Kim Su Min, Baek Eseul, Ryu Ki Hyun, Choi Sun Hee
Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, Seoul 139-774, Republic of Korea.
Plant Virus GenBank, Seoul Women's University, Seoul 139-774, Republic of Korea; Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, Seoul 139-774, Republic of Korea.
Virus Res. 2016 Sep 2;223:43-51. doi: 10.1016/j.virusres.2016.06.019. Epub 2016 Jun 27.
Cymbidium mosaic virus (CymMV)-induced expressed sequence tag (EST) clones from Chenopodium amaranticolor were identified. CymMV was mechanically inoculated onto C. amaranticolor, and local lesion symptoms were observed. Inoculated leaves were collected on serial days post inoculation (dpi) to identify activated or suppressed genes. mRNA isolation and suppression subtractive hybridization (SSH) were then performed to identify differentially expressed genes related to the local lesion response. Fifty-three ESTs, including genes related to defense and stress responses (e.g., lipoxygenase, jasmonate-induced protein, and heat shock protein), were generated. In addition, a large proportion of the ESTs were found to be involved in photosynthesis, as determined by their functional categories. Expression levels of several EST genes were observed using quantitative real-time reverse transcription-polymerase chain reaction, and the evaluated genes showed varying levels of expression during the experimental period. In this study, differentially expressed sequences via SSH were identified from CymMV-infected C. amaranticolor, and profiling and annotation were carried out to determine the expression pattern of CymMV and its interaction with C. amaranticolor.
已鉴定出从苋色藜中获得的由建兰花叶病毒(CymMV)诱导的表达序列标签(EST)克隆。将CymMV机械接种到苋色藜上,并观察到局部病斑症状。在接种后的连续几天收集接种叶片,以鉴定激活或抑制的基因。然后进行mRNA分离和抑制性消减杂交(SSH),以鉴定与局部病斑反应相关的差异表达基因。产生了53个EST,包括与防御和应激反应相关的基因(例如脂氧合酶、茉莉酸诱导蛋白和热休克蛋白)。此外,根据其功能类别发现很大一部分EST参与光合作用。使用定量实时逆转录-聚合酶链反应观察了几个EST基因的表达水平,评估的基因在实验期间表现出不同的表达水平。在本研究中,从感染CymMV的苋色藜中鉴定出通过SSH差异表达的序列,并进行了分析和注释,以确定CymMV的表达模式及其与苋色藜的相互作用。