It has been reported that squash leaf curl China virus(SLCCNV)infects some Cucurbitaceae crops except for melon(Cucumis melo L.).A new disease of melon exhibiting severe leaf curl and dwarfing was observed in Hainan P...It has been reported that squash leaf curl China virus(SLCCNV)infects some Cucurbitaceae crops except for melon(Cucumis melo L.).A new disease of melon exhibiting severe leaf curl and dwarfing was observed in Hainan Province of China.In this study,the pathogen was identified as SLCCNV through biological and molecular characterization.The isolate(SLCCNV-HN)possess a bipartite genome,DNA-A(HM566112.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062251.1)pumpkin and SLCCNV-Hn61(AM260205.1)squash isolates from China,whereas DNA-B(HM566113.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062252.1).Phylogenetic analyses based on the full-length SLCCNV-HN DNA-A and-B sequences indicated that SLCCNV-HN melon isolate is clustered with SLCCNV-Hn pumpkin,SLCCNV-Hn61 and SLCCNV-SY squash isolates from southern China,forming an independent cluster.Infectious clone of SLCCNV-HN was constructed and the melon plants were inoculated and the infection rate is 100%,the systemic symptoms in melon showed identical to those of melon plants infected in fields.Additionally,melon plants transmission of this virus by Bemisia tabaci with a transmission rate of 95%(19/20)showed leaf curl and dwarf symptoms 15 days post transmission,thereby fulfilling Koch’s postulates.Analysis of genomic organization and phylogenetic trees indicated that SLCCNV-HN melon isolate belongs to the Begomovirus genus.To the best of our knowledge,this is the first characterization of meloninfecting SLCCNV through its genome,infectious clone and transmission.展开更多
人博卡病毒1(human bocaparvovirus 1,HBoV1)为感染人并引起疾病的两种细小病毒之一。其感染2-5岁婴幼儿,能引起轻度或重度急性呼吸道疾病,严重时可危及生命。HBoV1基因组末端含末端反向重复序列(repeat the sequence in reverse, ITR)...人博卡病毒1(human bocaparvovirus 1,HBoV1)为感染人并引起疾病的两种细小病毒之一。其感染2-5岁婴幼儿,能引起轻度或重度急性呼吸道疾病,严重时可危及生命。HBoV1基因组末端含末端反向重复序列(repeat the sequence in reverse, ITR),为病毒基因组复制所必需,但是难以进行PCR扩增合成。本研究通过分步合成末端ITR及分子克隆方法成功构建HBoV1的全长感染性克隆pSKHBoV1。经转染HEK293细胞后,分别从重要非结构蛋白的表达、病毒RNA转录后修饰与加工、病毒基因组复制水平以及子代病毒粒子基因组鉴定等方面,证实构建的感染性克隆在转染HEK293细胞后能够进入正常的复制周期并具有拯救出病毒粒子的潜力,这为后续研究HBoV1的复制增殖、病毒与宿主互作关系以及病毒疫苗的研发奠定了基础。展开更多
The China foot-and-mouth virus (FMDV) iso-late OH/CHA/99 was isolated from swine, which was unable to infect bovine thyroid cells in vitro or to cause typical dis-ease in bovines following intradermal inoculation in t...The China foot-and-mouth virus (FMDV) iso-late OH/CHA/99 was isolated from swine, which was unable to infect bovine thyroid cells in vitro or to cause typical dis-ease in bovines following intradermal inoculation in the tongue. To enhance antigenicity, replication, maturation and pathogenicity studies of OH/CHA/99, an infectious full- length cDNA clone, designated pBlFMDV, was prepared. The in vitro and in vivo biological properties of the virus derived from pBlFMDV were studied by analyzing antigenicity, plaque morphology and virulence in pigs. The results showed that the virus derived from pBlFMDV had the same biologi-cal properties as the parent strain OH/CHA/99; the full- length infectious cDNA clone, pBlFMDV, will be very useful in studies of the antigenicity, virulence, pathogenesis, matu-ration and replication of FMDV.展开更多
基金supported by the National Natural Science Foundation of China (31701941 and 31401810)the grants from the earmarked fund for China Agriculture Research System (CARS-26-13)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences (ASTIP) (CAAS-ASTIP-2018-ZFRI-08)
文摘It has been reported that squash leaf curl China virus(SLCCNV)infects some Cucurbitaceae crops except for melon(Cucumis melo L.).A new disease of melon exhibiting severe leaf curl and dwarfing was observed in Hainan Province of China.In this study,the pathogen was identified as SLCCNV through biological and molecular characterization.The isolate(SLCCNV-HN)possess a bipartite genome,DNA-A(HM566112.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062251.1)pumpkin and SLCCNV-Hn61(AM260205.1)squash isolates from China,whereas DNA-B(HM566113.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062252.1).Phylogenetic analyses based on the full-length SLCCNV-HN DNA-A and-B sequences indicated that SLCCNV-HN melon isolate is clustered with SLCCNV-Hn pumpkin,SLCCNV-Hn61 and SLCCNV-SY squash isolates from southern China,forming an independent cluster.Infectious clone of SLCCNV-HN was constructed and the melon plants were inoculated and the infection rate is 100%,the systemic symptoms in melon showed identical to those of melon plants infected in fields.Additionally,melon plants transmission of this virus by Bemisia tabaci with a transmission rate of 95%(19/20)showed leaf curl and dwarf symptoms 15 days post transmission,thereby fulfilling Koch’s postulates.Analysis of genomic organization and phylogenetic trees indicated that SLCCNV-HN melon isolate belongs to the Begomovirus genus.To the best of our knowledge,this is the first characterization of meloninfecting SLCCNV through its genome,infectious clone and transmission.
文摘人博卡病毒1(human bocaparvovirus 1,HBoV1)为感染人并引起疾病的两种细小病毒之一。其感染2-5岁婴幼儿,能引起轻度或重度急性呼吸道疾病,严重时可危及生命。HBoV1基因组末端含末端反向重复序列(repeat the sequence in reverse, ITR),为病毒基因组复制所必需,但是难以进行PCR扩增合成。本研究通过分步合成末端ITR及分子克隆方法成功构建HBoV1的全长感染性克隆pSKHBoV1。经转染HEK293细胞后,分别从重要非结构蛋白的表达、病毒RNA转录后修饰与加工、病毒基因组复制水平以及子代病毒粒子基因组鉴定等方面,证实构建的感染性克隆在转染HEK293细胞后能够进入正常的复制周期并具有拯救出病毒粒子的潜力,这为后续研究HBoV1的复制增殖、病毒与宿主互作关系以及病毒疫苗的研发奠定了基础。
文摘The China foot-and-mouth virus (FMDV) iso-late OH/CHA/99 was isolated from swine, which was unable to infect bovine thyroid cells in vitro or to cause typical dis-ease in bovines following intradermal inoculation in the tongue. To enhance antigenicity, replication, maturation and pathogenicity studies of OH/CHA/99, an infectious full- length cDNA clone, designated pBlFMDV, was prepared. The in vitro and in vivo biological properties of the virus derived from pBlFMDV were studied by analyzing antigenicity, plaque morphology and virulence in pigs. The results showed that the virus derived from pBlFMDV had the same biologi-cal properties as the parent strain OH/CHA/99; the full- length infectious cDNA clone, pBlFMDV, will be very useful in studies of the antigenicity, virulence, pathogenesis, matu-ration and replication of FMDV.
