An efficient system to produce transgenic plants via cyclic leave-originated secondary somatic embryogenesis in Rosa rugosa

Abstract

An efficient and reproducible Agrobacteriummediated transformation system via repetitive secondary somatic embryogenesis was developed for Rosa rugosa 'Bao white'. Somatic embryogenesis was induced from in vitro-derived unexpanded leaflet explants on MS medium supplemented with 4.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 0.05 mg/L Kinetin and 30 g/L glucose. Secondary somatic embryos were successfully proliferated via cyclic secondary somatic embryogenesis on MS medium containing 1.0 mg/L 2,4-D, 0.01 mg/L 6-benzyladenine and 45 g/L glucose under light intensity of 500-1,000 lux. The highest germination rate (86.33 %) of somatic embryos was observed on 1/2-strength MS medium containing 1.0 mg/L BA. Relying on the repetitive secondary somatic embryogenesis and A. tumefaciens strain EHA105 harboring the binary vector pBI121, a stable and effective Agrobacterium-mediated transformation pattern was developed. The presented transformation protocol, in which somatic embryo clumps at globular stage (0.02-0.04 g) were infected by Agrobacterium for 60 min and co-cultivated for 2 days, and then selected under a procedure of 3 steps, were confirmed to be optional by GUS histochemical assay and Southern blot analysis. The procedure described here will be very useful for the introgression of desired genes into R. rugosa 'Bao white' and the molecular analysis of gene function.

RrMYB5‐ and RrMYB10‐regulated flavonoid biosynthesis plays a pivotal role in feedback loop responding to wounding and oxidation in Rosa rugosa

“…For transformation, the CDSs of RrMYB5 and RrMYB10 were recombined into pCAMBIA 2300s and then transformed into A. tumefaciens EHA105. The Rosa rugosa and tobacco transformation was performed as previously described by us (Ning et al ., ; Xing et al ., ). After 2‐month culture, somatic embryos of R. rugosa were sampled to analyse gene expression and measure the content of PAs.…”

“…Rosa rugosa is a commercially important ornamental plant cultivated worldwide, whose populations suffer from herbivorous insects, powdery mildew and blackspot. These stresses negatively impact Rosa plant growth, flower quality and field performance (Xing et al ., ). Wounding stress induces the accumulation of important bioactive compounds in wounded tissues (Ioannidi et al ., ; Torres‐Contreras et al ., ).…”

A conservative pathway for coordination of cell wall biosynthesis and cell cycle progression in plants

“…Agrobacterium tumefaciens EHA105 and GV3101 were transformed with the resulting plasmids. The A. thaliana , R. rugosa and tobacco transformations were performed as previously described (Ning et al ., 2012; Wang et al ., 2017; Xing et al ., 2014). After 6–8 months of tissue culture, gene expression was analyzed in the germinated shoots.…”

Indução e histologia de embriões somáticos primários e secundários do híbrido Phalaenopsis Classic Spotted Pink (Orchidaceae)

“…Esse comportamento está associado ao fato da luz promover a regeneração do embrião em plantas, logo fatores como intensidade e qualidade (da luz) alteram as concentrações endógenas de fitoreguladores, espessura dos tecidos e atuam na síntese de pigmentos (Xing et al, 2014).…”