Activity levels and expression of antioxidant enzymes in the ascorbate-glutathione cycle in artificially aged rice seed
Activity levels and expression of antioxidant enzymes in the ascorbate-glutathione cycle in artificially aged rice seed
复制标题
人工老化水稻种子抗坏血酸-谷胱甘肽循环中抗氧化酶的活性水平和表达
DOI：
10.1016/j.plaphy.2014.03.006
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发表时间：
2014
影响因子：
6.5
通讯作者：
Xinxiong Lu
中科院分区：
文献类型：
--
作者：
Guangkun Yin;Xia Xin;Chao Song;Xiaoling Chen;Jinmei Zhang;Shuhua Wu;Ruifang Li;Xu Liu;Xinxiong Lu
活性氧是种子劣变的主要原因。为了研究老化种子中活性氧的清除系统，我们采用Western印迹、实时定量逆转录聚合酶链反应、高效液相色谱和抗氧化酶活性分析等方法对人工老化的水稻种子进行了分析。11）。老化种子在50 °C下储存1、9或17个月，增加了超氧自由基和过氧化氢的水平，并将发芽率分别从99%降低到92%、55%和2%。超氧化物歧化酶（SOD），谷胱甘肽还原酶（GR）和脱氢抗坏血酸还原酶（达尔）的活性水平在老化种子没有变化。与此相反，过氧化氢酶（CAT），抗坏血酸过氧化物酶（APX），和单脱氢抗坏血酸还原酶（MDHAR）的活性水平显着下降，在老化的种子，过氧化氢酶和胞质抗坏血酸过氧化物酶蛋白的表达。转录本积累分析表明，水稻胚中每种抗氧化酶类型的特定表达模式是复杂的。总体而言，大多数基因的表达下调，沿着其蛋白质表达。此外，抗坏血酸和谷胱甘肽的量的减少与清除酶活性的降低在老化的水稻胚。我们的数据表明，抗氧化系统的抑制，特别是CAT 1，APX 1和MDHAR 1的表达减少，可能是负责活性氧在人工老化的种子胚中的积累，导致种子活力的丧失。
Reactive oxygen species are the main contributors to seed deterioration. In order to study scavenging systems for reactive oxygen species in aged seed, we performed analyses using western blotting, real-time quantitative reverse-transcription polymerase chain reaction, high-performance liquid chromatography, and antioxidant enzyme activity analyses in artificially aged rice seeds (Oryza sativaL. cv.wanhuano.11). Aging seeds by storing them at 50 °C for 1, 9, or 17 months increased the superoxide radical and hydrogen peroxide levels and reduced the germination percentage from 99% to 92%, 55%, and 2%, respectively. The activity levels of superoxide dismutase (SOD), glutathione reductase (GR), and dehydroascorbate reductase (DHAR) did not change in aged seeds. In contrast, the activity levels of catalase (CAT), ascorbate peroxidase (APX), and monodehydroascorbate reductase (MDHAR) were significantly decreased in aged seeds, as were the expression of catalase and cytosolic ascorbate peroxidase protein. Transcript accumulation analysis showed that specific expression patterns were complex for each of the antioxidant enzyme types in the rice embryos. Overall, the expression of most genes was down-regulated, along with their protein expression. In addition, the reduction in the amount of ascorbate and glutathione was associated with the reduction in scavenging enzymes activity in aged rice embryos. Our data suggest that the depression of the antioxidant system, especially the reduction in the expression of CAT1, APX1 and MDHAR1, may be responsible for the accumulation of reactive oxygen species in artificially aged seed embryos, leading to a loss of seed vigor.