Generation of marker‐free transgenic hexaploid
wheat
via an Agrobacterium‐mediated co‐transformation strategy in commercial Chinese
wheat
varieties
ABCD PBi
Generation of marker‐free transgenic hexaploid
wheat
via an Agrobacterium‐mediated co‐transformation strategy in commercial Chinese
wheat
varieties
Author:
Wang, Ke
;
Liu, Huiyun
;
Du, Lipu
;
Ye, Xingguo
Subjects:
Agrobacterium
;
Agrobacterium - genetics
;
Bar gene
;
Biosecurity
;
China
;
Corn
;
co‐transformation
;
Crosses, Genetic
;
Cultivars
;
Deoxyribonucleic acid
;
DNA
;
DNA Methylation
;
DNA sequencing
;
Efficiency
;
Gene Silencing
;
Genes
;
Genetic engineering
;
Genetic Markers
;
Genetic modification
;
Genetic transformation
;
Genetically engineered organisms
;
Genetically modified organisms
;
Genetically modified plants
;
Genomes
;
Genotypes
;
GUS gene
;
hexaploid
wheat
;
Hybrids
;
Laboratories
;
marker‐free transgenic plants
;
Methylation
;
Plants, Genetically Modified
;
Plasmids
;
Polyploidy
;
Progeny
;
Promoter Regions, Genetic
;
Proteins
;
Sorghum
;
Southern blotting
;
Soybeans
;
Tobacco
;
Transformation, Bacterial
;
transgene silencing
;
Transgenic plants
;
Triticum - genetics
;
Triticum aestivum
;
two independent T‐DNA vectors
;
Wheat
Is Part Of:
Plant biotechnology journal, 2017-05, Vol.15 (5), p.614-623
Notes:
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
These authors contributed equally to this work.
Description:
Summary Genotype specificity is a big problem lagging the development of efficient hexaploid
wheat
transformation system. Increasingly, the biosecurity of genetically modified organisms is garnering public attention, so the generation of marker‐free transgenic plants is very important to the eventual potential commercial release of transgenic
wheat
. In this study, 15 commercial Chinese hexaploid
wheat
varieties were successfully transformed via an Agrobacterium‐mediated method, with efficiency of up to 37.7%, as confirmed by the use of Quickstix strips, histochemical staining, PCR analysis and Southern blotting. Of particular interest, marker‐free transgenic
wheat
plants from various commercial Chinese varieties and their F1 hybrids were successfully obtained for the first time, with a frequency of 4.3%, using a plasmid harbouring two independent T‐DNA regions. The average co‐integration frequency of the gus and the bar genes located on the two independent T‐DNA regions was 49.0% in T0 plants. We further found that the efficiency of generating marker‐free plants was related to the number of bar gene copies integrated in the genome. Marker‐free transgenic wheat plants were identified in the progeny of three transgenic lines that had only one or two bar gene copies. Moreover, silencing of the bar gene was detected in 30.7% of T1 positive plants, but the gus gene was never found to be silenced in T1 plants. Bisulphite genomic sequencing suggested that DNA methylation in the 35S promoter of the bar gene regulatory region might be the main reason for bar gene silencing in the transgenic plants.
Publisher:
England: John Wiley & Sons, Inc
Language:
English