH3OH-assimilation Pathway and Enhances Its Ability to Assimilate CH3OH |
| LIU Lei, SUN Zhen, SONG Zhong-bang, XIAO Su-qin, CHEN Li-mei |
| Faculty of Live Science and Engineering, Chenggong Campus, Kunming University of Science and Technology, Kunming 650500, China |
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Abstract 3-hexulose-6-phosphate synthase (HPS) and 6-phosphate-3-hexuloisomerase (PHI) are two key enzymes in a HCHO-assimilation pathway in methylotroph. It has demonstrated that overexpression of HPS/PHI fusion protein in chloroplasts of geranium installs a photosynthetic HCHO-assimilation pathway and thereby enhanced the ability of transgenic plants to assimilate and detoxify formaldehyde. Alcohol oxidase (AOD1) is the first key enzyme in the methanol metabolic pathway in methylotrophic yeasts. AOD catalyzes oxidation of methanol to formaldehyde. AOD1 or/and HPS-PHI were over-expressed in chloroplasts of tobacco to generate AOD1 (AO), HPS-PHI (AB) over-expression line as well as AOD1 and HPS-PHI (AA) simultaneous over-expression line. The transgenic and wild-type tobacco (WT) was treated with 2 mmol/L and 6 mmol/L 13CH3OH. 13C-NMR analysis showed that the accumulation of H13COOH was maximum in AO trangenic plants. There is no significant difference in the generation of [U-13C] glucose (Gluc) and [U-13C] fructose (Fruc) between AO and WT lines. However, the production of [U-13C] glucose (Gluc) and [U-13C] fructose (Fruc) in AA and AB lines was significantly higher than that in AO and WT lines. Moreover, the formation of the two metabolites in AA line was higher than in AB line. These differences were more significant in 6mmol/L 13CH3OH treated-plants than in 2 mmol/L 13CH3OH-treated plants. These results suggested that overexpression of AOD1 in chloroplasts enhanced the ability of tobacco to oxidize methanol to formaldehyde, while simultaneous overexpression of AOD1 and HPS-PHI could successfully created a photosynthetic CH3OH-assimilation pathway, thus enhanced ability of tobacco to assimilate CH3OH as sugars.
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Received: 12 October 2013
Published: 25 December 2013
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