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A narrow genetic base has hindered improvement of Brassica juncea (AjAjBjBj). In this study, large-scale genomic components were introduced from diploid ancestor species into modern B. juncea using a digenomic hexaploid strategy. The hexaploids AjAjArArBjBj and AjAjBjBjBnBn were first developed from B. juncea × B. rapa (ArAr) and B. juncea × B. nigra (BnBn), and then crossed with dozens of B. nigra and B. rapa, respectively. Both types of hexaploid showed high pollen fertility and moderate seed set throughout the S1 to S3 generations, and could be crossed with diploid progenitor species under field conditions, in particular for the combination of AjAjBjBjBnBn × B. rapa. Thirty AjAr Bn Bj-type and 31 Aj Ar BnBj-type B. juncea resources were generated, of which the AjAr Bn Bj type showed higher fertility. Of these new-type B. juncea resources, 97 individual plants were genotyped with 42 simple sequence repeat markers, together with 16 current B. juncea accessions and 30 hexaploid plants. Based on 180 polymorphic loci, the new-type B. juncea resources and current B. juncea were separated clearly into distinct groups, with large genetic distance between the new-type B. juncea resources and current B. juncea. Our study provides a novel approach to introducing large-scale genomic components from diploid ancestor species into B. juncea.

Phenotypic and genotypic characters a source of superior character in the framework of plant breeding programs, especially in increasing local food yields and productivity. In this study, local cultivars of garlic from Eban, North Central Timor (TTU) were used. A total of 15 phenotypic characters were observed , and threeISSR primers were used to identify the phenotypic and genotypic characters. The phenotype characterization showed the similarities in most of the characters observed. The tuber characters showed some differences in tuber diameter (2.5 cm and 1.9 cm), the number of cloves (14 and 10 cloves), tuber weight (11.28 g and 5.18 g), weight of cloves (0.89 g and 0.45 g) for local garlic from Saenam and Fatuneno, respectively. A total of ninepolymorphic bands and 17 monomorphic bands from threeprimers Inter Simple Sequence Repeat (ISSR) were detected. The coefficient of similarity of two groups based on genotypic characters was 0.65% - 1%. This study concluded that the two local Eban garlic varieties were different based on morphology and ISSR data. However, phenotypically there were differences in the size of the tubers and cloves, the number of cloves per tuber, and the weight of the cloves.

Genetic studies through molecular markers proved important to find out the genetic diversity of canola. In this study, 50 lines of canola were used to find the polymorphism using 15 SSR primers and investigated the genetic diversity, PIC values, frequency-based genetic distance, and allelic frequencies. Mean gene diversity, frequency-based genetic distance, and PIC values were 0.8777, 0.233 and 0.8666, respectively for the canola lines. A good range of genetic diversity was found among studied canola lines with value 85.91% polymorphism. Maximum and minimum genetic distances among 50 lines were 1 and 0.26, respectively. Accessions ACC-26068, ACC-24241, ACC-24244, ACC-24233, ACC-24423 and ACC-24224 have maximum genetic distance. Accessions ACC-24879 and ACC-24169 had minimum genetic distance i.e., 0.26. Dendrogram based on genetic distances showed four main clusters that were further dividing into several sub-clusters. The primers utilized in the present study, were valuable to identify different accessions of canola to find the variability present. This variability will be helpful to initiate the breeding program with their molecular genetic basis.

To analyze the genetic diversity of 9 species of Clematis from 31 different populations, we extracted DNA by the improved CTAB method, used ISSR-PCR for amplification, and then selected 9 primers with clear amplified bands from amongst 220 primers. A total of 127 clear bands were amplified, of which 126 were polymorphic bands, yielding a ratio of 99.2%. The polymorphism information index (PIC) of the primers ranged from 0.9326 to 0.9649. The Nei’s genetic diversity index (H) was 0.2750, the total gene diversity (Ht) was 0.2845, and the genetic differentiation coefficient (Gst) was 0.6696, indicating high genetic differentiation among populations of Clematis. After cluster analysis, the 31 Clematis populations were divided into 3 categories. Principal coordination analysis (PCoA) of 9 Clematis species then showed that the genetic relationship between samples of the same Clematis germplasms was closer than that of samples from the same region. The mantel test revealed a significant positive correlation between genetic distance and geographical distance among the populations. The population clustering results are broadly consistent with the clustering graphs of UPGMA and PCoA. We can conclude the polymorphism of the 9 primers is good, and that the genetic diversity of 31 Clematis populations is rich. Individual Clematis germplasms are closely related and will gather together preferentially.

Micronutrient deficiency is a widespread food-related health problem around the world. The present study was conducted to evaluate a set of 63 advanced breeding lines of bread wheat (Triticum aestivum L.) for grain iron (GFe) and grain zinc (GZn) concentrations, and to characterise the germplasm set via simple sequence repeat (SSR) markers (both genic and random). Substantial variation was found for both micronutrients. GFe concentration ranged from 28.9 to 67.4 mg kg–1 and GZn from 26.3 to 56.6 mg kg–1. Molecular characterisation with six genic and 20 random SSR markers detected 168 alleles with an average of 3.170 alleles per locus. Analysis of genotypic data based on division into two subpopulations revealed 165 alleles with an average of 3.113 alleles per locus in the low GFe–GZn subpopulation, whereas in the high GFe–GZn subpopulation, 149 alleles with an average of 2.811 alleles per locus were detected. Genic SSRs detected a higher average number of alleles (3.273 alleles per locus) than random SSRs (3.143 alleles per locus). Hierarchical clustering using genic markers alone clustered the whole germplasm set into two distinct groups: one possessing low GFe–GZn genotypes, the other with high GFe–GZn genotypes. Study of marker–trait associations (MTAs) identified seven new MTAs (six for GZn and one for GFe) and validated one MTA for GZn concentration. The promising genotypes and MTAs identified during the study will prove useful in wheat bio-fortification programs in the future.

Mulberry (Morus spp.) is a cross-pollinating and highly hybridized plant of which productivity are greatly varied in different varieties. Accurate distinction among mulberry varieties and understanding of phylogenetic relationship among them would be crucial for the development of sericulture. We have analysed molecular distinction among four mulberry species and varieties cultivated in DPR Korea by using nuclear ribosomal internal transcribed spacer (ITS) sequences and inter simple sequence repeat (ISSR) markers. ITS sequences doesn’t represent a remarkable interspecific distinction among four mulberry species used in our study, suggesting that it could not be employed to identify them. ISSR analysis using 16 random primers generated 158 different markers ranging from 100 to 4000 bp in size. The results showed the inter-specific genetic variation (55.34%) was slightly higher than intra-specific genetic variation (44.66%), with comparatively low average number of migrants per generation (Nm) among populations (0.3886). Using ISSR primers selected in this study, in the future, the suitable breeding strategy might be established in raising of elite mulberry varieties on the basis of interspecific hybridization.

To reveal the genetic diversity and genetic relationships of China’s Bergenia germplasm, 28 Bergenia accessions from different regions in China were analyzed by 24 intersimple sequence repeat (ISSR) markers. The results showed that 318 sites were amplified in all germplasm, including 307 polymorphic sites, and the percentage of polymorphic sites was 96.54%. Cluster analysis showed that the 28 accessions were divided into three categories, with a similarity coefficient of 0.5475. Bergenia purpurascens was clustered into one category; B. scopulosa was clustered into one category; and B. tianquaninsis, B. emeiensis, B. stracheyi, and B. crassifolia were clustered into one category. The results of the cluster analysis indicated that the 28 accessions were not completely classified by origin. Using the ISSR marker technique to analyze the phylogenetic relationship of Bergenia germplasm is helpful for identifying valuable resources and providing a theoretical basis for the selection of breeding parents.

Genetic diversity among ten maize genotypes (seven inbred lines and three testers) was investigated using ten simple sequence repeats (SSRs). Primers (bnlg128, bnlg1839, Umc1117, bnlg1144, and bnlg1152) generated the highest number of bands (4 bands) for inbred lines while the primer bnlg128 showing the highest number of bands (3 bands) for testers. The primer bnlg128 shows the highest effective number of alleles (ne) for inbred lines and testers. Among the studied primer bnlg1839 in inbred lines and primer bnlg128 in testers showed the maximum polymorphism information content (PIC) and the greatest diversity. Using UPGMA cluster analysis, the seven inbred lines were grouped under three clusters, while grouped the testers under two clusters. Most of the inbred lines which were derived from the same source population were grouped in the same cluster based on the SSRs DNA markers, indicating high genetic differentiation among their source populations. Results showed that the SSRs were informative in detecting genetic differences among the maize inbred lines and testers, as exhibited by the high average of Shannon’s information index (I), Nei’s expected heterozygosity (Nei’s), and PIC. The results suggest that the studied genotypes are diverse and may be utilized for further breeding programs.

Aquarium fish industry has become a capacity for job creation and income generation. Aquarium fish uses for decoration and creating tranquility in the environment and beautification in public places. This study was conducted to investigate genetic variation between different populations of various varieties of guppy ornamental aquarium fish using inter simple sequence repeat (ISSR) markers in Iran for the first time. DNA was extracted from twelve strains. Polymerase chain reaction was performed using seven ISSR primers. GenAlex 6.501 and XLSTAT software was used to analyze data. Results of analysis of molecular variance showed that 54% of observed polymorphism belongs to variance among populations and 46% corresponds to diversity between populations. The highest and the lowest polymorphism percentage belonged to (GAG)5GC (90%) and (AG)8C (41%) markers, respectively. Percentage of polymorphic loci ranges from 32.31% to 86.15%. The results of phylogenetic tree showed that this tree was divided into 2 major clusters. Cluster I included only 2 populations and cluster II contained other 10 studied populations. It is suggested that this marker system was found to be efficient in discriminating each genotype at the molecular level and can be used for genetic diversity analysis for aquarium guppy fish because a greater understanding of the potential of the species is necessary for supporting long-term genetic improvement.

Cowpea is a leguminous crop that has received widespread attention due to its high nutritional value. However, it is prone to losing some of its nutritional content due to the long cooking process. In this study, fourteen cowpea genotypes were evaluated for their chemical properties before and after cooking, along with the effect of different cooking treatments on the cooking time, considered as the main indicator of the cooking quality. Moreover, the correlation between molecular markers (simple sequence repeat (SSR) and intersimple sequence repeat (ISSR)) and the cooking time of cowpea genotypes was determined. The obtained results showed significant differences between all the investigated genotypes before and after the cooking procedure, reflecting significant genotypic and heritability estimates. Kareem 7 and Greenish Black Balady genotypes showed the shortest cooking time. Microwave's treatment manifested the shortest cooking time compared with the other treatments, which appeared as a new approach to improve the cooking quality of cowpea seeds. Spearman's rank correlation showed that the calculated values were smaller than the tabulated value at 0.05, reflecting the existence of a rank correlation between SSR/ISSR‐PCR banding products and the cooking time of cowpea genotypes. Such a study appeared to be a new approach, particularly in Egypt for the proper selection of the optimal cowpea seed on the basis of its cooking quality., Such a study appeared to be a new approach, particularly in Egypt for the proper selection of the optimal cowpea seed on the basis of its cooking quality.

Monilinia fructicola, causal agent of brown rot of stone fruits, is an economically important problem worldwide. Six of the sequence tagged microsatellite sites developed for M. fructicola were used to genotype 68 M. fructicola isolates, which included isolates from three cities in Turkey (n = 42) that were compared to isolates from the USA (n = 15) and Italy (n = 11). AMOVA indicated a significant differentiation among samples from the three different countries. Samples from Turkey represented with nine haplotypes indicating a low diversity according to these markers. Samples from the USA with 12 haplotypes showed the highest genetic diversity values among the sample group from the three countries. However, all the samples from Italy were found as a single haplotype with those markers. Based on Nei’s genetic distance measurements, the single genotype from Italy was distinct from the others and samples from Turkey and the USA were genetically closer to each other, which also presented on the principal coordinate plot. While informative, these results suggest that ascertainment bias in marker development may limit the power of these markers when applied to populations of M. fructicola from other locations in the world. Knowledge on genetic diversities and comparative analysis provides valuable insight for recent changes and movements in pathogen populations which is important for the disease management.

As a medicinal and edible plant, the plum (Prunus mume Sieb. et Zucc.) plays an important role in the treatment of diseases and human health. However, the genetic diversity and genetic composition of medicinal plum species cultivated in China remain incompletely clear, hampering the accurate identification. Here, the genetic diversity and population structure of 40 medicinal plum varieties in China were analyzed using polymorphic simple sequence repeat (SSR) molecular markers, and the lineages compared. SSR allelic analysis was followed by a STRUCTRE analysis. The mean observed heterozygosity (HO) and polymorphism information content were 0.74 and 0.69, respectively. The cumulative probability of identity was very low with a value of 7.17 × 10−19. STRUCTURE analysis provided evidence for three genetic groups. Further, a genetic fingerprint consisting of four pairs of SSR primers has been successfully developed to accurately distinguish 40 varieties of medicinal plum. This study provides reference data which can be used to assist the breeding, quality evaluation, and development of medicinal plum varieties.

Over 450 potato varieties and landraces are preserved as in vitro cultures by the Plant Biotechnology department of Estonian Crop Research Institute. This collection contains germplasm from various countries as well as old Estonian landraces. These potato accessions are periodically grown in the field for verification; however, this is not always sufficient to validate varieties and, in many cases, the true origin or identity of old landraces is not known. A reliable and cost-effective method for fingerprinting Estonian potato germplasm needed to be implemented. Simple sequence repeat (SSR) markers are known as robust and efficient markers for the identification of tetraploid potato. Here, fingerprinting results are presented for 103 genotypes from the Estonian potato collection using 8 SSR markers. It was shown that as few as three markers were sufficient to yield unique fingerprints for the given set of samples. SSR markers were used to estimate the allele copy number with varying success. Additionally, the reproducibility of four of these markers was assessed in two different laboratories, and repeatable results were observed. It was verified that the putative landrace ‘Kuldkera’ has the same genetic fingerprint as the variety ‘Ando’, whilst an expected clone of ‘Early Rose’ is a completely different genotype. Implementation of genetic fingerprinting and these results is of high importance for further potato genebank management and Estonian breeding programme.

Two cultivars of local rice, namely Mentik Wangi and Mentik Susu, have been grown around west to south flank of Merapi volcano. They are highly valued for their good taste and cooking characteristics. An attempt to introduce drought-tolerant rice varieties has been conducted by crossing them with two sources, namely Kasalath and Bluebonnet. Molecular-assisted selection using a set of SSR markers was applied to check the F1 and F2 generation trueness and segregation. This research was conducted in the facilities of Department of Agronomy, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta. The research was a field experiment arranged in a completely randomized design, consisting of Mentik Wangi, Mentik Susu, Bluebonnet, and Kasalath, as well as their crossings’ F1 and F2 plants as treatments. The SSR markers used are RM72, RM228, RM518, and RM20(A). Polymorphism test of the parents showed that polymorphism exists between local and donor parents, thus these markers were considered eligible for the F1 and F2 tests. The heterozygous individuals of F1 were 75% for Bluebonnet × Mentik Wangi, 44.4% for Bluebonnet × Mentik Susu, and 46.7% for Mentik Wangi × Bluebonnet; all were consistent in every primer used. We could not confirm that the F2 populations showed segregation pattern that followed Mendelian segregation in some crosses due to too small size of the sample. Heterozygous individuals in F2 showed the differences pattern for each marker, indicating that the location of the SSR markers were far from each other in the genome.

Eastern filbert blight (EFB), caused by Anisogramma anomala, is a fungal disease threatening the european hazelnut (Corylus avellana) industry in the Willamette Valley of Oregon. The pathogen is endemic to the eastern United States where it causes little damage to the wild Corylus americana but causes severe cankers on most cultivars of the commercially important european hazelnut. The host genetic resistance in ‘Gasaway’ is conferred by a dominant allele at a single locus on linkage group 6 (LG6), and resistance from several other sources has been mapped to the same region. Some fungal isolates can overcome ‘Gasaway’ resistance, prompting a search for other sources of resistance. Resistance from other sources has been mapped to LG2 and LG7, for which additional simple sequence repeat (SSR) markers would facilitate marker-assisted selection (MAS). In this study, an in silico approach was used to develop new polymorphic SSR markers in the EFB resistance regions on LG2 and LG7. Starting with a search of 17 contigs of the ‘Jefferson’ genome sequence, 45 new polymorphic SSR markers were developed, characterized, and placed on the linkage map. The new SSR markers had an average of 10.18 alleles per locus, and average values for expected heterozygosity, observed heterozygosity, polymorphism information content, and frequency of null alleles of 0.72, 0.65, 0.68, and 0.068, respectively. Of the 42 new polymorphic SSRs segregating in the mapping population, 24 were on LG2, 12 were on LG7, and six were placed on other LGs. The new and previously developed SSR markers were used to study six new sources of EFB resistance, four from Russia and two from Crimea. Six resistant selections were crossed with susceptible selections, resulting in 7 progenies. Phenotyping for disease response revealed that segregation in progenies of the two Moscow selections (#2 and #27), one Russian selection (OSU 1187.101), and one Crimean selection (H3R12P62) fit the 1:1 segregation ratio expected for control of resistance by a dominant allele at a single locus; but in progenies of the other Russian selection (OSU 1166.123) and the other Crimean selection (H3R07P11), there was an excess of resistant seedlings. Correlation of disease scores and alleles at SSR loci indicated that resistance from three Russian selections (Moscow selections #2 and #27 and OSU 1166.123) and the Crimean selection H3R12P62 was on LG7, while resistance from Russian selection OSU 1187.101 was on LG2. Resistance from Crimean selection H3R07P11 was not correlated with markers on LG6, or LG2, or LG7. These sources and new SSR markers will be useful in MAS and the pyramiding of resistance genes in the breeding of new EFB-resistant cultivars.

Macrophomina phaseolina (Tassi) Goid is one of the most important soil-borne fungi that affects sesame and causes charcoal rot disease, with a great economic challenge to sesame growers worldwide. Pathogenic and molecular characterizations of eleven M. phaseolina isolates collected from different geographical regions in Egypt were carried out to determine the pathogenicity and genetic diversity. Pathogenicity tests showed a pathogenic variability among the isolates. The identification of M. phaseolina isolates was confirmed by a specific primer and analyzed for genetic diversity using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. The correlation between RAPD analysis of M. phaseolina isolates and their pathogenicity were estimated. Cluster analysis placed the isolates in two distinct clusters and exhibited clear correlation with their pathogenicity. Analysis of ISSR profiles revealed distinct genetic diversity among isolates and showed different clusters according to the geographical regions, in which close geographic origins tend to group nearly. The present study clearly demonstrated that M. phaseolina isolates which were obtained from different geographical regions were highly variable. RAPD and ISSR markers were suitable to reflect the genetic diversity among the studied isolates and could help in DNA finger printing which can be used in future breeding programs of sesame.

Chinese wild Vitis is a useful gene source for resistance to biotic and abiotic stresses, although there is little research on its genetic diversity and structure. In this study, nine simple sequence repeat (SSR) markers were used to assess the genetic diversity and genetic structure among 100 Vitis materials. These materials included 77 indigenous accessions representing 23 of 38 wild Vitis species/cultivars in China, 18 V. vinifera cultivars, and the five North American species V. aestivalis, V. girdiana, V. monticola, V. acerifolia, and V. riparia. The SSR loci used in this study for establishing an international database (Vitis International Variety Catalogue) revealed a total of 186 alleles in 100 Vitis accessions. The mean values for the gene diversity (GD) and polymorphism information content (PIC) per locus were 0.91 and 0.90, respectively, which indicates that the discriminatory power of the markers is high. Based on the genetic distance data, the 100 Vitis accessions were divided into five primary clusters by cluster analysis, and five populations by structure analysis; these results indicate these Chinese wild grapes were more genetically close to European grapes than to North American species. In addition, the clustering patterns of most accessions correlated with the geographic distribution. An analysis of molecular variance (AMOVA) revealed that 3.28%, 3.27%, and 93.46% of the variance occurred between populations, between individuals within populations, and between individuals within the entire population, respectively. In addition, we identified three previously undescribed accessions (Wuzhi-1, MZL-5, and MZL-6) by cluster analysis. Our results reveal a high level of genetic diversity and variability in Vitis from China, which will be helpful in the use of genetic resources in future breeding programs. In addition, our study demonstrates that SSR markers are highly suitable for further genetic diversity analyses of Chinese wild grapes.

Plectranthus edulis (Vatke) Agnew is an underutilized and neglected ancient Ethiopian indigenous annual tuber crop. Very limited research has been done to improve this crop especially in the area of molecular genetic diversity. Therefore, this study was aimed to assess the genetic diversity of this crop using ISSR markers. Fourteen ISSR primers were screened, and four polymorphic and reproducible primers were selected. These primers generated a total of 46 scorable bands of which 42 were polymorphic. Primer UBC-880 showed 100% polymorphism, and a unique band in some individuals was observed. Within-population diversity ranged from 41.30% to 65.22% with overall percentage of polymorphic loci of 91.30%, Nei’s gene diversity of 0.18–0.30 with overall diversity of 0.33, Shannon information index of 0.25–0.39 with overall index of 0.48, and 74.23% within-population variation was detected. Sodo Zuria, Gojam-II and Areka populations showed the highest diversity, whereas the lowest was exhibited by Nekemte. According to Jaccard’s pairwise similarity coefficient, Ambo and Sodo Zuria were most related populations exhibiting 0.648 similarity coefficient, and Gojam-I and Sodo Zuria were the most distantly related populations with similarity coefficient of 0.489. Wolaita and Wollega populations showed the highest genetic diversity so that these populations should be considered as the primary sites in designing conservation strategies for this crop.

Panax bipinnatifidus Seem. is a medical plant which has been used as treatment for medical diseases and health maintenance for aged people. The species was listed among Critically Endangered (CR) plants in 2007 Vietnam’s Red Data Book. Inter simple sequence repeat (ISSR) markers were first used to investigate genetic diversity of naturally distributed P. bipinnatifidus which was classified by 2 age groups (the older (>6 years old) and the younger (≤6 years old)). 30 samples were collected from 3 different locations in Lai Chau and Lao Cai (two provinces in the North of Vietnam). The results showed that there were 17 polymorphic primers and 148 bands at length of 150 to 1500 bp were created from the 17 selected primers. The number of bands in each primer ranged from 6 to 12 bands with a mean value of 8.7. It should be noted that the genetic diversity of P. bipinnatifidus was fairly high (PPB = 89.2%; He = 0.251). The genetic diversity among the older group (PPBo = 72.30%; Heo = 0.210) was lower than that of the younger (PPBy = 77.03%; Hey = 0.251). The two investigated groups had average gene differentiation (GST = 0.093) with gene distance of Dyo = 0.028 while gene flow between them was moderate (Nm = 2.438). These findings showed that P. bipinnatifidus was endangered; however, it can still grow in the future under proper protection and development thanks to its high genetic diversity.

Identification of zygotic and nucellar seedlings of Harumanis mango by morphological approach is impossible or hard to be performed. Therefore, this research were to identify zygotic or nucellar seedlings from Harumanis mango polyembryonic seeds through simple sequence repeat (SSR) molecular markers in relation with germination sequence and vigour of seedlings for source of true-to-type cutting material. The results showed that there were 3 seedlings per seed on average. The result from the molecular analysis of 136 seedlings (45 seeds) showed four zygotic seedlings in seeds 1, 15, 29 and 43 representing 8.9% of seeds evaluated. Most of the zygotic seedlings were found towards the end of germination sequence except for seed number 15 and the rest were considered as nucellar seedlings. Based on the fitted logistic regression, the predicted sequence to obtain 90% nucellar seedlings is 5.47. This means that, the germination sequence of less than 6 has 90% chance of getting nucellar seedlings compared to zygotic seedlings. Morphological characters such as number of leaves, stem diameter and leaf area could also be used as references with germination sequence. The results showed that there were significant (p

Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.

The experiment have been done on the winter farming season 2020 in one of the farms that belongs to Faris company in Basrah governorate, the technique ISSR have been used to study the genetic distance for twenty one isolates of snake melon Cucumis melo var. flexuosus species. The variations between amplified samples have been revealed after running them on a gel of agarose which have been previously stained by ethidium bromide. Five primers which gave varied product on the agarose have been selected. Those five primers produced 713 bands, both primers UBC 813 and UBC 815 showed the higher numbers of bands reached to 177 while the primer UBC862 showed the least numbers of bands (100) and the bands which showed multi-variations showed (46) bands, and the results primers amplification unique bands their number reached to (14) and five of those bands belong to the primer UBC842 while the primer UBC862 produced three bands while the primer UBC807 did not produced any bands. While it has shown 100% polymorphisms with the primers 813, 815, 842 and 862, and the least polymorphism percentage have shown with the primer UBC807 reached 75%. According to the efficiency primers, the highest efficiency percentage shown with the primer UBC813 and 815 reached to 24.82% and the least percentage shown 14.02% by the primer UBC862. Cluster analysis showed the effect on the variance of the studied cultivars.

Registration of crop varieties under Indian PPV & FR Act requires establishment of their Distinctiveness, Uniformity and Stability (DUS). At present, DUS of crop varieties are being established using morphological traits. DNA marker assay-based DUS testing would complement that based on morphological traits. Four markers, from a total of 220 tested SSR markers were polymorphic among two advanced breeding lines and two elite germplasm accessions (used as candidate varieties), and two released varieties (used as reference varieties) of dolichos bean. The four polymorphic simple sequence repeat (SSR) markers were used to establish distinctness of candidate varieties from reference varieties based on alleles unique to candidate varieties. Uniformity of candidate varieties were established using two criteria, (1) per cent homozygous plants (> 95%) and (2) SSR-homozygous loci ratio (HLR) (> 95%).The candidates which were found uniform at tested SSR loci were deemed stable. The candidate varieties were distinct from reference varieties at two of the four tested polymorphic SSR markers, as evident by the presence of alleles unique to candidate varieties. One hundred per cent homozygous plants and SSR-HLR within all the candidate varieties at all the four tested SSR marker loci suggested uniformity and deemed-stability of candidate varieties. The study demonstrated the utility of SSR markers to establish DUS of ABL/elite germplasm accessions in dolichos bean.

Niger plant (Guizotia abyssinica), exhibits phenotypic plasticity in different environments. There is need to assess its genetic diversity since guizotia species has a high number of species which may be confused amongst themselves. To achieve this, inter simple sequence repeat (ISSR) markers were used to estimate genetic diversity among 12 wild populations of Niger plant from Moiben sub-county. Total genomic DNA was extracted as per the cetyltrimethylammonium bromide (Ctab) method and subjected to ISSR analysis using 20 primers. None of the primers produced unique banding patterns. ISSR data were used to calculate a squared-euclidean distance matrix. All the twenty primers (100%) gave polymorphic bands thus they were all considered for further analysis. The allele frequency of all the primers was below 0.95 indicating that they were all polymorphic in character. Gene diversity was high ranging from 0.3550 to 0.7337 with a mean value of 0.6302. The ISSR based upgma clustering produced four clusters. Niger plant within Moiben sub-county was found to be genetically diverse though heterozygosity was not noticed. The study recommends further analysis of Niger plant so as to form a basis for further development of the plant.

Genetic purity is conventionally performed through grow-out-test (GOT) with morphological characters. Simple sequence repeat (SSR) markers are independent of G X E interaction. Herewith, 16 high yielding varieties of rice were analysed using 55 SSR markers for DNA fingerprinting and identification of genetic impurities: 14 were found to be polymorphic and amplified 48 alleles with an average of 3.43 alleles per each primer pair. The number of alleles amplified ranged from 2 to 6 and the size of the PCR products amplified from these 14 primer pairs ranged from 80 to 450 bp with polymorphic information content (PIC) from 0.14 (RM 346) to 0.99 (RM 5900). PICs at 0.5 or higher are highly informative SSR markers for genetic studies and the study reported (7/14) markers to be highly informative based on PIC values. Besides, the values of effective multiplex ratio, marker index and resolving power for the selected polymorphic primer pairs disclosed that the SSR markers used in the study are highly informative and could be potentially used for distinctness, uniformity and stability (DUS) testing, genetic purity analysis and DNA fingerprinting of rice varieties. Economic analysis of grow-out-test and SSR marker based genetic purity showed that the GOT process incurs Rs. 5.8 (INR) per seed (USD 0.07), while DNA-based marker analysis incurs Rs. 53.75 (INR) or $ 0.71 (USD).

Cross-amplification is a cost-effective method to extend the applicability of SSR markers to closely related taxa which lack their own sequence information. In the present study, 35 SSR markers developed in four oak species of Europe, North America and Asia were selected and screened in five species of the western Himalayas. Fifteen markers were successfully amplified in Quercus semecarpifolia, followed by 11 each in Q. floribunda and Q. leucotrichophora, 10 in Q. glauca, and 9 in Q. lana-ta. Except two primer pairs in Q. semecarpifolia, all were found to be polymorphic. Most of the positively cross-amplified SSRs were derived from the Asian oak, Q. mongolica. The genoty-ping of 10 individuals of each species with positively cross-amplified SSRs displayed varied levels of polymorphism in the five target oak species, viz., QmC00419 was most polymorphic in Q. floribunda, QmC00716 in Q. glauca and Q. lanata, QmC01368 in Q. leucotrichophora, and QmC02269 in Q. semecarpifolia. Among five oak species, the highest gene diversity was depicted in Q. lanata and Q. semecarpifolia with expected heterozygosity (He = 0.72), while the minimum was recorded for Q. leucotrichophora and Q. glauca (He = 0.65). The SSRs validated here provide a valuable resource to carry out further population genetic analysis in oaks of the western Himalayas.

Einkorn (Triticum monococcum L. ssp. monococcum) is an ancient diploid wheat species with many useful traits and used as a wheat gene discovery model. In this research, a total of 41 diploid and tetraploid wheat accessions were tested using simple sequence repeat (SSR) markers. A total of 33 genotypes of T. monococcum along with four genotypes each of tetraploid wheat (T. dicoccon and T. durum) were used as plant material. The analysis utilized 10 polymorphic markers, including a total number of 41 alleles with an average frequency of 4.1 alleles per locus during exploration of the level of genetic variations. Various diversity analyses, which are the effective number of alleles (Ne), gene diversity (h), Polymorphic Information Content (PIC), and Shannon's information index (I), were performed for 10 'A' genome wheat SSR markers. The results showed a narrow variation in einkorn genotypes, supported by Analysis of Molecular Variance (AMOVA), with 66% maximum variation in all genotypes. The structure analysis divided the whole germplasm into two populations. A dendrogram was constructed to determine the genetic similarities using the unpaired group method with arithmetic averages (UPGMA), which separated tetraploid wheat from other genotypes/accessions. Principal Coordinate Analysis (PCoA) co-supported the clustering of UPGMA and structure by differentiating the diploid and tetraploid wheat. These findings will help understand the genetic relationships among these wheat accessions and their use in breeding programs in the future works.

Identifying the accurate boundaries of a species is critical to have a better perspective of any biological studies. Therefore, species delimitation is a subject of extensive part of studies in the framework of biology. Erodium species possess significant pharmacological and biological activities. The whole plant was used as astringent and haemostatic in uterine and other bleeding Therefore, due to the importance of these plant species, we performed a molecular data analysis for this species. For this study, we used 60 randomly collected plants from 5 species in five provinces. Amplification of genomic DNA using 10 primers produced 52 bands, of which 50 were polymorphic (98.48%). The obtained high average PIC and MI values revealed high capacity of ISSR primers to detect polymorphic loci among Erodium species. The genetic similarities of five collections were estimated from 0.77 to 0.91. According to Inter-Simple sequence repeats (ISSR) markers analysis, E. cicutarium and E. malacoides had the lowest similarity and the species of E. malacoides and E. oxyrrhynchum had the highest similarity. The aims of present study are: 1) can ISSR markers identify Erodium species, and 2) to investigate the species inter-relationship? The present study revealed that ISSR markers can identify the species.

This study investigated simple sequence repeat (SSR) marker associated with fiber fineness in ramie (Boehmeria nivea L. Gaudich). Firstly, association analysis was carried out using 107 ramie acces...

Tamarix is an important genus of Tamarieaceae family, known for its medicinal importance. Its taxonomy is tricky, and a few molecular makers are available for species identification. The present study was designed to evaluate the genetic diversity among 21 Tamarix specimens collected from Thal desert, Punjab, Pakistan. Ten Inter-Simple Sequence Repeats (ISSR) and six Simple Sequence Repeats (SSR) markers were used. Results revealed that ISSR markers produced 131 bands, of which 116 were polymorphic (88.5%) with a mean Polymorphic Information Content (PIC) value of 0.34. The SSR markers yielded 30 amplified bands, of which 24 were polymorphic (80.28%) with mean PIC value of 0.35. Cluster and factorial analysis revealed a high level of genetic variability among tested Tamarix ecotypes. Dissimilarity index indicated a genetic variability among tested specimens ranging from 0.00 to 0.77. The ISSR analysis reflected that Tamarix specimens stemming from Kalurkot and Bhakkar region revealed the highest dissimilarity level with respect to the rest of the specimens.

Sugarcane brown rust, caused by Puccinia melanocephala, is one of the main diseases of sugarcane in China. The identification and discovery of new resistance genes have important theoretical and practical significance for preventing outbreaks of brown rust and ensuring the sustainable production of sugarcane. To screen for polymorphic simple-sequence repeat (SSR) molecular markers for localization of brown rust resistance genes, we used two populations that are suitable for genetic linkage map construction and mapping of new resistance genes to construct resistant and susceptible genetic pools. We then screened 449 pairs of primers to identify polymorphic SSR markers in the parental lines and the resistant/susceptible genetic pools. The results showed that 25 pairs of primers directed amplification of polymorphic DNA fragments between the parents of the cross combination ‘Yuetang 03-393’ × ‘ROC 24’, and 16 pairs of primers amplified polymorphic fragments between the parents of the cross combination ‘Liucheng 03-1137’ × ‘Dezhe 93-88’. Four pairs of primers (SMC236CG, SCESSR0928, SCESSR0636 and SCESSR2551) amplified polymorphic DNA fragments between the parental lines and the resistant/susceptible genetic pools in ‘Yuetang 03-393’ × ‘ROC 24’. The results of this study will establish a solid foundation for the mapping of new brown rust resistance genes, genetic linkage map construction and the development of closely-associated molecular markers in sugarcane.

Panax vietnamensis var. langbianensis is a new variety from Lam Vien plateau of Vietnam. In this study, inter simple sequence repeat (ISSR) markers were employed to investigate the genetic diversity and variability of 115 individuals belonging to two naturally distributed populations of this variety, which classified by habitat. Genetic diversity at the taxon level was high (HeT = 0.284 and PPBT = 97.2 %). The result showed lightly higher genetic diversity in population in Lac Duong region (HeLD= 0.228 and PPBLD = 81.5 %) as compared to those located in Dam Rong region (HeDR= 0.213 and PPBDR = 79.4 %). The interpopulation gene differentiation was high (GST Total = 0.221) with the genetic distance among populations was DLD-DR = 0.191. Gene flow within populations was Nm = 0.8793. In Lac Duong population, the genetic diversity of older group (HeLD O = 0.233; PPBLD O = 77.1%) was higher than of younger group (HeLD Y = 0.214; PPBLD Y = 72.4 %) and the intergroup gene differentiation was GSTDL = 0.0205 with the genetic distance between these two group was DLD O-Y = 0.0061 showed the risk of reduction in genetic diversity. In Dam Rong population, the genetic diversity of older group (HeDR O = 0.204; PPBDR O = 75.2 %) was equal to younger group (HeDR Y = 0.209; PPBDR Y = 72.7 %) and the intergroup gene differentiation was GSTDR = 0.0304 with the genetic distance between them was DDR O-Y = 0.01393 showed the stability in genetic diversity. Data for genetic diversity and variation from this study can be used to further investigate and protect this variety for conservation and development purposes and for sustainable exploiting and use of these valuable natural resources.

In this study, the molecular profiles of five soybean cultivars (Burangrang, Gema, Grobogan, Panderman, and Sinabung) exhibiting salinity resistance were elucidated. The DNA profiles of the five cultivars were found to differ based on simple sequence repeat (SSR), insertion-deletion polymorphism (InDel)-QS080465, and sequence characterized amplified region (SCAR)-QS08064 markers. Three distinct SSR profiles¾Satt-243, Satt-294, and Satt-308¾and the SCAR-QS08064 marker were only observed in the Grobogan cultivar, whereas the InDel-QS080465 marker was only successfully amplified from the Burangrang, Gema, and Grobogan cultivars. The results indicate that the Grobogan cultivar is the most tolerant soybean cultivar, followed by the Burangrang and Gema cultivars. Results were consistent with those from genetic similarity analysis, which showed that Grobogan is genetically more similar to Burangrang and Gema compared to Sinabung and Panderman. In conclusion, the five soybean cultivars have different molecular profiles that are related to their resistance to salinity. SSR markers, InDel QS080465-152, and SCAR QS08064-383 are molecular markers specific to salinity-resistant cultivars.

Sargassum is a common macroalgae found across the world, especially in the warm temperate regions. It is also known as brown seaweed. Sargassum is known for its medicinal and industrial applications. In the present study Sargassam samples were collected from three different coastal regions of Egypt in the Northern coast, Marsa Matrough and Hurghada. In addition to morphological parameters of identification, the collected species were differentiated and further characterized by Inter Simple Sequence Repeat (ISSR), and 18 S rDNA techniques. This led to the identification of four different species, namely Sargassum subrepandum var. rueppellii, S. platycarpum, S. latifolium, S. subrepandum var. massauense from the areas of collection. Four different ISSR primers used enabled differentiation of total polymorphism to the order of 33.24% among them. The phylogenetic relationship resulted in similarity between Sargassum subrepandum var. rueppellii and S. subrepandum var. massauense.

As espécies do gênero Passiflora são amplamente distribuídas no território brasileiro, sendo algumas delas utilizadas na indústria, nos setores de ornamentação, farmacêutico e alimentício. Concomitante ao interesse econômico, o gênero, possui importância ecológica, devido à grande diversidade de espécies silvestres. Desta forma, o conhecimento molecular do maracujazeiro traz benefícios para os programas de melhoramento e conservação dessas espécies. Com finalidade de produzir dados e gerar subsídios em estudos genéticos de maracujazeiro, foram caracterizados e selecionados 23 iniciadores ISSR (Inter Simple Sequence Repeat) para 8 espécies de Passiflora spp. e 12 cultivares (BRS). Foi utilizado gel de agarose a 2% e luz ultravioleta para amplificação e visualização dos produtos amplificados. Os iniciadores ISSR apresentaram resultados satisfatórios, dentre os 23 iniciadores, 16 obtiveram valores acima de 60% de eficiência na classificação como bom (presença de 2 marcas ou mais). Em decorrência dos resultados apresentados, pode-se atestar que, tais iniciadores são adequados para estudos de polimorfismo, estrutura e diversidade genética em espécies do gênero Passiflora, além de, auxiliar programas de conservação e melhoramento.

Soybean [Glycine max (L.) Merr.] responded differently to storage stress factors. This research work aimed at assessing genetic potentials of fifteen soybean varieties to storage stress via accelerated aging technique and grouping them based on their levels of tolerance to storage stress using simple sequence repeat (SSR). Tolerance of the seed of 15 soybean varieties to storage stress was assessed by subjecting them to accelerated aging for 0, 6, 12, and 24 hours at 42 °C temperature and 100 % relative humidity (RH), after which their quality was assessed to determine their tolerance to storage stress. Same varieties were also stored for 6 months under ambient environment at 65 ± 5 % R.H and 25 ± 2 °C and they were genotyped with 19 simple sequence repeat (SSR) markers. The varieties were grouped on the basis of their levels of tolerance to storage stress. The principal components analysis (PCA) showed that germination rate index (GRI) and germination index (GI) were the major indices responsible for the significant variation in the seedling vigor characters. TGX1835-10E and TGX1448-2E were identified as varieties with good storage ability and therefore recommended for storage improvement in soybean breeding programs. Six SSR markers (Satt 565, Satt 175, Satt 281, Satt 600, Satt 160 and Satt 281) were identified as candidate markers for detection of alleles for tolerance to storage stress.

A set of nine Simple Sequence Repeat (SSR) loci, approved by the ECPGR Prunus working group, are proposed as a standard set for genotyping European plum accessions. These loci show sufficient reliability in spite of problems caused by hexaploidy. Polymorphism in the loci is high and enables differentiation between unique plum accessions as well as analyses of genetic grouping and overall genetic structure. A set of seven reference accessions are described. A compiled dataset with allelic information for 165 accessions is presented. Genetic structure reveals three different K-values (2, 4 and 9) demonstrating a major dichotomy between Prunus insititia-related accessions and cultivars belonging to Prunus domestica sensu stricto, as well as differentiation among minor subgroups defined by pomological traits and geographical origin.

Erianthus arundinaceus possesses high biomass, vigor, ratoonability and resistance to biotic and abiotic stresses and is considered to be an ideal species for improving sugarcane. In this study, a genome survey sequence was performed in E. arundinaceus, and SSR markers were developed based on a genome survey sequence. A total of 167.36 Gb raw data were obtained and were assembled into 3161 Mb genome sequences, including 15,238,738 scaffolds. The genome size was estimated to be approximately 3.23 Gp based on kmer analysis. In total, 36,616 SSR motifs were identified and characterized, including 60.69% dinucleotide, 34.27% trinucleotide, 3.34% tetranucleotide, 0.98% pentanucleotide and 0.71% hexanucleotide repeats. AG/CT repeat motifs were the most abundant, and GC/CG repeat motifs were somewhat rare, accounting for 54.16% and 2.69% of dinucleotide repeat motifs, respectively. A total of 365 SSR loci were obtained by 20 SSR primer pairs amplified in 20 clones of E. arundinaceus and its related species, and 188 SSR loci were specific to E. arundinaceus. Genetic similarity coefficient analysis by UPMGA methods clustered 20 clones of E. arundinaceus and its related species into two groups. Five clones of E. arundinaceus clustered separately with the remaining clones. These results may establish a foundation for further research investigating the E. arundinaceus genome and identifying genuine hybrids between sugarcane and E. arundinaceus.

Current cultivation and breeding strategy based on selections from local and wild vegetation does not meet the hazelnut demand. This situation would benefit from the development of high-yielding cu...

Simple Sequence Repeat (SSR) or microsatellite markers have become extremely useful tools in genetic identification and variability studies. To date around one hundred of these markers have been developed in the olive tree (Olea europaea L. subsp. europaea) and the majority of them are di- or tri-nucleotide. The analysis of the numerous articles recently published on plant microsatellite markers, shows however a higher relevance of markers with long core repeat motifs than those with di- or tri-nucleotide repeat motifs. This work presents a new set of highly informative and polymorphic tetra- and hexa-nucleotide SSR markers. The newly designed SSR markers have been employed to genotype the 36 olive cultivars of the core collection from Worldwide Olive Germplasm Bank of Cordoba, Spain (WOGBC). This article describes the allelic profiles of the 36 olive cultivars for a set of ten SSR markers and also a protocol for a multiplex polymerase chain reaction (PCR). The designed multiplex PCR can lead to an optimal amplification of up to five different SSR markers in the same reaction tube. Finally, the use of only four of the set of ten SSR markers employed, allows the differentiation of any cultivar of the WOGBC core collection from the remaining 35 cultivars.

Dioscorea, commonly known as yam, is an economically, socially, and culturally-relevant tuber cropin many tropical countries. This study aimed to characterize the genetic diversity of Dioscorea alata L. and related species from Colombia (42 D. alata L., 6 D. bulbifera L., 3 D. rotundata Poir., and 3 D. trifida L. f.) through seven inter- simple sequence repeat (ISSR) markers. The plant material was collected in five departments of Colombia. The seven ISSR generated 164 bands that showed high polymorphism (83.62%). Nei-Li genetic diversity coefficient (0.66) and Shannon information index (0.2636) revealed a high level of genetic diversity. The 54 genotypes were grouped into six groups according to geographic location but not by species. We found moderate genetic differentiation (Gst = 0.13) and high gene flow (Nm = 2.7367). The analysis of molecular variance (AMOVA) showed higher variation (86%) within groups than among groups (14%). Our results suggest that the high genetic diversity in yam in Colombia can be exploited in future work on crop improvement.

In this study, a total of 20 simple sequence repeat (SSR) markers were used to study the genetic diversity and population structure among 60 accessions of eggplant. Out of 20 SSR markers, 15 were f...