Medicinal Plants
Negin Afsharzadeh; Majid Azizi; Leila Samiei
Abstract
Introduction Hypericum perforatum is a perennial plant that has been used in traditional medicine. H. perforatum have several types of medicinal compounds including antiviral compounds, antioxidants, flavonoides and also has valuable compounds such as Hypericin, Hyperforin, Pseudo-hypericin and xanthones ...
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Introduction Hypericum perforatum is a perennial plant that has been used in traditional medicine. H. perforatum have several types of medicinal compounds including antiviral compounds, antioxidants, flavonoides and also has valuable compounds such as Hypericin, Hyperforin, Pseudo-hypericin and xanthones that have effect on human physiology. Aerial parts of H. perforatum are dotted with dark glands that appear as black or red nodules. Black glands were known as localization of secondary metabolites. as a consequence of the commercial potential of this species attempts have been at plant improvement by application of in vitro culture methods. Among seedling explants of H. perforatum, it appears that roots are superior for shoot regeneration. It is generally accepted that explants source is an important factor r successful establishment of tissue culture in many cases. Production of secondary metabolites via plant cell and tissue culture yields various advantage es, including standardization and quality. These criteria are also valid for the main economically important chemical in st. John’s wort, namely hypericin, pseudo hypericin and hyperforin. The aim of this study was to evaluate the effectof some tissue cultures on plant Callogenesis, regeneration and also, study the effect of cytokinin and auxin on rooting rate and shoot multiplication. Materials and Methods: This research included two experiments; first experiment plan was a completely randomized in the form of a factorial. Second experiment plan was completely ranom.first part of experiment : this part were conducted with two explants, leaf and shoot ,maintained in light and dark condition. Shoot explants were derived from sterile seedlings that was obtained from seeds were cultured on MS medium. Seeds were decontaminated by NaClO 20% (V/V) for 20 min and were washed with sterile deionized water. Leaf explants were derived from seedlings in the in vivo condition and decontaminated by NaClO 20% (V/V) for 20 min then washed with sterile deionized water. Both of explants cultured on MS media supplemented with BA(3 and 4 mg/l) and 2,4-D (0, 1 and 2.5 mg/l). Callogenesis and regeneration was measured after 4 weeks.Second part of experiment: shoot of indirect regeneration for rooting study, were cultured on ½ MS media supplemented with BA (0 and 5/0 mg/l) and IBA (0 and 1 mg/l). Proliferation, shoot and root length were measured after 4 and 8 weeks.Results and Discussion:Effects of the factors on first part of experiment; calluses of shoot and leaf explants were induced after 4 weeks. Shoot explants Medium supplemented with 4 mg/l BA and 2.5 mg/l 2, 4-D showed 95% Callogenesis. Leaf explants Medium supplemented with 3 mg/l BA and 2.5 mg/l 2, 4-D showed 98% Callogenesis. Shoot explants Medium supplemented with BA 3 mg/l showed 62% regeneration and leaf explants Medium supplemented with BA 4 mg/l showed 8% regeneration.For second part of experiments; root induction on half strength medium without hormone and medium supplemented with 0.5 mg/l BA and 1 mg/l IBA had highest rooting frequencies. Average of root length was registered 5.25 cm. half strength medium supplemented with 0.5 mg/l BA had 100% and also, medium with 0.5 mg/l BA and 1 mg/l IBA had 86% shoot multiplication and were not appeared any roots. Average Shoot length on medium without hormone and medium supplemented with 0.5 mg/l BA and 1 mg/l IBA was registered 6.24 cm and in media with 0.5 mg/l BA and 1 mg/l IBA was registered 1.8 cm. based on result of this experiments, the concentration levels of the two hormone BA and 2, 4-D in the induction of calli formation and regeneration of the H. perforatum have been effective. In the second experiment, hormone BA, in the absence of IBA did not cause rooting and increased the degree of shoots and ultimately proliferation was effective. Also, in treatment with 0.5 mg/l BA and 1 mg/l IBA, although the concentration of cytokinin was less than auxin, rooting was not done. conclusionThe goal of this study was to introduce the suitable medium for Callogenesis and regeneration of H. perforatum for production and breeding aims. Tissue and cell culture methods are used for various purposes. The result of each section can be used effectively in research topics, including corrective tests, processes for increasing secondary metabolites, as well as increasing commercial products. Therefore, by introducing efficient and effective methods, it is possible to increase the production of plants by micropropagation in less time.
Maedeh Aghdaei; Seyyed Hossein Nemati; Leila Samiei; Ahmad Sharifi
Abstract
Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South ...
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Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South American countries, including Bolivia, Colombia, Ecuador and Peru, as well as in countries such as New Zealand and Australia. Pepino is propagated by seed, cutting, and tissue culture methods. Most pepino cultivars are sexually fertile and produce viable seeds, but their seeds have poor germination and high level of heterozygosis causing to highly variable plants. Both mentioned negative aspects have limited the mass production of this plant through seed. In this case, stem cutting is used as the most common way of propagating pepino led to transmission of viral diseases and increasing propagation costs as two main limiting factors of pepino propagation. So, micropropagation systems are a promising tool to produce disease-free clonal plant material with low costs. Therefore, the present study was aimed to assess the effect of different media and plant growth regulators on micropropagation traits of pepino.
Materials and Methods: Three separate experiments were carried out in institute of plant sciences of Ferdowsi University of Mashhad in 2016. Pepino seeds were bought from company of Plant World Seed, UK, were cultivated on MS medium. Grown plants were used as source of providing explants. Four mediums, including MS, ½ MS, SH and B5 were used to determine the best culture medium for shoot regeneration of pepino using single node explant. A factorial experiment was conducted based on a completely randomized design. Some growth properties such as number of shoots, shoot length, number of roots, root length, leaf number and leaf length were evaluated after two and four weeks. In proliferation experiment, MS medium was compared with MS supplemented with different concentrations of BA (0.5, 1 and 2 mg L-1) and Kin (0.5, 1 and 2 mg L-1) applied as combined treatments, and also BA used alone at concentrations of 2, 4 and 6 mg L-1 that was conducted based on a completely randomized design. For rooting of explants, an experiment was conducted based on a completely randomized design containing of two concentrations of IBA (at 0.3 and 0.6 mg L-1) and three concentrations of NAA (at 0.3, 0.6 and 0.9 mg L-1) in MS medium. Some growth properties including root number and length, root density and root quality were evaluated after four weeks
Results and Discussion: Results indicated that micropropagation rate of pepino was affected by culture medium type. The highest shoot length, number of root, root length and leaf number were obtained in MS medium, although statistically there was no significant difference between MS and ½ MS media. The highest number of shoots and leaf length were observed in MS medium, which led to a significant difference with other media (½ MS, SH and B5). Overall, Based on obtained results MS medium was the best culture medium for micropropagation of pepino using single node. In the proliferation experiment, the highest shoot and leaf number and plant color were obtained with using 2 mg L-1 BA + 1 mg L-1 Kin, whereas the highest shoot length and leaf length were observed in the 1 mg L-1 BA + 2 mg L-1 Kin and 1 mg L-1 BA+1 mg L-1 Kin treatments, respectively. Increasing in concentration of BA up to 2 mg L-1 in combination with Kin had a positive effect on shoot proliferation, while applying BA at concentration 2, 4 and 6 mg L-1 alone led to decrease in proliferation. Results obtained from rooting experiment showed that the highest root number, root density and root quality were obtained using IBA at the concentration of 0.6 mg L-1, whereas the highest root length was observed by applying IBA at concentration of 0.3 mg L-1, which led to a significant difference with other treatments. Furthermore, results indicated that the effect of IBA on rooting of pepino microshoots was more than NAA.
Conclusion: Generally, the best results were obtained by MS medium, 2 mg L-1 BA with 1 mg L-1 Kin for shoot proliferation, and IBA at concentration of 0.6 mg L-1 for the rooting of pepino nodal segments.
Shadi Attar; Gholamhossein Davarynejad; Leila Samiee; Mohammad Moghadam
Abstract
Introduction: Persian walnut (Juglans regia L.), belonging to the Juglandaceae family, has its natural origin in the mountainous regions of central Asia and especially northern forests of Iran. Most walnut genotypes are seedling and sexually reproduced. Conducting studies on the genetic structure of ...
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Introduction: Persian walnut (Juglans regia L.), belonging to the Juglandaceae family, has its natural origin in the mountainous regions of central Asia and especially northern forests of Iran. Most walnut genotypes are seedling and sexually reproduced. Conducting studies on the genetic structure of these genotypes to identify, select and maintain their genetic resources is important. Identifying and collecting local varieties of fruit trees is considered as the first step on the path of breeding programs and lack of information regarding plants genetic characteristics causes the breeding work to be done slowly. Various methods have been used for studying genetic diversity and determining the genetic relationship between European and Asian varieties of walnut and identifying commercial walnut varieties, among which we can mention: Morphologic indices, Alozyme, Isozym, RFLP, RAPD, AFLP and ISSR markers. ISSR molecular marker was used in order to investigate genetic diversity of some genotypes of Persian walnut (Juglans regia L.) in Mashhad orchards. .
Materials and methods: To begin with, about 56 walnut trees from 4 orchards in Mashhad (Esteghlal (1), Golestan (2), Alandasht (3) and Emam Reza (4)) were selected and tagged from 2014 to 2016. In the spring of 2014 with the beginning of trees growth and opening of leaves, a number of leaves from each genotype were collected. After DNA extraction, the quality of samples by agarose gel (1 percentage) and electrophoresis method and quantity of them via spectrophotometer device at 260 and 280 nm wavelengths were determined. First, 24 primers of ISSR marker were prepared and after initial evaluation on 5 random genotypes, 9 primers with high polymorphism and repeatability were selected for further investigation. For PCR reaction, Amplicon kit (code 180 301, made in Denmark) was used. Gel electrophoresis images of primers that produced polymorphic bands with suitable resolution were analyzed manually. After scoring the bands, in which 0 used for showing the absence of a band and 1 showing the presence of a band in each sample, 1 and 0 numbers were changed to matrix by using NTedit software. Genetic similarities of samples were calculated by using NTSYSpc software, SIMQUAL method and DICE similarity coefficient. Dendrogram by cluster analysis was drawn by using UPGMA method. Principle coordinate analysis (PCO) was performed using the NTSYS software and grouping samples were evaluated in a two-dimensional plot.
Results and discussion: Results showed that from 9 primers in total, 118 bands amplified were in 300 to 3000 base pairs, while 29 bands were polymorphic. The number of amplified fragment for each primer was different so that UBC 844 (14 bands) had the highest and UBC 890 (8 bands) had the lowest amplified bands. The average amplified fragments for each primer was 9.83. The percentage of polymorphic in various primers was different. Maximum polymorphism (80%) of the primers was observed in UBC830. In general, according to the percentage of polymorphic bands, low adjustment to any changes in the environment was indicated. This can be used as an indicator to determine the value of diversity and genetic erosion. In genotypes cluster analysis, clustering was performed based on Dice similarity coefficient and UPGMA method, and 10 clusters were formed. ISSR molecular marker somewhat revealed genetic diversity among walnuts genotypes, whereas the genetic diversity was lower than expected. In general, by reviewing the findings in other parts of the world about walnut genetic diversity and comparing them with the results of this study, despite existing high genetic diversity among walnuts in many areas, some reports of low genetic diversity among walnuts populations have been published and unfortunately in recent years these reports has made more attention. Based on the results of several studies reporting low genetic diversity among walnuts, the following factors can be effective in this problem: natural disasters, human impacts such as deforestation and selection and propagation of superior genotypes, and sometimes walnuts self - pollination. In this respect, there is concern that if this trend of decreasing genetic diversity in the walnut population continues, this invaluable crop will be in danger of extinction. So we should think about a remedy. Finally, this investigation can be used as a start for conducting more researches in the region to maintain and manage this valuable crop germplasm and maximize genetic diversity for performing breeding programs in the future.
Amirhassan Hosseini; Ali Tehranifar; Leila Samiei; Mahmood Shoor; Farshid Memariani
Abstract
Introduction: Allium is one of the greatest genera in Amarylidaceae family with more than 900 species distributed in northern hemisphere from temperate region to tropical and subtropical areas. There are more than 135 Allium species in Iran, many of which have the ornamental potential and can be exploited ...
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Introduction: Allium is one of the greatest genera in Amarylidaceae family with more than 900 species distributed in northern hemisphere from temperate region to tropical and subtropical areas. There are more than 135 Allium species in Iran, many of which have the ornamental potential and can be exploited in floriculture industry. Northeast of Iran with more than 35 species especially in Razavi and North Khorassan provinces is regarded as one of the main centers of diversity of Alliums. The species of Allium have the great potential to be used either as cut flower and potted plant or as outdoor plant in urban landscape. There are indeed a rich germplasm of Allium in Iran with diverse characteristics of flower colour, size and form. For the utilization of the wild species, it is necessary to identify and fully assess their main morphological and phenotypic characteristics as the initial step. Morphological markers are one of the commonly used tools which are applied in phenotypic evaluation, taxonomic classification and collection management of plant species. The aim of present study was to evaluate morphological traits and genetic diversity of some of the native Allium species of Razavi khorassan in Iran.
Materials and Methods: In this study the Allium species were collected from diverse regions of Khorassan, Iran and were cultured as a collection in Research Center for Plant Sciences (RCPS) of Ferdowsi University of Mashhad in a completely randomized block design. Following the establishment of the Allium species, 29 qualitative and 13 quantitative traits were recorded during the growing season in two successive years. Analysis of variance and comparison of all traits were performed using JMP8 software and simple correlation and factor analysis was calculated using SPSS software. Principal component analysis as well as maximum variance was also performed on the collected data.
Results and Discussion: The results exhibited that the species and genotypes were significantly different in their morphological characteristics. Some of the traits like scape length, leaf colour, flower diameter and length and width of perianth were more diverse among the species and consequently were considered as the most discriminating traits whereas some qualitative traits like perianth segment form, ovary form and perianth tip form remain uniform amongst the species. There are number of studies indicating the existence of high genetic diversity in phenotypic characteristics of some species of Allium like garlic, onion and shallot .The traits having the higher coefficient of variance bring the opportunity for the wider range of selection. Correlation analysis revealed the significant correlations among some of the studied traits. High positive correlation observed among leaf length with scape length, scape diameter with perianth length, leaf width with scape diameter and scape diameter with flower diameter. In garlic, similar positive correlation was reported on leaf length and width with scape length. Considering the quantitative traits, various parameters like environmental conditions are also effective in correlation between traits along with the corresponding genetic controlling. Factor analysis is used to identify the most important factors involving in discriminating among the species and genotypes. Based on the results, the 42 morphological characters were classified into five main factors which could justify 91.62% of total variance. Characters like inflorescence diameter, scape diameter, flower form, pedicel length, filament length and ovary and style length were placed in the first factor and expressed 30.03% of the variation. The second factor that could explain 24.9% of variation was included scape form and length, leaf length and pedicel colour. The selected discriminating traits of this study were similar to the ones of Allium roseum which had been earlier introduced by Zammuri et al (2009). The first two factors explained the highest amount of diversity while the 3rd to fifth factors could define 16.10%, 12.265% and 9.19% of variance, respectively. Traits like scape length, leaf form and colour and inflorescence diameter are amongst the main factors for the selection of ornamental plants.
Conclusion: In overall, substantial valuable information obtained for some of the native species of Alliums of Iran for the first time. This information can help in selection of desired species for specific landscape designing however characterization of the native germplasm can also be a very beneficial task in future breeding program as well as the conservation strategies for preserving the endangered species.
Shadan Khorshidi; Gholamhossein Davarynejad; Leila Samiei; Mohammad Moghaddam
Abstract
Introduction: Most deciduous trees need low temperature to break flower bud dormancy. One of the most important abiotic stresses is low temperature which limits production of temperate fruits. Pear production has been considerably reduced in recent years. Important pear cultivars show different levels ...
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Introduction: Most deciduous trees need low temperature to break flower bud dormancy. One of the most important abiotic stresses is low temperature which limits production of temperate fruits. Pear production has been considerably reduced in recent years. Important pear cultivars show different levels of resistance to cold. Cold compatibility followed by resistance increase is controlled genetically and contains several mechanisms which lead to production of different metabolites such as: polypeptides, amino acids and sugars. The object of this research was to evaluate the frost resistance of different ‘Dare Gazi’ genotypes and other pear cultivars in Mashhad climate condition.
Materials and Methods: This study was conducted to investigate the frost resistance of 23 ‘Dare Gazi’ pear genotypes and nine other cultivars include: ‘William’s’, ‘Bell de june’, ‘Spadona’, ‘Koshia’, ‘Domkaj’, ‘Torsh’, ‘Sebri’ and ‘Tabrizi’. Plant material contained vegetative and reproductive buds of one-year-old shoot samples which were collected from 25-year old trees on March 2014, four days after winter cold (-6.6 °C) in three directions of trees and sent to the laboratory. Frost damages of vegetative and reproductive buds were investigated based on visual observations (%), electrolyte leakage (EC) and proline content. EC was measured with a Metrohm 644 digital conductivity meter and proline content was measured based on Bates et al. (1973) method, using acid ninhydrin. The experiment was performed on completely randomized experimental design with three replications. Statistical analysis was carried out using MSTAT-C and Excel software. Mean values were compared using the least significance difference test (LSD) at 1% levels. Cluster analysis was conducted by SPSS 16 program.
Results and Discussion: Highest EC of reproductive buds was observed in ‘Dare Gazi’ 10, 19, ‘Tabrizi’ and ‘Torsh’ whereas ‘Dare Gazi’ 8, 18 and ‘Bell de June’ had the lowest EC. Based on visual observations, the least percentage of damaged reproductive buds was observed in ‘Dare Gazi’ 22 while ‘William’s’ suffered from frost at the highest damage level (96%). Göndör and Tóth (1998) studied 13 pear cultivars by microscopic observations of flower buds and found that ‘Packham's Triumph’ was relatively resistant under Hungarian ecological conditions. Honty et al. (2008) reported that Kaiser was the most sensitive pear cultivar to low temperatures during endodormancy and ecodormancy. Khorshidi et al. (2014) described that pear reproductive buds of ‘Ghodumi’ were the most tolerant. Considering the vegetative buds, ‘Dare Gazi’ 19 had the highest EC (74.47 %) which was not significantly different from ‘Dare Gazi’ 10, 20 and 3 whereas the lowest one was found in ‘William’s’(24.75%). The highest percentage of healthy vegetative buds was found in ‘Dare Gazi’ 1 (50%) which did not show a significant difference with ‘Dare Gazi’ 12, 5 and 7. ‘Tabrizi’ was the most sensitive and had most damaged vegetative buds (95.99%). Khorshidi et al. (2014) observed the least damaged vegetative buds in ‘Dare Gazi’ and the most damaged buds in ‘Boheme’ and ‘Ghodumi’. Palonen and Buszard (1997) mentioned that hardiness of woody tissue of apples did not seem related to flower bud hardiness. The highest proline content of reproductive buds was found in ‘Dare Gazi’ 20 (21.28 µmol g-1FW) and the lowest content observed in ‘Dare Gazi’ 2 (0.1 µmol g-1 FW). Young (1977) described that increase in proline was not correlated well with relative cold hardiness of citrus rootstocks. Data did not show any significant correlation between EC and proline content. Barka and Audran (1997) studied grape buds and shoots and reached a high negative correlation between proline content and frosttolerance. Yelonsky (1979) indicated that accumulation of proline was not correlated to cold hardiness. The results of present experiment were in agreement with Duncan and Jack (1987) findings which reported that increase in proline was not always correspondent to cold resistance. Based on the cluster analysis, the genotypes laid in two distinct groups. First group included ‘Dare Gazi’1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 17, 18, 19, 20, 23 and ‘Sebri’, ‘Domkaj’, ‘Bell de june’ and ‘Koshia’ and second group include ‘Dare Gazi’ 11,16, 21 and ‘William’s’, ‘Spadona’ and ‘Torsh’. Frost damage, EC and proline content were higher in the first group compared to second group.
Conclusion: Cosidering‘DareGazi’ genotypes, no correlation was found between proline content and frost damage rate. This shows that morphological differences among ‘Dare Gazi’ genotypes could be due to the existing of genetic variation of these genotypes or they are different clones of one cultivar.
