Auxin-cytokinin synergism in vitro for producing genetically stable plants of Ruta graveolens using shoot tip meristems

Abstract

An efficient micropropagation protocol was developed for Linn. using shoot tip meristems derived from a 4-month-old field grown plant. shoot regeneration and proliferation was accomplished on Murashige and Skoogs (MS) semi-solid medium in addition to different doses of cytokinins viz.6- benzyl adenine (BA), Kinetin (Kn) or 2-isopetynyl adenine (2iP), singly or in combination with auxins viz. indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA). Highest regeneration frequency (27.6%) was obtained on (MS) medium composed of BA (10 µM) with maximum number (9.4) of shoots and 4.3 cm shoot length after 4 weeks of incubation. Among various combinations tried best regeneration frequency (71%) of multiple shoot formation with highest number (12.6) of shoots per shoot tip explants were achieved in MS medium augmented with a combination BA (10.0 µM) and NAA (2.5 µM) after 4 weeks of incubation. The optimum frequency (97%) of rhizogenesis was achieved on half-strength MS medium having 0.5 µM IBA after 4 weeks of incubation. Tissue culture raised plantlets with 5-7 fully opened leaves with healthy root system were successfully acclimatized off in Soilrite™ with 80% survival rate followed by transportation to normal soil under natural light. Genetic stability among raised progeny was evaluated by ISSR and RAPD markers. The entire banding pattern revealed from regenerated plants was monomorphic to the donor. The present protocol provides an alternative option for commercial propagation and fruitful setting up of genetically uniform progeny for sustainable utilization and germplasm preservation.

Optimizing micropropagation conditions for a recalcitrant ninebark (Physocarpus opulifolius L. maxim.) cultivar

“…Low concentration of NAA stimulated the cytokinininduced proliferation in ninebark. Similarly, a synergetic influence of BA and NAA was noticeable in Ruta graveolens when the combinations of the most favorable concentration of BA with variant concentrations of NAA were tested: supplementation of NAA resulted in a better shoot proliferation rate and the best regeneration (Faisal et al 2018). In Juglans regia the best proliferation results were obtained with 1 mg•L −1 of BA plus 0.01 mg•L −1 of IBA (Kepenek and Kolagasi 2016).…”

“…Cytokinins are adenine derivatives capable of inducing cell divisions in tissue cultures, initiating shoots and stimulating their proliferation and further growth, both of which are critical for in vitro multiplication efficiency. Exogenously applied cytokinins are more active in the presence of an auxin (Perveen et al 2011;Faisal et al 2018;Nowakowska et al 2019). Frequently used cytokinins are of natural origin such as zeatin (Zea), the 2-isopenthenyladenine (2iP), kinetin (Kin), and 6-benzyladenine (BA) or synthetic cytokinins such as thidiazuron (TDZ) (Pierik 1997).…”

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“…MS medium augmented with different concentration (0.5, 2.5, 4.5, 6.5 or 8. IBA and NAA (0.05, 0.10, 0.50, 1.00 or 1.50 µM) or different concentration (10,15,20,25,30,35,40 or 45 μM) of AgNO3 were used for shoot proliferation and multiplication. Cultures were transferred onto fresh media after every 21 days.…”

The use of tissue cultures in the mass production of Heuchera‘Silver Scrolls’

“…During micropropagation, cytokinins act synergistically with other growth regulators, such as, for example, auxins, which are indispensable in rhizogenesis (Faisal et al 2018). Martin (2003 compared the efficiency of naphtylacetic acid (NAA), indolil-3-butyric acid (IBA) and indolil--3-acetic acid (IAA) in the in vitro rooting of Rotula aquatica Lour.…”