Morphogenetic and chemical stability of long-term maintained<i>Agrobacterium</i>-mediated transgenic<i>Catharanthus roseus</i>plants

Verma, Priyanka; Sharma, Abhishek; Khan, Shahanavaj; Mathur, Anmol; Shanker, Karuna

doi:10.1080/14786419.2014.940348

Cited by 19 publications

(8 citation statements)

References 32 publications

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“…Although spontaneous deletion of T-DNA (Hänisch et al 1990;Taneja et al 2010), somaclonal variation (Sevón et al 1998) and diminished expression of transgene (Verma et al 2015) among the Ri-transformed plants are reported, in our study, even after long-term culture in laboratory conditions (4 years), and after transfer to soil (1 year), the transgenes in different B. monnieri plant lines are found to be stably integrated and expressed.…”

Section: Discussioncontrasting

confidence: 40%

“…High bacoside accumulation capacity remains the significant attribute of the crypt-transformed plant lines. Retention of the ability of higher secondary metabolite accumulation in transgenic plants over non-transformed plants has been reported in P. graveolens (Saxena et al 2007), T. indica (Roychowdhury et al 2013a), and Catharanthus roseus (Verma et al 2015).…”

Section: Discussionmentioning

confidence: 96%

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Effects associated with insertion of cryptogein gene utilizing Ri and Ti plasmids on morphology and secondary metabolites are stable in Bacopa monnieri-transformed plants grown in vitro and ex vitro

Paul

Sarkar

Jha

2015

Plant Biotechnol Rep

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Bacopa monnieri (Linn.) Wettst. (family Scrophulariaceae), a therapeutically important perennial herb, was transformed to study the stability of the effects associated with the insertion of a gene encoding b-cryptogein, a proteinaceous elicitor, either alone or in addition to the T-DNA genes in long-term in vitro culture as well as after transfer to the greenhouse. Plant lines BmAr-IXcrypt obtained via transformation by LBA 9402-crypt (pRi1855?pBin19??crypt) showed integration and expression of rolA, rolB, rolC, and rolD genes after 4 years of maintenance in vitro. The plant line BmAt-ncrypt obtained via transformation with A. tumefaciens strain LBA4404-crypt (harboring pBin19??crypt) as well as plant line BmAr-IXcrypt showed stable integration and expression of nptII gene and crypt gene even after transfer to greenhouse. The clones of Ri-crypt-transformed plants differed morphologically from Ri-transformed plants in that the typical ''hairy root syndrome'' was not observed in plants expressing crypt gene in the presence of rol genes. Except plant line BmAr-IXcrypt, none of the other transgenic plant lines showed any alteration in floral morphology and onset of flowering. The crypt-transformed plant lines (BmAr-IXcrypt and BmAt-ncrypt), maintained under long-term culture, showed significantly higher (p B 0.05) amount of bacoside production in vitro (1.66-to 2.05-fold, with respect to non-transformed plants). In addition, accumulation of all the four bacosides was significantly higher (p B 0.05) in crypt-transformed plants as compared to non-transformed plants grown for 1 year in greenhouse. Thus, the crypt-transformed plant lines of B. monnieri may be considered as the potential source for sustainable bioproduction of bacosides.

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Section: Discussioncontrasting

confidence: 40%

Section: Discussionmentioning

confidence: 96%

Effects associated with insertion of cryptogein gene utilizing Ri and Ti plasmids on morphology and secondary metabolites are stable in Bacopa monnieri-transformed plants grown in vitro and ex vitro

Paul

Sarkar

Jha

2015

Plant Biotechnol Rep

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“…They were showed that alkaloid vindoline accumulation of the transgenic plants were two times more compared to non-transformed ones. Their alkaloid profile was almost constant during 5 years [39]. In this study, suspension cells originated from both long-term and short-term cultured calli were negatively affected by 24 hour elicitation.…”

Section: Initiation Of Cell Suspension Cultures and Elicitormentioning

confidence: 56%

Identification and Production of Phenolic Nicotiflorin in Astragalus Chrysochlorus Callus

Turgut-Kara¹

2018

FARMACIA

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Astragalus chrysochlorus Boiss. & Kotschy (2n = 16) is one of the rare Turkish endemic species and it is listed in the Red Data Book of Turkish Plants as endangered. This species has been used traditionally for its wound healing properties and a crude ethanol extract prepared from the roots exhibits antioxidant and cytotoxic activities. In this study, a detailed phytochemical analysis was performed on A. chrysochlorus calli that resulted in the isolation of a major constituent. The purified molecule's structure elucidation was completed by spectral methods [nuclear magnetic resonance (NMR) and mass spectrometry (MS)], which revealed a rarely encountered the flavonoid in the Astragalus genus, nicotiflorin. In order to increasing nicotiflorin content in the callus cultures, the effects of culturing time and elicitor treatment were investigated. The HPLC analyses showed that the maximal production of nicotiflorin occurred with long-term cultured (13 years old) callus as 4775 µg/g dry weight (DW), whereas it was 132 µg/g DW for short-term cultured (2 months old) ones. Then, the 24 h treatment of the yeast extract that was used as biotic elicitor had negative effect on the production of nicotiflorin. The data obtained from this study could be significant for the mass production of nicotiflorin from long term in vitro cultured A. chrysochlorus callus. RezumatAstragalus chrysochlorus Boiss. & Kotschy (2n = 16) este una dintre speciile endemice din Turcia și este încadrată în lista roșie ca fiind pe cale de dispariție. Această specie a fost folosită în mod tradițional pentru proprietățile sale de vindecare a rănilor, iar un extract etanolic a fost preparat din rădăcini și prezintă activitate antioxidantă și citotoxică. În acest studiu s-a efectuat o analiză fitochimică detaliată pe A. chrysochlorus calli care a dus la izolarea unui constituent major. Elucidarea moleculei purificate a fost completată prin metode spectrale (rezonanță magnetică nucleară (NMR) și spectrometrie de masă (MS)), care au evidențiat flavonoidul nicotiflorin. Pentru a crește conținutul de nicotiflorin în culturile de calus, s-au investigat efectele timpului de cultivare și tratamentul elicitor. Analizele HPLC au arătat că producția maximă de nicotiflorin a apărut la calusul de lungă durată (13 ani), 4775 µg/g DW, în timp ce acesta a fost de 132 µg/g DW pentru culturile de scurtă durată (cu vârsta de 2 luni).

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“…The success of genetic engineering depends on genetic and chemical stability of transgenic plants in subsequent generations ( Verma et al, 2015 ). The copy number of integrated gene affects the expression level and genetic stability in transgeniclines.…”

Section: Discussionmentioning

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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Morphogenetic and chemical stability of long-term maintainedAgrobacterium-mediated transgenicCatharanthus roseusplants

Cited by 19 publications

References 32 publications

Effects associated with insertion of cryptogein gene utilizing Ri and Ti plasmids on morphology and secondary metabolites are stable in Bacopa monnieri-transformed plants grown in vitro and ex vitro

Effects associated with insertion of cryptogein gene utilizing Ri and Ti plasmids on morphology and secondary metabolites are stable in Bacopa monnieri-transformed plants grown in vitro and ex vitro

Identification and Production of Phenolic Nicotiflorin in Astragalus Chrysochlorus Callus

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

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