Effects of Anti-ethylene Compounds on Vitrification and Genome Fidelity of Stevia rebaudiana Bertoni

Document Type : Regular issue (Original Article)

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Central Laboratory of Genetic Engineering, Botany and Microbiology Department, Faculty of Science, Sohag University, 82524 Sohag, Egypt

Abstract

SUBCULTURE repeats was essential prerequisite for mass micropropagation in stevia but it accompanied by vitrification of the cultured shoots. Increase the number of shoots/explant without vitrification was detected when nodal segments of in vitro obtained shoots were subcultured twice, three weeks each, on Murashige and Skoog (MS) medium with 0.25mg/L BAP. From the third subculture, shoot multiplication decreased and vitrification increased as the number of subcultures increased; where 40% of the formed shoots of 10th subculture were vitrified. Decrease of water potential by polyethylene glycol (PEG) increased the incidence of shoot vitrification but shortened the period of subculture from three to two weeks delayed its appearance up to sixth subcultures. Also, application of anti-ethylene agents [AgNO3, CoCl2 or salicylic acid (SA)] decreased vitrification problems up to six subcultures. The best results were obtained by 1.7mg/L AgNO3 where verification was reduced without significant reduction in the number of shoots/explant and shoot growth. The equivalent concentration of AgNO3 in nano-particles form (AgNPs) showed a drastic increase in shoot vitrification (20%). When shoots of the 1st, 4th, 7th and 10th subcultures were subjected to genome amplification using RAPD or ISSR primers, the registered polymorphism under the influence of 1.7mg/L AgNO3 was lower than that of 1.7mg/L AgNPs; both of them were lower than that of shoots cultured on anti-ethylene free medium (control).

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