Abstract
The effect of different concentrations of NaCl (6, 8 and 10 dS/m) on growth and alkaloids production in 21 days-old callus of Egyptian henbane (Hyoscyamu muticus L.) was studied. Results showed that 10 dS/m of NaCl increased fresh and dry weight by 4.1 and 2.8 fold more than control, respectively. Two fold Total alkaloids increased by 2 fold at 10 dS/m compared to control or wild leaves. In contrary, wild leaves had the highest amount of hyosyamine (2.8 mg/g DW) followed by 6 dS/m stressed-callus (1.14 mg/g DW) as detected by HPTLC. Peroxidase and superoxide dismutase activity were increased by 2.6 and 2.3 fold, respectively in highly salt stressed callus than control while catalase activity (CAT) was decreased. Anthocyanin concentration was increased 3 fold in 10 dS/m of NaCl stressed callus compared to control. Glutathione content was increased by 33.8% under low level of NaCl more than control. Reduction of photosynthetic pigments under all NaCl levels led to decrease the total carbohydrate and protein content. Protein band with molecular weight 109 KDa was a unique band in 10dS/m of NaCl treated callus with high expression of protein bands with MW 44, 38 and 32. Histologically, callus cells under high level of NaCl had amoeboid shape and start in senescence with formation of lysiogenous intercellular spaces.
Highlights
Salt stress is an important abiotic elicitor affecting plant growth, development, morphogenesis and the accumulation of secondary metabolites such as phenols, terpenes and alkaloids (Naik and Alkhayri, 2016; Selmar, 2008; Haghighi et al, 2012)
Water content (WC) of callus was calculated using fresh weight (FW) and dry weight (DW) values according to Henson et al (1981)
Determination of Hyoscyamine concentration: High-Performance Thin-Layer Chromatography (HPTLC) was performed on 20 cm ×10 cm HPTLC silica gel 60 F254 plates (Merck) with a mobile phase consisting of chloroform : methanol : acetone : aqueous ammonia (25%) 75 : 15 : 10 : 1.6 (v/v/v/v) (Jaremicz et al, 2014)
Summary
Salt stress is an important abiotic elicitor affecting plant growth, development, morphogenesis and the accumulation of secondary metabolites such as phenols, terpenes and alkaloids (Naik and Alkhayri, 2016; Selmar, 2008; Haghighi et al, 2012). NaCl stressor had stimulating or inhibiting effect on growth or synthesis of natural products according to its concentration, exposure period and plant species. In this respect, low concentration of NaCl (2000 ppm) enhanced the production of secondary metabolites in Mentha longifolia in vitro cultures but high concentration (6000 ppm) inhibited growth and yellowing cells (El-shennawy et al, 2017). The present study was carried out to investigate the effect of salt stress by different concentration of NaCl on callus growth, histological structure, biochemical parameters, and alkaloid production from Hyoscyamu muticus L
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