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In vitro effect of recombinant amaranth cystatin (AhCPI) on spore germination mycelial growth stress response and cellular integrity of Aspergillus niger and Aspergillus parasiticus

机译:重组a菜半胱氨酸蛋白酶抑制剂(AhCPI)对黑曲霉和寄生曲霉的孢子萌发菌丝生长应激反应和细胞完整性的体外影响

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摘要

The inhibitory effect of recombinant amaranth cystatin (AhCPI) on the spore germination and growth of the mycotoxigenic fungus Aspergillus parasiticus and Aspergillus niger was investigated. AhCPI showed a concentration-dependent antifungal activity against both fungi. Differential effects were observed when fungi were treated with cystatin in two developmental stages. When AhCPI was added to young mycelium cultures of A. niger, it had a dramatic effect on mycelial growth compared with old mycelium cultures. On the contrary, there was no differential effect of AhCPI addition to either old or young mycelium of A. parasiticus. Furthermore, electron microscopic observations showed that cystatin caused important effects at the level of cell morphology and organelle integrity of both fungi. Additionally, A. parasiticus spores treated with AhCPI presented sensitivity to oxidative, osmotic and ionic stresses; in opposition, under same conditions, A. niger did not show sensitivity to any stressful agent. These results suggest that AhCPI antifungal activity might be related with damage to cell integrity, affecting the survival of the fungi. In addition, our evidences showed that fungal species respond dissimilarly to cystatin; however, such disparities can be used to the control of unwanted fungi.
机译:研究了重组a菜半胱氨酸蛋白酶抑制剂(AhCPI)对真菌毒素寄生曲霉(Aspergillus parasiticus)和黑曲霉(Aspergillus niger)的孢子萌发和生长的抑制作用。 AhCPI对两种真菌均表现出浓度依赖性的抗真菌活性。当在两个发育阶段用半胱氨酸蛋白酶抑制剂处理真菌时,观察到了不同的作用。当将AhCPI添加到黑曲霉的年轻菌丝培养物中时,与旧菌丝培养物相比,它对菌丝生长具有显着影响。相反,AhCPI添加到寄生曲霉的旧菌丝或幼丝菌丝中都没有差异作用。此外,电子显微镜观察表明,半胱氨酸蛋白酶抑制剂对两种真菌的细胞形态和细胞器完整性均具有重要作用。另外,用AhCPI处理的寄生曲霉孢子对氧化,渗透和离子胁迫表现出敏感性。相反,在相同条件下,黑曲霉对任何压力因素均不敏感。这些结果表明,AhCPI抗真菌活性可能与细胞完整性受损有关,从而影响了真菌的存活。此外,我们的证据表明真菌种类对半胱氨酸蛋白酶抑制剂的反应不同。但是,这种差异可用于控制有害真菌。

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