Department of Life Sciences, Tzu Chi University, Hualien 97004, Taiwan.
Institute of Medical Sciences, Tzu Chi University, Hualien 97004, Taiwan.
Int J Mol Sci. 2018 Jun 23;19(7):1854. doi: 10.3390/ijms19071854.
The chloroplast relies on proteins encoded in the nucleus, synthesized in the cytosol and subsequently transported into chloroplast through the protein complexes Toc and Tic (Translocon at the outer/inner membrane of chloroplasts). A Tic complex member, Tic55, contains a redox-related motif essential for protein import into chloroplasts in peas. However, Tic55 is not crucial for protein import in . Here, a -knockout mutant of was characterized for Tic55 localization, its relationship with other translocon proteins, and its association with plant leaf senescence when compared to the wild type. Individually darkened leaves (IDLs) obtained through dark-induced leaf senescence were used to demonstrate chlorophyll breakdown and its relationship with plant senescence in the -knockout mutant. The IDLs of the -knockout mutant contained higher chlorophyll concentrations than those of the wild type. Our microarray analysis of IDLs during leaf senescence identified seven senescence-associated genes (SAGs) that were downregulated in the -knockout mutant: , , , , , , and . Real-time quantitative PCR confirmed the reliability of microarray analysis by showing the same expression patterns with those of the microarray data. Thus, Tic55 functions in dark-induced aging in by indirectly regulating downstream SAGs expression. In addition, the expression of four NAC genes, including , , , and of IDL treated -knockout mutant appeared to be downregulated. Yeast one hybrid assay revealed that only promoter region can be bound by MYB108, suggesting that a MYB-NAC regulatory network is involved in dark-stressed senescence.
叶绿体依赖于核编码的蛋白质,这些蛋白质在细胞质中合成,然后通过 Toc 和 Tic(叶绿体内外膜的转运体)蛋白复合物运输到叶绿体中。Tic 复合物的一个成员 Tic55 包含一个与氧化还原相关的基序,对于豌豆中蛋白质向叶绿体的输入是必需的。然而,Tic55 对于蛋白质在 中的输入并不是至关重要的。在这里,我们对 进行了 -敲除突变体的特征分析,包括 Tic55 的定位、与其他易位子蛋白的关系,以及与野生型相比与植物叶片衰老的关系。通过黑暗诱导叶片衰老获得的个体暗化叶片 (IDL) 用于证明叶绿素的分解及其与 -敲除突变体植物衰老的关系。-敲除突变体的 IDL 中含有比野生型更高浓度的叶绿素。我们对叶片衰老过程中 IDL 的微阵列分析鉴定出七个与衰老相关的基因 (SAGs) 在 -敲除突变体中下调:,,,,,, 和 。实时定量 PCR 通过显示与微阵列数据相同的表达模式,证实了微阵列分析的可靠性。因此,Tic55 通过间接调节下游 SAGs 的表达在 中发挥作用黑暗诱导的衰老。此外,四个 NAC 基因(包括 IDL 处理的 -敲除突变体中的 、 、 和 )的表达似乎下调。酵母单杂交分析表明只有 启动子区域可以被 MYB108 结合,这表明一个 MYB-NAC 调控网络参与了黑暗胁迫诱导的衰老。