The WRKY transcription factor gene family is one of the unique gene families in plants.It plays an important role in response to abiotic stresses such as cold and drought,hormone signal transduction,regulation of bios...The WRKY transcription factor gene family is one of the unique gene families in plants.It plays an important role in response to abiotic stresses such as cold and drought,hormone signal transduction,regulation of biosynthesis,leaf senescence seed germination,etc.However,little information is available about WRKY transcription factors in Verbena bonariensis.In this study,70 VbWRKY genes were identified from the whole genome.The phylogenetic analysis of the WRKY gene family in V.bonariensis and Arabidopsis shows that the WRKY genes in V.bonariensis can be divided into three groups:I,II,and III,which contain 13,47,and 10 members,respectively.Group II can be further divided into five subclasses:IIa(5),IIb(10),IIc(18),IId(6),and IIe(8).Conservative motif analysis showed that 64 proteins encoded by the VbWRKY gene had conserved motifs 1,2 and 3,and the same subclass motif elements were approximately the same.The collinearity analysis showed that there were 44 homologous gene pairs among the VbWRKYs,and these homologous gene pairs may have the same function.Promoter sequence analysis showed that the VbWRKY gene has multiple cis-acting elements,including not only cis-acting elements related to low-temperature and light responses,but also cis-acting elements related to hormone regulation,Among them,most VbWRKY genes contain response elements about low-temperature,and 30 VbWRKY genes contain low-temperature response elements(LTR),and 61 VbWRKY genes contain abscisic acid response elements(ABRE),indicating that VbWRKY plays a crucial role in plant growth and abiotic stress.According to the expression of VbWRKY in the cold stress and different tissues transcriptome,70 VbWRKY genes played their respective roles in various tissues and stages to regulate plant growth,Also,some of them participated in the process of cold stress tolerance,52 VbWRKYs showed significant differences in expression under cold stress,and 37 VbWRKY genes were up-regulated under cold stress.9 VbWRKY genes were selected for quantitative real-time PCR(qRT-PCR)analysis under low-temperature stress,and the results showed that all 9 genes were upregulated under low-temperature stress.Ultimately,the present study provides a comprehensive analysis of the predicted V.bonariensis WRKY genes family,which provided a theoretical basis for the study of low-temperature resistance and growth and development of V.bonariensis.展开更多
Objective:To examine the action of the effective component,4'-methylether-scutellarein,from Verbena officinalis L.(VOL)on the proliferation and apoptosis of human choriocarcinomaJAR cells.Methods:Cell proliferatio...Objective:To examine the action of the effective component,4'-methylether-scutellarein,from Verbena officinalis L.(VOL)on the proliferation and apoptosis of human choriocarcinomaJAR cells.Methods:Cell proliferation was measured by MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyl tetrasodium bromide,MTT]assay and the incorporation of tritiated thymidine(~3H-TdR).Apoptosis of cell was evaluated by flow cytometry(FCM)and the characteristic apoptoticDNA ladder by agarose gel electrophoresis,and the morphological changes of apoptotic JAR cellswere observed under fluorescence microscopy and electron microscopy(EM).Expressions of ap-optosis proteins,poly(ADP-ribose)polymerase(PARP)and caspase-3,-8,and -9 were deter-mined with Western blot.Results:The effective component from VOL inhibited the proliferation of JAR cells in a dose-and time-dependent manner.The treated cell cycle was arrested in S phase and an apoptotic peakwas found in S phase using FCM analysis.A typical DNA ladder appeared in the treatment groupwhen analyzed by agarose gel electrophoresis.Using fluorescence microscopy,the percentage ofapoptotic cell was 0.9%,6%,and 14% after treatments of 10,20,and 40 mg·L^(-1) of the effec-tive component,respectively,for 48 h.Typical apoptotic changes,such as condensed chromatinand presence of apoptotic bodies,were observed under EM.Treatment with effective componentfor 48 h and 72 h also induced protein expression of PARP and caspase-3,-8,and -9 as seen byWestern blot.Conclusions:The effective component from VOL inhibits cell proliferation and induces apop-tosis in human choriocarcinoma JAR cells.展开更多
基金funded by the National Natural Science Foundation of China(32160722)the Key Research Project of Guizhou Provincial Science and Technology Projects(QKHJC-ZK[2023]ZD-006).
文摘The WRKY transcription factor gene family is one of the unique gene families in plants.It plays an important role in response to abiotic stresses such as cold and drought,hormone signal transduction,regulation of biosynthesis,leaf senescence seed germination,etc.However,little information is available about WRKY transcription factors in Verbena bonariensis.In this study,70 VbWRKY genes were identified from the whole genome.The phylogenetic analysis of the WRKY gene family in V.bonariensis and Arabidopsis shows that the WRKY genes in V.bonariensis can be divided into three groups:I,II,and III,which contain 13,47,and 10 members,respectively.Group II can be further divided into five subclasses:IIa(5),IIb(10),IIc(18),IId(6),and IIe(8).Conservative motif analysis showed that 64 proteins encoded by the VbWRKY gene had conserved motifs 1,2 and 3,and the same subclass motif elements were approximately the same.The collinearity analysis showed that there were 44 homologous gene pairs among the VbWRKYs,and these homologous gene pairs may have the same function.Promoter sequence analysis showed that the VbWRKY gene has multiple cis-acting elements,including not only cis-acting elements related to low-temperature and light responses,but also cis-acting elements related to hormone regulation,Among them,most VbWRKY genes contain response elements about low-temperature,and 30 VbWRKY genes contain low-temperature response elements(LTR),and 61 VbWRKY genes contain abscisic acid response elements(ABRE),indicating that VbWRKY plays a crucial role in plant growth and abiotic stress.According to the expression of VbWRKY in the cold stress and different tissues transcriptome,70 VbWRKY genes played their respective roles in various tissues and stages to regulate plant growth,Also,some of them participated in the process of cold stress tolerance,52 VbWRKYs showed significant differences in expression under cold stress,and 37 VbWRKY genes were up-regulated under cold stress.9 VbWRKY genes were selected for quantitative real-time PCR(qRT-PCR)analysis under low-temperature stress,and the results showed that all 9 genes were upregulated under low-temperature stress.Ultimately,the present study provides a comprehensive analysis of the predicted V.bonariensis WRKY genes family,which provided a theoretical basis for the study of low-temperature resistance and growth and development of V.bonariensis.
文摘Objective:To examine the action of the effective component,4'-methylether-scutellarein,from Verbena officinalis L.(VOL)on the proliferation and apoptosis of human choriocarcinomaJAR cells.Methods:Cell proliferation was measured by MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyl tetrasodium bromide,MTT]assay and the incorporation of tritiated thymidine(~3H-TdR).Apoptosis of cell was evaluated by flow cytometry(FCM)and the characteristic apoptoticDNA ladder by agarose gel electrophoresis,and the morphological changes of apoptotic JAR cellswere observed under fluorescence microscopy and electron microscopy(EM).Expressions of ap-optosis proteins,poly(ADP-ribose)polymerase(PARP)and caspase-3,-8,and -9 were deter-mined with Western blot.Results:The effective component from VOL inhibited the proliferation of JAR cells in a dose-and time-dependent manner.The treated cell cycle was arrested in S phase and an apoptotic peakwas found in S phase using FCM analysis.A typical DNA ladder appeared in the treatment groupwhen analyzed by agarose gel electrophoresis.Using fluorescence microscopy,the percentage ofapoptotic cell was 0.9%,6%,and 14% after treatments of 10,20,and 40 mg·L^(-1) of the effec-tive component,respectively,for 48 h.Typical apoptotic changes,such as condensed chromatinand presence of apoptotic bodies,were observed under EM.Treatment with effective componentfor 48 h and 72 h also induced protein expression of PARP and caspase-3,-8,and -9 as seen byWestern blot.Conclusions:The effective component from VOL inhibits cell proliferation and induces apop-tosis in human choriocarcinoma JAR cells.
