Isolation and Characterization of a Topramezone-Resistant 4-Hydroxyphenylpyruvate Dioxygenase from Sphingobium sp. TPM-19.,Journal of Agricultural and Food Chemistry

当前位置： X-MOL 学术 › J. Agric. Food Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Isolation and Characterization of a Topramezone-Resistant 4-Hydroxyphenylpyruvate Dioxygenase from Sphingobium sp. TPM-19.
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-01-14 , DOI: 10.1021/acs.jafc.9b06871
Bin Liu ₁ , Qian Peng ₁ , Mengyao Sheng ₁ , Haiyan Ni ₂ , Xiang Xiao ₃ , Qing Tao ₃ , Qin He ₁ , Jian He ₁

Affiliation

Topramezone is a 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor. Due to its broad-spectrum, high efficiency, and low toxicity, topramezone is a candidate herbicide for the construction of genetically modified (GM) herbicide-resistant crops. In the present study, we screened a topramezone-resistant isolate Sphingobium sp. TPM-19 and cloned a topramezone-resistant HPPD gene (SphppD) from this isolate. SpHPPD shared the highest similarity (53%) with an HPPD from Vibrio vulnificus CMCP6. SpHPPD was synthesized in Escherichia coli BL21(DE3) and purified to homogeneity using Co2+-affinity chromatography. SpHPPD was found to be a monomer. The Km and kcat of SpHPPD for 4-hydroxyphenylpyruvate (4-HPP) were 82.8 μM and 15.0 s-1, respectively. SpHPPD showed high resistance to topramezone with half maximal inhibitory concentration (IC50) and Ki values of 5.2 and 2.5 μM, respectively. Additionally, SpHPPD also showed high resistance to isoxaflutole (DKN) (IC50: 8.7 μM; Ki: 6.0 μM) and mesotrione (IC50: 4.2 μM; Ki: 1.3 μM) and moderate resistance to tembotrione (IC50: 2.5 μM; Ki: 1.0 μM). The introduction of the SphppD gene into Arabidopsis thaliana enhanced obvious resistance against topramezone. In conclusion, this study provides a novel topramezone-resistant HPPD gene for the genetic engineering of GM herbicide-resistant crops.

中文翻译：

鞘氨醇单胞菌耐托吡酮的4-羟基苯丙酮酸双加氧酶的分离与鉴定。TPM-19。

Topramezone是一种4-羟苯基丙酮酸双加氧酶（HPPD）抑制剂。由于托普拉zone具有广谱，高效和低毒性的特点，因此它是构建抗转基因（GM）抗除草剂作物的候选除草剂。在本研究中，我们筛选了耐托普美的菌株鞘氨醇单胞菌。TPM-19并从该分离物中克隆了耐托普兰的HPPD基因（SphppD）。SpHPPD与来自创伤弧菌CMCP6的HPPD具有最高的相似性（53％）。SpHPPD在大肠杆菌BL21（DE3）中合成，并使用Co2 +亲和色谱纯化至均一。发现SpHPPD是单体。SpHPPD对于4-羟基苯丙酮酸（4-HPP）的Km和kcat分别为82.8μM和15.0 s-1。SpHPPD表现出对pramezone的高抗性，最大抑菌浓度（IC50）的一半和Ki值分别为5.2和2.5μM。此外，SpHPPD还显示出对异恶氟丁酮（DKN）（IC50：8.7μM; Ki：6.0μM）和甲基磺草酮（IC50：4.2μM; Ki：1.3μM）的高耐药性和对替康三酮的中等耐药性（IC50：2.5μM; Ki：1.0）（μM）。将SphppD基因导入拟南芥中明显增强了对托普美的抗性。总之，这项研究为抗转基因除草剂的农作物的基因工程提供了一种新的抗托普拉酮抗性的HPPD基因。将SphppD基因导入拟南芥中明显增强了对托普美的抗性。总之，这项研究为抗转基因除草剂的农作物的基因工程提供了一种新的抗托普拉酮抗性的HPPD基因。将SphppD基因导入拟南芥中明显增强了对托普美的抗性。总之，这项研究为抗转基因除草剂的农作物的基因工程提供了一种新的抗托普拉酮抗性的HPPD基因。

更新日期：2020-01-14

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南