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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.

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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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Exendin-4 is a human Glucagon-Like Peptide-1 (GLP-1) analogue, resistant to DiPeptidyl Peptidase (DPP), which activates the GLP-1 receptor, increases insulin secretion, and improves glycemic control. In this study, Exendin-4 (EX4) was fused to Cholera Toxin B subunit (CTB) and transiently expressed in tobacco leaves. The sequence of the Ex4 fused to CTB subunit gene, with BamHI and SacI restriction enzymes sites at the beginning of CTB and at the end of EX4 gene. After codon optimization, the sequence was synthesized and cloned in pUC57 plasmid. The recombinant vectors were transformed into Escherichia coli strain DH5α. The pUC57-CTB-EX4 construct was digested with BamHI and SacI restriction enzymes, cloned into pBI121 expression binary vector, and transferred into tobacco leaves through agroinfiltration. Transcription of the Ex4 fused to cholera toxin B subunit gene in leaves was confirmed by RT-PCR analysis. After agroinfiltration, the protein was extracted from treated leaves, and ELISA test was performed using anti-CTB antibody. The production of recombinant protein was approved by ELISA test in transformed leaves.

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The objective of this study was to assess changes in proline, Glycine Betaine (GB), photosynthetic pigment concentration, Membrane Stability Index (MSI), and grain yield of wheat cultivars induced by Supplemental Irrigation (SI) in the terminal growth stages. Two field experiments with four irrigation levels (rainfed, SI at booting stage, SI at booting and anthesis stages, SI at booting, anthesis, and grain filling stages) and five cultivars (Rejaw, Sardari, Homa, Azar2, and Sirwan) were conducted during the 2015-2017 growing seasons. Results indicated that, overall, SI increased cultivars' chlorophyll concentration, MSI, and grain yield. Proline and GB highly accumulated under drought stress, but rapidly decreased after SI; the severity of the decrease in proline concentration was more remarkable. Sardari and Azar2 cultivars exhibited greater MSI, proline, and GB accumulation during drought stress and more rapid recovery from drought. Our results suggest that the ability of the wheat cultivars to maintain functions during drought and recover after SI during the terminal growth stage is essential for determining final grain yield.