‘Lvbao-701’ is a cultivar of Chinese flowering cabbage(Brassica parachinensis) that exhibits low cadmium(Cd) accumulation and high Cd tolerance.In this study, this cultivar was compared with a high-Cd accumulating c...‘Lvbao-701’ is a cultivar of Chinese flowering cabbage(Brassica parachinensis) that exhibits low cadmium(Cd) accumulation and high Cd tolerance.In this study, this cultivar was compared with a high-Cd accumulating cultivar, ‘Chixin-4’, to characterize the mechanisms influencing Cd accumulation in B. parachinensis. Root cell walls were isolated by dissolving the cytoplasm with an organic solvent, and root Cd and phytochelatin(PC) contents were analyzed. In addition, a PC synthase gene fragment was cloned and expressed under Cd stress conditions. The proportions of Cd bound to root cell walls were higher in the ‘Lvbao-701’ plants(68.32%–76.80%) than in the ‘Chixin-4’ plants(35.36%–54.18%) after exposure to Cd stress. The proportions of Cd bound to root cell walls measured using cell walls isolated with a non-destructive method were higher than those obtained using a conventional method that required grinding and centrifugation. Exposure to Cd stress induced the PC production and resulted in higher PC contents in the ‘Lvbao-701’ plants than in the ‘Chixin-4’ plants. Cloning and expression analysis of a B. parachinensis PC synthase cDNA fragment indicated that PC synthase gene expression was induced by Cd and occurred mainly in the roots of both ‘Lvbao-701’ and ‘Chixin-4’ plants. However, the PC synthase gene expression level was higher in the‘Lvbao-701’ roots than in the ‘Chixin-4’ roots. Therefore, a higher abundance of Cd in the root cell walls of ‘Lvbao-701’ and up-regulated PC production in the roots are probably the main reasons why ‘Lvbao-701’ exhibits lower Cd translocation to the shoots and higher tolerance to Cd than ‘Chixin-4’.展开更多
White spot syndrome virus(WSSV)is one of the most virulent pathogens in shrimp aquaculture.An in-depth understanding of the surface protein-protein interplays between WSSV and host is the key to further elucidating th...White spot syndrome virus(WSSV)is one of the most virulent pathogens in shrimp aquaculture.An in-depth understanding of the surface protein-protein interplays between WSSV and host is the key to further elucidating the pathogenesis mechanism.In this study,VP39A was identified as a novel cellular binding protein of WSSV by reversely conducting the biotinylation based affinity chromatography(BBAC)method.The result of indirect immunofluorescence assay,competitive ELISA,and in-vivo neutralization assays show that VP39A is vital to the cellular entry and infection of WSSV.Na^(+),K^(+)-ATPase alpha subunit(NKAα),a trans-membrane subunit of complex Na^(+),K^(+)-ATPase,was captured as the VP39A binding protein with pull-down assay.Co-localization of NKAαwith VP39A in shrimp hemocytes was confirmed by immunofluorescence and co-immunoprecipitation studies.In addition,blocking with anti-NKAαantibody can reduce the binding of WSSV to the gill cellular membrane proteins.These results collectively demonstrate that VP39A plays an important role in the viral adsorption via its interaction with NKAα,which could be an effective target for drug design.展开更多
基金This study was supported by the National Natural Science Foundation of China(No.41371308),the Science and Technology Supporting Project of China(No.2015-BAD05B05)the Research Project of the Education Department of Guangdong Province,China(No.2013-CXZDA007).
文摘‘Lvbao-701’ is a cultivar of Chinese flowering cabbage(Brassica parachinensis) that exhibits low cadmium(Cd) accumulation and high Cd tolerance.In this study, this cultivar was compared with a high-Cd accumulating cultivar, ‘Chixin-4’, to characterize the mechanisms influencing Cd accumulation in B. parachinensis. Root cell walls were isolated by dissolving the cytoplasm with an organic solvent, and root Cd and phytochelatin(PC) contents were analyzed. In addition, a PC synthase gene fragment was cloned and expressed under Cd stress conditions. The proportions of Cd bound to root cell walls were higher in the ‘Lvbao-701’ plants(68.32%–76.80%) than in the ‘Chixin-4’ plants(35.36%–54.18%) after exposure to Cd stress. The proportions of Cd bound to root cell walls measured using cell walls isolated with a non-destructive method were higher than those obtained using a conventional method that required grinding and centrifugation. Exposure to Cd stress induced the PC production and resulted in higher PC contents in the ‘Lvbao-701’ plants than in the ‘Chixin-4’ plants. Cloning and expression analysis of a B. parachinensis PC synthase cDNA fragment indicated that PC synthase gene expression was induced by Cd and occurred mainly in the roots of both ‘Lvbao-701’ and ‘Chixin-4’ plants. However, the PC synthase gene expression level was higher in the‘Lvbao-701’ roots than in the ‘Chixin-4’ roots. Therefore, a higher abundance of Cd in the root cell walls of ‘Lvbao-701’ and up-regulated PC production in the roots are probably the main reasons why ‘Lvbao-701’ exhibits lower Cd translocation to the shoots and higher tolerance to Cd than ‘Chixin-4’.
基金Supported by the National Natural Science Foundation of China(Nos.31402322,31472299)the Tianjin Research Program of Application Foundation and Advanced Technology(Nos.15JCQNJC46400,15JCZDJC33800)+1 种基金the Tianjin Mariculture Modern Agriculture Industrial Technology System Innovation Team Project(No.ITTMRS2022006)the Tianjin Model Worker Innovation Team Project and Tianjin Normal University Doctoral Research Foundation(No.52XB1412)。
文摘White spot syndrome virus(WSSV)is one of the most virulent pathogens in shrimp aquaculture.An in-depth understanding of the surface protein-protein interplays between WSSV and host is the key to further elucidating the pathogenesis mechanism.In this study,VP39A was identified as a novel cellular binding protein of WSSV by reversely conducting the biotinylation based affinity chromatography(BBAC)method.The result of indirect immunofluorescence assay,competitive ELISA,and in-vivo neutralization assays show that VP39A is vital to the cellular entry and infection of WSSV.Na^(+),K^(+)-ATPase alpha subunit(NKAα),a trans-membrane subunit of complex Na^(+),K^(+)-ATPase,was captured as the VP39A binding protein with pull-down assay.Co-localization of NKAαwith VP39A in shrimp hemocytes was confirmed by immunofluorescence and co-immunoprecipitation studies.In addition,blocking with anti-NKAαantibody can reduce the binding of WSSV to the gill cellular membrane proteins.These results collectively demonstrate that VP39A plays an important role in the viral adsorption via its interaction with NKAα,which could be an effective target for drug design.
