Optimization of transient gene expression system in Gerbera jemosonii petals

Abstract

Low transformation efficiency and long generation time for production of transgenic Gerbera jemosonii plants leads to vulnerable gene function studies. Thus, transient expression of genes would be an efficient alternative. In this investigation, a transient expression system for gerbera petals based on the Agrobacterium infiltration protocol was developed using the reporter genes β-glucuronidase (gus) and green florescence protein (gfp). Results revealed the incapability of using the gfp gene as a reporter gene for transient expression study in gerbera flowers due to the detection of green fluorescent color in the non-infiltrated gerbera flower petals. However, the gus reporter gene was successfully utilized for optimizing and obtaining the suitable agroinfiltration system in gerbera flowers. The expression of GUS was detectable after three days of agroinfiltration in gerbera cultivars "Express" and "White Grizzly" with dark pink and white flower colors, respectively. The vacuum agroinfiltration protocol has been applied on the cultivar "Express" for evaluating the transient expression of the two genes involved in the anthocyanin pathway (iris-dfr and petunia-f3' 5'h), which is responsible for the color in flowers. In comparison to the control, transient expression results showed change in the anthocyanin pigment in all infiltrated flowers with color genes. Additionally, blue color was detected in the stigma and pollen grains in the infiltrated flowers. Moreover, blue colors with variant intensities were observed in produced calli during the routine work of stable transformation with f3' 5'h gene.

Genetic engineering of novel yellow color african violet (Saintpaulia ionantha) produced by accumulation of Aureusidin 6-O-glucoside

“…After injection of gene constructs, they conducted to produce delphinine anthocyanin in petals of Gerbera flower (up to 44 and 75%, correspondingly), resulted in color change of petals into blue. In a similar research, the transfer of gene construct containing F3ʹ5ʹH and DFR gene from respective Petunia hybrida and Iris to the petals of Gerbera jamesonii caused a change in the level of anthocyanins among the injected petals and turned the stigma and pollen into blue [34]. Shang et al [35] investigated the effect of RNAi agroinflitration of chalcone synthase gene on the A. majus petals.…”

In planta Female Flower Agroinfiltration Alters the Cannabinoid Composition in Industrial Hemp (Cannabis sativa L.)

“…In planta flower agroinfiltration has been performed for the study of the flower petal color in ornamental plants (Shang et al, 2007 ; Hussein et al, 2013 ; Ratanasut et al, 2014 ; Fresquet-Corrales et al, 2017 ; Zeinipour et al, 2018 ). Unlike previous studies on in planta flower agroinfiltration, we performed agroinfiltration on female hemp flowers because it produces a high amount of cannabinoids in stalked glandular trichomes: a cellular metabolite factory (Tanney et al, 2021 ), which is abundant on the calyces and bracts in the female flower.…”

Transient Expression of Biologically Active Anti-rabies Virus Monoclonal Antibody in Tobacco Leaves

“…In this study, for producing human anti-rabies MAb SO57, agroinfiltration technique was employed as a preferred technique for transient expression in tobacco leaves ( 22 , 28 ). We used codon adaptation index of the tobacco plant to optimize antibody-coding genes (i.e., heavy and light chains) and utilized the RNA silencing suppressor strategy to enhance the expression of the cloned genes.…”