Cellular apoptosis of hemocytes from Dendrolimus tabulaeformis Tsai et Liu larvae induced with the secondary metabolites of Beauveria brongniartii (Sacc.) Petch.
Cellular apoptosis of hemocytes from Dendrolimus tabulaeformis Tsai et Liu larvae induced with the secondary metabolites of Beauveria brongniartii (Sacc.) Petch.
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DOI：
10.1371/journal.pone.0071600
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Yang Q
中科院分区：
文献类型：
--
作者：
Fan J;Xie Y;Xue J;Zhang Y;Yang Q
为研究昆虫病原真菌次级代谢产物对宿主昆虫血细胞免疫功能的影响， 布氏白僵菌以三种浓度（5.5、55和550 µg/mL）使用，松毛虫的4龄幼虫 以油松毛虫为寄主。在生物测定中，通过注射用SMC溶液接种幼虫。用荧光显微镜、透射电镜和流式细胞仪观察幼虫血细胞的凋亡。FM结果显示，在处理组中，幼虫血细胞在处理后6 h通过辐射不均匀的Kelly荧光表现出早期凋亡症状，并且在处理后12 h通过辐射不均匀的橙子荧光表现出晚期凋亡症状。透射电镜下观察到与细胞凋亡有关的超微结构变化：细胞核肥大，核膜轻微褶皱，染色质浓缩，线粒体嵴消失或紊乱，大部分细胞出现泡状突起，胞质内出现纤维状聚集物。在处理后6 h通过FCM检测幼虫血细胞的凋亡; SMC 5.5、55和550 µg/mL处理组的早期凋亡细胞百分比分别为11.93%、13.10%和18.42%。晚期凋亡在给药后12 h首次出现; SMC 55 µg/mL给药组在给药后24 h凋亡率最高，为36.54 ± 4.37%。细胞凋亡率与SMC浓度及作用时间呈正相关。这些结果表明， B。布氏杆菌能够攻击 D .研究结果表明，真菌次级代谢产物可作用于宿主的免疫系统，并诱导细胞凋亡。
To investigate the effect of the secondary metabolites of entomopathogenic fungus on the hemocyte immunity of host insect, the secondary metabolite complex (SMC) of Beauveria brongniartii was used in three concentrations (5.5, 55, and 550 µg/mL), and the 4th instar larvae of the pine caterpillar Dendrolimus tabulaeformis were employed as host insects. The larvae were inoculated with the SMC solutions by injection in bioassays. Apoptosis of the larval hemocytes was observed using fluorescence microscopy (FM), transmission electron microscopy (TEM), and flow cytometry (FCM). The FM results showed that in the treated groups, larval hemocytes exhibited symptoms of early apoptosis at 6 h post-treatment by radiating a non-uniform kelly fluorescence and exhibited symptoms of late apoptosis at 12 h post-treatment by radiating a non-uniform orange fluorescence. Under TEM, the following ultra-structural changes associated with apoptosis of the larval hemocytes were observed in the treated groups: the nuclei were hypertrophied, slight folds were on the nuclear envelope, the chromatin became concentrated, the mitochondrial cristae disappeared or were disorderly, most cells developed blebs, and fibrillar aggregation appeared and accumulated in the cytoplasm. Apoptosis of the larval hemocytes was detected by FCM at 6 h post-treatment; the percentage of early apoptotic cells in the SMC 5.5, 55, and 550 µg/mL treatment groups were 11.93%, 13.10%, and 18.42%, respectively. Late apoptosis first occurred at 12 h post-treatment; the highest rate of apoptosis was 36.54 ± 4.37% at 24 h post-treatment in the SMC 55 µg/mL treatment group. In general, the cellular apoptosis rate was positively correlated with the SMC concentration and the time post-treatment. These results indicate that secondary metabolites of B . brongniartii are able to attack the hemocytes of D . tabulaeformis larvae and induce cellular apoptosis, thereby providing new evidence that secondary metabolites of mycopathogens can act on host immune systems.
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