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Abstract: The effect of five topolins (meta-Topolin= mT; meta-Topolin riboside= mTR; meta-Methoxy topolin=MemT; meta-Methoxy topolin riboside=MemTR and 6-(meta-methoxy)-9-(tetrahydropyran-2-yl)-topolin=MemTTHP) on the photosynthetic pigments and leaf structures of micropropagated ‘Williams’ bananas was compared with the commonly used benzyladenine (BA). Surface-decontaminated explants were cultured for 70 d on modified Murashige and Skoog (MS) basal medium and supplemented with 10, 20 or 30μM cytokinins (CKs). At 10 d intervals, the photosynthetic pigments were quantified via spectrophotometric methods for 7 cycles. Generally, the maximum pigment content was attained between 40 and 50 d. The control plantlets had the highest pigment content (1150μg/g FW). Among the CKs, 10μM MemTTHP generally had the best pigment stimulatory effect at the same period. After 40 d, scanning electron microscopy (SEM) of the foliar surface showed that the stomata density was highest in 10μM MemTTHP-treated and lowest in 10μM MemTR-treated plantlets. The stomatal structure and pore area also varied with the type and concentration of CK added. Generally, prolonging culture duration as well as increasing CK concentrations reduced the pigment content. However, the drastic breakdown in chlorophyll pigments beyond 50 d was slightly inhibited by the presence of mT, mTR, MemTTHP and BA compared to the control. The CK-treated plantlets at equimolar concentration had comparable chlorophyll a/b and total chlorophyll/carotenoid ratios after 10 d; probably as an adaptive measure. At the end of the current study, 10μM mT and mTR plantlets remained green as reflected by the higher total chlorophyll/carotenoid ratio as well as by the visual observations. A well-developed photosynthetic apparatus enhances the survival of in vitro plantlets during the acclimatization stage. Current findings provide some insight into the role of meta-topolins on photosynthetic parameters in vitro, which inevitably partly contributed to the better acclimatization capability of meta-topolin-regenerants. [Copyright &y& Elsevier]

Highlights: [•] A total of 47 endogenous cytokinin-forms grouped into 9 classes were quantified. [•] Tissue-cultured plants had more endogenous cytokinin levels than acclimatized plants. [•] 9-Glucosides were the major metabolites in tissue-cultured and acclimatized plants. [Copyright &y& Elsevier]

Abstract: Merwilla plumbea (Lindl.) Speta is an important medicinal plant widely used in traditional medicine. We evaluated the effect of five cytokinins [benzyladenine (BA), 2-isopentenyladenine (2iP), meta-topolin (mT), meta-topolin riboside (mTR), and meta-methoxy-9-tetrahydropyran-2-yl-topolin (MemTTHP)] on the level of phenolic acids and antioxidant activity of M. plumbea during the tissue culture and acclimatization stages. Two cytokinins (mT and mTR) significantly improved the antioxidant activity of tissue culture plantlets while the control plantlets were better after acclimatization. Using UPLC–MS/MS, the levels of hydroxybenzoic and hydroxycinnamic acid derivatives (phenolic acids) varied significantly during tissue culture and acclimatization, depending on the cytokinin and plant part analyzed. Vanillic acid (24.9μgg−1) detected in underground parts of tissue culture plants supplemented with BA was the most abundant phenolic acid detected. The current findings indicate that the phytochemicals together with the bioactivity during in vitro propagation of M. plumbea is influenced by the cytokinin type used and the stage of plant material collection. [Copyright &y& Elsevier]