Involvement of reactive oxygen species and Ca2+ in the differential responses to low-boron in rapeseed genotypes
Involvement of reactive oxygen species and Ca2+ in the differential responses to low-boron in rapeseed genotypes
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活性氧和Ca2参与油菜基因型对低硼的差异反应
DOI：
10.1007/s11104-017-3337-3
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发表时间：
2017-07
期刊：
影响因子：
4.9
通讯作者：
Fangsen Xu
中科院分区：
文献类型：
--
作者：
Ting Zhou;Yingpeng Hua;Fangsen Xu
缺硼显著抑制植物的生长发育。油菜（Brassica napus L.）对维生素B缺乏非常敏感。活性氧（ROS）和Ca2+在植物对环境胁迫的响应中起着关键作用。我们的目的是鉴定B高效基因型“QY10”和B低效基因型“W10”对B缺乏的Ca2+通量和ROS爆发的差异，并建立与低B诱导的细胞死亡有关的Ca2+和ROS信号通路。采用组织化学、细胞化学和生物化学方法研究了植物和悬浮细胞系统中活性氧的产生；采用无创微检测技术（NMT）检测K+和Ca2+外排；测定了活性氧产生基因的表达和抗氧化酶的活性，并利用活性氧清除剂和Ca2+通道抑制剂分别表征了活性氧和Ca2+在低b环境下的作用。细胞死亡是缺B条件下油菜籽生长受到抑制的主要原因。低b诱导O2−积累，其分布与植物根系细胞死亡区相似。‘W10’的O2−产量增长明显强于‘QY10’。H2O2的变化趋势与O2−相似，但不太显著。脂质过氧化、离子泄漏和K+外排的增强表明，低b通过诱导氧化损伤导致细胞死亡，特别是在‘ W10 ’中。O2−清除剂预处理可提高细胞活力。低b诱导Ca2+内流，作用于ROS的上游。不同基因型油菜的氧化损伤差异不是由抗氧化酶决定的，而是由活性氧生成酶决定的。低b诱导Ca2+内流，然后刺激ROS爆发，最终导致细胞死亡。本研究丰富了我们对ROS和Ca2+参与油菜基因型对B缺乏的差异反应的理解。
Boron (B) deficiency significantly inhibits plant growth and development. Oilseed rape (Brassica napus L.) is highly susceptible to B deficiency. Reactive oxygen species (ROS) and Ca2+ play pivotal roles in plant responses to environmental stresses. We aim to identify the differential Ca2+ fluxes and ROS bursts of a B-efficient genotype ‘QY10’ and a B-inefficient genotype ‘W10’ to B deficiency, and establish a signalling pathway involving Ca2+ and ROS implicated in the low-B-induced cell death. Under both plant and suspension cell systems, the ROS production was investigated histochemically, cytochemically and biochemically; K+ and Ca2+ effluxes were assayed using the Non-invasive Micro-test Technology (NMT); the expression of ROS-producing genes and the activity assays of antioxidant enzymes were tested, and the ROS scavengers and Ca2+ channel inhibitors were used to characterize the roles of ROS and Ca2+ in response to low-B, respectively. The cell death was mainly responsible for rapeseed growth inhibition under B deficiency. Low-B induced O2− accumulation, whose distribution was similar to the cell death regions in the plant roots. The increase in O2− production was much stronger in ‘W10’ than in ‘QY10’. The change trend of H2O2 was similar to that of O2−, whereas less significant. The enhancement of lipid peroxidation, ion leakage and K+ efflux indicated that low-B caused cell death through the induction of oxidative damages, particularly in ‘W10’. Pretreatment with O2− scavenger increased the cell viabilities. Low-B induced Ca2+ influx, which worked upstream of ROS. It was not the antioxidant enzymes but the ROS-generating enzymes that determined the differential oxidative damages in rapeseed genotypes. Low-B induced Ca2+ influx, which then stimulated the ROS burst and eventually caused cell death. The present study enriches our understanding of the involvement of ROS and Ca2+ in the differential responses to B deficiency in rapeseed genotypes.
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