Biotechnology and genetics
Mohammad Ramezani Kaporchali; Mohammad Reza Safarnejad; Abolghasem Ghasemi; Naser Farrokhi
Abstract
Burkholderia gladioli pv. gladioli (Bgg) is the causative agent of saffron bacterial rot. Accurate identification methods will play an important role in disease control management. Flagellum is the main commodity of the bacteria that facilitates its movement. Furthermore, it is of particular importance ...
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Burkholderia gladioli pv. gladioli (Bgg) is the causative agent of saffron bacterial rot. Accurate identification methods will play an important role in disease control management. Flagellum is the main commodity of the bacteria that facilitates its movement. Furthermore, it is of particular importance in colonization and pathogenicity. The fliC gene is one among five other genes encoding flagellin, the protein that makes the flagella in bacteria. In this study, the flagellin genes of 11 bacterial species were selected and the phylogenetic tree analysis divided them into plant and animal bacteria groups. Electron microscope images were taken from the bacterial flagella and its length was up to nine microns with 400-600 nm in diameter; as observed in both Lophotrichous and Monotrichous forms. fliC was amplified and analyzed for the first time by polymerase chain reaction for the two isolates of bacterial pathogen of saffron. Specific primers were designed and used that managed to separate Bgg from other bacteria, and helped in distinguishing the infected saffron corms from the healthy. Electron microscopy of the flagellum was illustrative of different types of Bgg. Investigating and studying the fliC gene can be useful in diagnosis of the causative agent of saffron corm rot disease.
Biotechnology and genetics
Ali Mohammad Mohit-Ardakani; Seyed Ebrahim Seifati; Ali Izanloo; Zahra Karimi-bekr
Abstract
In recent years, the use of saffron spice is increasing due to its medicinal properties and effective components. In addition to increasing crop production, increasing the quantity and quality of secondary metabolites is considered one of the most important objectives of plant breeding programs. On the ...
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In recent years, the use of saffron spice is increasing due to its medicinal properties and effective components. In addition to increasing crop production, increasing the quantity and quality of secondary metabolites is considered one of the most important objectives of plant breeding programs. On the other hand, the saffron plant has low genetic diversity due to the nature of vegetative propagation and induction of variation through mutagenesis is a suitable tool to achieve the desired diversity. Gamma radiation is one of the most widely used physical mutagens in plant breeding programs. In order to investigate the effect of gamma-ray on some growth and phytochemical indices of saffron plants, an experiment was conducted in an unbalanced complete randomized design with three treatments (Control, 15 and 18 Gy) at Yazd University. In this experiment, 15 and 18 Gy doses of gamma irradiation (Co60) were applied at Karaj Nuclear Agricultural Research Institute to induce mutations on collected saffron corms from Qaen, South Khorasan in August 2017. In this study, picrocrocin, safranal and crocin were measured in a 1% aqueous solution at 254, 330 and 440 nm wavelengths, respectively. Based on the results of this experiment, the effect of irradiation on picrocrocin and crocin was significant (P <0.001) and the observed amounts of these compounds (93.77 and 263.02) at the dose of 15 Gy were significantly higher than 18 Gy (92.34 and 262.73), while in terms of safranal content, the difference between radiation levels was not significant. Leaf area index and harvest index were also significant and showed the highest amount of these two indices at 15 Gy dose (3.06 and 0.14). In this study, although gamma radiation treatment could not significantly change the number of daughter corms between different levels of radiation and control, a significant reduction in relative chlorophyll content and an average weight of corms were observed compared to the control (without radiation). In general, the results of this study showed that in phytochemical traits as well as harvest index and leaf area, 15 Gy had a significant advantage over plants without radiation, but at higher levels of gamma radiation, probably due to degradation of genetic content and its effect on some biochemical parameters, 18 Gy plants failed to show acceptable results.
Biotechnology and genetics
ali darvishian; Farhad Nazarian-Firouzabadi; Mostafa Darvishnia; Ahmad Ismaili
Abstract
Saffron (Crocus sativus L.) is the most valuable spice in the world. Due to the lack of genomic information, saffron breeding has encountered many problems. To this end, generating and collecting genetic data using Next Generation Sequencing (NGS) techniques is crucial for saffron traditional and molecular ...
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Saffron (Crocus sativus L.) is the most valuable spice in the world. Due to the lack of genomic information, saffron breeding has encountered many problems. To this end, generating and collecting genetic data using Next Generation Sequencing (NGS) techniques is crucial for saffron traditional and molecular breeding programs. Molecular markers, especially powerful co-dominance markers such as simple sequence repeats (SSRs) markers play an important role in breeding projects. In the present study for the first time, SSR markers related to genes associated with F. oxysporum disease of corms were identified following a RNA-Seq based transcriptomic approach. To this end, total RNA was extracted from mature corms and RNA-Seq was performed based on the Novaseq6000 platform. The De-novo assembly was performed with Trinity software and the MISA search tool was used to identify SSRs. Based on the results of this study, 357028 transcripts were identified. A total of 70060 transcripts were identified to contain SSR sequences. BLAST algorithm analysis revealed that the highest similarity between saffron SSRs was found with that of rice and Arabidopsis. Among the identified unigenes, 18846, 23988 and 10969 genes were identified in UNIPROT, Nr and GO databases, respectively, in which 10375 unigenes were common to all databases. Due to the high priority of saffron in Iran as a strategic crop plant, any genetic information, including mining SSRs, is of great importance in studying genetic diversity, constructing genetic maps, linkage and QTLs analysis in saffron future breeding programs.
Biotechnology and genetics
Mohammadreza Rezaei; Hamidreza sharifi; Alireza Seifi
Abstract
Crocus sativus is a triploide plant and propagating by vegetative propagation. Therefore, trait segregation and genetic diversity are limited in this plant. EST-SSR markers have some priority, for example co-dominant inheritance, locus specific and highly polymorphic against all other markers. Due to ...
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Crocus sativus is a triploide plant and propagating by vegetative propagation. Therefore, trait segregation and genetic diversity are limited in this plant. EST-SSR markers have some priority, for example co-dominant inheritance, locus specific and highly polymorphic against all other markers. Due to the availability of transcriptome data, it is possible to develop EST-SSR markers and polymorphism studies in saffron. Development of EST-SSR markers in C. sativus make it possible to study genetic diversity and molecular polymorphism in different genotypes. In order to develop EST-SSR marker for C. sativus, we downloaded public available C. sativus RNA-seq data. Quality control and preprocessing of raw reads were done using FastQC and Trimmomatic tools, respectively. We performed de novo transcriptome assembly using RNA-Bloom. CD-HIT-EST was used in order to reduce redundancy in transcriptome assembly. The assembly quality was evaluated using the BUSCO software and completeness of transcriptome assembly was 90%. After achieving to high quality transcriptome assembly of C. sativus, EST-SSRs were identified by MISA tool. The EST-SSRs primers were designed using Primer3. 35459 SSR-containing sequences were detected and primer pairs were designed for them. Ten EST-SSR primer pairs were randomly selected to amplify C. sativus DNA. Seven pairs of the primers (70%) generated clear and reproducible bands with the expected size. These EST-SSR markers can be functional and useful for C. sativus genetic studies.
Biotechnology and genetics
Mohammad Zakiaghl; Soroor Khorramdel; Alireza Koocheki; Jafar Nabati; Ahmad Nezami; Amin mirshamsi kachki; Abdollah Mollafilabi; Parviz Rezvani Moghaddam; Mehdi Nassiri Mahallati
Abstract
Sustainable development of saffron production as one of the major and strategic goals of Iranian agriculture requires a well-defined and organized plan. Over the past half century saffron cultivation has increased 34-fold in Iran, more than any other crop. But the average yield did not follow this development ...
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Sustainable development of saffron production as one of the major and strategic goals of Iranian agriculture requires a well-defined and organized plan. Over the past half century saffron cultivation has increased 34-fold in Iran, more than any other crop. But the average yield did not follow this development and is reducing with slight negative slope; so, the average saffron production in Iran has reduced from 5.76 kg/ha in 1973 to 3.42 kg/ha in 2017. It is also estimated that there is a 90-70% yield gap in Iranian saffron farms. Saffron pests and diseases such as saffron mite (Rhizoglyphus robini), saffron corm rot, saffron dry rot (Burkholderia gladioli) and saffron viral diseases are also responsible for quantitative and qualitative reduction of saffron yields in farms. Saffron is propagated by its daughter corms, so increasing the yield of saffron is directly related to the quality of mother corm seed. This paper describes the criteria’s needs to generate and certify prebasic, mother corm and seed corm classes of saffron for standard saffron corm schedule. Production of standard pathogen-free seed corms as the only means of saffron propagation in nature is a prerequisite for any planning to increase saffron yield. Undoubtedly, application of standard free pathogen corms along with other ecological and physiological indices of saffron plant will increase yield and improve Sustainable development indicators.
Biotechnology and genetics
Tofigh Taherkhani; Rasool Asghari Zakaria; Mansoor Omidi; Nasser Zare; mahboubeh taherkhani
Abstract
Saffron (Crocus sativus) as an important medicinal and economical plant of Iran is rich in flavor, aroma and color, along with medicinal properties in addition to nutritional benefits. The effect of chitosan on Crocin and Safranal amounts as two important medicinal components and expression of their ...
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Saffron (Crocus sativus) as an important medicinal and economical plant of Iran is rich in flavor, aroma and color, along with medicinal properties in addition to nutritional benefits. The effect of chitosan on Crocin and Safranal amounts as two important medicinal components and expression of their controlling genes in suspension culture of saffron was subjected as the aim of this study. For this purpose, saffron bulbs were cultured in ½ MS medium being treated with 100 and 150 mg/l of chitosan under cell suspension medium and callus optimal growth conditions. Samples were taken at 24 and 72 hours after the application of treatment in 3 replications. Measurement of secondary metabolites was done with HPLC and analysis of genes’ expression was performed with real-time PCR. The results showed that after the use of 100 and 150 mg/l of chitosan and after 24 and 72 hours, the two CsLYC and CsGT-2 genes expression significantly increased. Also, the results showed that Safranal and Crocin levels by the use of chitosan are significantly different at both harvesting times, so that 150 mg/l at harvest time of 72 hours after application of the treatment had the highest amount of Crocin and Safranal. Usage of chitosan as a bio-stimulant in the growth of medicinal and economic plants of saffron increased the amount of valuable secondary metabolites in the cell suspension culture of the plant.
Biotechnology and genetics
Seid Mohammad Alavi-Siney; Jalal Saba; Seyyed Siamak Alavikia; Mohammad Reza Azimi
Abstract
In order to investigate the relationship between quantitative traits and ISSR markers, an experiments were carried out under farm and laboratory conditions at Zanjan University. In this experiment, 20 primers of ISSR marker were used and the agronomic traits (including flower number, fresh weight of ...
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In order to investigate the relationship between quantitative traits and ISSR markers, an experiments were carried out under farm and laboratory conditions at Zanjan University. In this experiment, 20 primers of ISSR marker were used and the agronomic traits (including flower number, fresh weight of Stigma, fresh weight of flower, dry weight of Stigma, dry weight of flower, stigma length, saffron yield, corm number, feresh and dry weight of corm, number of leaf, leaf length, leaf width, leaf area, leaf dry weight, biomass, harvest index), physiological traits (transpiration rate, stomatal conductance, photosynthesis) and secondary metabolites (picocrocin, Safranal and Crocin) were measured during the growing season. Three primers of total did not amplified. 17 ISSR primers amplified 133 loci among 20 saffron ecotypes, with an average of 7.82 loci per primer. The highest number of alleles were for the I-8 primer (15 alleles).
Biotechnology and genetics
Fariba Afkhami; Nasser Zare; Rasool Asghari; Mohammad Mehdizadeh; Behnam Firoozi
Abstract
Saffron is one of the most important pharmaceutical plants in the world due to its valuable secondary metabolites. The aim of this study is to investigate the different factors on callus induction and growth in saffron corm explants. In the first experiment, saffron corms surface were sterilized and ...
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Saffron is one of the most important pharmaceutical plants in the world due to its valuable secondary metabolites. The aim of this study is to investigate the different factors on callus induction and growth in saffron corm explants. In the first experiment, saffron corms surface were sterilized and were excised to equal segments, then they were treated with ultrasound and then they were cultured on MS medium supplemented with 2 mg.L-1 auxin (NAA and 2,4-D) and 0.5 mg.L-1 Kinetin. In the second experiment, the effect of temperature, light and chitosan were evaluated. The results of analysis of variance showed that there were significant differences (P≤0.05) among temperature, light, chitosan as well as ultrasound treatments in terms of callus induction percentage and fresh weight of callus. Callus induction and growth on MS medium containing 2 mg.L-1 NAA + 0.5 mg.L-1 Kin was higher than those containing 2 mg.L-1 2,4-D +0.5 mg.L-1 Kin. In MS medium containing 2,4-D which had low callus induction and callus growth rate, utilization of ultrasound stimulated callus induction and especially it stimulated callus growth from saffron corm explants. In addition, in MS medium containing 2,4-D, utilization of 0.25 g.L-1 chitosan stimulated callus induction and increased callus induction of saffron corm explants. However, increasing chitosan concentration from 0.25 mg.L-1 to 0.75 g.L-1 decreased callus induction and callus growth, while, in MS medium containing NAA, which had efficient callus induction and growth, utilization of these treatments reduced callus induction and callus growth from saffron corm explants. In other words, the effect of ultrasound and chitosan on response of saffron explants in vitro cultures was used, depending on the type of auxin used in composition of the culture medium. Generally, the highest percentage of callus induction occurred on MS medium supplemented with 2 mg.L-1 NAA + 0.5 mg.L-1 Kin and incubated at 25 ºC in the dark, which could be suitable for in vitro culture and gene transfer studies in saffron.
Biotechnology and genetics
Noraddin Hosseinpour Azad
Abstract
The existence of isoform genes in plants led to the creation of the different types of isomeric metabolites like these glycosylated forms. This study was conducted to investigate the identification and heterologous expression of Saffron stigmas Glycosyl transferase coding isoform gene during the pollination ...
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The existence of isoform genes in plants led to the creation of the different types of isomeric metabolites like these glycosylated forms. This study was conducted to investigate the identification and heterologous expression of Saffron stigmas Glycosyl transferase coding isoform gene during the pollination stage. The results have been obtained by sequencing and bioinformatics analysis of an isolated gene from saffron genomes by degenerate oligo’s revealed that the gene is in1283 bp length and belongs to CsUGT protein family which has Apo- plastic secretion in the cell. to evaluate the enzymatic function, firstly the isolated sequence was sub-cloned under arabinose induce promoter in pThio-UGT expression vector by Gibson assembly technique, then the recombinant vector transformed into BL21-pGro7 bacteria which were able to express chaperon proteins with EL& ES subunits. Followed by destructing the bacteria cell wall via ultrasound, the pellet was suspended by PBS solution and then the soluble proteins were extracted by boiling method. Finally, the protein electrophoresis by SDS pages10% was showed that the recombinant protein of CsUGT expressed correctly in bacteria with 69/5 kDa molecular weight. The gained results in this project could be applied to determine the breeding's strategies to improve qualitative and quantitative traits such as color and aroma in saffron.
Biotechnology and genetics
Ghasem Najari; Khoshnood Nourollahi
Abstract
Alternaria leaf spot caused by Alternaria alternata is one of the most important plant diseases in world. In order to determine genetic diversity, 50 samples were collected from saffron farms of different regions in Razavi Khorasan province (Kashmar, Torbatjam, Torbathaidaryeh, Mahvellat, Bardeskan) ...
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Alternaria leaf spot caused by Alternaria alternata is one of the most important plant diseases in world. In order to determine genetic diversity, 50 samples were collected from saffron farms of different regions in Razavi Khorasan province (Kashmar, Torbatjam, Torbathaidaryeh, Mahvellat, Bardeskan) and South Khorasan province (Qaen, Saraian, Brjand). A. alternata isolates were isolated with Potato Dextrose Agar media, Molecular test was carried out with a set of five pairs of SSR primers after purification and identification of isolates. A total of 22 alleles were produced by SSR primers with an average of 11.1 alleles in all populations. The highest amounts of alleles were related to locus Mavellat with 14 alleles and lowest of Saraian, Birjand and Qaen with 10 alleles. A Comparison of genetic diversity parameters in eight population showed that Torbatjam population has the highest genetic diversity but lower values were estimated for Qaenat. The highest and lowest genetic distance was detected between Qaen-Mahvellat (0.328) and Bardaskan-Mahvellat (0.054), respectively. Based on dendrogram of populations revealed two distinct groups, one group contained Qaen and the other Birjand, Mavellat, Saraian, Bardaskan, Kashmar, Torbatjam and Torbat hidaryeh. Analysis of molecular variance showed that 71 percent of the genetic diversity belongs to within populations and 29% is located among populations. There was the high genetic similarity between isolates from different regions. Results in this study will be useful in breeding for saffron resistant cultivars and developing necessary control measures.
Biotechnology and genetics
Mohammad Ali Behdani; Ali Izanloo
Abstract
Saffron is the most valuable spice in the world. It is genetically a monomorphic clone. However, differences in phenotype and quality have been reported. The most important agro-economically phenotypic variation is the appearance of flowers with more than three stigmas. The main objective of this study ...
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Saffron is the most valuable spice in the world. It is genetically a monomorphic clone. However, differences in phenotype and quality have been reported. The most important agro-economically phenotypic variation is the appearance of flowers with more than three stigmas. The main objective of this study was to study the genetic variability of saffron clones with more than 3 stigmas using SSR and ISSR molecular markers. In this research, saffron clones with more than three stigmas were collected along with the corm and the root from Saffron fields of Qaen and Sarayan, South Khorassan province, then transferred as a whole to the Biotechnology Laboratory of the Faculty of Agriculture, the University of Birjand. The number of stigmas in each flower was counted. Genomic DNA was extracted according to the CTAB method from leaves of the flower with more than three stigmas. Flower with more than three stigmas was larger and had more petals than ordinary ones. The most frequent number of flowers with more than three stigmas was related to four and five stigmas with 38%. Six-spike samples with a frequency of 14% were observed. Among the collected samples, only one specimen with seven stigmas was observed. Of the 48 tested ISSR primers on the bulk of DNA, only 16 primers had amplified bands and selected. The results of agarose gel electrophoresis for ISSR primers amplified the bands ranged from 100 to 1000 bp. By examining the bands formed for ISSR primers, no significant polymorphism was observed between different clones of saffron. Therefore, based on this marker system, no sign of genetic diversity was observed between clones with different number of stigmas. Among the tested SSR markers, 10 primer pairs showed amplified band among the clones. The results of correlation analysis based on Spearman correlation coefficient showed that there was no statistically significant correlation between microsatellite marker alleles and number of stigmas.
Biotechnology and genetics
Maryam Vahedi; Seyed Alireza Salami; Majid Shokrpour; Hassan Rezadoost
Abstract
Saffron (Crocus sativus L.) belonging to Iridaceae family as a source of apocarotenoids is one of the most valuable spices and medicinal plants in the world. Because of the large size and high complexity of saffron genome, its sequencing remains a challenge. The arrival of next-generation sequencing ...
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Saffron (Crocus sativus L.) belonging to Iridaceae family as a source of apocarotenoids is one of the most valuable spices and medicinal plants in the world. Because of the large size and high complexity of saffron genome, its sequencing remains a challenge. The arrival of next-generation sequencing technologies (NGS) has allowed the rapid and efficient development for RNA sequencing. De novo assembly of transcriptome from short-read RNA-Seq data provides a great resource for the study of species without a reference genome. De novo assembly of the transcriptome has some unique challenges, particularly in the case of plants, which possess a large amount of paralogs, orthologs, homoeologs and isoforms. In this research, we attempted to compare the performance of de novo assembly tools including BinPacker, Bridger, Oases-Velvet and Trinity through consideration of a quality metrics such as N50 length, the total number of contigs and alignment scores. The results of these analyses revealed that assembly using Bridger had a superior performance for saffron transcriptome, Oases suffered from relatively high chimera rates and redundancies which causes genes family with high similarity assembled into one transcript, Trinity performs worse than Bridger in the increase of false positives. Our comparison study will assist researchers in selecting a well-suited assembler and offer essential information for the improvement of existing assemblers.
Biotechnology and genetics
Sara Sabertanha; BARAT ALI FAKHERI; Nafiseh Mahdinezhad; Zohre Alizadeh
Abstract
The present research was conducted to investigate the effect of water stress and silver nanoparticles on morphological traits of some saffron ecotypes. The experiment was done in form of split plot factorial in a randomized complete block design format in three replications at research farm of Birjand ...
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The present research was conducted to investigate the effect of water stress and silver nanoparticles on morphological traits of some saffron ecotypes. The experiment was done in form of split plot factorial in a randomized complete block design format in three replications at research farm of Birjand University with two levels including full and low water irrigation on 10 ecotypes of saffron and at three levels of silver nanoparticles including control treatment (distille water), 55 and 110 ppm. The traits such as germination date (germination), the number of germinated corm, the number of tillers, the number of leaves, leaf length, the pods length and leaf width were measured. The best date of emergence and the largest number of paws in complete irrigation treatment was observed under distilled water treatment (control). The highest number of corm germinated was observed in irrigated conditions at 55 ppm of silver nanoparticles in the Qaeen ecotype. The highest leaf length was observed under drought stress in the treatment with distilled water (control). The largest number of leaves and leaf width were observed under drought stress at 55 ppm of silver nanoparticles. According to the obtained results of this study, some of the traits at the conditions of drought stress under the silver nanoparticles with a concentration of 55 ppm and some of the traits in the full irrigation and lack of use of silver nanoparticles had the best performance, which it could be said that probably, in stress dryness, the amount of ethylene increases. Some of the obtained results of this study can be related to the effect of silver in preventing ethylene activity.
Biotechnology and genetics
Khadijeh Bagheri; Pejman Azadi; Mitra Gholami; Massoud Mir Masoumi
Abstract
Saffron (Crocus sativus L.) is the world's most expensive spice. Moreover, it is important since it contains various drug metabolites. Saffron is a triploid (2n=3X=24) and sterile plant and it does not have any viable seeds. Because of the sterility, classical breeding of this plant is limited. Developing ...
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Saffron (Crocus sativus L.) is the world's most expensive spice. Moreover, it is important since it contains various drug metabolites. Saffron is a triploid (2n=3X=24) and sterile plant and it does not have any viable seeds. Because of the sterility, classical breeding of this plant is limited. Developing an efficient callus induction protocol is studied for two reasons, i.e. molecular breeding and the production of secondary metabolites. In order to provide a suitable callus induction protocol, establishment of corms was considered. Five different treatments were applied to sterilization of corms. Thin cell layer explants with approximately 1 mm thickness and typical explants with approximately 1 cm thickness were prepared from sterilized corms. For callus induction, different explants were planted in MS medium containing different concentration of 2, 4-D, BAP and NAA. Then, they were incubated in dark conditions at 20 ±20C for 3 months. The results showed that the use of Benomyl fungicide, followed by surface sterilization using sodium hypochlorite (2.5 %) was the best sterilization treatment. The highest survival rates of explants (90%) were observed in this treatment and all explants were free of contamination. The highest amount of callus induction (75%) was obtained in MS medium supplemented with 2mg/l NAA and 0/5 mg/l BAP from thin cell layer of basal corm. The same result was observed with 1mg/l 2, 4-D from typical explants of basal corm. The results obtained from this study show that the thin cell layer explants are suitable explants because of the high amount of callus formation and the advantages for gene transfer studies. The results showed that the use of Benomyl fungicide, followed by surface sterilization using sodium hypochlorite (2.5 %) was the best sterilization treatment. The highest survival rates of explants (90%) was observed in this treatment and all explants were free of contamination. The highest amount of callus induction (75%) was obtained in MS medium supplemented with 2mg/l NAA and 0/5 mg/l BAP from thin cell layer of basal corm. The same result was observed with 1mg/l 2,4-D from typical explants of basal corm. Our results showed that the thin cell layer explants are suitable explant because of high amount of callus formation and advantages for gene transfer studies.
Biotechnology and genetics
faezeh gharari
Abstract
Saffron is grown for using its flower and red color stigma. This plant has many uses in the food industry, paints and medicine. Due to its high monetary worth, identification of genes involved in the flowering is very important. In order to better understand the molecular mechanisms involved in the onset ...
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Saffron is grown for using its flower and red color stigma. This plant has many uses in the food industry, paints and medicine. Due to its high monetary worth, identification of genes involved in the flowering is very important. In order to better understand the molecular mechanisms involved in the onset of flowering, Gene and gene expression of Pistillata-Like MADS box (PIC2) were studied. The Gene sequences have high homology with other family members of the MADS-box family (one of the transcription factors control the expression of proteins flowers). In this research, the gene was studied using bioinformatics resources. Primers were designed for amplification of the gene and then the total DNA and RNA were extracted from the leaves of saffron. cDNA synthesis was performed by the reverse transcriptase enzyme and it was used as a template for amplification of PIC2 gene by PCR reaction. The reaction was amplified to a fragment with length 900 bp from cDNA and another fragment with length 2100 bp from genomic DNA. This gene encodes a protein that includes 210 amino acids. By analyzing the molecular structure of protein transcripts based on modeling of the gene homology, the spatial figure showed that they have a regular structure.
Biotechnology and genetics
Majid Shokrpour; Zeinab Abedi; Siamak Kalantari; Seyed Alireza Salami
Abstract
Saffron (Crocus sativus L.) is one of the most valuable medicinal and spice herbs in the world. In spite of the ancient cultivation history in Iran, there are limited breeding studies on the plant due to its vegetative reproduction. In order to evaluation genetic diversity of Iranian saffron germplasm, ...
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Saffron (Crocus sativus L.) is one of the most valuable medicinal and spice herbs in the world. In spite of the ancient cultivation history in Iran, there are limited breeding studies on the plant due to its vegetative reproduction. In order to evaluation genetic diversity of Iranian saffron germplasm, sixty-five different saffron accessions from the main cultivation areas in Khorasan including Torbat heidarieh, Gonabad, Mahvelat, Ghaenat and Ferdows were collected and were studied by molecular markers. The used RAPD and ISSR primers produced 43 and 122 polymorphic markers loci, respectively, and totally 165 markers with average of 7.5 markers by each primer, totally. Diversity index ranged from 0.36 to 0.7 with average of 0.23. Also, marker index with the average of 0.16 varied in the range of 0.2 to 0.7. The accessions from Ghaenat and Mahvelat had the maximum (83.03%) and the minimum (52.73%) polymorphism, respectively. Grouping the studied saffron accessions using cluster analysis displayed four distinct groups which had little correspondence to their collection areas, while clustering for the main cultivation areas had relatively good correspondence to their geographical distances. So, it is expected to have nearly approaching improvements of qualitative and quantitative yields via the selection of superior clones of saffron. Key words: Saffron, Molecular variation, Germplasm, RAPD, ISSR, Khorasan region, clustering .
Biotechnology and genetics
Fereshteh Babaee; Zahra Tahmasebi; Hasan Feyzi; Arash Fazeli
Abstract
Iran is one of the important distribution centers of medicinal species, including plant saffron (Crocus sativus L.). The presence or absence of genetic diversity in common native and commercial cultivars of saffron of the country has always been one of the important questions for researchers of this ...
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Iran is one of the important distribution centers of medicinal species, including plant saffron (Crocus sativus L.). The presence or absence of genetic diversity in common native and commercial cultivars of saffron of the country has always been one of the important questions for researchers of this filed. In the present study, varioussaffronsamplesfrom different regions of Iran, including seventeen cultivated samplesfrom North Khorasan, Khorasan Razavi, South Khorasan, Lorestan and Ilam, eight wild types (C. haussknechtii) from Lorestan, Kermanshah and Ilam and one wild type (C. cancellatuse) from Kermanshah were collected and then their genetic diversity was obtained using random amplified polymorphism DNA (RAPD) marker. A total number of 161 DNA bands were produced by eight primers with an average of 14.3 bands; the primer Oligo 340 produced the most number of bands. The polymorphism percentage mean was 73.77%. Cluster analysis using UPGMA method divided the samplesinto four groups. In this grouping, there was a minimum distance between saffron populations collected from Kermanshah and Ilam. In other words, these two populations had the maximum genetic similarity with each other. The maximum similarity was observed between the samplescollected from Saleh-Abad and Lomar in Ilam and the minimum similarity was observed between ecotypes collected from Bisotoon in Kermanshah and Torbat-jam in the Khorasan Razavi province. In most cases, samples of different species were divided into different groups such that it can be said that this Genus has a great diversity. Although there are some samples of wild species (C. haussknechtii) that were with saffron crop species in a group in which there exists many molecular Genetics similarities between these species that is indicative of a common genetic background.
Biotechnology and genetics
mahdi bayat; reza amirnia; mahdi tajbakhsh; bahattin tanyolac
Abstract
In order to study genetic diversity and some of the quality and agronomic traits in saffron, six cultivated ecotypes were collected from different regions of the Khorasan province (Mashhad, Torbat-Jam, Gonabad, Torbat-Heydarieh, Qaen and Birjand) and they were evaluated during two cropping seasons in ...
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In order to study genetic diversity and some of the quality and agronomic traits in saffron, six cultivated ecotypes were collected from different regions of the Khorasan province (Mashhad, Torbat-Jam, Gonabad, Torbat-Heydarieh, Qaen and Birjand) and they were evaluated during two cropping seasons in 2011-13 in the research farm of the agricultural faculty, Urmia University, Urmia-Iran. The farming results showed that there were significant differences between the ecotypes in the studied agronomic and qualitative traits. Thus,, the cluster analysis grouped ecotypes Torbat-Heydarieh, Mashhad and Torbat-Jam in a cluster and ecotypes Birjand, Qaen and Gonabad in the other cluster. The results of mean comparison also showed that ecotypes Torbat-Heydarieh and Mashhad were the best saffron ecotypes in the Urmia condition. On the other hand, the genetic results showed that 22 of 30 SSR primers (73%) detected a total of 44 polymorphic alleles (average 2). The average of Shannon index (I), genetic diversity (h) and polymorphic information content (PIC) were estimated to be 0.688, 0.424 and 0.402, respectively. These results not only clarified the existence of genetic diversities among saffron ecotypes, but also, the genetic diversities were proven with other analyses such as analysis of molecular variance. The results of cluster analysis of Darwin grouped each ecotype of Birjand, Mashhad and Gonabad in a cluster and three ecotypes. Torbat-Heydarieh, Qaen and Torbat-Jam were placed in the same cluster because of their high genetic similarity. This grouping was confirmed by principal coordinate analysis. It was notable that high genetic similarities were estimated between saffron ecotypes (65-90%). Overall, the results of this research not only introduced microsatellite molecular markers as useful tools for evaluating genetic diversity and grouping of saffron ecotypes, but also confirmed that saffron is not a monomorphic plant and we can identify useful genetic diversities for breeding programs.
Biotechnology and genetics
Abbas Safarnejad; Seyedeh Bibi Leyla Alamdari; Hadi Darroudi; Marzieh Dalir
Abstract
Saffron (Crocus sativus; Iridaceae) is an important economic and medicinal crop in Iran. The saffron that is an herbaceous triploid geophyte is used mainly as a source of secondary metabolites having aromatic and medicinal value. The plant develops annually from buds on the mother corm – i.e. a ...
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Saffron (Crocus sativus; Iridaceae) is an important economic and medicinal crop in Iran. The saffron that is an herbaceous triploid geophyte is used mainly as a source of secondary metabolites having aromatic and medicinal value. The plant develops annually from buds on the mother corm – i.e. a thickened stem - which acts as a resting, perennating storage organ. In this research in vitro propagation of Crocus sativus through direct and indirect methods has been studied. Corm slices as explants were sterilized and transferred to MS media with different concentrations of plant growth regulators. The results showed that use of HgCl2 0.15% for 20 minutes is the best treatment for sterilization. The results of ANOVA of direct regeneration indicated that the most corm formation was observed in 2 mg/l BAP. Maximum callus induction was achieved on MS supplemented with 1 mg/l 2,4-D + 2 mg/l BAP. As results indicate the most number of embryos were obtained in MS medium containing 0.15 mg/ l NAA. Also this medium was suitable for germination of embryo. Corm formation was only observed in MS supplemented with1 mg/l 2, 4-D + 2 mg/l BAP in the indirect method. The corm was transferred to JF pot for adaptation and then it was transferred to the pot and soil for growing.
Biotechnology and genetics
Mohsen Sajjadi; Maghsoud Pazhouhandeh
Abstract
Saffron (Crocus sativus L.) is one of the medicinal plants that contain active components and medicinal materials. Tissue culture of saffron can improve the quality and quantity of the saffron product, increase its export and the farmers’ income. In this study, 36 different types of hormone combinations ...
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Saffron (Crocus sativus L.) is one of the medicinal plants that contain active components and medicinal materials. Tissue culture of saffron can improve the quality and quantity of the saffron product, increase its export and the farmers’ income. In this study, 36 different types of hormone combinations in the dark and 9 different treatments of hormone combinations in cold (4°C), using different saffron explants (bulb, leaf, scales around leaf and distal parts of the leaf) were studied in tissue culture. To investigate the growth of corms, the callus formation and the regeneration rate, three replications for each treatment were used and the length of shoot (cm), the callus formation percentage and the regeneration percentage were measured and statistical analysis was performed. Among the types of explants, only explants from bulbs produced the callus on MS medium containing 2 mg.l-1 BAP and 1 mg.l-1 IBA in both the dark and cold conditions. The highest percentage of regeneration was obtained in MS medium with hormonal composition of 0.3 mg.l-1 TDZ, 1 mg.l-1 BAP, 2 mg.l-1 IBA and 0.01 mg.l-1 GA3 in the cold conditions.
Agriculture
Fatemeh Fani Yazdi; Naser Mahdavi Shahri; Khadije Nejad Shahrokhabadi; Amin Fani Yazdi; Saeed Jahedi Pour
Abstract
Saffron (Crocus sativusL.), a native plant from Iran and especially of Khorasan region, has a specific place for people's diet. According to several studies about effects of saffron and because there are no specific studies on the effects of saffron aqueos extract on the development of Drosophila melanogaster ...
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Saffron (Crocus sativusL.), a native plant from Iran and especially of Khorasan region, has a specific place for people's diet. According to several studies about effects of saffron and because there are no specific studies on the effects of saffron aqueos extract on the development of Drosophila melanogaster as an animal model, this subject evaluated in conversion of larvae to pupa and pupa to mature of fruit flies. 5 pairs of 3-day-old wild D. melanogaster were transferred to every culture plate containing different concentrations of saffron aqueos extract in order to intercross and oviposition and were brought out after 8 hrs. The percent of larvae to pupa transitionand pupa to mature conversion, were evaluated in all concentrations. The obtained data were evaluated statistically using SAS software and the mean of data were compared using Tukey test with minimum significance level of pD. elanogaster depends on dose proportionately. It means that saffron has some effects on the percentage of conversion of larvae to pupa, pupa to mature in low volumes and increased consuming concentrations of saffron may have inverse effects on that.
Processing, food industry and biochemistry
Nasrin Moshtaghi; Elham Hamidi; Abdolreza Bagheri; Ahmad Sharifi; Abbas Hemati Kakhki
Abstract
Saffron (Crocus sativus L.) is the most valuable food additive in the world which little production and high price of it caused some adulterations such as plant and chemical material similar to saffron. There are several methods for detecting these fraudulent based on morphological and chemical tests ...
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Saffron (Crocus sativus L.) is the most valuable food additive in the world which little production and high price of it caused some adulterations such as plant and chemical material similar to saffron. There are several methods for detecting these fraudulent based on morphological and chemical tests but they are not effective in some cases. In this research a novel molecular method based on ITS-2 marker is introduced. A common forward primer based on 5.8s rDNA for all plant frauds such as safflower, corn stigmas, pomegranate, turmeric and capsicum slices was designed then specific reverse primers based on ITS-2 for any frauds have been designed for polymerase chain reaction. Related ITS-2 bands were amplified in any adulterations in saffron. Specific primer for camel and cow meet fibers was designed based on cytochrome b gene and could amplified the related bands. Multiplex PCR with all of these primers could amplify all of the bands related to any adulterations. Furthermore, using 20% polyacrylamide gel lead to good segregation of bands. This method can be used successfully for detection of low percentage (1%) of fraudulent in saffron. So this marker can be used efficiently for detection of these frauds in commercial saffron.
Biotechnology and genetics
Akbar Tavakkoli; Ali Sorooshzade; Majid Ghorbani Javid
Abstract
The objectives of this research were investigated the effects of removal of lateral and main buds in different corm size on vegetative traits and yield of saffron. The research was conducted as factorial based on a randomized complete block design with three replications at the Research field of Faculty ...
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The objectives of this research were investigated the effects of removal of lateral and main buds in different corm size on vegetative traits and yield of saffron. The research was conducted as factorial based on a randomized complete block design with three replications at the Research field of Faculty of Agriculture, Tarbiat Modares University in Tehran-Iran, during growing season of 2012-2013. The first factor was corm size with two levels of corm weight (2-4 and 6-8 g) and the second factor was buds removal with nine levels included without bud removal, remove all lateral buds, remove all buds except main buds and one lateral bud, remove all buds except main buds and two lateral buds, remove all buds except main buds and three lateral buds, remove all buds except one lateral bud, remove all buds except two lateral buds, remove all buds except three lateral buds, remove all buds except four lateral buds. In this research, vegetative traits in the first year and reproductive traits in the second year were investigated. The results showed that by removing main bud from corm leaf length and root length were decreased. Also, there was most root length by removing all lateral buds. Overall, the results showed that bud removal could be produce large corm but main bud should not eliminated. The most appropriate the number of buds that could be remaining is 4 large buds on saffron corm.
Biotechnology and genetics
Masoumeh Aliakbari; Rouhollah Shamloo-Dashtpagerdi; Esmaeil Ebrahimie
Abstract
Functional genomics methods such as Expressed Sequenced Tag (EST) analysis have provided possibilities for identification, expression analysis and study of transcripts involved in metabolic and regulatory networks. In order to identify of genome orientation and to determine gene networks involved in ...
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Functional genomics methods such as Expressed Sequenced Tag (EST) analysis have provided possibilities for identification, expression analysis and study of transcripts involved in metabolic and regulatory networks. In order to identify of genome orientation and to determine gene networks involved in the evolution of saffron stigma, 6202 EST sequences from mature saffron stigma were analyzed. After initial trimming, sequences clustering and assembling resulted in 910 unigenes (604 Contigs and 304 Singleton). BLAST X revealed that 570 unigene had significant hit among the Arabidopsis protein database, whereas the remaining unigenes displayed no significant match with the any hit. Classifying and gene enrichment analysis of unigenes, put them into 31 distinct functional groups, where 12 groups of them were statistically significant at α=0.01. Gene network of high represented Contigs (which had greater than 20 transcripts), showed that there is a complex gene interaction in mature saffron stigmas. Results revealed that jasmonic acid signalling pathway and its transcription factors such as MYB21 and Zinc fingers play a key role in regulating of stigma primary and secondary metabolism, especially in metabolism of carotenoids (as the most important saffron metabolites). The genes identified in this study could be good candidates for manipulating the evolution and metabolism of saffron stigma.
