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
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
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.
