Gorgan Branch 2 Assistant professor in Plant Physiology -Islamic Azad University of Iran -Gorgan Branch 3 Full Professor in Plant Physiology -Islamic

ABSTRACT The present study aimed at evaluation of putrescine treatment on growth factors of soybean (Glycine max L.) under drought stress induced by polyethylene glycol (PEG). Soybean seeds were planted and irrigated with five solutions (control, drought, PEG, Put, and PEG-Put). Then, growth factors (length and weight) of seedlings, root, shoot, stem, hypocotyl, first internode, and plant were measured. Statistical analyses were performed by one-way ANOVA through Duncan Test at p≤0.05 in SPSS (Version 21) in three iterations. Graphs were drawn in Excel Software (Microsoft Office, 2010). The results obtained from the present study showed that putrescine could not have considerable effect to alleviate adverse effect of drought stress caused by PEG in soybean

IN- VITRO CULTURE OF ARTEMISIA AUCHERI BOISS. ON FOUR DIFFERENT TISSUE CULTURE MEDIA FOR COMPARATIVE CYTOTOXIC EFFRCTS AND GROWTH

Objective: Artemisia   genus is   an important medicinal plant in Iran.  The effect of four different culture media was investigated on growth and cytotoxic production of Artemisia aucheri in callus culture after 20th subculture. Methods: Callus induction was initiated from seedling on a Murashige and Skoog (MS) basal medium containing different concentrations of vitamins and combinations of growth regulators. Cytotoxicity of methanolic extract of callus culture grown on different culture media was assessed using the brine shrimp assay. Result: Different concentrations and combinations of phytohormones had no significant influence on callus fresh weight, callus dry weight and percentage of callus dry matter. However, cytotoxic production seems depend on the lowest level of growth index (1.94 ± 0.12) because from among the four methanolic extracts of the four isolated cultures of A. aucheri tested on MS medium supplemented with high level of 6-benzylaminopurine (BAP), naphthalene acetic acid (NAA) and thiamine-HCl was found to have the cytotoxicity (LC50 < 1000µg/ml) using brine shrimp lethality assay. Methanolic extracts of callus culture on MS medium containing kinetin (Kin), indole-3-acetic (IAA) and 2,4-dichlorophenoxy acetic acid (2,4,D) had the highest level of growth index (4.31 ± 0.56) and  no cytotoxic effect was observed. Conclusion: In the establishment of cell culture, high level of BAP, NAA and thiamine-HCl were the best plant growth regulators in supporting the cytotoxic production. This is the first report on the use of phytohormones on the establishment of cell cultures in cytotoxic production of A. aucheri.KEYWORDS: Artemisia aucheri, Artemia franciscana, cytotoxic, tissue culture, methanol extract, growth inde

Phytochemical and Morphological Evidences for Shikonin Production by Plant Cell Cultures of Onosma sericeum Willd

ABSTRACT Shoot regeneration, callus growth, and biosynthesis of shikonin in callus cultures of Onosma sericeum were examined. Plant tissue culture was used as an alternative method for increasing the production of shikonin, a secondary metabolite. The isolated cultures were subjected to abiotic factors such as light, plant growth regulators, and nutritional factors. Identification was carried out by High- Performance Liquid Chromatography (HPLC) after 10th subculture. Nodal explants were incubated in Murashige and Skoog (MS) medium along with different combination of growth hormones. Shoot regeneration from calli were achieved on MS basal medium supplemented with 3 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l Naphthalene acetic acid (NAA) under light cycle. Shikonin was formed in dark culture. Calli grown on MS (ammonium ion-free) medium supplemented with 3 mg/l BAP and 0.5 mg/l NAA contained the maximum shikonin level (15.26 µg/mg DW). Minimum shikonin content (9.85 µg/mg DW) was observed in calli cultured on MS (ammonium ion-free) medium supplemented with 3 mg/l BAP and 0.5 mg/l indole-3-acetic acid (IAA). In establishing cell culture, the ammonium ion, and light cycle inhibited shikonin formation. This is the first report on the establishment of isolated cultures of O. sericeum for shikonin production and callus growth