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表达反义铁氧还蛋白-NADP(H)还原酶转录本的转基因烟草植株对光氧化损伤的敏感性增加。

Transgenic tobacco plants expressing antisense ferredoxin-NADP(H) reductase transcripts display increased susceptibility to photo-oxidative damage.

作者信息

Palatnik Javier F, Tognetti Vanesa B, Poli Hugo O, Rodríguez Ramiro E, Blanco Nicolás, Gattuso Martha, Hajirezaei Mohammad-Reza, Sonnewald Uwe, Valle Estela M, Carrillo Néstor

机构信息

Instituto de Biología Molecular y Celular de Rosario (IBR), Universidad Nacional de Rosario, Suipacha 531, S2002-LRK Rosario, Argentina.

出版信息

Plant J. 2003 Aug;35(3):332-41. doi: 10.1046/j.1365-313x.2003.01809.x.

Abstract

Ferredoxin-NADP(H) reductase (FNR) catalyses the final step of the photosynthetic electron transport in chloroplasts. Using an antisense RNA strategy to reduce expression of this flavoenzyme in transgenic tobacco plants, it has been demonstrated that FNR mediates a rate-limiting step of photosynthesis under both limiting and saturating light conditions. Here, we show that these FNR-deficient plants are abnormally prone to photo-oxidative injury. When grown under autotrophic conditions for 3 weeks, specimens with 20-40% extant reductase undergo leaf bleaching, lipid peroxidation and membrane damage. The magnitude of the effect was proportional to the light intensity and to the extent of FNR depletion, and was accompanied by morphological changes involving accumulation of aberrant plastids with defective thylakoid stacking. Damage was initially confined to chloroplast membranes, whereas Rubisco and other stromal proteins began to decline only after several weeks of autotrophic growth, paralleled by partial recovery of NADPH levels. Exposure of the transgenic plants to moderately high irradiation resulted in rapid loss of photosynthetic capacity and accumulation of singlet oxygen in leaves. The collected results suggest that the extensive photo-oxidative damage sustained by plants impaired in FNR expression was caused by singlet oxygen building up to toxic levels in these tissues, as a direct consequence of the over-reduction of the electron transport chain in FNR-deficient chloroplasts.

摘要

铁氧还蛋白-NADP(H)还原酶(FNR)催化叶绿体中光合电子传递的最后一步。利用反义RNA策略降低转基因烟草植株中这种黄素酶的表达,结果表明,在光照受限和饱和条件下,FNR均介导光合作用的限速步骤。在此,我们表明这些FNR缺陷型植株异常容易受到光氧化损伤。在自养条件下生长3周后,还原酶活性尚存20%-40%的植株样本会出现叶片漂白、脂质过氧化和膜损伤。这种效应的程度与光照强度和FNR缺失程度成正比,并伴有形态变化,包括异常质体的积累以及类囊体堆叠缺陷。损伤最初局限于叶绿体膜,而核酮糖-1,5-二磷酸羧化酶(Rubisco)和其他基质蛋白仅在自养生长数周后才开始减少,同时NADPH水平部分恢复。将转基因植株暴露于适度的高辐射下,会导致光合能力迅速丧失,叶片中单线态氧积累。收集到的结果表明,FNR表达受损的植株遭受的广泛光氧化损伤是由于这些组织中单线态氧积累到有毒水平所致,这是FNR缺陷型叶绿体中电子传递链过度还原的直接后果。

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