Abstract
Effects of sodium nitroprusside (SNP; nitric oxide donor) treatment on the enhancement of secondary metabolites production, oxidative stress mediators (O2-.) accumulation and antioxidant defense enzymes of Ginkgo biloba callus culture was investigated. On one hand, the obtained data showed a highly metabolic modification of chemical constituents, PAL activity and various antioxidant defense enzymes (APX, SOD), which gradually increased in response to SNP treatments. On the other hands the high NO levels significantly increased the accumulation of various oxidative burst of O2-.. MS basal medium supplemented with casein hydrolase (500 mg/L), NAA and BA at equal concentration (0.5 mg/L) recorded the highest number of regenerated shoots (4.81 cm) and shoot height (4.96 cm) as well as root number (2.25 cm) and root length (4.5 cm). The highest survival (40 %) was shown in acclimatization on the mixture containing sand, peat moss and vermiculite (1: 1: 1, v/v/v), which significantly confirmed and reflected the variation in survival percentage. Meanwhile, higher treatment (500 μM) of NO positively enhanced secondary products accumulation of total tannins, saponins, phenols and total flavonoids in G. biloba callus 
 culture.
Highlights
Ginkgo biloba has been known as a popular remedy in traditional Chinese medicine for over 4000 years, and it has been a common herbal medicine in Europe since the 1730’s
Establishment of Ginkgo callus culture Callus formation occurred on young leaves excised from G. biloba tree
The results demonstrate that nitric oxide (NO) signaling is involved in reactive oxygen species (ROS) induced secondary products production of G. biloba cell suspension culture
Summary
Ginkgo biloba has been known as a popular remedy in traditional Chinese medicine for over 4000 years, and it has been a common herbal medicine in Europe since the 1730’s. SNP at high concentrations stimulated catharanthine formation of Catharantuse roseus cells, but inhibited the growth of the cells (Xu et al, 2009). It stimulated production of hypericin by Hypericum perforatum. The cells were significantly increased after elicitor treatment by NO, achieving 4.2-fold higher over the control (Xu et al, 2005) Antioxidant defense enzymes such as superoxide distumase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR) and monodehydroascorbate reductase (MDAR) are the systems designed to minimize the concentrations of superoxide and hydrogen peroxide. Many previous studies had reported the presence of NO in the plant kingdom and its involvement in growth, development and defense responses (Beligni and Lamattina 1999). This study was conducted to investigate the molecular role of nitric oxide in secondary products production in G. biloba cell suspension culture
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