Overexpressing the Multiple-Stress Responsive Gene At1g74450 Reduces Plant Height and Male Fertility in Arabidopsis thaliana.
Overexpressing the Multiple-Stress Responsive Gene At1g74450 Reduces Plant Height and Male Fertility in Arabidopsis thaliana.
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DOI：
10.1371/journal.pone.0140368
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Harberd NP
中科院分区：
文献类型：
--
作者：
Visscher AM;Belfield EJ;Vlad D;Irani N;Moore I;Harberd NP
已知拟南芥中的一个基因子集响应于广泛的不同环境胁迫因子而上调。然而，并不是所有这些基因的特点，尚未就其功能。本研究采用转基因敲除、过表达和报告基因等方法，试图阐明拟南芥中5个未知的多重胁迫应答基因的生物学功能。所选基因具有以下基因座标识符：At 1g 18740、At 1g 74450、At 4g 27652、At 4g 29780和At 5g 12010。首先，鉴定每个基因座的T-DNA插入敲除系，并筛选改变的表型。未发现这些线与野生型Col-0在视觉上有任何不同。其次，对于每个开放阅读框产生35 S驱动的过表达系。对这些转基因株系的分析表明，过表达At 1g 74450 ORF的株系的表型发生了改变。过表达多重胁迫响应基因At 1g 74450的植物高度发育不良，雄性育性降低。来自发育阶段12-13的花的花药的亚历山大染色显示，与野生型Col-0和At 1g 74450敲除系相比，有活力的花粉不存在或减少。有趣的是，胁迫对作物生产力的影响在发育阶段（如雄配子体发育）最为严重。然而，环境胁迫诱导的雄配子体改变的分子因素和调控网络仍然是未知的。我们的研究结果表明，At 1g 74450基因提供了一个潜在的多重环境压力，株高和花粉发育之间的联系。此外，钌红染色分析表明At 1g 74450可能会影响种子内种皮粘液层的组成。最后，At 1g 74450的C-末端GFP融合蛋白显示定位于胞质溶胶。
A subset of genes in Arabidopsis thaliana is known to be up-regulated in response to a wide range of different environmental stress factors. However, not all of these genes are characterized as yet with respect to their functions. In this study, we used transgenic knockout, overexpression and reporter gene approaches to try to elucidate the biological roles of five unknown multiple-stress responsive genes in Arabidopsis. The selected genes have the following locus identifiers: At1g18740, At1g74450, At4g27652, At4g29780 and At5g12010. Firstly, T-DNA insertion knockout lines were identified for each locus and screened for altered phenotypes. None of the lines were found to be visually different from wildtype Col-0. Secondly, 35S-driven overexpression lines were generated for each open reading frame. Analysis of these transgenic lines showed altered phenotypes for lines overexpressing the At1g74450 ORF. Plants overexpressing the multiple-stress responsive gene At1g74450 are stunted in height and have reduced male fertility. Alexander staining of anthers from flowers at developmental stage 12–13 showed either an absence or a reduction in viable pollen compared to wildtype Col-0 and At1g74450 knockout lines. Interestingly, the effects of stress on crop productivity are most severe at developmental stages such as male gametophyte development. However, the molecular factors and regulatory networks underlying environmental stress-induced male gametophytic alterations are still largely unknown. Our results indicate that the At1g74450 gene provides a potential link between multiple environmental stresses, plant height and pollen development. In addition, ruthenium red staining analysis showed that At1g74450 may affect the composition of the inner seed coat mucilage layer. Finally, C-terminal GFP fusion proteins for At1g74450 were shown to localise to the cytosol.