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一种氧化还原介导的对差异表达外部线粒体NADPH脱氢酶的转基因森林烟草中茎抽薹的调控。

A redox-mediated modulation of stem bolting in transgenic Nicotiana sylvestris differentially expressing the external mitochondrial NADPH dehydrogenase.

作者信息

Liu Yun-Jun, Nunes-Nesi Adriano, Wallström Sabá V, Lager Ida, Michalecka Agnieszka M, Norberg Fredrik E B, Widell Susanne, Fredlund Kenneth M, Fernie Alisdair R, Rasmusson Allan G

机构信息

Department of Cell and Organism Biology, Lund University, SE-22362 Lund, Sweden.

出版信息

Plant Physiol. 2009 Jul;150(3):1248-59. doi: 10.1104/pp.109.136242. Epub 2009 May 8.

Abstract

Cytosolic NADPH can be directly oxidized by a calcium-dependent NADPH dehydrogenase, NDB1, present in the plant mitochondrial electron transport chain. However, little is known regarding the impact of modified cytosolic NADPH reduction levels on growth and metabolism. Nicotiana sylvestris plants overexpressing potato (Solanum tuberosum) NDB1 displayed early bolting, whereas sense suppression of the same gene led to delayed bolting, with consequential changes in flowering time. The phenotype was dependent on light irradiance but not linked to any change in biomass accumulation. Whereas the leaf NADPH/NADP(+) ratio was unaffected, the stem NADPH/NADP(+) ratio was altered following the genetic modification and strongly correlated with the bolting phenotype. Metabolic profiling of the stem showed that the NADP(H) change affected relatively few, albeit central, metabolites, including 2-oxoglutarate, glutamate, ascorbate, sugars, and hexose-phosphates. Consistent with the phenotype, the modified NDB1 level also affected the expression of putative floral meristem identity genes of the SQUAMOSA and LEAFY types. Further evidence for involvement of the NADPH redox in stem development was seen in the distinct decrease in the stem apex NADPH/NADP(+) ratio during bolting. Additionally, the potato NDB1 protein was specifically detected in mitochondria, and a survey of its abundance in major organs revealed that the highest levels are found in green stems. These results thus strongly suggest that NDB1 in the mitochondrial electron transport chain can, by modifying cell redox levels, specifically affect developmental processes.

摘要

胞质中的烟酰胺腺嘌呤二核苷酸磷酸(NADPH)可被植物线粒体电子传递链中存在的一种钙依赖性NADPH脱氢酶NDB1直接氧化。然而,关于胞质NADPH还原水平的改变对生长和代谢的影响,我们所知甚少。过表达马铃薯(Solanum tuberosum)NDB1的林烟草(Nicotiana sylvestris)植株表现出早花抽薹,而对同一基因进行反义抑制则导致抽薹延迟,进而使开花时间发生相应变化。该表型依赖于光照强度,但与生物量积累的任何变化无关。虽然叶片中的NADPH/NADP(+) 比值未受影响,但遗传修饰后茎中的NADPH/NADP(+) 比值发生了改变,且与抽薹表型密切相关。对茎进行代谢谱分析表明,NADP(H) 的变化影响了相对较少但至关重要的代谢物,包括2-酮戊二酸、谷氨酸、抗坏血酸、糖类和己糖磷酸酯。与表型一致,NDB1水平的改变也影响了SQUAMOSA和LEAFY类型的假定花分生组织特征基因的表达。在抽薹过程中茎尖NADPH/NADP(+) 比值明显下降,这进一步证明了NADPH氧化还原参与茎的发育。此外,在马铃薯线粒体中特异性检测到了NDB1蛋白,对其在主要器官中的丰度进行的调查显示,绿色茎中的含量最高。因此,这些结果有力地表明,线粒体电子传递链中的NDB1可通过改变细胞氧化还原水平,特异性地影响发育过程。

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