Production of transgenic Aralia elata regenerated from Agrobacterium rhizogenes-mediated transformed roots

Spontaneous shoot regeneration was observed from Agrobacterium rhizogenes-induced hairy roots of Plumbago indica when these were incubated in liquid MS medium for a period of 3 weeks under continuous light. Insertion of the rolB gene in putative transformed plants was confirmed by PCR and sequencing. Transformed plants grown for a period of 1 week on solid MS medium containing 0.5 mg l -1 6-benzyladenine and then transplanted to growth regulator-free medium showed better overall growth than control plants. Transformed plants had a higher root bio-biomass and an increased plumbagin content relative to non-transformed plants.

Section: Resultsmentioning

confidence: 99%

Regeneration of transformed plants from hairy roots of Plumbago indica

Gangopadhyay

Chakraborty

Bhattacharyya

et al. 2010

“…Several differences were observed in the morphology and biomass of SD-and HR-R. glutinosa plants, most notably, a considerable increase in root biomass of the HR-plants. This is one of the most commonly observed properties of pRi-transformed regenerants, which was found also in such transgenic plants as Catharanthus roseus (Choi et al 2004), T. indica (Chaudhuri et al 2006), Aralia elata (Kang et al 2006) and P. indica (Gangopadhyay et al 2010). The increase is due to the influence of the introduced rol genes on hormonal synthesis and sensitivity.…”

Section: Discussionmentioning

confidence: 53%

Iridoid and phenylethanoid glycoside production and phenotypical changes in plants regenerated from hairy roots of Rehmannia glutinosa Libosch.

Piątczak

Kuźma

Skała

et al. 2015

Spontaneous shoot regeneration was obtained from the hairy roots of Rehmannia glutinosa cultured in liquid woody plant medium for 4 weeks in darkness after induction by Agrobacterium rhizogenes strain A4. These shoots were proliferated on solid Murashige and Skoog (MS) medium in 300 mL glass jars and in liquid MS medium in a 5 L nutrient sprinkle bioreactor. These media were supplemented with 0.1 mg L -1 of indole-3-acetic acid (IAA) and 1.0 mg L -1 of 6-benzylaminopurine. All shoots became rooted on MS agar medium containing IAA or indole-3-butyric acid within 4 weeks. The plantlets were then transferred into soil and grew in a greenhouse. The transgenic nature of the plants was confirmed by polymerase chain reaction (PCR) and reverse-transcriptase PCR analyses. The pRi-regenerated transformants demonstrated a few phenotypical alterations. Production of six iridoid (catalpol, aucubin, loganin, catalposide, harpagide, harpagoside) and two phenylethanoid (verbascoside and isoverbascoside) glycosides was determined in shoots cultured in vitro and leaves and roots of 6-and 12-month-old plants by ultra high performance liquid chromatography. The shoot culture was able to synthesize low amounts of catalpol, verbascoside and isoverbascoside. Leaves and roots of pRi-regenerants accumulated similar (harpagoside, isoverbascoside), higher (catalposide, aucubin, harpagide, verbascoside) or lower (catalpol, loganin) levels of the metabolites in comparison with untransformed R. glutinosa plants and their content varied with the age of the plants and the organ analyzed. However, the increase in shoot and root biomass of transformed plants resulted in a significantly greater yield of all analyzed compounds.

“…Moon et al (1998) induced somatic embryogenic callus from the winter buds of 10-year-old trees incubated on Murashige and Skoog (MS) medium basal salts (Murashige and Skoog 1962) and containing 2,4-dichlorophenoxyacetic acid (2,4-D) for a period of 1 month. Kang et al (2006) induced embryogenic callus from hairy roots cultured under the same conditions as described above. Amemiya and Mochizuki (2002) obtained somatic embryogenic callus from leaflet petioles in early spring after culture on MS medium with 1.0 mg/l 2,4-D and 0.5 mg/l 6-benzyladenine (BA) for 1 month.…”

Section: Shmentioning

confidence: 99%

Rapid and repetitive plant regeneration of Aralia elata Seem. via somatic embryogenesis

JinLing

Tan

et al. 2010

In this work, we established a rapid and repetitive plant regeneration system for Aralia elata Seem. via primary and secondary somatic embryogenesis. Primary somatic embryogenesis was induced using leaf disks, petiole, and root segments, individually cultured for 5 weeks on Schenk and Hildebrandt (SH) (1972) medium with 0-5.0 mg/l indolebutyric acid (IBA). Our investigation demonstrated that optimal IBA concentrations of 3.0, 2.0, and 0.3 mg/l resulted in 100% somatic embryogenesis rates and averages of 11.3, 10.0, and 8.6 somatic embryos per explant for leaf disks, petiole, and root segments, respectively. The primary somatic embryos were used to conduct secondary somatic embryogenesis and the following treatments, in a gradient series, were examined: 0.3-4.0 mg/l IBA, 10-70 g/l sucrose and 0.2-3.0 mg/l abscisic acid (ABA). The results indicated that IBA was more effective than sucrose and ABA, and 3.0 mg/l IBA was the most suitable concentration for secondary somatic embryogenesis. Histological preparations indicated a multi-cellular origin of secondary somatic embryos and different morphological developmental stages during secondary somatic embryogenesis. Primary and secondary somatic embryos germinated readily and developed into normal plantlets after 2 weeks in woody plant medium (WPM, Lloyd and McCown 1980) with 20 g/l sucrose. At 4-5 cm in length, plantlets were transferred to soil (1:1 v/v of peat moss and sand) and the survival rate was 89% after 4 weeks under greenhouse conditions. This system provides a viable contribution to A. elata gene transformation, breeding and regeneration.