نشانگر مولکولی

Taxol is one of the most valuable effective substances found in some plants, which is used in chemotherapy against breast, stomach, ovary, liver, lung, cervix and pancreas cancers. Hazel leaf (Corylus avellana L.) is one of the plants in which taxol has been proven. The natural forests of Ardabil, Arsbaran and Miane regions are important areas of hazelnut germplasm. The purpose of this research is to investigate and group the subpopulation samples of hazelnuts in these regions based on the SCoT genetic marker and compare the samples of a cluster using principal component analysis in terms of the amount of taxol present in hazelnut leaves and the phytochemical characteristics of the fruit. Current research was done in 2018-2019. In this research, 78 samples of native trees were selected from the natural forests of Fandalo, Ardabil, Mianeh, and Arsbaran (according to the geographic distance of the samples of each region, the population of that region was considered) and the geographic location of each region was recorded with GPS. Quantitative and qualitative characteristics of hazelnuts were collected based on the size, geographic characteristics and possible diversity among the hazelnut stands in the northwest of the country and based on the description of hazelnuts of the deputy research and identification of plant varieties, Ministry of Agricultural Jihad. In this research, based on phenotypic diversity, subpopulation samples (single tree) were selected, and DNA extraction was done from the leaf part by CTAB method. After measuring the quality of DNA in the absorption spectrum of 260 and 280 nm in the nanodrop, using SCoT specific primers, the PCR reaction was performed. Then, electrophoresis was performed for the presence or absence of markers (zero and one) and the data were evaluated using POP Gene v1.32 software. From each group, depending on the number of samples, at least two samples were selected and the amount of taxol in their leaves was extracted and measured. Selected samples of hazelnut fruits were selected and the amount of antioxidant properties, phenols, fatty acids and the amount of some mineral elements were measured. Based on the presence and absence of SCoT genetic bands in the genome of selected hazelnut samples from different regions, principal component analysis (PCOA) was performed. Correlation (Mantel's correlation coefficient) between the similarity coefficient based on the grouping of genotypes using the SCoT marker and the similarity coefficients using geographic characteristics data was calculated as 0.54, which was significant (Pvalue<.05). Examination of the amount of genetic information content (PIC) based on the used SCoT primers showed that the amount of PIC was different in the groups separated based on PCoA. between genotypic groups in terms of protein content, fatty acid content, fiber, carbohydrate, total phenol, palmitic acid. There was a significant difference in the probability levels of 0.05 and 0.01, and no significant difference was seen in terms of other traits. The comparison of the averages of hazelnut genotypic groups showed that in terms of protein content, percentage of fatty acids, amount of antioxidant property and palmitic acid, the highest average was related to the genotypes of group 1. The values of the similarity coefficients in the grouping based on the SCOT indicator and the similarity coefficients obtained from the results showed that the correlation between the calculated coefficients is 0.76. Evaluation of gene flow (Nm) and fixation index (Fst) within the populations and genotypes of hazelnut showed the highest amount of gene flow Nm and the lowest amount of Fst in all populations (Fst=0.55 and Nm=0.7) They showed that the same trend was observed in the groups as well, although the amount of Nm coefficient in the genotypes of group 1 was (Nm=81). The amount of inter-population heterozygosity (Dst) was estimated to be 0.15. Since the study of phytochemical traits always requires time and conducting numerous tests, the existence of reliable and reproducible markers is necessary to find genotypes with high phytochemical and taxol characteristics. The low value of the gene flow parameter (Nm) showed that the studied populations have a high tendency to differentiate. The existence of high genetic diversity within populations makes it possible to implement breeding programs. The high genetic similarity between the populations, the study of the SCoT indicator (the highest 0.97, the lowest 0.81) showed that the populations belong to the same origin and genetic background.

Heterodera schachtii, the sugar beet cyst nematode (BCN), is a limiting factor in most of the beet-growing regions of Iran. Considering this disease's importance and spread, generating resistant cultivars is one of the most important methods to reduce its damage. Achieving accurate and fast resistance assessment methods facilitates the development of resistant cultivars. This research aims to compare the response of twenty domestic and foreign sugar beet genotypes to BCN under greenhouse conditions and with the presence of the MN3 molecular marker, which is linked to the BCN resistance gene. Analysis of variance showed a significant difference among genotypes for resistance to BCN in greenhouse conditions. Using the MN3 molecular marker and greenhouse survey, no susceptible plants were observed in genotype No. F- 21238. Hybrids No. 34781 and 34791 demonstrated the highest resistance (80 and 78% of plants were resistant) under greenhouse conditions and good resistance (80 and 89% of plants were resistant) to this pathogen evaluating the MN3 marker. The results showed that to save time and more accuracy in selecting resistant genotypes, it is better to evaluate genotypes with MN3 molecular marker, and then the chosen genotypes are assessed to determine the level of resistance to this pathogen in greenhouse conditions.

Genetic diversity is crucial for plant breeding and results in natural evolution of the biological systems. Among various Aegilops species, Ae. tauschii is the primary wild relative of bread wheat, contributing the D genome to cultivated wheat. The objective of this research was to explore the allelic diversity of D genome microsatellite loci in 89 Ae. tauschii accession originated from various regions of Iran, Turkey, Afghanistan, Armenia, Sweden, and Azerbaijan and to evaluate the effectiveness of 32 pairs of mir-SSR microsatellite markers in segregating Ae. tauschii accessions for identification and use in germplasm management and breeding programs. 

Seeds from 89 Ae. tauschii accessions were sown in pots, and DNA was extracted from young leaves using the CTAB method. Allelic amplification was performed using 32 pairs of mir-SSR microsatellite primers. Band patterns were scored based on the presence/absence of bands and allele sizes. Statistical analysis was conducted using GenAlex and NTSYS software. 

From the 32 primer pairs studied, 31 pairs with suitable band patterns were selected, producing 104 alleles, of which 91 were polymorphic. The number of alleles per primer pairs, ranged from 2 to 5, with an average of 3.35 alleles per pair. The average percentage of polymorphism for the primers was 88.91%. The polymorphic information content (PIC) for different primer pairs ranged from 0.52 to 0.95, with an average of 0.81. Specific alleles were identified for each population, and 13 specific alleles were found in the 14 Ae. tauschii studied populations with the Mazandaran population showing the highest number of specific alleles. Cluster analysis grouped the accessions into five main clusters, although the 14 populations were not distinguishable. 

The results demonstrated the effectiveness of mir-SSR markers in detecting polymorphisms across the studied populations, revealing high genetic diversity and differentiation, particularly in the Mazandaran population. Cluster analysis indicated significant genetic mixing or similar geographical conditions among accessions in the same group. Overall, the mir-SSR markers used in this study exhibited high polymorphism and could distinguish populations and their allelic diversity. These markers are valuable tools for investigating genetic diversity in other breeding programs.

Next-generation sequencing (NGS) technology offers an opportunity to analyze microsatellite molecular markers linked to terpenoid pathway genes. Microsatellite molecular markers are a powerful way to understand genetic diversity. In this study, the transcriptome sequencing of the Citrullus colocynthis medicinal plant was used for the first time to identify microsatellite markers.

In this research, the data of the transcriptome sequencing study of the fruit of the C. colocynthis medicinal plant were used in which 83,807 de novo assembly sequences were prepared. The assembled transcripts of C. colocynthis plant were analyzed to determine the frequency and distribution of microsatellites. We searched for microsatellite markers using Krait v1.5.1, a robust and ultra-fast identification tool with a user-friendly graphical interface for genome-wide screening of microsatellites. Also, specific primers for microsatellite markers identified by Krait software were designed and saved as Fasta files. For the functional annotation and gene similarity comparison, the microsatellites of the C. colocynthis transcriptome against the microsatellites of the nucleotide sequences of the five reference genomes include Uniprot, Arabidopsis thaliana (Atha), Citrullus lanatus subsp. 97103 (Clan), Momordica charantia (Mcha) and Citrullus lanatus subsp. Charleston Gray (WCG) were blasted with E-Value less than 10e-5. In other functional interpretation, the sequences of unigenes containing microsatellites were uploaded to the KEGG automatic interpretation server (http://www.Genome.Jp/kegg/kaas) and interpreted using specific KO (KEGG Orthology) identifiers. The execution method of KO specific identifiers was based on the Single-directional Best Hit method. According to KO assignment, the information of the metabolic pathways related

The use of heterosis in hybrid rice breeding aims to increase productivity, with hybrid varieties yielding 20 to 30 percent more than conventional varieties. Maintaining genetic purity in hybrid seed production is critical to prevent contamination and selfing, especially in cytoplasmic male sterile lines. This study aimed to identify microsatellite markers to distinguish hybrid rice parental lines and to assess seed purity and characterize the morphology of F1 hybrid rice. Initial trait assessments, including outcrossing levels and sterility stability, are critical for selecting preferred male sterile lines. Eighteen Iranian and exotic cytoplasmic male sterile lines and one thermosensitive genetic male sterile line, were evaluated for various traits in a randomized complete block design. PCR-based molecular markers were used to assess purity, revealing significant trait differences among lines. Selection and prioritization of superior lines such as Fajr A, Neda A and IR58025 A was based on fertility percentage, panicle yield and other traits. The Dasht B line with favorable traits such as proper height and panicle length proved to be a promising candidate for seed production. Molecular tests with PCR-based markers effectively discriminated between male sterile and conservation lines. The drrcms marker showed promise in distinguishing cytoplasmic male sterility lines from different sources, offering potential for genetic purity evaluation and infection testing of seed parents. These results point to valuable alternatives for maintaining genetic integrity in hybrid rice production.

Basil (Ocimum basilicum) is one of the most important medicinal as well as vegetables plants that are cultivated and consumed worldwide One of the essential aspects of plant breeding programs is to study the correlation between DNA polymorphisms and phenotypic trait diversity. The purpose of this research is to investigate the genetic diversity of five basil cultivars under drought stress conditions using SCoT markers and traits, and also to analyze the relationship between traits and markers by stepwise regression. In this research, five basil genotypes were studied under drought stress in the form of a split plot based on a completely randomized design with three replications in pots and under greenhouse conditions. The main plot included drought stress in two levels (normal and drought stress) and the sub plot factor included genotype (5 levels) and their morphological and physiological traits were evaluated. Also, their genomic DNA was extracted from the leaves, and the genotypic diversity of the genotypes was investigated based on eight SCoT primers, and finally, the relationship between the traits and markers was determined by stepwise regression. Correlation of traits in two conditions showed that leaf yield had a positive and significant correlation with traits of plant height and total chlorophyll. The percentage of trait changes under stress conditions showed that root length, chlorophyll a and total chlorophyll traits had the greatest decrease and cluster analysis based on them placed the genotypes in three groups and the traits in three groups. Eight primers amplified a total of 101 polymorphic bands and ScoT1 produced the most bands with 17 polymorphic bands. Cluster analysis by UPGMA and dice similarity criterion based on SCoT data were placed five basil genotypes in three groups. The results of regression analysis showed that in normal and drought stress conditions, 15 and 12 markers (alleles) had a significant relationship with the studied traits, respectively. Selection based on molecular markers provides a rapid method for breeding programs. The obtained genetic information of markers in this study showed their important role. Therefore, in addition to traits, it is possible to select superior genotypes and high value populations in breeding programs. The findings showed that certain markers are associated with multiple traits and emphasized the critical importance of this trait in plant breeding for simultaneous improvement of multiple traits. Insights from this study on markers have potential for application in breeding programs.

Knowledge on genes effect and action (additive/dominance) is one of the necessities to achieve cultivars with high performance and quality. Estimating the breeding value (additive effect) can be done thanks to molecular markers through best linear unbiased prediction (BLUP). In the present study, 100 oilseed sunflower genotypes were evaluated based on the 10×10 lattice design during two crop years of 1392-1393 under normal and drought stress (irrigation limitation) conditions. The breeding value of 13 traits in 78 genotypes out of 100 was estimated due to having genotyping data with SSR and Retrotransposon based markers in each one of normal and drought stress (irrigation limitation) conditions through BLUP. For this purpose, the kinship matrix was calculated by SSR and Retrotransposon based markers data. According to total ranks of breeding values of all studied traits estimated by molecular data of both markers, in normal conditions, genotypes 47, 11, 8 and 35 and under drought stress (irrigation limitation) conditions, genotypes 8, 11 and 35 showed the highest breeding value. Based on SSR markers data in normal conditions; genotypes 76, 36, 34 and 41 and based on Retrotransposon based markers data; genotypes 61, 78, 72 and 52, and in drought stress (irrigation limitation) conditions based on SSR markers data; genotypes 76, 38, 34, 29 and 70 and based on Retrotransposon based markers data; genotypes 16, 71, 78 and 61 showed the lowest breeding value. Considering both studied conditions and all studied traits and both molecular markers information, genotypes 8, 11 and 35 with high breeding value are introduced as desirable parents for breeding programs.

Fusarium wilt (FW) caused by F. oxysporum f. sp. ciceris causes extensive damage to chickpea in many parts of the Iran. The technique of Marker-assisted backcrossing (MABC) is one of the effective and accurate methods in transferring specific genes such as disease resistance genes in a short time in self-pollination plants. Hashem is one of Iranian chickpea cultivars with high yield, plant height and tall stems which its cultivation is limited due to susceptibility to the fusarium wilt. Therefore, Purpose of this study was to introgression resistance to fusarium wilt from Ana cultivar, as a donor, to Hashem as a recurrent and susceptible cultivar using molecular marker-assisted backcrossing.

This research was conducted during five cropping seasons (2018-2023). Crossing was done between the selected parents of chickpea cultivar Ana (as a donor parent) and Hashem (as a recurrent parent) and the F1 progeny was backcrossed with Hashem to produce the BC1F1 generation. By using three backcrosses and one rounds of selfing, BC3F2 progeny was obtained. Foreground selection (FGS) was conducted with four markers (TA59, TA96, TR19, and CaM1402) linked to FW resistance genes. Background selection (BGS) was employed with evenly distributed 16 (Ana × Hashem) SSR markers in the chickpea genome to select plant(s) with higher recurrent parent genome (RPG) recovery. Finally, the selected lines of BC3F2 generation were phenotypically evaluated in terms of Fusarium disease resistance.

In first year, from the crossing of two parents, 20 real F1 plants were obtained and backcrossed with Hashem to produce BC1F1 plants. Based on results of foreground selection (FGS), was undertaken using four markers (TA59, TA96, TR19, and CaM1402) linked to resistance genes, 6 BC1F1 plants contained 4 resistant alleles and participated in the second backcrossing to produce BC2F1 plants. In the following, from a total of 52 BC2F1 plants 12 BC2F1 plants contained 4 resistant alleles, for background selection (BGS) to observe the recovery of recurrent parent genome using 16 SSRs, At this stage based on the BGS results 5 plant, with the highest background recovery, selected and backcrossed with recurrent parent to produce BC3F1 plants. Followed by cycles of, 6 plants in BC3F1 containing resistant genes and most similar to the recurrent parent were selected. The identified plants were selfed to obtain 6 BC3F2 lines which were screened phenotypically for resistance to fusarium wilt. Finally, 12 BC3F2 lines were obtained which led to identification of 3 highly resistant lines of Hashem with FWR gene introgressed in them. Also, in this study using Meta-QTL analysis region associated with genetic resistance to different race of fusarium wilt were identified on chromosomes 2, 4, 5 and 6.

Chickpea, cultivated in rainfed and irrigated fields across many countries, faces biological stresses such as Ascochyta blight caused by the pathogenic fungus Ascochyta rabiei. This disease poses a severe threat to chickpea fields, resulting in substantial annual damage in various countries. Identifying resistance sources through molecular methods, specifically genomic tracking of the resistance trait, holds the potential to expedite and enhance control measures effectively. The use of the Amplification Refractory Mutation System (ARMS) method, involving functional point mutations and resistance-related alleles, proves highly efficient in advancing selection programs targeted at combating Ascochyta blight.

The fragment harboring the SNP18-Pos57723 point mutation within the GSh118 gene sequence underwent amplification and sequencing using PSh118-F/R specific primers. This process utilized DNA extracted from both resistant (MCC113) and susceptible (ILC263) chickpea genotypes. The identification of alleles associated with resistance and sensitivity was established. Distinct differentiating primers were formulated for resistant (PSh118-Fc and PSh118-R) and sensitive (PSh18-Fg and PSh18-R) genotypes. The specific annealing temperature for each reaction was determined through a temperature gradient analysis. The efficacy of the designed primers in distinguishing between resistant and sensitive genotypes was assessed by conducting PCR and comparing their electrophoresis patterns.

The presence of alleles linked to GSh18-2773C resistance and GSh18-2773g sensitivity was confirmed in resistant and sensitive chickpea genotypes, respectively. The specific primers PSh18-Fc and PSh18-R, designed for detecting the resistance allele, successfully amplified the expected 330 bp fragment in the resistant chickpea genotype MCC133. The study results affirmed the accurate amplification of alleles using these designated primers.

In the present study, the ARMS method, known for its high sensitivity, specificity, rapidity, cost-effectiveness, multiplexing capabilities, compatibility with high throughput methods, and non-destructive nature, was employed to differentiate chickpea genotypes resistant to Ascochyta blight. The ARMS method's effectiveness in selecting resistance sources among various plant lines and cultivars has been demonstrated through allele tracking related to the resistance gene in prior studies. The results of this study indicated that the ARMS method accurately distinguishes chickpea genotypes resistant and sensitive to Ascochyta blight based on alleles associated with the GSh18-2773 position's point mutation. Thus, employing specific primers for the resistance allele is recommended in molecular selection studies of chickpea sources resistant to Ascochyta blight.

The study of genetic diversity with the help of molecular markers is the main precondition and an important step in the improvement of vegetable plants, including tomato and is the basis for the effective use of heterosis and the protection of genetic pools. Several studies have been conducted using molecular markers between different tomato lines, but there is little studies in Iran. Based on this, the present study intends to estimate the genetic distance between a numbers of tomato lines using ISSR molecular markers.

In the present study, 12 tomato lines were investigated using microsatellite primers (ISSR) at Sari University of Agricultural Sciences and Natural Resources. Polymerase chain reaction (PCR) was performed by ISSR primers and PCR products fractioned on 1.5% using agarose gel electrophoresis then the bands were analysed.

The primers used showed acceptable polymorphism (85.28) and suitable score able banding patterns. The highest and lowest polymorphism information content (PIC) with values of 0.40 and 0.22 were related to ISSR2 and ISSR10 primers, respectively. The ISSR2 primer was more efficient than other primers in identifying and classifying the studied lines. The degree of genetic similarity varied from 0.23 to 0.95 between the studied lines and lines Fanal and Harlekyn had the most similarity and lines CP with Bibor and after that SP with Bibor showed greatest genetic distances. Results of cluster analysis and evaluation of quantitative characterestics of studied lines showed that genotypes 4, 5, 6 and 8 placed in first group had better yield and earliness comparing other genotypes.   

Lines CP, SP and Bibor had a great genetic distance from each other and hybridization between CP and SP with Bibor as well as utilizing of selected genotypes in first group is recommended for next breeding programs in tomato. The ISSR2 primer has useful information in distinguishing the studied lines than other primers and its useful in breeding programs of tomato.

Parrotia persica, a native species of the Hyrcanian forests, is distributed from the west to the east of the forests of northern Iran. This study investigated the genetic diversity of Parrotia persica using molecular information from populations collected from three habitats in Gilan, four habitats in Mazandaran, and three habitats in Golestan. Using the Amplified Fragment Length Polymorphism (AFLP) marker, 826 bands were obtained, with more than 90% of the bands being polymorphic. The mean content of polymorphic information (PIC) and marker index (MI) were calculated to be 0.24 and 22.4, respectively. Cluster analysis based on the Jaccard similarity coefficient and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) algorithm revealed a wide variety in the collected samples. Based on the Jaccard similarity coefficient, the range of genetic similarity varied from 0.26 to 0.44. The results of the molecular analysis of variance (AMOVA) showed that 21% of the total genetic diversity in Parrotia persica populations was due to genetic diversity between the populations of the three provinces of Golestan, Mazandaran, and Guilan. Genetic diversity within the populations of the habitats accounted for 79%. It can be concluded that there is no difference between the populations based on the diversity of molecular information, but there is diversity within the populations. The diversity among the evangelical populations seems to be more influenced by the environment than genetically. However, for a more accurate conclusion, it is suggested that different ecotones of the population be examined with molecular markers.

Molecular science has an important role in the conservation and stability of animal species. Biology conservasion is essential to identify populations decline to implement adequate restoration programs. Despite molecular developments, there are many studies in this field to better understanding molecular merkers.

In this study, the genetic markers were examined that were used in more than 100 Persian and English articles.

Sources were selected from studies in the field of conservation biology, biology, molecular ecology, molecular markers and other related topics to provide a clearer model of each marker usage in the field of conservation biology.

In general, each molecular marker is not limited to one case, but there is a clear relationship between the characteristics of each marker and the maximum usage in many cases.

However, selecting the adequate marker in a wildlife conservation study the following steps are suggested: 1) study the species 2) Check the problems scale 3) Characteristics of molecular markers. Also, considering the mutation rate and the variability degree among nuclear and mitochondrial markers has a role in selecting of adequate marker. But in order to obtain accurate genetic and ecological information in conservation biology, the principles of population genetics and molecular evolution require more comprehensive education; By comparing molecular results with other sciences and quality control of the sequenced gene, more reliable results were obtained in the field of genetics and conservation.

Kelussia odoratissima Mozaff, is one of the most important medicinal plants in Iran and is facing the risk of extinction. Keluss is not only used as a valuable herb in its raw form but also possesses numerous therapeutic properties in traditional medicine and modern herbal drug combinations from a pharmacological perspective. However, the lack of information on its genetic diversity and the mechanisms of important metabolite production in this plant pose challenges for genetic research on this species. In the present study, with the aim of developing high-throughput EST-SSR markers, the assembly process of short reads obtained through next generation sequencing technology was employed. A total of 7575 microsatellite loci were identified in 6388 unigenes. Among these markers, dinucleotide motifs followed by trinucleotide and mononucleotide motifs exhibited the highest abundance. Blast analysis of the sequences containing microsatellites revealed that 79.74% of the transcripts had at least one record in the non-redundant protein database. Further investigations involving transcription factors and functional annotations indicated that most of the microsatellite-containing unigenes are involved in metabolic pathways and the biosynthesis of secondary metabolites. The results showed that these markers assist in the analysis of genetic diversity and metabolic pathways of this plant, information that can be crucially important for the conservation and sustainable utilization of this valuable plant.

Production of high-quality seeds to stabilize crop yield is an important challenge for breeders. One of the most important answers to this challenge is to clarify the molecular mechanisms associated with seed vigor characteristics. Functional proteins of Cupin superfamily are among the molecules in signaling pathway. Previous research has shown that in maize, a storage protein similar to the functional Cupin superfamily protein called ZmGLP is effective in seed germination. However, in the previous experiments, suitable indicators were not used to assess seed vigor and its relationship with field establishment. So, it is needed to study the performance of ZmGLP in predicting field emergence to complete the previous research.

An experiment was performed on 14 samples of commercial inbred maize lines. In this experiment, in addition to the laboratory evaluation of seed germination, field indices of physiological seed quality including the percentage of seedling emergence in the field, time to 50% seedling emergence, time to 90% seedling emergence, seedling dry weight, seedling height and coefficient of variation of seedling height was also assessed. In the polymerase chain reaction, two pairs of primers (CF / CR primers and IDF / IDR primers) were used to identify the DNA sequence of the Cupin.

The results show that the seeds were different in terms of physiological quality. The lowest percentage of germination in laboratory was related to K1264/1, while the lowest physiological quality of seeds in field indices was observed in K1263/17. The molecular test confirmed the presence of the desired allele at the InDel9 site of vigor-related genes in the three samples of B73, K1264/1, and K1264/5-1, but no amplification band of the InDel9 site was observed in all K1263/17 seed samples. Due to the fact that line K1264/1, which had the lowest germination percentage in the laboratory, had an amplification band at this related site to vigor, it is not enough to rely on the results of the laboratory germination test to investigate the relationship between this gene and seed vigor. The field emergence test and seed vigor test that have a good prediction of field emergence must be used in these studies.

According to the results of this experiment, molecular tests with functional markers based on Indel9 can be used to accelerate the evaluation of vigor, especially when the breeder is breeding a new line or hybrid. It is a useful, rapid, and effective molecular method to predict seed emergence in the field and screen the lines to ensure the genetic strength of the germination of the lines, especially in the temperate germplasms of corn. Finally, it is necessary to determine the threshold of low vigor during seed quality investigation in different cultivars, and relationship between the presence or absence InDel9 site should be considered in future research.

Highlights:

1- The feasibility of using molecular markers to determine the seed vigor of corn lines in the field was studied and optimized for the first time.
2- The results of physiological quality assessment of seeds in the field for the studies related to the relationship between molecular markers and seed vigor were exploited for the first time.
3- The Indel9 site and molecular markers related to seed vigor in the field were introduced.

The PCR-RFLP method was used for the first time in Iran to evaluate and determine haplotype relationships and population structure in 14 tea genotypes from three countries: Iran, India (Assam) and Japan. Three universal chloroplast primers (HK, CS and B1B2) and four restriction enzymes (HinfI, AluI, PstI and BglI) were used in this study and agarose gel was used to separate the bands. The B1B2 primer could not be used in this study due to the production of several bands. Also, the DT primer produced a monomorphic state after cutting, and only the fragments obtained from the HK primer and HinfI enzyme showed a polymorphic state. The mutations identified by the primer-cutting enzyme HK-HinfI were deletion-addition mutations in the range of 20-50 bp, which grouped the samples into five haplotypes (H1, H2, H3, H4 and H5). The distribution of samples in haplotype groups did not correspond to the geographical distribution, and there was no specific haplotype of a sample series. According to the results, it can be said that the PCR-RFLP method is a suitable method for investigating the genetic diversity of tea genotypes at the organ level.

Wheat is the most important cereal crops; it is a stable diet for more than one third of the world population. DNA markers play the most important role in diversity due to the relative ease in their generation and elimination of the influence of environment. The objective of this research were study of genetic diversity of bread wheat cultivars using RAPD markers and efficiency of these markers in grouping and distinguishing wheat cultivars based DNA fingerprint. 

In this study 42 wheat cultivar was evaluated with using 20 RAPD markers. The amplified fragment profiles were visually scored for presence (1) and absence (0) of bands and entered in a binary matrix. 

The results showed that, RAPD primers detected 132 fragments and 88 of them (66/67%) were polymorphic. The amplified DNA fragments varied in size from <100bp to 3000bp. The number of polymorphic fragments primer ranged from 2-7 with an average of 4/4. RAPD68 and RAPD28 each whit 7 polymorphic bands showed the highest amount of polymorphism. Primer RAPD68 were able to distinguish the cultivars omid and azadi, primer OPB08 cultivars Chenab. Sepahan, Salyasonez and primers TIBMBD17 and TIBMBB09 cultivar Atrac from other cultivars. Cluster analysis using Jaccard matrix and UPGMA method was performed, wheat genotype clustered in seven distinct groups, and the genetic similarity values ranged from 0.74 to 0.94. sistan and gasparo showed the most genetic similarity (94%). Atrac, Azadi and vrinac were clustered as outliers. Analysis of molecular variance (AMOVA) determined 1% inter group diversity and 99% intra group diversity for studied genotypes. 

The results of this research showed that there was not genetic variation among bread wheat cultivars, that can be due to the low number of primers and cultivars under investigation. Suggested to use other primers such as SSR, which shows more diversity.

The genetic diversity of wild species associated with the early barley gene pool is crucial for exploitation in breeding programs. Barley (H. vulgar) and its ancestor (H. spontaneum) are excellent and economical model systems for genetic research, exploration and exploitation. In the study of the genetic diversity of 114 native barley populations collected from the west of the country, the EST-SSR molecular marker has been used to aim at the degree of genetic diversity of these native genotypes and evaluate the efficiency of these native genotypes and evaluate Is. There is genetic diversity in genotypes.

First, seeds were cultured in pots for DNA extraction. DNA extraction was performed by modified CTAB method for each genotype. Polymerase chain reaction (PCR) was performed in a volume of 20 μl. The PCR product was injected in 4% agarose gel with 1% TBE reaction buffer and Safe View dye to show the strips. In order to perform statistical analysis, the data were prepared in the form of a matrix and the parameters related to primers and populations were evaluated.

The number of amplified alleles varied from 2 to 4 alleles for markers and the studied primers reproduced a total of 40 alleles in the studied genotypes with an average of 2.85 per marker. The average percentage of polymorphisms for the studied primers was 96.42%. The content of polymorphic information (PIC) ranged from 0.397 to 0.189. The results showed that there is the highest level of diversity in the populations of Lorestan province and the lowest level of diversity in the populations of Kurdistan province. The dendrogram obtained from the cluster analysis of genotypes was divided into 14 groups, which according to the grouping of genetic diversity were partially adapted to the geographical distribution. Also, the results of the analysis to the original coordinates were somewhat consistent with the cluster analysis.

The results showed that there was good diversity among the studied populations. Also, suitable polymorphisms were observed among the primers. The primers GBM1221 with 3 alleles, GBM1461 with 3 alleles and SCSSR04163 with 4 alleles, which had the best polymorphism, are introduced as superior primers for use in future atmospheric research. The presence of high genetic diversity in natural populations of H. spontaneum wild barley indicates that the germplasm of this plant can be preserved in natural habitats. This diversity is also a valuable resource for identifying molecular markers informative for different phenotypic traits.

In rice, the use of male sterility is a prerequisite for the commercial exploitation of heterosis. Two well established male sterility systems in rice are cytoplasmic genetic male sterility (CMS), a three-line system, and environmentally sensitive genic male sterility (EGMS). EGMS has two types of mechanisms: PGMS and TGMS. TGMS lines are sterilized when they are at a temperature above 25-30°C during the cluster and flowering stages. In order to accelerate the development of TGMS lines in various genetic fields, marker-assisted selection (MAS) can increase the speed and accuracy of this transmission. The aim of this study was to identify the chromosomal location of the gene controlling the sterility trait TGMS and the SSR marker associated with this gene in Nemat TGMS cultivar.

 For this study, Nemat TGMS cultivar was crossed with Fajr cultivar to obtain F2 and BC1 populations for genetic studies. After DNA extraction from 142 genotypes and PCR, the results observed in F1, F2 and BCF1 generations clearly indicated that the TGMS trait in TGMS is controlled by a recessive gene. For this study 14 SSR markers were used to determine the marker correlated with TGMS gene. The frequency of recombination between markers and TGMS locus was estimated using maximum likelihood, assuming that all individuals were completely sterile in terms of TGMS homozygous location. Linkage between marker loci and TGMS QTL was tested by chi square (χ2) test and LOD score.

Among the markers, RM110 and RM29 primers had a high correlation with TGMS gene (5.74 and 11.63 cM, respectively). Various markers have been reported by researchers for the TGMS gene on chromosome 2.

The use of mutation in sterilization (TGMS) and variation in cultivar genotype has led to different markers being reported even for a particular gene. The use of markers that have a high correlation with this trait (MAS) can facilitate the transfer of TGMS gene to other cultivars.