为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制,本文报道利用Make Your Own"Mate&Plate" Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA,采用SMART法反转录合成双链c...为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制,本文报道利用Make Your Own"Mate&Plate" Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA,采用SMART法反转录合成双链cDNA,经靳,酶切并去除短片段之后,与经同样酶切的pGADT7-SfiI载体连接,利用电转法将重组载体转化到大肠杆菌宿主细胞中,获得初级cDNA文库,最后以初级文库100万克隆为基数扩增,得到扩增文库并提取质粒。结果得到库容量大于2.0X10。的初级文库,检测表明文库cDNA插入片段长度主要分布在700—2000bp,文库重组率为87.5%。结果表明,该文库质量较好,可用于后续酵母双杂交互作蛋白筛选试验,本研究为开展病毒与寄主互作的研究奠定基础。展开更多
Objective:Chilling tolerance of salicylic acid(SA) in banana seedlings(Musa acuminata cv.,Williams 8818) was investigated by changes in ultrastructure in this study.Methods:Light and electron microscope observation.Re...Objective:Chilling tolerance of salicylic acid(SA) in banana seedlings(Musa acuminata cv.,Williams 8818) was investigated by changes in ultrastructure in this study.Methods:Light and electron microscope observation.Results:Pretreatment with 0.5 mmol/L SA under normal growth conditions(30/22 °C) by foliar spray and root irrigation resulted in many changes in ultrastructure of banana cells,such as cells separation from palisade parenchymas,the appearance of crevices in cell walls,the swelling of grana and stromal thylakoids,and a reduction in the number of starch granules.These results implied that SA treatment at 30/22 °C could be a type of stress.During 3 d of exposure to 7 °C chilling stress under low light,however,cell ultrastructure of SA-pretreated banana seedlings showed less deterioration than those of control seedlings(distilled water-pretreated).Conclusion:SA could provide some protection for cell structure of chilling-stressed banana seedling.展开更多
文摘为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制,本文报道利用Make Your Own"Mate&Plate" Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA,采用SMART法反转录合成双链cDNA,经靳,酶切并去除短片段之后,与经同样酶切的pGADT7-SfiI载体连接,利用电转法将重组载体转化到大肠杆菌宿主细胞中,获得初级cDNA文库,最后以初级文库100万克隆为基数扩增,得到扩增文库并提取质粒。结果得到库容量大于2.0X10。的初级文库,检测表明文库cDNA插入片段长度主要分布在700—2000bp,文库重组率为87.5%。结果表明,该文库质量较好,可用于后续酵母双杂交互作蛋白筛选试验,本研究为开展病毒与寄主互作的研究奠定基础。
基金Project (No. 30471112) supported by the National Natural Science Foundation of China
文摘Objective:Chilling tolerance of salicylic acid(SA) in banana seedlings(Musa acuminata cv.,Williams 8818) was investigated by changes in ultrastructure in this study.Methods:Light and electron microscope observation.Results:Pretreatment with 0.5 mmol/L SA under normal growth conditions(30/22 °C) by foliar spray and root irrigation resulted in many changes in ultrastructure of banana cells,such as cells separation from palisade parenchymas,the appearance of crevices in cell walls,the swelling of grana and stromal thylakoids,and a reduction in the number of starch granules.These results implied that SA treatment at 30/22 °C could be a type of stress.During 3 d of exposure to 7 °C chilling stress under low light,however,cell ultrastructure of SA-pretreated banana seedlings showed less deterioration than those of control seedlings(distilled water-pretreated).Conclusion:SA could provide some protection for cell structure of chilling-stressed banana seedling.