Establishment of a transformation system in close relatives of wheat under the assistance of TaWOX5  

作　　者：Yanan Chang Junxian Liu Chang Liu Huiyun Liu Huali Tang Yuliang Qiu Zhishan Lin Ke Wang Yueming Yan Xingguo Ye 

机构地区：[1]Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China [2]Beijing Key Laboratory of Plant Gene Resources and Biotechnology for Carbon Reduction and Environment Improvement/College of Life Science,Capital Normal University,Beijing 100048,China [3]College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193,China [4]State Key Laboratory of Plant Cell and Chromosome Engineering/Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China

出　　处：《Journal of Integrative Agriculture》2024年第6期1839-1849,共11页农业科学学报（英文版）

基　　金：supported by grants from the National Natural Science Foundation of China (31971946);the Technology Innovation Program of the Chinese Academy of Agricultural Sciences, China (2060302-2-23 and ASTIP-2060302-2-19).

摘　　要：Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature embryos.Transgenic plants were efficiently generated.During the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar protein.The transformation efficiency of T.monococcum genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic T.monococcum and hexaploid triticale plants.In the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR genomes.Furthermore,the transgene was almost stably inherited in the next generation by Mendel’s law.The findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.

关 键 词：Triticum monococcum hexaploid triticale RYE TaWOX5 AGROBACTERIUM transformation efficiency 

分 类 号：S512.1[农业科学—作物学]