The dissolution of polyhedra of Mythimna separata nuclear polyhedrosis. virus by digestive fluid (pH11. 03) collected from the 5th instar M. separata larvae was studied in vitro. Observations were made at timed interv...The dissolution of polyhedra of Mythimna separata nuclear polyhedrosis. virus by digestive fluid (pH11. 03) collected from the 5th instar M. separata larvae was studied in vitro. Observations were made at timed intervals using phase contrast microscopy and scanning electron microscopy. Under phase contrast microscopy, the polyhcdra lost their refrigence by 5 minute exposure to the digestive fluid. After exposure to the fluid for 30 minutes, all of the PIBs were dissolved. Chages of the PIBs were also observed under scanning electron microscopy, after 5 minute exposure to the fluid, damaged PIBs and PIB-derived debris were seen. After 30 minute exposure, only remains of PIBs were found. The effect of M. separata digestive fluid on the infectivity of Ms NPV was examined by nconatcs bioassay. The results indicated that virions from Ms NPV-PIBs were rapidly inactivated after 15 minute exposure to digestive fluid and all of virions were non-infectious.展开更多
【目的】探究家蚕Bombyx mori肠道特异表达的金属羧肽酶基因BmMCP 18的功能及对外源病毒侵染的抵抗机制。【方法】构建敲除BmMCP18的家蚕BmMCP18KO(C18KO),对C18KO和对照组野生型家蚕(C18KOC)的5龄幼虫中肠以及感染家蚕核型多角体病毒(B...【目的】探究家蚕Bombyx mori肠道特异表达的金属羧肽酶基因BmMCP 18的功能及对外源病毒侵染的抵抗机制。【方法】构建敲除BmMCP18的家蚕BmMCP18KO(C18KO),对C18KO和对照组野生型家蚕(C18KOC)的5龄幼虫中肠以及感染家蚕核型多角体病毒(Bombyx mori nuclear polyhedrosis virus,BmNPV)的家蚕BmMCP18KO(C18KOV)和对照野生型(C18KOCV)的5龄幼虫中肠进行转录组测序;分析差异表达基因并进行GO功能注释和KEGG通路富集;利用qRT-PCR验证相关基因表达。【结果】与对照组C18KOC相比,C18KO有75个基因上调表达,303个基因下调表达。与对照组C18KOCV相比,C18KOV有96个基因上调表达,57个基因下调表达。C18KOC vs C18KO比较组差异表达基因显著富集的GO条目为细胞外空间、细胞表面、肽交联和突触靶标识别等。C18KOCV vs C18KOV比较组差异表达基因富集最显著的GO条目为跨膜转运蛋白活性。C18KO 5龄幼虫中肠转录组中免疫通路、碳水化合物和能量代谢通路相关基因的表达比C18KOC的显著下调,包括Toll和Imd通路及MAPK通路相关基因,而泛素介导的蛋白质水解通路相关基因表达比C18KOC的显著上调。C18KOV 5龄幼虫中肠转录组中的能量代谢基因的表达比C18KOCV的显著上调。qRT-PCR验证结果表明转录组数据可靠。【结论】BmMCP18的功能涉及家蚕中肠的细胞识别、免疫调节和能量代谢,可能是通过参与中肠细胞免疫反应、能量和物质供应,从而影响蚕体对外源病原侵染的抵抗能力。展开更多
基金The project is supperted by National Natural Science Fundation of China
文摘The dissolution of polyhedra of Mythimna separata nuclear polyhedrosis. virus by digestive fluid (pH11. 03) collected from the 5th instar M. separata larvae was studied in vitro. Observations were made at timed intervals using phase contrast microscopy and scanning electron microscopy. Under phase contrast microscopy, the polyhcdra lost their refrigence by 5 minute exposure to the digestive fluid. After exposure to the fluid for 30 minutes, all of the PIBs were dissolved. Chages of the PIBs were also observed under scanning electron microscopy, after 5 minute exposure to the fluid, damaged PIBs and PIB-derived debris were seen. After 30 minute exposure, only remains of PIBs were found. The effect of M. separata digestive fluid on the infectivity of Ms NPV was examined by nconatcs bioassay. The results indicated that virions from Ms NPV-PIBs were rapidly inactivated after 15 minute exposure to digestive fluid and all of virions were non-infectious.
文摘【目的】探究家蚕Bombyx mori肠道特异表达的金属羧肽酶基因BmMCP 18的功能及对外源病毒侵染的抵抗机制。【方法】构建敲除BmMCP18的家蚕BmMCP18KO(C18KO),对C18KO和对照组野生型家蚕(C18KOC)的5龄幼虫中肠以及感染家蚕核型多角体病毒(Bombyx mori nuclear polyhedrosis virus,BmNPV)的家蚕BmMCP18KO(C18KOV)和对照野生型(C18KOCV)的5龄幼虫中肠进行转录组测序;分析差异表达基因并进行GO功能注释和KEGG通路富集;利用qRT-PCR验证相关基因表达。【结果】与对照组C18KOC相比,C18KO有75个基因上调表达,303个基因下调表达。与对照组C18KOCV相比,C18KOV有96个基因上调表达,57个基因下调表达。C18KOC vs C18KO比较组差异表达基因显著富集的GO条目为细胞外空间、细胞表面、肽交联和突触靶标识别等。C18KOCV vs C18KOV比较组差异表达基因富集最显著的GO条目为跨膜转运蛋白活性。C18KO 5龄幼虫中肠转录组中免疫通路、碳水化合物和能量代谢通路相关基因的表达比C18KOC的显著下调,包括Toll和Imd通路及MAPK通路相关基因,而泛素介导的蛋白质水解通路相关基因表达比C18KOC的显著上调。C18KOV 5龄幼虫中肠转录组中的能量代谢基因的表达比C18KOCV的显著上调。qRT-PCR验证结果表明转录组数据可靠。【结论】BmMCP18的功能涉及家蚕中肠的细胞识别、免疫调节和能量代谢,可能是通过参与中肠细胞免疫反应、能量和物质供应,从而影响蚕体对外源病原侵染的抵抗能力。
