Xinjiang Production and Construction Corps Key Laboratory of Protection and Utilization of Biological Resources in Tarim Basin/College of Life Science, Tarim University, Alar, 843300, Xinjiang, PR China; Engineering Laboratory of Chemical Resources Utilization in South Xinjiang of Xinjiang Production and Construction Corps/College of Life Science, Tarim University, Alar, 843300, Xinjiang, PR China.
College of Animal Science, Tarim University, Alar, 843300, Xinjiang, PR China.
Int J Biochem Cell Biol. 2018 Oct;103:35-44. doi: 10.1016/j.biocel.2018.08.001. Epub 2018 Aug 3.
Verticillium wilt, caused by Verticillium dahliae, is a cancer of cotton which affects cotton quality and yield in China. In our previous study, a novel anti-Verticillium dahliae protein TUBP1 was obtained from Bacillus axarquiensis. Then, we have systematically studied the anti-V. dahliae activity and the pore formation action of TUBP1 protein on V. dahliae membrane. In present study, we provide detailed whether TUBP1 protein induced mitochondrial damaged and mitochondria-mediated apoptotic cell death in V. dahliae. In V. dahliae cells exposed to the TUBP1 protein, the mitochondrial dehydrogenases, FF-ATPase, malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) activities were reduced and reactive oxygen species (ROS), which is a major cause of apoptosis, were increased. The results demonstrated that mitochondria dysfunction and ROS-induced oxidative stress caused the release of apoptotic factors. The following cellular changes, which are characteristic of apoptosis, were measured including mitochondrial membrane potential (MMP), Cytochrome c (Cyt C) release, metacaspase activation, phosphatidylserine (PS) exposure, and DNA condensation and fragmentation. The results showed that an important feature of apoptosis, MMP, was caused by ROS. Significantly, cyt c was released, which is a factor in metacaspase activity after treatment with the TUBP1 protein. Number of stained cells with activated intracellular metacaspases exposed to TUBP1 protein was increased in a concentration-dependent manner. We also showed that in the early and late stages of apoptosis, the effects of the TUBP1 protein were mediated by PS and DNA fragmentation and condensation in the plasma membrane, respectively. There turned out that the TUBP1 protein led to mitochondria-mediated apoptotic cell death in V.dahliae. The results of this investigation indicated that TUBP1 stain or protein is a potent candidate against V.dahliae infections in crop species.
黄萎病是由大丽轮枝菌引起的一种棉花癌症，会影响中国棉花的质量和产量。在我们之前的研究中，从阿克萨拉氏芽孢杆菌中获得了一种新型抗大丽轮枝菌蛋白 TUBP1。然后，我们系统地研究了 TUBP1 蛋白对大丽轮枝菌膜的抗大丽轮枝菌活性和孔形成作用。在本研究中，我们详细研究了 TUBP1 蛋白是否诱导大丽轮枝菌中的线粒体损伤和线粒体介导的细胞凋亡。在暴露于 TUBP1 蛋白的大丽轮枝菌细胞中，线粒体脱氢酶、FF-ATP 酶、苹果酸脱氢酶（MDH）和琥珀酸脱氢酶（SDH）活性降低，活性氧（ROS）增加，ROS 是细胞凋亡的主要原因。结果表明，线粒体功能障碍和 ROS 诱导的氧化应激导致凋亡因子的释放。随后测量了细胞凋亡的几个特征性变化，包括线粒体膜电位（MMP）、细胞色素 c（Cyt C）释放、半胱天冬酶激活、磷脂酰丝氨酸（PS）暴露以及 DNA 凝聚和片段化。结果表明，凋亡的一个重要特征是 ROS 引起的 MMP。重要的是，在用 TUBP1 蛋白处理后，细胞色素 c 被释放，这是半胱天冬酶活性的一个因素。暴露于 TUBP1 蛋白的细胞中，具有激活的细胞内半胱天冬酶的细胞数量呈浓度依赖性增加。我们还表明，在细胞凋亡的早期和晚期，TUBP1 蛋白的作用分别通过质膜中的 PS 和 DNA 片段化和凝聚来介导。事实证明，TUBP1 蛋白导致大丽轮枝菌中的线粒体介导的细胞凋亡。这项研究的结果表明，TUBP1 染色剂或蛋白是作物中大丽轮枝菌感染的有效候选物。
