Development of a fast and efficient root transgenic system for functional genomics and genetic engineering in peach

Abstract

Peach is an economically import fruit crop worldwide, and serves as a model species of the Rosaceae family as well. However, peach functional genomics studies are severely hampered due to its recalcitrance to regeneration and stable transformation. Here, we report a fast and efficient Agrobacterium rhizogenes-mediated transformation system in peach. Various explants, including leaf, hypocotyl and shoot, were all able to induce transgenic hairy roots, with a transformation efficiency of over 50% for hypocotyl. Composite plants were generated by infecting shoots with A. rhizogenes to induce transgenic adventitious hairy roots. The composite plant system was successfully used to validate function of an anthocyanin-related regulatory gene PpMYB10.1 in transgenic hairy roots, and two downstream genes, PpUFGT and PpGST, were strongly activated. Our stable and reproductive A. rhizogenes-mediated transformation system provides an avenue for gene function assay, genetic engineering, and investigation of root-rhizosphere microorganism interaction in peach. open Scientific RepoRtS | (2020) 10:2836 | https://doi.

Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants

“…Chimeric plants affect the segregation of the transgene to the subsequent generation and reduce the efficiency of recovering stable transgenic lines (Das Bhowmik et al ., 2019). Reporter genes, such as uidA (encoding GUS) (Das Bhowmik et al ., 2019), GFP (Dutt et al ., 2007) and Ds‐RED (Xu et al ., 2020), are often used to determine the uniformity of gene expression on the regenerated shoots for chimera formation. In this study, the formation of chimerism was evaluated for all the transgenic events generated using binary vector pPHP86170 (Figure S1(a)) containing the proDMMV:TagRFP as the visual marker and pPHP92782 (Figure S1(c)) containing the proGM‐EF1A2:Ds‐RED as the visual marker.…”

Creation of Borer Pests Resistance Genetically Engineering Peach (Prunus Persica L.) Plants by Cloning cry1Ab Gene

“…The expected 2035 bp cry1Ab fragment was ampli ed only from genomic DNA isolated from the kanamycin resistant peach plants compared to nontransformed peach plants. The PCR has the potential to become a widespread tool to con rm the presence of a foreign gene such as Vis1 in tomato , GmbZIP2 in soybean (Yang et al 2020) and PpMYB10.1 in peach (Xu et al 2020) in transgenic plant tissues. In the current study out of 60 plants examined from kanamycin resistant peach regenerated plants only 17 gave positive results.…”

Genetic Transformation in Peach (Prunus persica L.): Challenges and Ways Forward

“…Honda and Moriguchi [ 19 ] described a protocol for transient gene expression analysis using protoplasts isolated from immature peach fruits. Xu et al [ 22 ] developed an A. rhizogenes -mediated transformation method to generate transgenic hairy roots from peach shoots to produce composite peach plants with transgenic roots and non-transgenic shoots. They proposed this method for studying root–rhizosphere microorganism interactions in peach and as a method for clonal propagation.…”

“…As reviewed by Giri and Narasu [ 89 ], adventitious shoot regeneration can occur directly from transgenic roots or by moving them to regeneration medium. As recently shown, the transgenic hairy root phenotype has been induced in different peach explants such as leaves, hypocotyls, and shoots using A. rhizogenes strain MSU440 [ 22 ]. The main goal of this study was to optimize a reproducible A. rhizogenes -mediated transformation protocol for gene function and genetic engineering studies in peach.…”