Although the use of stable transformation technology has led to great insight into gene function,its application in high-throughput studies remains arduous.Agro-infiltration have been widely used in species such as Ni...Although the use of stable transformation technology has led to great insight into gene function,its application in high-throughput studies remains arduous.Agro-infiltration have been widely used in species such as Nicotiana benthamiana for the rapid detection of gene expression and protein interaction analysis,but this technique does not work efficiently in other plant species,including Arabidopsis thaliana.As an efficient high-throughput transient expression system is currently lacking in the model plant species A.thaliana,we developed a method that is characterized by high efficiency,reproducibility,and suitability for transient expression of a variety of functional proteins in A.thaliana and 7 other plant species,including Brassica oleracea,Capsella rubella,Thellungiella salsuginea,Thellungiella halophila,Solanum tuberosum,Capsicum annuum,and N.benthamiana.Efficiency of this method was independently verified in three independent research facilities,pointing to the robustness of this technique.Furthermore,in addition to demonstrating the utility of this technique in a range of species,we also present a case study employing this method to assess protein–protein interactions in the sucrose biosynthesis pathway in Arabidopsis.展开更多
Differentiation processes in the primary root meristem are controlled by several signaling pathways that are regulated by phytohormones or by secreted peptides. Long-term maintenance of an active root meristem require...Differentiation processes in the primary root meristem are controlled by several signaling pathways that are regulated by phytohormones or by secreted peptides. Long-term maintenance of an active root meristem requires that the generation of new stem cells and the loss of these from the meristem due to differentiation are precisely coordinated. Via phenotypic and large-scale transcriptome analyses of mutants, we show that the signaling peptide CLE40 and the recep- tor proteins CLV2 and CRN act in two genetically separable pathways that antagonistically regulate cell differentiation in the proximal root meristem. CLE40 inhibits cell differentiation throughout the primary root meristem by controlling genes with roles in abscisic acid, auxin, and cytokinin signaling. CRN and CLV2 jointly control target genes that promote cell differentiation specifically in the transition zone of the proximal root meristem. While CRN and CLV2 are not acting in the CLE40 signaling pathway under normal growth conditions, both proteins are required when the levels of CLE40 or related CLE peptides increase. We show here that two antagonistically acting pathways controlling root meristem differentiation can be activated by the same peptide in a dosage-dependent manner.展开更多
基金supported by funding from the Max Planck Society(A.R.F.and Y.Z.)the European Union’s Horizon 2020 project PlantaSYST SGA-CSA no.739582 under FPA no.664620(A.R.F.and Y.Z.).
文摘Although the use of stable transformation technology has led to great insight into gene function,its application in high-throughput studies remains arduous.Agro-infiltration have been widely used in species such as Nicotiana benthamiana for the rapid detection of gene expression and protein interaction analysis,but this technique does not work efficiently in other plant species,including Arabidopsis thaliana.As an efficient high-throughput transient expression system is currently lacking in the model plant species A.thaliana,we developed a method that is characterized by high efficiency,reproducibility,and suitability for transient expression of a variety of functional proteins in A.thaliana and 7 other plant species,including Brassica oleracea,Capsella rubella,Thellungiella salsuginea,Thellungiella halophila,Solanum tuberosum,Capsicum annuum,and N.benthamiana.Efficiency of this method was independently verified in three independent research facilities,pointing to the robustness of this technique.Furthermore,in addition to demonstrating the utility of this technique in a range of species,we also present a case study employing this method to assess protein–protein interactions in the sucrose biosynthesis pathway in Arabidopsis.
文摘Differentiation processes in the primary root meristem are controlled by several signaling pathways that are regulated by phytohormones or by secreted peptides. Long-term maintenance of an active root meristem requires that the generation of new stem cells and the loss of these from the meristem due to differentiation are precisely coordinated. Via phenotypic and large-scale transcriptome analyses of mutants, we show that the signaling peptide CLE40 and the recep- tor proteins CLV2 and CRN act in two genetically separable pathways that antagonistically regulate cell differentiation in the proximal root meristem. CLE40 inhibits cell differentiation throughout the primary root meristem by controlling genes with roles in abscisic acid, auxin, and cytokinin signaling. CRN and CLV2 jointly control target genes that promote cell differentiation specifically in the transition zone of the proximal root meristem. While CRN and CLV2 are not acting in the CLE40 signaling pathway under normal growth conditions, both proteins are required when the levels of CLE40 or related CLE peptides increase. We show here that two antagonistically acting pathways controlling root meristem differentiation can be activated by the same peptide in a dosage-dependent manner.
