论文部分内容阅读
最近的研究证明在叶绿体内过量表达甲基营养细菌RuMP中固定HCHO的关键酶6-磷酸己酮糖合成酶(HPS)和6-磷酸果糖异构酶(PHI)的融合蛋白HPS-PHI可以在转基因天竺葵中构建一条甲醛光合同化途径从而提高植物对甲醛的同化和脱毒能力。甲醇氧化酶(AOD1)是甲基营养型酵母甲醇代谢途径的第一个关键酶,可催化甲醇氧化为甲醛。在烟草叶绿体中过量表达AOD1或/和HPS-PHI产生过量表达AOD1(AO)、HPS-PHI(AB)、同时过量表达AOD1和HPS-PHI(AA)的转基因烟草。用2和6mmoL/L~(13)CH_3OH处理3种转基因烟草和野生型烟草(WT),~(13)CNMR分析表明AO植株中积累的H~(13)COOH最多,[U-~(13)C]Gluc和[U-~(13)C]Fruc的生成量在AO和WT间无显著差异,在AA和AB中的产生量显著高于AO和WT,且在AA中的生成量又高于AB,这种差异在6mmol/L~(13)CH_3OH处理植株中比2 mmol/L更为明显。这些结果证实在叶绿体中过表达AOD1增加烟草氧化甲醇为甲酸的能力,同时过量表达AOD1和HPS-PHI在烟草中成功地构建一个甲醇光合同化途径,提高烟草同化甲醇为糖类物质的能力。
Recent studies have demonstrated that the fusion protein HPS-PHI, which is a key enzyme that immobilizes HCHO in methylotrophic bacteria RuMP, overexpressing chlorophyll-6-phosphate hexose phosphate synthase (HPS) and 6-phosphate fructose isomerase (PHI) Genetically modified geranium creates a formalin photo-assimilation pathway that enhances plant assimilation and detoxification of formaldehyde. Methanol oxidase (AOD1) is the first key enzyme in methanol metabolism of methylotrophic yeast and catalyzes the oxidation of methanol to formaldehyde. Overexpression of AOD1 or / and HPS-PHI in tobacco chloroplasts resulted in transgenic tobacco that overexpressed AOD1 (AO), HPS-PHI (AB), overexpressing both AOD1 and HPS-PHI (AA). Three transgenic tobacco and wild type tobacco (WT) were treated with 2 and 6mmoL / L ~ (13) CH_3OH. The ~ (13) CNMR analysis showed that the accumulation of H ~ (13) ) C] Gluc and [U- ~ (13) C] Fruc had no significant difference between AO and WT, and the production in AA and AB was significantly higher than that in AO and WT, and the production in AA Higher than AB, the difference was more obvious than that of 2 mmol / L in 6 mmol / L ~ (13) CH_3OH treatment plants. These results demonstrate that overexpression of AOD1 in chloroplasts increases the ability of tobacco to oxidize methanol to formic acid while overexpressing AOD1 and HPS-PHI successfully establish a methanol photo-assimilation pathway in tobacco to increase the ability of tobacco to assimilate methanol to carbohydrates.