An effector-reporter system is a powerful tool used to study cellular signal transduction,but this technique has been traditionally used in protoplasts.A similar system to study cellular signal transduction in fruits ...An effector-reporter system is a powerful tool used to study cellular signal transduction,but this technique has been traditionally used in protoplasts.A similar system to study cellular signal transduction in fruits has not yet been established.In this study,we aimed to establish an effector–reporter system for strawberry fruit,a model nonclimacteric fruit.We first investigated the characteristics of transient gene expression in strawberry fruits and found marked variation in gene expression levels among individual fruits,and this variation has complicated the establishment of a technical system.To overcome this difficulty,we investigated a sampling strategy based on a statistical analysis of the activity pattern of four different reporters(GUS,GFP,FLuc,and RLuc)among individual fruits and combinations of pairs of reporters(GUS/GFP and RLuc/FLuc).Based on an optimized sampling strategy,we finally established a step-by step protocol for the effector/reporter assay.Using FaMYB10 and FaWRKY71 as the effectors and GUS driven by the FaCHS promoter as the reporter,we demonstrated that this effector/reporter system was practical and reliable.This effector/reporter technique will contribute to an in-depth exploration of the signaling mechanism for the regulation of strawberry fruit ripening.展开更多
基金the National Key Research and Development Program(SQ2019YFD100078)the National Natural Science Foundation of China(Grants 31672133 and 31872086)+2 种基金the National Key Research and Development Program(2019YFD1000200)the Construction of Beijing Science and Technology Innovation and Service Capacity in Top Subjects(CEFF-PXM2019_014207_000032)the Beijing Municipal Natural Science Foundation(Grant No.6171001).
文摘An effector-reporter system is a powerful tool used to study cellular signal transduction,but this technique has been traditionally used in protoplasts.A similar system to study cellular signal transduction in fruits has not yet been established.In this study,we aimed to establish an effector–reporter system for strawberry fruit,a model nonclimacteric fruit.We first investigated the characteristics of transient gene expression in strawberry fruits and found marked variation in gene expression levels among individual fruits,and this variation has complicated the establishment of a technical system.To overcome this difficulty,we investigated a sampling strategy based on a statistical analysis of the activity pattern of four different reporters(GUS,GFP,FLuc,and RLuc)among individual fruits and combinations of pairs of reporters(GUS/GFP and RLuc/FLuc).Based on an optimized sampling strategy,we finally established a step-by step protocol for the effector/reporter assay.Using FaMYB10 and FaWRKY71 as the effectors and GUS driven by the FaCHS promoter as the reporter,we demonstrated that this effector/reporter system was practical and reliable.This effector/reporter technique will contribute to an in-depth exploration of the signaling mechanism for the regulation of strawberry fruit ripening.
