.png){kind=icon}
返回
Robust soybean leaf agroinfiltration
.png){kind=icon}
PRE
AI
DOI：10.1007/s00299-024-03245-4.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Key messageA robust agroinfiltration-mediated transient gene expression method for soybean leaves was developed. Plant genotype, developmental stage and leaf age, surfactant, and Agrobacterium culture conditions are important for successful agroinfiltration.AbstractAgroinfiltration of Nicotiana benthamiana has emerged as a workhorse transient assay for plant biotechnology and synthetic biology to test the performance of gene constructs in dicot leaves. While effective, it is nonetheless often desirable to assay transgene constructs directly in crop species. To that end, we innovated a substantially robust agroinfiltration method for Glycine max (soybean), the most widely grown dicot crop plant in the world. Several factors were found to be relevant to successful soybean leaf agroinfiltration, including genotype, surfactant, developmental stage, and Agrobacterium strain and culture medium. Our optimized protocol involved a multi-step Agrobacterium culturing process with appropriate expression vectors, Silwet L-77 as the surfactant, selection of fully expanded leaves in the VC or V1 stage of growth, and 5 min of vacuum at - 85 kPa followed by a dark incubation period before plants were returned to normal growth conditions. Using this method, young soybean leaves of two lines-V17-0799DT, and TN16-5004-were high expressors for GUS, two co-expressed fluorescent protein genes, and the RUBY reporter product, betalain. This work not only represents a new research tool for soybean biotechnology, but also indicates critical parameters for guiding agroinfiltration optimization for other crop species. We speculate that leaf developmental stage might be the most critical factor for successful agroinfiltration.
Keyword:
Transient gene expression
Soybean
Agroinfiltration
Reporter gene
RUBY
Betalain
期刊
IF:
4.5
论文数:
5.3K
被引数:
1.3W
机构
引用论文
Role of the host cell cycle in the Agrobacterium-mediated genetic transformation of Petunia: Evidence of an S-phase control mechanism for T-DNA transfer.png)
PLANTA
PLANTAIF3.8
PREAI
分享收藏
Acetosyringone treatment duration affects large T-DNA molecule transfer to rice callusBMC BIOTECHNOLOGY
OAAI
BMC BIOTECHNOLOGYIF3.4
OAAI分享收藏Engineering of soybean mosaic virus as a versatile tool for studying protein-protein interactions in soybeanSCIENTIFIC REPORTSOAAI
SCIENTIFIC REPORTSIF3.9
OAAI分享收藏Susceptibility to Agrobacterium tumefaciens and cotyledonary node transformation in short-season soybeanPLANT CELL REPORTS
PLANT CELL REPORTSIF4.5
PREAI
分享收藏分享收藏
查看更多内容