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Background: One of the most effective strategies to increase phytochemicals production in plant cultures is elicitation. In the present study, we studied the effect of abiotic and biotic elicitors on the growth, key biosynthetic genes expression, antioxidant capacity, and phenolic compounds content in Rhizobium (Agrobacterium) rhizogenes-induced hairy roots cultures of Ficus carica cv. Siah. Methods: The elicitors included methyl jasmonate (MeJA) as abiotic elicitor, culture filtrate and cell extract of fungus Piriformospora indica as biotic elicitors were prepared to use. The cultures of F. carica hairy roots were exposed to elicitores at different time points. After elicitation treatments, hairy roots were collected, and evaluated for growth index, total phenolic (TPC) and flavonoids (TFC) content, antioxidant activity (2,2-diphenyl-1-picrylhydrazyl, DPPH and ferric ion reducing antioxidant power, FRAP assays), expression level of key phenolic/flavonoid biosynthesis genes, and high-performance liquid chromatography (HPLC) analysis of some main phenolic compounds in comparison to control. Results: Elicitation positively or negatively affected the growth, content of phenolic/flavonoid compounds and DPPH and FRAP antioxidant activities of hairy roots cultures in depending of elicitor concentration and exposure time. The maximum expression level of chalcone synthase (CHS: 55.1), flavonoid 3′-hydroxylase (F3'H: 34.33) genes and transcription factors MYB3 (32.22), Basic helix-loop-helix (bHLH: 45.73) was induced by MeJA elicitation, whereas the maximum expression level of phenylalanine ammonia-lyase (PAL: 26.72) and UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT: 27.57) genes was obtained after P. indica culture filtrate elicitation. The P. indica elicitation also caused greatest increase in the content of gallic acid (5848 µg/g), caffeic acid (508.2 µg/g), rutin (43.5 µg/g), quercetin (341 µg/g), and apigenin (1167 µg/g) phenolic compounds. Conclusions: This study support that elicitation of F. carica cv. Siah hairy roots can be considered as an effective biotechnological method for improved phenolic/flavonoid compounds production, and of course this approach requires further research. [ABSTRACT FROM AUTHOR]

The current study investigated the effects of gamma irradiation on biochemical parameters and secondary metabolite accumulation in Summer Savory under field conditions. The dry seeds of Summer Savory (with a moisture content of 12%) were exposed to gamma radiation at the doses of 20, 40, 60, 80, and 100 Gy. Non-irradiated seeds (0 Gy) were used as control. Our findings showed that gamma radiation at low doses (20–40 Gy) had no effect on biochemical parameters and secondary metabolites accumulation in S. hortensis. These parameters are steadily and significantly increased by raising gamma irradiation doses from 40 to 100 Gy. The highest amount of chlorophyll a and b, carotenoids, anthocyanin, and total phenolic and flavonoid content were observed in 80 and 100 Gy treatments. Plants exposed to 80 and 100 Gy treatments accumulated the maximum amounts of rosmarinic acid and caffeic acid, respectively. Furthermore, the analysis of S. hortensis essential oil revealed that gamma radiation significantly alters its components. Carvacrol, α-Pinene, and α-Thujene levels raised dramatically compared to control with an increase in gamma irradiation dose from 20 to 100 Gy, while Thymol and α-Terpinene levels lowered. Our results showed that treatment of Summer Savory seeds with gamma radiation at 80 and 100 Gy doses could significantly be raised biochemical parameters and secondary metabolites accumulation under field conditions. The current study showed that gamma irradiation could be used as a pre-sowing elicitor to improve the quantity and quality of phytochemicals in Summer Savory. [ABSTRACT FROM AUTHOR]

Introduction Summer savory (Satureja hortensis L.) belonging to the Lamiaceae family, is used as a spice and traditional herb in Iran. This plant has shown antispasmodic, antidiarrheal, antioxidant, sedative and antimicrobial activities. The basic requirement for plant breeding programs is a germplasm diversity that provides necessary facilities for breeding species with desirable features. Therefore, accurate identification of genotypes is considered as a prerequisite in this manner. Phenotypic Materials and Methods In this research, 20 accessions of S. hortensis were chosen from Iran, Bulgaria, Germany, Czech, Georgia, Syria, Hungary, Poland, Italy and Uzbekistan. Some morphological and phytochemical characteristics such as internode number, shoot number, number of days to germination, collar diameter, fresh weight of aerial part, bract fresh weight, flower number, peduncle length, 1000- seeds weight, flower fresh and dry weight and essential oil components were studied. The Results and Discussion Based on the results of morphological studies, internode number (11.03), shoot number (18.58), 1000-seeds weight (925.97 mg) high variable Uzbekistan accessions were best in this traits. Maximum flower number (26.34) and flower fresh weight (26.14 mg) were obtained from Bulgaria accession. Essential oil amount was higher in Karaj accession. Correlation coefficients among traits showed that fresh weight of aerial part had the highest correlation with flower fresh weight (0.97). Cluster analysis based on Euclidean distance divided the accessions into four major groups in which the third and fourth groups had valuable characteristics. The Bulgaria accession had the highest y-Terpinene and Borneol. The Karaj accession was the best in carvacrol and delta-3-carene content. Clustering from essential oil component assigned accessions into five clusters. It can be suggested that there is a considerable genetic variation among S. hortensis accessions. The main purpose of this study was to determine genetic diversity among S. hortensis accessions from Iran and other parts of the world. Significant differences were found among the accessions in almost every parameter measured. The range of vegetative and flower values obtained in this study was more than those reported in previous researches about S. hortensis. With increasing shoot and leaf number, Iran appears to be the main place to trap sunlight for photosynthesis and consequently provides the condition to produce flowers, seeds and secondary metabolites. Thus, Uzbekistan and Bulgaria accessions having high values of leaf and flower can also be considered for cultivation and breeding programs. Conclusion In this study, agro-morphological and essential oil properties of S. hortensis accessions originated from Iran and some Asian and European countries were evaluated. The study revealed a high level of diversity in morphological and phytochemical traits among S. hortensis accessions. This needs a breeding approach during the domestication process to obtain homogenous cultivars with suitable agricultural properties. [ABSTRACT FROM AUTHOR]

Objective Many secondary metabolites produced by plants are used in manufacturing drugs and new treatments, but are too difficult, expensive or impossible to prepared by chemical synthesis. Biotechnology progresses have provided a huge source of new chemical compounds of plant origin to various applications. Purslane (Portulaca oleracea L.) is one of the valuable medicinal plants that World Health Organization (WHO) called it Global Panacea. Hairy root induction by Agrobacterium rhizogenes is an effective method for production of secondary metabolites. Materials and methods In this study, hairy roots of Purslane were produced using Agrobacterium rhizogenes strains A4 and ATCC15834. The presence of the rolB gene in the hairy roots was confirmed by PCR using rolB gene specific primers. The effect of the type of strain (A4 and ATCC15834), explant (cotyledon, hypocotyl and seedling) and also different concentrations of Iron oxide nanoparticles (0, 20, 40 and 6 mg l-1) were examined as abiotic elicitors on the hairy root induction, growth and increase of secondary metabolites of hairy roots. Results The results showed that the highest hairy root induction (80 percent) was observed in the A4 strain, seedling explants and 40 mg l-1 the iron oxide nanoparticles. The greatest hairy root length was obtained by 60 mg l-1 the iron oxide nanoparticles (4.17 cm) and also ATCC15834 strain, seedling explants and 40 mg l-1 the iron oxide nanoparticles were superior for highest phenolic and flavonoid content (4.92 and 0.48 mg g DW) and also 2,2-diphenyl-1-picrylhydrazyl (52.59%) and ferric reducing antioxidant power (60.37(Mol g-1 DW)). Hairy root lines showed significant differences in growth rate and biomass production and the largest biomass production was exhibited by G line. Conclusions Overall, the strain and explant types and iron oxide nanoparticles had considerable effects on the hairy roots induction secondary metabolites production. The results indicated significant increases in hairy root induction and medicinal compounds by A4 and ATCC15834 strains, seedling explant and 40 mg l-1 the iron oxide nanoparticles. [ABSTRACT FROM AUTHOR]

• The successful Rhizobium (Agrobacterium)mediated hairy root (HR) induction in Ficus carica L. was confirmed by the presence of rol B gene in the HR lines. • The effects of cell extract and medium culture filtrate (two elicitors) derived from axenic culture of Piriformospora indica on some phenolic compounds and expression level of some genes in F. carica HR cultures was investigated in this study. • Depending on concentration and treatment time, P. indica elicitation induced to change biomass, phenolic compounds accumulation and antioxidant activity of HRs. • P. indica elicitation improved the expression rate of PAL, CHS, UFGT, MYB3 genes and increased the synthesis of apigenin, quercetin, rutin and phenolic acids (gallic, caffeic, chlorogenic, coumaric, rosmarinic and cinnamic acids) in F. carica elicited-HRs. Ficus carica L. is an important source of phenolic and flavonoid compounds with valuable pharmaceutical application across various diseases. The current study was carried out to investigate the influence of Piriformospora indica elicitation on growth, production of phenolic compounds, antioxidant capacity, and expression level of flavonoid biosynthetic pathway genes in hairy root (HR) cultures of F. carica. The maximum improvement in accumulation of phenolic compounds was observed when HR culture of Ficus carica L. was exposed to 2% culture filtrate of P. indica for 72 h: gallic acid (80.5- fold), caffeic acid (26.2-fold), coumaric acid (4.5-fold), and cinnamic acid (60.1-fold), apigenin (27.6-fold) and rutin (5.7-fold). While the highest levels of chlorogenic acid (4.9-fold) and quercetin flavonoid (8.8-fold) were obtained after 48 h elicitation with culture filtrate and cell extract of P. indica at 6% (v/v), respectively. The analysis of biosynthetic genes revealed that the exposure to fungal elicitors resulted in up-regulation of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT) and MYB3 transcription factor. This study shows the potential of P. indica as an efficacious elicitor for enhancing the secondary metabolites production by F. carica HRs. [ABSTRACT FROM AUTHOR]

BACKGROUND Ficus carica L., an ancient source of food and medicines, is rich in valuable nutritional and secondary compounds with antioxidant, antimicrobial, and anticancer effects. The present study is the first attempt to examine hairy root (HR) induction of F. carica (Sabz and Siah) by inoculating the 3‐week‐old shoots and leaves with different strains of Agrobacterium rhizogenes and also to investigate methyl jasmonate (MeJA) elicitation of HRs to produce a fast and high‐yield production method for secondary metabolites. RESULTS: The maximum transformation rate (100%) was achieved by inoculating the shoots with Agrobacterium rhizogenes strain A7. Siah HRs elicited with 100 and 200 μmol L−1 MeJA and Sabz HRs with 100 μmol L−1 MeJA showed the highest total phenolic content. The highest flavonoid content was 3.935 mg QE g−1 DW in Siah HRs treated with 200 μmol L−1 MeJA and 2.762 mg QE g−1 DW in Sabz HRs treated with 300 μmol L−1 MeJA. The 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging capacity and ferric reducing antioxidant power (FRAP) value of HRs were affected by MeJA treatments. Methyl jasmonate elicitation also significantly enhanced the content of six phenolic acids (gallic acid, caffeic acid, chlorogenic acid, coumaric acid, rosmarinic acid, and cinnamic acid) and three flavonoids (rutin, quercetin, and apigenin). Thymol, a monoterpene phenol, was the main HR compound detected in gas chromatography mass spectrometry (GC–MS) analysis of the essential oils. CONCLUSION: Induction of HRs and elicitation of F. carica HRs by MeJA resulted in a significant increase in the production of important phenolic compounds and a significant increase in antioxidant capacity. © 2020 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Highlights • In this study a reliable gene transfer system via modifying the co-cultivation medium using different strains of Agrobacterium rhizogenes and explants were established in C. intybus for the first time. • Different co-cultivation media including MS basal medium and MS medium lacking KNO 3 , NH 4 NO 3 , KH 2 PO 4 , CaCl 2 and MgSO 4 respectively was used. • Observed an increase in hairy roots induction percentage, biomass accumulation and secondary metabolites content by all of strains. • Maximum hairy roots induction occurred by A 4 strain when a MS medium without KNO 3 was used. • The high frequency of hairy roots induction with ATCC11325 and ATCC15834 strains was achieved when KH 2 PO 4 was removed of MS medium. Abstract Transformed hairy roots technology provides new opportunities for mass production of pharmaceutical metabolites. Hairy roots culture has been considered as an alternative method for producing medicinal biomolecule s in plants. Chicory (Cichorium intybus L.) is a medicinal herb from Asteraceae family. It contains many important metabolites including chicoric acid, inulin, scoline, coumarin, and flavonoids. In this study, for the first time, a reliable gene transfer system via modifying co-cultured medium using different strains of Agrobacterium rhizogenes and explants were established for C. intybus. Different co-culture media were used for this purpose, including MS basal medium and MS medium without either of KNO 3 , NH 4 NO 3 , KH 2 PO 4 , CaCl 2 , and MgSO 4. The results showed increases in hairy roots induction percentage, biomass production, and phenolic content of the hairy roots. Maximum hairy root induction percentage obtained by using A 4 strain when a KNO 3 -free MS medium was used. High frequency of hairy root induction percentage with ATCC11325 and ATCC15834 strains was achieved when MS medium had KH 2 PO 4 removed. In all of the experiments, very low hairy root induction was observed when NH 4 NO 3 was removed from co-culture MS medium. Molecular confirmation of transgenic hairy roots was done with PCR using gene-specific primers for rol B gene. Total phenols, flavonoids, anthocyanins and chicoric acid contents of hairy root clones and the control (untransformed) roots were also determined. The results indicated significant increases in total metabolites content of hairy root clones induced by A 4 , ATCC11325 and ATCC15834 strains with some macronatrient–free co-culture MS media and also directly-regenerated shoots from hairy roots were spontaneous without using any plant growth regulators. [ABSTRACT FROM AUTHOR]

Landraces of spinach in Iran have not been sufficiently characterised for their morpho-agronomic traits. Such characterisation would be helpful in the development of new genetically improved cultivars. In this study 54 spinach accessions collected from the major spinach growing areas of Iran were evaluated to determine their phenotypic diversity profile of spinach genotypes on the basis of 10 quantitative and 9 qualitative morpho-agronomic traits. High coefficients of variation were recorded in some quantitative traits (dry yield and leaf area) and all of the qualitative traits. Using principal component analysis, the first four principal components with eigen-values more than 1 contributed 87% of the variability among accessions for quantitative traits, whereas the first four principal components with eigen-values more than 0.8 contributed 79% of the variability among accessions for qualitative traits. The most important relations observed on the first two principal components were a strong positive association between leaf width and petiole length; between leaf length and leaf numbers in flowering; and among fresh yield, dry yield and petiole diameter; a near zero correlation between days to flowering with leaf width and petiole length. Prickly seeds, high percentage of female plants, smooth leaf texture, high numbers of leaves at flowering, greygreen leaves, erect petiole attitude and long petiole length are important characters for spinach breeding programmes. [ABSTRACT FROM AUTHOR]

Interest in growing winter spinach (Spinacia oleracea L.) in Iran is increasing due to its good nutritional potential returns relative to other vegetable crops. The objectives of this research were to investigate the interrelationships among different traits of spinach and to evaluate different Iranian spinach landraces with application of the genotype × trait (GT) biplot methodology in visualizing research data. 81 spinach landraces were grown during 2-years according to randomized complete block design with four replications. Ranking of the genotypes based on the ideal entry revealed that genotypes G1, G20, G7, G8, G9, G27, G49 G70 and G79 were higher in the measured traits and could be good candidates for improving most of the measured traits. Ranking of traits for the leaf yield showed that petiole diameter, petiole length, leaf numbers at flowering, 1000-seed weight and root dry weight were the most discriminating traits which influence spinach leaf yield at both years. There were 9 winning genotypes and 4 which-won-where patterns at the first year while there were 8 winning genotypes and 4 which-won-where patterns at the second year. As a result, the findings from our study are as follows: (i) traits leaf numbers at flowering, leaf length, leaf width, leaf area, petiole diameter and petiole length could be as selection indices for spinach leaf yield improvement, (ii) genotypes G1, G20, G7, G8, G9, G27, G49 G70, and G79 were the most favorable and is thus recommended for commercial release or incorporating in breeding programs; (iii) the GT biplot method can be used to identify superior genotypes in other crops and in other parts of the world. [ABSTRACT FROM AUTHOR]

This research uses path analysis to determine the interrelationships among seed yield and 16 related Morphological traits. Eighty-one garden cress accessions from IPK (Department of Leibniz Institute of Plant Genetics and Crop Plant Research) were grown in two growing seasons (2012-13) to determine the important components of seed yield. Observations were recorded on 20 other canola traits. Correlation coefficient analysis revealed that seed yield was positively correlated with all traits except plant height (PH) in the first year and except main axis length (MAL) and PH in the second year. Sequential path analysis (SPA) identified the thousand-seed weight (TSW), number of siliques per plant (NSP) and height of first silique (HFS) as important first order traits influenced seed yield in first year. Plant height, NSP, and the TSW were important first-order traits that influenced seed yield in the second year. This indicates that breeding programs should be based on these traits for further improvement of the garden cress. All direct effects were significant, as indicated by bootstrap analysis. The results suggest that TSW and NSP could be used as a selection criterion in selecting for increased seed yield in garden cress. [ABSTRACT FROM AUTHOR]

The development of genotypes, which can be adapted to a wide range of diversified environment, is the ultimate goal of plant breeders in a crop improvement program. In this study, several stability methods were used to evaluate the genotype by environment interaction (GE) in 11 lentil ( Lens culinaris Medik) genotypes. The genotypes were evaluated for grain yield at 4 different locations for 3 years in semi arid areas of Iran. The testing locations have different climatic and edaphic conditions providing the conditions necessary for the assessment of stability. A combined analysis of variance, stability statistics, rank correlations among stability statistics and yield stability statistic were determined. Significant differences were detected between genotypes and their GE interactions. Different univariate stability parameters were used to determine stability of the studied genotypes. The level of association among the parameters was assessed using Spearman's rank correlation. The different stability statistics which measured the different aspects of stability was substantiated by rank correlation coefficient. Rank-correlation coefficients between yield and some stability parameters were highly significant. Genotypes mean yield (Mean) was significantly correlated to the Lin and Binns stability parameter PI ( r = 0.93* *í) and desirability index Di ( r = 0.89* *í). A principal component analysis based on rank correlation matrix was performed for grouping the different stability parameters studied. In conclusion, based on most stability parameters, the genotypes G2, G5 and G9 were found to be the most stable. Results from rank correlation and principal component analysis showed that the stability variance (σi 2) was strongly correlated with Wricke's ecovalance, stability parameters of Plaisted and Peterson, and Plaisted. [ABSTRACT FROM AUTHOR]

• In this study, a reliable gene transfer system using different strains of Agrobacterium rhizogenes and explants were established in Trigonella foenum graecum L. • Different strains of Agrobacterium rhizogenes and various levels of Iron oxide and Zinc oxide nanoparticles were used. • Observed an increase in hairy roots induction percentage, biomass accumulation and secondary metabolites content by ATCC11325 strain. • The Maximum trigonelline content of hairy roots was achieved when treated Zinc oxid (2x) nanoparticles. Fenugreek (Trigonella foenum-graecum L.) is a rich source of important medicinal metabolites. This plant belongs to the Fabaceae family that grows anywhere around the world. It is extensively used in medicine and as a food additive. Several secondary metabolites are detected in fenugreek, especially a valuable alkaloid called trigonelline. Enhancing the production of secondary metabolites is possible through the application of modern biotechnological techniques such as culturing hairy roots. In the present study, different strains of Agrobacterium rhizogenes (ATCC11325, ATCC15834, A 4 , A 7 , A 13 , and K599) were used for hairy root induction from fenugreek cotyledon explants. Then, the effect of various levels (x, 2x, and 4x concentrations of B5 medium) of iron oxide and zinc oxide nanoparticles were evaluated on growth, total phenolic, flavonoids, and trigonelline content of hairy roots. The growth curve analysis revealed an increase in the fresh and dry weight of treated hairy roots, as compared with the control. The highest levels of fresh (1.49 g) and dry weight (1.23 g) were obtained in hairy roots which were induced by the ATCC11325 strain and were treated with Zn (2x). The results indicated that using Zn nanoparticles on hairy roots can lead to higher levels of trigonelline, compared with the control group (i.e. transformed hairy roots without NPs). [ABSTRACT FROM AUTHOR]

• The studied accessions of Satureja hortensis were diverse based on morphological and phytochemical traits. • The Bulgaria accession had the highest plant height, dry yield, leaf length and also thymol and ɣ-terpinene contents. • The highest correlation coefficient was observed between internode number and internode length. • Thymol, γ-terpinene, delta-3-carene, carvacrol, myrcene and camphene were the major components of essential oils. • Morphological and phytochemical variability of Satureja hortensis L. accessions: an effective opportunity for industrial production. Summer savory (Satureja hortensis L.) as an endemic species identified in Iran and other countries is widely used in traditional medicine. Several isolated molecules of this species are used to elaborate antifungals and can also be used in the pharmaceutical industry. This investigation was performed to evaluate the levels of variabilities of some agro-morphological characteristics and essential oil constituents of S. hortensis accessions originated from Iran, Bulgaria, Germany, Czech, Georgia, Syria, Hungary, Poland, Italy, Greece, and Uzbekistan. The different properties of accessions such as the highest plant fresh, dry weight, plant height, leaf length, internode length, and main branch number were found in the Bulgaria. The maximum amount of essential oil content was obtained in Uzbekistan accessions (0.75 %). The Bulgaria accession had the highest thymol and ɣ-terpinene contents, while the Karaj accession was the best in carvacrol and delta-3-carene contents. The highest correlation coefficient was observed between the plant's internode number and internode length (r = 0.99) and the plant height had a positive correlation with traits influencing plant yield. The accessions were classified into five main categories according to morphological properties and most of the accessions originated from each geographical site placed in the same class. The findings of this research can be used for sketching efficient breeding programs for this species. [ABSTRACT FROM AUTHOR]

The current study investigates the effects of humic acid concentration, its application methods as well as nitrogen ratios on the morphological, yield and biochemical characteristics of strawberry cv. Paros, during 2014–2015 in Ardabil Province, Iran. The study has been conducted within the framework of a double split plot randomized complete block design with three replications and density of 10 plants per replications. Different nitrogen levels (50, 100 and 150 kg ha−1) were examined in the main plots while application methods of humic acid (foliar and soil) and its different concentrations (0, 2, 4 and 6 kg ha−1) were considered in the subplots and sub-sub plots, respectively. The results of the analysis of variance (ANOVA) indicated that the interaction effects of different concentrations of nitrogen, humic acid and its application methods are significant on most of the measured variables at 5% probability level. Mean comparisons showed that the highest values for the leaf area, fruit yield, chlorophyll a, carotenoids and titratable acidity are associated with the combined treatments of nitrogen and humic acid with concentrations of 100 kg ha−1 and 4 kg ha−1, respectively. In three-way interactions, the highest shoot and root biomass and total chlorophyll content were obtained from the application of 100 kg ha−1nitrogen and 4 kg ha−1 foliar humic acid, while the highest nitrogen and protein contents of leaf and root were achieved upon applying 2 kg ha−1 foliar humic acid and 150 kg ha−1 nitrogen. Moreover, the highest amounts of ascorbic acid and total soluble solids were found at a concentration level of 100 kg ha−1 nitrogen along with applying 2 and 6 kg ha−1 foliar and soil humic acids, respectively. Finally, the best results of most of the studied traits were observed at 100 kg ha−1 nitrogen and 4 kg ha−1 foliar humic acid application. [ABSTRACT FROM AUTHOR]

Nowadays, approximately 5.8% in adult population around the world are suffering by diabetes. It can be caused by an increase in risk factors such as being overweight. Also it has been estimated that the number of patients will be doubled in near future and the demands for insulin hormone will be growing up by 3 to 4 % annually. Therefore, it's necessary to develop new methods for hormone production with high rate of capacity in future. By advanced technology of transgenic DNA, the transgenic plants are introduced as an attractive system for expression and production of many kinds of pharmaceutical proteins. In this study, we investigated transfer of Human Proinsulin Gene into the Cucumber (Cucumissativus L.). Transgenic cucumber could be a great prospect for future source of eatable insulin pharmaceutical drugs to be taken by patients.Agrobacterium tumefaciensstrain LBA4404 carrying proinsulin genes with CaMV 35S promoter was used for the transformation purpose. The transgenic plants were analyzed by PCR, RT-PCR, SDS-PAGE, Dot blot and Electrochemiluminescence techniques. Production of proinsulin in cucumber could be a great prospect in molecular farming of human proinsulin. [ABSTRACT FROM AUTHOR]

Investigation of native accessions of spinach (Spinacia oleracea L.) would be aid in the development of new genetically improved varieties, so in this research 121 spinach landraces, collected from the various spinach growing areas of Iran, were evaluated to determine their diversity using several agro-morphological traits. High coefficients of variation (CV) were recorded in fresh yield, leaf area and dry yield. Using principal component (PC) analysis, the first three PCs with eigenvalues more than 0.9 contributed 80.56% of the variability among accessions. The first PC was related to leaf yield performance (fresh and dry yields, leaf numbers at flowering and lateral branches) while the PC2 was related to leaf characteristic (leaf width, petiole length, petiole diameter and leaf area). The third PC was related to seed characteristic (seed yield and 1000-seed weight) and was named as seed property component. The 121 spinach landraces were grouped into six clusters using cluster analysis. Each cluster had some specific characteristics of its own and the clusters I and II were clearly separated from clusters III and V and also from clusters IV and VI. The studied accessions are an important resource for the generation of a core collection of spinach in the world. The results of present research will support tasks of conservation and utilization of landraces in spinach breeding programs. [ABSTRACT FROM AUTHOR]

Nanotechnology is an emerging field of science widely exploited in agriculture in recent years. In this investigation, application of nanotechnology in agriculture via application of some nano-particles (nano-iron and nano-silicon) have been investigated in seed priming of dragonhead. Seeds were subjected to prehydration treatments by factor nano-silicon dioxide as; (S1) 0 mM or distilled water, (S2) 1 mM concentration and (S3) 2 mM concentration and and factor nano-iron oxide as; (F1) 0 Mm or distilled water, (F2) 1 mM concentration and (F3) 2 mM concentration. Germination percent, root fresh weight, shoot fresh weight, root length, shoot length, dry weight of the seed residue, root dry weight and shoot dry weight were measured. Analysis of variance showed significant variation for the main effect of nano-silicon dioxide as well as nano-iron dioxide in root length and dry weight of the seed residue. The interaction effect of nano-silicon x nano-iron priming treatments were significant in all of the measured traits except germination percentage and root fresh weight. The highest germination percentage was recorded in S2-F3, S3-F1 and S3-F3 while the root fresh weight was high in S2-F3 and treatments S1-F1 following to S2-F3 and S3-F2 produced the highest shoot fresh weight. Also, S2-F3 has the highest root length (16.1 cm) and the highest shoot length (18.4 cm). The best treatment combination suitable for obtaining of high values of germination characteristics of dragonhead was identified as S2-F3 (1 mM nano-silicon dioxide plus 2 mM nano-iron dioxide). [ABSTRACT FROM AUTHOR]

Different expression systems such as bacteria and mammalian cells have been used to produce pharmaceutical proteins. In recent years, the use of plants as bioreactors offers efficient and economical systems in recombinant protein production. Furthermore, because of the large number of plastid copies in plants, chloroplast engineering functions as an effective method to increase recombinant protein expression. Because the commercially available insulin for treatment does not contain C-peptide, which is of great importance for type 1 diabetic patients, the current study introduces the human proinsulin gene fused with protein A into the tobacco chloroplast genome using the biolistic method. To achieve homoplasmy, three rounds of selection and regeneration of transforming cells were performed on the medium that contained spectinomycin antibiotic and hormones. The PCR analysis indicated the presence of the proinsulin gene in transplastomic plants. The reverse-transcription PCR analysis confirmed the expression of the proinsulin-protein A fusion at the transcription level. Immunoblot assays of leaf-derived protein extracts confirmed that the target gene expression is up to 0.2% of the total soluble protein. Our study showed that protein A fusion is not as efficient as other reported fusions. The transplastomic plants were also confirmed for homoplasmy using Southern blot analysis. [ABSTRACT FROM AUTHOR]

For overcoming interspecific incompatibility, protoplast combination method is a proper procedure for making a new plant with desired traits. For this purpose, protoplast preparation is a first and important step. Hence, experiments were conducted to evaluate various combinations of cellulose, pectinase and their treatment times on protoplast production and protoplast viability in Lilium ledebeourii Bioss. The results of experiment revealed that the protoplast yield was significantly affected by different treatment levels. Cellulase at 4% gave the highest numbers of protoplasts at 3.71x105 protoplast/g FW. Pectinase at 1% gave the highest numbers of protoplast. For treatment times, the highest yield of protoplast was with leaf explants treated for 24 h. Analysis of variance indicated that concentration, time and three-way interaction of cellulase, pectinase and time were significant at p<0.01. Cellulase at 4% and pectinase at 0.2% for 24 h gave the highest viability. Interactions of cellulase x pectinase, cellulase x time, pectinase x time and cellulase x pectinase x treatment time were significant at P≤0.05 for protoplast number. The highest and lowest protoplast numbers were produced in media containing 4% cellulase and 1% pectinase for 24 h (6.65×105 protoplast/g FW) and 1% cellulase and 0.2% pectinase for 12 h, respectively. It's concluded that, the best treatment for isolation of Lilium protoplast was 4% cellulase and 1% pectinase for 24 h. [ABSTRACT FROM AUTHOR]