Direct shoot bud organogenesis and plant regeneration from pre-plasmolysed leaf explants in Catharanthus roseus

Verma, Priyanka; Mathur, Anmol

doi:10.1007/s11240-011-9936-4

Cited by 25 publications

(11 citation statements)

References 37 publications

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“…IBA has been effective in the induction of adventitious roots in regenerated shoots of many plant species including Catharanthus roseus (Verma and Mathur 2011). However, in this study, the elongated in vitro-derived shoots of B. sinuspersici did not develop roots on media containing IBA.…”

Section: Discussioncontrasting

confidence: 74%

Multiple shoot induction and plant regeneration of the single-cell C4 species Bienertia sinuspersici

Northmore

Zhou

Chuong

2011

Plant Cell Tiss Organ Cult

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Direct organogenesis provides an efficient means of propagating plants in vitro without the necessity of growing callus cultures. Shoot regeneration was achieved from bud explants of Bienertia sinuspersici on Murashige and Skoog's media supplemented with 6-benzyladenine (BA). The frequency of regeneration varied based on the concentration of BA used. The highest rate of new shoot induction was observed when explants were cultured on media containing 4.4 lM BA after three weeks. Shoots were successfully elongated on media containing gibberellic acid with the highest shoot elongation observed at 2.9 lM. Elongated shoots developed adventitious roots in media without plant growth regulator and were subsequently transplanted to soil. Light microscopic analysis of cross sections of leaves from regenerated shoots showed similar intracellular cytoplasmic compartmentalization of organelles in chlorenchyma cells compared to those observed in leaves grown under greenhouse conditions. Moreover, Western blot analysis detected high expression of key C 4 photosynthetic enzymes, suggesting the C 4 cycle is maintained in the single cell system. We have developed a procedure for whole plant regeneration via direct organogenesis in B. sinuspersici.

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Section: Discussioncontrasting

confidence: 74%

Multiple shoot induction and plant regeneration of the single-cell C4 species Bienertia sinuspersici

Northmore

Zhou

Chuong

2011

Plant Cell Tiss Organ Cult

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“…In vitro grown hypocotyl and nodal explants were used for generation of G(G)PPS and G(G)PPS + GES transgenic lines, respectively. While G(G)PPS transformants were generated by indirect regeneration using hypocotyl explants according to Wang Q. et al (2012) , plants co-expressing G(G)PPS + GES were generated by direct regeneration using nodal explants following the method reported in Verma and Mathur (2011a) .…”

Section: Methodsmentioning

confidence: 99%

Terpene Moiety Enhancement by Overexpression of Geranyl(geranyl) Diphosphate Synthase and Geraniol Synthase Elevates Monomeric and Dimeric Monoterpene Indole Alkaloids in Transgenic Catharanthus roseus

2018

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Catharanthus roseus is the sole source of two of the most important anticancer monoterpene indole alkaloids (MIAs), vinblastine and vincristine and their precursors, vindoline and catharanthine. The MIAs are produced from the condensation of precursors derived from indole and terpene secoiridoid pathways. It has been previously reported that the terpene moiety limits MIA biosynthesis in C. roseus. Here, to overcome this limitation and enhance MIAs levels in C. roseus, bifunctional geranyl(geranyl) diphosphate synthase [G(G)PPS] and geraniol synthase (GES) that provide precursors for early steps of terpene moiety (secologanin) formation, were overexpressed transiently by agroinfiltration and stably by Agrobacterium-mediated transformation. Both transient and stable overexpression of G(G)PPS and co-expression of G(G)PPS+GES significantly enhanced the accumulation of secologanin, which in turn elevated the levels of monomeric MIAs. In addition, transgenic C. roseus plants exhibited increased levels of root alkaloid ajmalicine. The dimeric alkaloid vinblastine was enhanced only in G(G)PPS but not in G(G)PPS+GES transgenic lines that correlated with transcript levels of peroxidase-1 (PRX1) involved in coupling of vindoline and catharanthine into 3′,4′-anhydrovinblastine, the immediate precursor of vinblastine. Moreover, first generation (T1) lines exhibited comparable transcript and metabolite levels to that of T0 lines. In addition, transgenic lines displayed normal growth similar to wild-type plants indicating that the bifunctional G(G)PPS enhanced flux toward both primary and secondary metabolism. These results revealed that improved availability of early precursors for terpene moiety biosynthesis enhanced production of MIAs in C. roseus at the whole plant level. This is the first report demonstrating enhanced accumulation of monomeric and dimeric MIAs including root MIA ajmalicine in C. roseus through transgenic approaches.

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“…Following the direct shoot bud organogenesis procedure described earlier (Verma and Mathur 2011b), a total of 120 multiple shoot cultures were raised from pre-plasmolysed leaf explants of three C. roseus varieties namely cv.…”

Section: Plant Materialsmentioning

confidence: 99%

“…However, baring a few reports on genetically modifying C. roseus for over-expressing pathway genes at whole plant/tissue level of cellular differentiation (Choi et al 2004;Zarate and Verpoorte 2007;Verma and Mathur 2011a;Verma et al 2012b), the bulk of research in this area has largely been carried out with cell suspension and hairy root cultures that do not provide the requisite cellular complexity necessary for expressing the complete TIAs pathway. Authors presented the first report on the development of an efficient direct regeneration protocol via adventitious shoot bud organogenesis from leaf explants in C. roseus (Verma and Mathur 2011b). The developed protocol has also been found suitable for high frequency recovery of Agrobacterium tumefaciens-mediated transgenic plants of C. roseus (Verma and Mathur 2011a).…”

Section: Introductionmentioning

confidence: 99%

Tryptophan metabolism and evaluation of morphological, biochemical and molecular variations in a field grown plant population derived via direct adventitious shoot bud regeneration from pre-plasmolysed leaves of Catharanthus roseus

Verma

Khan

Mathur

et al. 2015

Plant Cell Tiss Organ Cult

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Sixty six plants raised via direct shoot bud organogenesis from pre-plasmolysed leaf explants of Catharanthus roseus were assessed under in vivo conditions for their physio-morpho traits, tryptophan metabolism, genetic fidelity and alkaloid profile. Morphologically all plants were parent like except the three morpho-types i.e. broad cup-shaped leaves, dwarf phenotype and enlarged pink corona. Vindoline was detected in most of the plants in substantial amount while ajmalicine, ajmaline and catharanthine were detected in few plants only. More than eightfold increased vindoline (0.08 % dry wt.) was showed by plant no. 1, 4 and 39. Except plant no. 54, which accumulated the highest alkaloid content of 3.09 % dry wt. with tryptophan content of 0.0283 % dry wt., majority of the plants showed a negative correlation between total alkaloid content and tryptophan accumulation and strong positive correlation between tryptophan content and 5-methyltryptophan tolerance. ISSR and RAPD profile of seventeen randomly selected, field established plants was generated by using 10 ISSR and 60 RAPD primers. In RAPD, a total of 753 bands were detected, out of which 624 (82.87 %) were monomorphic. In ISSR profiling, a total of 205 bands were detected out of which 200 bands were monomorphic (97.56 %) and only 5 bands were found to be polymorphic. Highly significant data for the monomorphic banding pattern across all the three genotypes (p \ 0.01) was observed. Principal components, AMOVA analysis and multivariate analysis using Nei and Li's coefficient further validate the monomorphism. Thirteen plants having superior traits were grown in Random Block Design in field. The relevance of physiological and epigenetic changes occurred in the light of morpho-types and alkaloid profile of directly regenerated plants during in vitro to in vivo acclimatization in C. roseus is discussed.

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Direct shoot bud organogenesis and plant regeneration from pre-plasmolysed leaf explants in Catharanthus roseus

Cited by 25 publications

References 37 publications

Multiple shoot induction and plant regeneration of the single-cell C4 species Bienertia sinuspersici

Multiple shoot induction and plant regeneration of the single-cell C4 species Bienertia sinuspersici

Terpene Moiety Enhancement by Overexpression of Geranyl(geranyl) Diphosphate Synthase and Geraniol Synthase Elevates Monomeric and Dimeric Monoterpene Indole Alkaloids in Transgenic Catharanthus roseus

Tryptophan metabolism and evaluation of morphological, biochemical and molecular variations in a field grown plant population derived via direct adventitious shoot bud regeneration from pre-plasmolysed leaves of Catharanthus roseus

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