Wi Soo Jin, Kim Soo Jin, Kim Woo Taek, Park Ky Young
Department of Biology, Sunchon National University, Sunchon, Chonnam, 540-742, Korea,
Planta. 2014 May;239(5):979-88. doi: 10.1007/s00425-014-2027-0. Epub 2014 Jan 30.
Using subtractive hybridization analysis, the S-adenosylmethionine decarboxylase (SAMDC) gene from Capsicum annuum was isolated and renamed CaSAMDC. We generated independent transgenic Arabidopsis (Arabidopsis thaliana) lines constitutively expressing a 35S::CaSAMDC construct. Drought tolerance was significantly enhanced in Arabidopsis T4 transgenic homozygous lines as compared to wild-type (WT) plants. The levels of main polyamines (PAs) were more significantly increased in CaSAMDC-overexpressing transgenic plants after 6 h of drought stress as compared to stressed WT plants. Basal transcription of polyamine oxidase (PAO) showed at a much higher level in unstressed-transgenic plants as compared to unstressed WT plants. However, the difference in PAO transcription level between WT and transgenic plants was reduced after drought stress. Cellular accumulation of reactive oxygen species (ROS) was significantly reduced following drought stress in transgenic Arabidopsis plants as compared to WT plants. These results were in agreement with additional observations that stress-induced ROS generation, as determined by qRT-PCR analysis of NADPH oxidase (RbohD and RbohF), was significantly suppressed while transcription of ROS-detoxifying enzymes was notably elevated in transgenic lines in response to drought stress. Further, ROS-induced transcription of the metacaspase II gene was remarkably inhibited in transgenic plants. Collectively, these results suggest that drought stress tolerance due to reduction of ROS production and enhancement of ROS detoxification can be attributed to elevation of PAs.
利用消减杂交分析,从辣椒中分离出S-腺苷甲硫氨酸脱羧酶(SAMDC)基因,并将其重新命名为CaSAMDC。我们构建了组成型表达35S::CaSAMDC构建体的独立转基因拟南芥株系。与野生型(WT)植株相比,拟南芥T4转基因纯合株系的耐旱性显著增强。在干旱胁迫6小时后,与受胁迫的野生型植株相比,过表达CaSAMDC的转基因植株中主要多胺(PA)的水平显著升高。与未受胁迫的野生型植株相比,未受胁迫的转基因植株中多胺氧化酶(PAO)的基础转录水平要高得多。然而,干旱胁迫后,野生型植株和转基因植株之间PAO转录水平的差异减小。与野生型植株相比,转基因拟南芥植株在干旱胁迫后细胞内活性氧(ROS)的积累显著减少。这些结果与其他观察结果一致,即通过对NADPH氧化酶(RbohD和RbohF)进行qRT-PCR分析确定,胁迫诱导的ROS生成在转基因株系中受到显著抑制,而ROS解毒酶的转录在干旱胁迫下显著升高。此外,ROS诱导的metacaspase II基因转录在转基因植株中受到显著抑制。总的来说,这些结果表明,由于ROS产生减少和ROS解毒增强而导致的耐旱性可归因于PA水平的升高。