Foxtail millet (Setaria italica L.) is a drought-tolerant millet crop of arid and semi-arid regions. Aldo-keto reductases (AKRs) are significant part of plant defence mechanism, having an ability to confer multiple st...Foxtail millet (Setaria italica L.) is a drought-tolerant millet crop of arid and semi-arid regions. Aldo-keto reductases (AKRs) are significant part of plant defence mechanism, having an ability to confer multiple stress tolerance. In this study, AKR1 gene expression was studied in roots and leaves of foxtail millet subjected to different regimes of PEG- and NaCl-stress for seven days. The quantitative Real-time PCR expression analysis in both root and leaves showed upregulation of AKR1 gene during PEG and salt stress. A close correlation exits between expression of AKR1 gene and the rate of lipid peroxidation along with the retardation of growth. Tissue-specific differences were found in the AKR1 gene expression to the stress intensities studied. The reduction in root and shoot growth under both stress conditions were dependent on stress severity. The level of lipid peroxidation as indicated by MDA formation was significantly increased in roots and leaves along with increased stress levels. Finally, these findings support the early responsive nature of AKR1 gene and seem to be associated at least in part with its ability to contribute in antioxidant defence related pathways which could provide a better protection against oxidative stress under stress conditions.展开更多
目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌E...目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌EscherichiacoliBL21(DE3)中进行重组和异丙基-β-D-硫代半乳糖苷(isopropylβ-D-thiogalactoside,IPTG)诱导表达,经镍亲和层析纯化并鉴定AKR18A2的酶学性质,构建德沃斯氏菌(Devosiasp.)来源的DON降解酶QDDH和AKR18A2双酶作用体系,实现DON的高效降解。结果来自鞘氨醇单胞菌属(Sphingomonas sp.)的呕吐毒素降解酶(AKR18A2)由343个氨基酸组成。该酶属于醛酮还原酶超家族,45℃和pH7.0为最适反应条件。AKR18A2可在24h内降解15.42%DON,而双酶(QDDH和AKR18A2)协同作用4h后对DON的降解率可提高至98.02%。结论本研究鉴定了一种新型DON降解酶AKR18A2,并首次创新建立双酶联用体系进一步实现DON的高效降解。展开更多
文摘Foxtail millet (Setaria italica L.) is a drought-tolerant millet crop of arid and semi-arid regions. Aldo-keto reductases (AKRs) are significant part of plant defence mechanism, having an ability to confer multiple stress tolerance. In this study, AKR1 gene expression was studied in roots and leaves of foxtail millet subjected to different regimes of PEG- and NaCl-stress for seven days. The quantitative Real-time PCR expression analysis in both root and leaves showed upregulation of AKR1 gene during PEG and salt stress. A close correlation exits between expression of AKR1 gene and the rate of lipid peroxidation along with the retardation of growth. Tissue-specific differences were found in the AKR1 gene expression to the stress intensities studied. The reduction in root and shoot growth under both stress conditions were dependent on stress severity. The level of lipid peroxidation as indicated by MDA formation was significantly increased in roots and leaves along with increased stress levels. Finally, these findings support the early responsive nature of AKR1 gene and seem to be associated at least in part with its ability to contribute in antioxidant defence related pathways which could provide a better protection against oxidative stress under stress conditions.
文摘目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌EscherichiacoliBL21(DE3)中进行重组和异丙基-β-D-硫代半乳糖苷(isopropylβ-D-thiogalactoside,IPTG)诱导表达,经镍亲和层析纯化并鉴定AKR18A2的酶学性质,构建德沃斯氏菌(Devosiasp.)来源的DON降解酶QDDH和AKR18A2双酶作用体系,实现DON的高效降解。结果来自鞘氨醇单胞菌属(Sphingomonas sp.)的呕吐毒素降解酶(AKR18A2)由343个氨基酸组成。该酶属于醛酮还原酶超家族,45℃和pH7.0为最适反应条件。AKR18A2可在24h内降解15.42%DON,而双酶(QDDH和AKR18A2)协同作用4h后对DON的降解率可提高至98.02%。结论本研究鉴定了一种新型DON降解酶AKR18A2,并首次创新建立双酶联用体系进一步实现DON的高效降解。
