Amini Saeed, Maali-Amiri Reza, Mohammadi Rahmat, Kazemi-Shahandashti Seyyedeh-Sanam
Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Karaj 31587-77871, Iran.
Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Karaj 31587-77871, Iran.
Plant Physiol Biochem. 2017 Feb;111:39-49. doi: 10.1016/j.plaphy.2016.11.011. Epub 2016 Nov 22.
We evaluated the effect of TiO nanoparticles (NPs) on cold tolerance (CT) development in two chickpea (Cicer arietinum L.) genotypes (Sel96Th11439, cold tolerant, and ILC533, cold susceptible) by using cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique during the first and sixth days of cold stress (CS) at 4 °C. Selective amplification by primer combinations generated 4200 transcript-derived fragments (TDFs) while 100 of them (2.62%) were differentially expressed. During CS, 60 differentially expressed TDFs of TiO NPs-treated plants were cloned and 10 of them produced successfully readable sequences. These data represented different groups of genes involved in metabolism pathways, cellular defense, cell connections and signaling, transcriptional regulation and chromatin architecture. Two out of 10 TDFs were unknown genes with uncharacterized functions or sequences without homology to known ones. The network-based analysis showed a gene-gene relationship in response to CS. Quantitative reverse-transcriptase polymerase chain reaction (qPCR) confirmed differential expression of identified genes (six out of 10 TDFs) with potential functions in CT and showed similar patterns with cDNA-AFLP results. An increase in transcription level of these TDFs, particularly on the first day of CS, was crucial for developing CT through decreasing electrolyte leakage index (ELI) content in tolerant plants compared to susceptible ones, as well as in TiO NPs-treated plants compared to control ones. It could also indicate probable role of TiO NPs against CS-induced oxidative stress. Therefore, a new application of TiO NPs in CT development is suggested for preventing or controlling the damages in field conditions and increasing crop productivity.
我们通过使用cDNA扩增片段长度多态性（cDNA-AFLP）技术，在4°C冷胁迫（CS）的第一天和第六天，评估了TiO纳米颗粒（NPs）对两种鹰嘴豆（Cicer arietinum L.）基因型（耐寒的Sel96Th11439和冷敏感的ILC533）耐寒性（CT）发育的影响。引物组合的选择性扩增产生了4200个转录本衍生片段（TDFs），其中100个（2.62%）差异表达。在冷胁迫期间，克隆了60个TiO NPs处理植物的差异表达TDFs，其中10个成功产生了可读序列。这些数据代表了参与代谢途径、细胞防御、细胞连接和信号传导、转录调控和染色质结构的不同基因组。10个TDFs中有2个是功能未知的基因，其序列与已知基因无同源性。基于网络的分析显示了对冷胁迫的基因-基因关系。定量逆转录聚合酶链反应（qPCR）证实了已鉴定基因（10个TDFs中的6个）在耐寒性方面具有潜在功能的差异表达，并且与cDNA-AFLP结果显示出相似的模式。与敏感植物相比，以及与对照植物相比，TiO NPs处理植物中这些TDFs转录水平的增加，特别是在冷胁迫的第一天，对于通过降低电解质渗漏指数（ELI）含量来发展耐寒性至关重要。这也可能表明TiO NPs对冷胁迫诱导的氧化应激的可能作用。因此，建议在耐寒性发育中对TiO NPs进行新的应用，以预防或控制田间条件下的损害并提高作物生产力。
