Obtaining transgenic plants is a common method for analyzing gene function.Unfortunately,stable genetic transformation is difficult to achieve,especially for plants(e.g.,soybean),which are recalcitrant to genetic tran...Obtaining transgenic plants is a common method for analyzing gene function.Unfortunately,stable genetic transformation is difficult to achieve,especially for plants(e.g.,soybean),which are recalcitrant to genetic transformation.Transient expression systems,such as Arabidopsis protoplast,Nicotiana leaves,and onion bulb leaves are widely used for gene functional studies.A simple method for obtaining transgenic soybean callus tissues was reported recently.We extend this system with simplified culture conditions to gene functional studies,including promoter analysis,expression and subcellular localization of the target protein,and protein-protein interaction.We also evaluate the plasticity of this system with soybean varieties,different vector constructs,and various Agrobacterium strains.The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-,vector-,and Agrobacterium strain-independent modes.We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.展开更多
Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herb...Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herbaceous plants,with notable applications in species such as Arabidopsis(Yin et al.,2024),soybean(Zhang et al.,2024),rice(Zhang et al.,2020),and Chinese cabbage(Li et al.,2021).However,its application in fruit trees is limited.This is primarily due to their long growth cycles and lack of rapid,efficient,and stable transgenic systems,which severely hinders foundational research involving plant genetic transformation(Mei et al.,2024).Furthermore,for subtropical fruit trees,the presence of recalcitrant seeds adds an extra layer of difficulty to genetic transformation(Umarani et al.,2015),as most methods rely on seed germination as a basis for transformation.展开更多
基金supported by the Transgenic Programs,China(2014ZX0800930B and 2016ZX08009-001)the National Natural Science Found of China(31371703)
文摘Obtaining transgenic plants is a common method for analyzing gene function.Unfortunately,stable genetic transformation is difficult to achieve,especially for plants(e.g.,soybean),which are recalcitrant to genetic transformation.Transient expression systems,such as Arabidopsis protoplast,Nicotiana leaves,and onion bulb leaves are widely used for gene functional studies.A simple method for obtaining transgenic soybean callus tissues was reported recently.We extend this system with simplified culture conditions to gene functional studies,including promoter analysis,expression and subcellular localization of the target protein,and protein-protein interaction.We also evaluate the plasticity of this system with soybean varieties,different vector constructs,and various Agrobacterium strains.The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-,vector-,and Agrobacterium strain-independent modes.We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.
基金funded by the Key-Area Research and Development Program of Guangdong Province(Grant No.2022B0202070002)the Guangxi Science and Technology Major Program(Grant No.GuikeAA23023007-2)+1 种基金the Guangdong Province Modern Agricultural Industry Technology System Innovation Team Construction Project(2024CXTD19)Guangdong Basic and Applied Basic Research Foundation(Grant No.2023A1515010303)。
文摘Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herbaceous plants,with notable applications in species such as Arabidopsis(Yin et al.,2024),soybean(Zhang et al.,2024),rice(Zhang et al.,2020),and Chinese cabbage(Li et al.,2021).However,its application in fruit trees is limited.This is primarily due to their long growth cycles and lack of rapid,efficient,and stable transgenic systems,which severely hinders foundational research involving plant genetic transformation(Mei et al.,2024).Furthermore,for subtropical fruit trees,the presence of recalcitrant seeds adds an extra layer of difficulty to genetic transformation(Umarani et al.,2015),as most methods rely on seed germination as a basis for transformation.
