Contribution of NADPH-cytochrome P450 Reductase to Azole Resistance in Fusarium oxysporum.
Contribution of NADPH-cytochrome P450 Reductase to Azole Resistance in Fusarium oxysporum.
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DOI：
10.3389/fmicb.2021.709942
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发表时间：
2021
影响因子：
5.2
通讯作者：
Wang L
中科院分区：
文献类型：
--
作者：
He D;Feng Z;Gao S;Wei Y;Han S;Wang L
镰刀菌对大多数抗真菌剂和杀菌剂表现出显著的内在耐药性，导致免疫功能低下患者的高死亡率。因此，为了有效的治疗和预防真菌感染和降低抗真菌耐药性，需要对抗真菌耐药性机制进行彻底的表征。在这项研究中，利用对常见抗真菌药物具有广泛抗性的尖孢镰刀菌（野生型）分离物，通过农杆菌介导的转化产生1,450个T-DNA随机插入突变体。抗真菌药敏试验结果显示一个突变体对唑类药物的敏感性增加。与抗性野生型相比，突变体对KTZ、ITC、VRC、POS和PCZ的mic均较低（分别为0.125、1、0.06、0.5和0.125μg/ml）。该突变体的T-DNA插入位点涉及两个相邻基因，一个编码功能未知的假设蛋白，另一个编码nadph -细胞色素P450还原酶，称为CPR1。为了证实这些基因参与了唑类药物易感性的改变，我们产生了独立缺失突变体，Cpr1缺失突变体表现出与T-DNA随机突变体相同的表型。Cpr1的缺失显著降低麦角甾醇水平。此外，下游Cyp51基因的表达受到影响，这可能是观察到的对唑类药物敏感性增加的原因。这些结果证实了Cpr1与尖孢菌对唑的敏感性之间的相关性。此外，该基因可用于改善抗真菌治疗。
Fusarium species exhibit significant intrinsic resistance to most antifungal agents and fungicides, resulting in high mortality rates among immunocompromised patients. Consequently, a thorough characterization of the antifungal resistance mechanism is required for effective treatments and for preventing fungal infections and reducing antifungal resistance. In this study, an isolate of Fusarium oxysporum (wild-type) with broadly resistant to commonly antifungal agents was used to generate 1,450 T-DNA random insertion mutants via Agrobacterium tumefaciens-mediated transformation. Antifungal susceptibility test results revealed one mutant with increased sensitivity to azoles. Compared with the resistant wild-type, the mutant exhibited low MICs to KTZ, ITC, VRC, POS, and PCZ (0.125, 1, 0.06, 0.5, and 0.125μg/ml, respectively). The T-DNA insertion site of this mutant was characterized as involving two adjacent genes, one encoding a hypothetical protein with unknown function and the other encoding the NADPH-cytochrome P450 reductase, referred as CPR1. To confirm the involvement of these genes in the altered azole susceptibility, the independent deletion mutants were generated and the Cpr1 deletion mutant displayed the same phenotypes as the T-DNA random mutant. The deletion of Cpr1 significantly decreased ergosterol levels. Additionally, the expression of the downstream Cyp51 gene was affected, which likely contributed to the observed increased susceptibility to azoles. These findings verified the association between Cpr1 and azole susceptibility in F. oxysporum. Furthermore, this gene may be targeted to improve antifungal treatments.
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镰刀菌的医学相关性。
DOI：
10.3390/jof6030117
发表时间：
2020-07-24
期刊：
Journal of fungi (Basel, Switzerland)
影响因子：
--
作者：
Hof H
通讯作者：
Hof H
影响因子：
3.2
作者：
Zhou, Yujun;Zhu, Yuanye;Zhou, Mingguo
通讯作者：
Zhou, Mingguo
影响因子：
6.7
作者：
Al-Hatmi AM;Meis JF;de Hoog GS
通讯作者：
de Hoog GS
藤黑镰刀菌抗苯并咪唑类杀菌剂的分子机制
影响因子：
2.1
作者：
Chen, Zihao;Gao, Tao;Chen, Changjun
通讯作者：
Chen, Changjun
影响因子：
4.9
作者：
Rispail, Nicolas;Di Pietro, Antonio
通讯作者：
Di Pietro, Antonio