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۱Dopamine Production in Hairy Root Cultures of Portulaca oleracea (Purslane) Using Agrobacterium rhizogenes
اطلاعات انتشار: Journal of Agricultural Science and Technology، شانزدهم،شماره۲، ۲۰۱۴ ، سال
تعداد صفحات: ۱۲
An efficient transformation system for the medicinal plant Portulaca oleracea was established using agropine–type Agrobacterium rhizogenes ATCC15834. Hairy roots were obtained directly from cotyledon leaves explants seven days after inoculation with the bacteria. The highest transformation efficiency was obtained from cotyledon leaves explants, and amounted to 53.3% within two weeks. Roots grew rapidly on solid growth regulator free ½ Murashige and Skoog medium and demonstrated characteristics of transformed roots such as fast growth and high lateral branching. Successful and stable transfer of rolB gene was illustrated by PCR using specific primers of the gene. The hairy roots showed an ability to synthesize natural and medicinal product, dopamine. Elicitation of dopamine production in P. oleracea hairy roots was tested using different concentrations of methyl jasmonate (0, 100, 150, 200 μM) and salicylic acid (0, 125, 250, 500 μM), added to the hairy root cultures during the late growth phase. The results showed that the various concentrations of the methyl jasmonate significantly increased the dopamine content, but, at concentration of 100 μM, its impact was the most pronounced. Salicylic acid had no significant influence on dopamine production in hairy roots of P. oleracea.

۲Molecular Cloning and Characterization of a Cyclotide Gene Family in Viola modesta Fenzl
اطلاعات انتشار: Journal of Agricultural Science and Technology، هفدهم،شماره۶، ۲۰۱۵، سال
تعداد صفحات: ۱۳
Cyclotides are small disulfide–rich proteins that have the unusual feature of a cyclic backbone. Cyclotides have a range of interesting biological activities and are found in a variety of tropical plants from the Rubiaceae, Violaceae, Cucurbitaceae and Fabaceae families. We have cloned and characterized cyclotides in Viola modesta, a Viola species native to western Asia, which was collected from the Kurdistan Province of Iran. Fifteen cyclotide sequences were obtained using homology based PCR strategy. Sequence analysis showed that 14 of them had continued open reading frames and showed high level of similarity to cyclotide genes from other species of the Violaceae. After analyzing the full endoplasmic reticulum signals of V. modesta cyclotides, two conserved sequences, AAFALPA and ATAFALP, were detected. Analysis of isolated cyclotide sequences showed that they all belonged to bracelet family and were separated into two subclasses. Phylogenetic analysis of cyclotide genes from V. modesta and other Viola species revealed that most V. modesta genes showed close relationship with their homologs from the Violaceae, while the V. modesta genes formed two separate clades. Transcription analysis by semi–quantitative RT–PCR revealed that Vmcyc1 and Vmcyc7 were differentially expressed in all tested tissues including roots, stems, leaves, flowers, seeds, peduncles, and capsules with the highest transcript level in the capsules.
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