Transient gene expression in western white pine using agroinfiltration  

作　　者：Zhenguo Ma Jun-Jun Liu Arezoo Zamany Holly Williams 

机构地区：[1]Canadian Forest Service,Natural Resources Canada,506 West Burnside Road,Victoria,BC V8Z 1M5,Canada

出　　处：《Journal of Forestry Research》2020年第5期1823-1832,共10页林业研究（英文版）

基　　金：supported by the CFS-GRDI fund。

摘　　要：A method for transient gene expression was developed for western white pine(WWP,Pinus monticola Dougl.ex D.Don)using reporter gene uidA encodingβ-glucuronidase(GUS).GUS was transiently expressed in cross sections of primary and secondary needles,cotyledons,and current and second year stems of WWP via vacuum-infiltration with Agrobacterium tumefaciens.Histochemical assays of cross sections of secondary needles showed stronger blue color indicating GUS expression at day 1 and 2 than on other days post agroinfiltration(dpa).GUS activity expressed inside WWP cells was confirmed using light microscopy.In fluorometric assays,GUS expression was high at 1 dpa and lasted until 4 dpa in detached secondary needles,while similarly high expression levels only lasted until 2 dpa in attached secondary needles then dropped significantly.Although the length of GUS-staining zones varied among different WWP organs and between growth and dormant seasons,all tested WWP tissues using the protocol had high levels of transient GUS expression.Thus,heterologous candidate genes or endogenous silencing can be expressed in various WWP tissues or organs using this agroinfiltration approach.The current protocol for efficient transient gene expression will aid functional genomics study of WWP and its pathogens and related conifer species.

关 键 词：AGROBACTERIUM AGROINFILTRATION Β-GLUCURONIDASE Transient gene expression Western white pine 

分 类 号：S791.27[农业科学—林木遗传育种]