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在叶绿体中过量表达AOD1和HPS-PHI创建光合甲醇同化途径增加烟草同化甲醇能力
  • ISSN号:1671-8135
  • 期刊名称:《中国生物工程杂志》
  • 时间:0
  • 分类:Q812[生物学—生物工程]
  • 作者机构:[1]昆明理工大学生命科学与技术学院生物工程技术研究中心,昆明650500
  • 相关基金:国家自然科学基金资助项目(30970263)
中文摘要:

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(AODl)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AODl或/和HPS.PHI产生过量表达AODl(AO)、HPS—PHI(AB)、同时过量表达AODl和HPS—PHI(AA)的转基因烟草。用2和6mmol/L 13CH3OH处理3种转基因烟草和野生型烟草(wT),13C—NMR分析表明AO植株中积累的H”COOH最多,[U-13C]Gluc和[U-13C]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 13CH,OH处理植株中比2mmol/L更为明显。这些结果证实在叶绿体中过表达AODl增加烟草氧化甲醇为甲酸的能力,同时过量表达AODl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。

英文摘要:

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 methylotrophie 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-13 C ] glucose ( Glue ) and [ U-13 C ] fructose ( Fruc ) between AO and WT lines. However, the production of [ U-13 C ] glucose (Glue) and [ U-13 C ] 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/Ll3CH3OH 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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期刊信息
  • 《中国生物工程杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中国科学院文献情报中心 中国生物技术发展中心 中国生物工程学会
  • 主编:张树庸
  • 地址:北京市中关村北四环西路33号
  • 邮编:100080
  • 邮箱:biotech@mail.las.ac.cn
  • 电话:010-82624544 82626611-6631
  • 国际标准刊号:ISSN:1671-8135
  • 国内统一刊号:ISSN:11-4816/Q
  • 邮发代号:82-673
  • 获奖情况:
  • 1991年中国科学院科技进步三等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:12959