Induction of hairy roots and plant regeneration from the medicinal plant Pogostemon Cablin

An efficient hairy root induction system for an important endangered medicinal plant, Dracocephalum kotschyi, was developed through Agrobacterium rhizogenesmediated transformation by modifying the co-cultivation medium using five bacterial strains, A4, ATCC15834, LBA9402, MSU440, and A13 (MAFF-02-10266). A drastic increase in transformation frequency was observed when a Murashige and Skoog medium lacking NH 4 NO 3 KH 2 PO 4, KNO 3 and CaCl 2 was used, resulting in hairy root induction frequencies of 52.3 %, 69.6 %, 48.6 %, 89.0 %, and 80.0 % by A4, A13, LBA9402, MSU440, and ATCC15834 strains, respectively. For shoot induction, hairy roots and unorganized tumors induced by strain ATCC15834 were placed on an MS media supplemented with 0.1, 0.25, 0.5, and 1 mg/l BA plus 0.1 mg/l NAA. The high frequency of shoot regeneration and number of shoot were obtained in the medium containing 0.25 mg/l BA and 0.1 mg/l NAA. Root induction occurred from the base of regenerated shoots on the MS medium supplemented with 0.5 mg/l IBA after 10 days.

Section: Shoot Regeneration From Hairy Root Culturessupporting

confidence: 87%

“…Putative transgenic regenerated plants from hairy roots could be formed directly or indirectly (via callus). He-Ping et al (2011) successfully reported adventitious shoot formation from Pogostemon cablin hairy roots. Also, regenerated shoots from the calli of hairy roots were formed in Malus baccata (Wu et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Hairy root induction and plant regeneration of medicinal plant Dracocephalum kotschyi

Sharafi

Sohi

Azadi

et al. 2014

“…Similar results have been drawn in another study on the bottle gourd that explains BA is essential for shoot bud formation (Saha et al 2007). In our experiments, shoot formation was not detected on the hormone-free medium, which is in consistence with the results obtained by Pogostemon cablin and Dracocephalum kotschyi (He-Ping et al 2011;Sharafi et al 2014), respectively. In summary, the insertion of native A. rhizogenes T-DNA into the O. vulgare genome worked successfully.…”

Section: Molecular Analysis Of Transgenic Hairy Rootssupporting

confidence: 92%

Efficient genetic transformation and regeneration system from hairy root of Origanum vulgare

Habibi

Silva

et al. 2016

Origanum vulgare L is commonly known as a wild marjoram and winter sweet which has been used in the traditional medicine due to its therapeutic effects as stimulant, anticancer, antioxidant, antibacterial, anti-inflammatory and many other diseases. A reliable gene transfer system via Agrobacterium rhizogenes and plant regeneration via hairy roots was established in O. vulgare for the first time. The frequency of induced hairy roots was different by modification of the co-cultivation medium elements after infection by Agrobacterium rhizogenes strains K599 and ATCC15834. ). The frequency of shoot generation (85.18 %) was achieved in medium fortified with 0.25 mg/L −1 of BA. Shoots were placed on MS medium with 0.25 mg/l IBA for root induction. Roots appeared and induction rate was achieved after 15 days.

“…The TL-DNA rolB gene does not increase auxin levels (Nilsson et al 1993) but increases the sensitivity of plant cells to the growth regulator (Cardarelli et al 1987;Shen et al 1988;Spanó et al 1988;Maurel et al 1991;Welander et al 1998), probably by enhanced binding of the auxin to plant cell membranes (Filippini et al 1994), consequently increasing the rooting efficiency. The changes in sensitivity and content of endogenous growth regulators in plant cells undoubtedly affected the response effect of plant cells to exogenous growth regulators (He-Ping et al 2011).…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, several workers have also used the liquid medium to establish the hairy root growth (He-Ping et al 2011;Pavlov et al 2005;Romero et al 2009;Sivanesan and Ryong Jeong 2009). Few previous studies support this idea that IBA could positively affect growth and secondary metabolism of various medicinal plants (Kim et al 2003;Baque et al 2010).…”

Section: Discussionmentioning

confidence: 99%

The influence of Agrobacterium rhizogenes on induction of hairy roots and ß-carboline alkaloids production in Tribulus terrestris L.

Sharifi

Sattari

Zebarjadi

et al. 2013

We have developed an efficient transformation system for Tribulus terrestris L., an important medicinal plant, using Agrobacterium rhizogenes strains AR15834 and GMI9534 to generate hairy roots. Hairy roots were formed directly from the cut edges of leaf explants 10-14 days after inoculation with the Agrobacterium with highest frequency transformation being 49 %, which was achieved using Agrobacterium rhizogenes AR15834 on hormone-free MS medium after 28 days inoculation. PCR analysis showed that rolB genes of Ri plasmid of A. rhizogenes were integrated and expressed into the genome of transformed hairy roots. Isolated transgenic hairy roots grew rapidly on MS medium supplemented with indole-3-butyric acid. They showed characteristics of transformed roots such as fast growth and high lateral branching in comparison with untransformed roots. Isolated control and transgenic hairy roots grown in liquid medium containing IBA were analyzed to detect ß-carboline alkaloids by High Performance Thin Layer Chromatograghy (HPTLC).Harmine content was estimated to be 1.7 μg g −1 of the dried weight of transgenic hairy root cultures at the end of 50 days of culturing. The transformed roots induced by AR15834 strain, spontaneously, dedifferentiated as callus on MS medium without hormone. Optimum callus induction and shoot regeneration of transformed roots in vitro was achieved on MS medium containing 0.4 mg L −1 naphthaleneacetic acid and 2 mg L −1 6-benzylaminopurine (BAP) after 50 days. The main objective of this investigation was to establish hairy roots in this plant by using A. rhizogenes to synthesize secondary products at levels comparable to the wild-type roots.