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Tamarix is the largest genus in the family Tamaricaceae with about 54 species. T. tetragyna var. meyeri and T. tetragyna var. deserti were previously reported from Iran. Due to the presence of overlapping features in the previously designed identification keys for the separation of these two varieties, it could be helpful to use other characters and character states derived from other sources, such as anatomical and molecular studies, in addition to find more effective morphological features for the separation of the two varieties. Therefore, the present study aimed to differentiate these two presumed varieties by multiple approaches using morphological, anatomical and molecular data. For morphological study, 6 quantitative and qualitative characters were examined in 12 samples of the two varieties. ANOVA analysis showed a significant difference between the studied characters. For anatomical study, 10 traits were examined in 4 samples of the two varieties, which also showed a significant difference. AMOVA analysis based on molecular studies using Scot marker showed a significant difference between the two varieties. The PCA biplot show the most variable traits in morphological and anatomical studies. For varieties delimitation, different clustering methods were drawn in all three studies. The results of this study showed that the use of appropriate diagnostic traits in morphological and anatomical studies as well as the use of molecular markers can be effective in showing the separated boundaries of the two varieties studied.

BackgroundRosa damascenaMill is a well-known species of the rose family. It is famous for its essential oil content. The aim of the present study was to assess the genetic diversity and population structure of a mini core collection of the Iranian Damask rose germplasm. This involved the use of universal rice primers (URP) and start codon targeted (SCoT) molecular markers.ResultsFourteen URP and twelve SCoT primers amplified 268 and 216 loci, with an average of 19.21 and 18.18 polymorphic fragments per primer, respectively. The polymorphic information content for URR and SCoT primers ranged from 0.38 to 0.48 and 0.11 to 0.45, with the resolving power ranging from 8.75 to 13.05 and 9.9 to 14.59, respectively. Clustering was based on neighbor-joining (NJ). The mini core collection contained 40 accessions and was divided into three distinct clusters, centered on both markers and on the combination of data.ConclusionCluster analysis and principal coordinate analysis were consistent with genetic relationships derived by STRUCTURE analysis. The findings showed that patterns of grouping did not correlate with geographical origin. Both molecular markers demonstrated that the accessions were not genetically diverse as expected, thereby highlighting the possibility that gene flow occurred between populations.

BackgroundEuphorbiais the largest genus in family Euphorbiaceae with a great biological and genetic diversity. The anti-trypanosomal activity of the crude extract of 15Euphorbiaspecies againstTrypanosoma brucei bruceiwas carried out. Furthermore, DNA fingerprinting of the tested species using ISSR and SCoT markers was also investigated.ResultsThe anti-trypanosomal activity of the 15Euphorbiaspecies revealed the highest activity ofE. officinarumL. andE. milliDes Moul. againstTrypanosoma brucei bruceiwith IC50values Euphorbiaspecies into two main groups, one contained two species and the other contained 13 species.ConclusionsSomeEuphorbiaspecies exhibited significant growth inhibitory activity towardTrypanosoma bruceistrain TC221. Results also indicated the suitability of both markers for genetic fingerprinting of the testedEuphorbiaspecies. To our knowledge, this is the first detailed comparison of the performance of two targeted DNA molecular markers (SCoT and ISSR) on the tested 15Euphorbiaspecies. The results guide future efficient use of these molecular markers in the genetic analysis ofEuphorbia.

Hedera helix L. is a specie that is used for its ornamental and medicinal properties widely. In spite of its very good biochemical characterization, the knowledge about the DNA variability is very limited and no DNA markers were used to analyses the genomic variability of the populations, up to date. In the present study, genetic diversity of 56 Hedera helix, individuals nine populations were studied using 10 Start Codon Targeted (SCoT) markers. High polymorphic bands (95.78%), polymorphic information content (0.25) and allele number (1.34) showed SCoT as a reliable marker system for genetic analysis in Hedera helix. At the species, the percentage of polymorphic loci [P] was 66.20%, Nei?s gene diversity [H] was 0.159, Shannon index [I] was 0.148 and unbiased gene diversity [UHe] was 0.56. Genetic variation within populations (70%) was higher than among populations (30%) based on analysis of molecular variance (AMOVA). We used SCoT molecular marker for our genetic investigation with the following aims: 1- Investigate genetic diversity both among and with date Hedera helix, 2-Identify genetic groups within these nine populations ivy, and 3-produce data on the genetic structure of date ivy populations. The results obtained revealed a high within-population genetic variability.

Persian hogweed (Heracleum persicum Desf. ex Fischer) is native to Iran but was introduced to Europe as an invasive plant. It has medicinal and edible values. Its fruits have been used to relieve flatulence, stomach aches as well as a flavoring as a digestive and an antiseptic. This medicinal plant forms many geographical populations in the country, particularly in mountainous regions; however, we have no data on the genetic structure and genetic diversity of this plant species. Such information can be used in gene pool identification and future genetic conservation and breeding programs. Therefore, the present study was performed with the following aims: 1- Provide data on genetic diversity of geographical populations, 2- Identify the potential gene pools for future genetic conservation and breeding programs. We used both Inter-simple sequence repeats (ISSR) and the Start Codon Targeted (SCoT) molecular markers as these are suitable molecular markers for genetic diversity investigations. There are highly polymorphic, reproducible, and relatively low cost markers. The present study revealed that both ISSR and SCoT markers produce data on the genetic variability and genetic affinity of the local populations. Both molecular markers revealed a good level of genetic variability within and among Heracleum persicum populations. Analysis of molecular variance (AMOVA) produced a significant differences between geographical populations for both markers. We could identify few ISSR as well as SCoT bands which can differentiate the studied populations. The moderate to a good level of genetic diversity we observed within each H. persicum may be due to cross pollination. In conclusion, we suggest using combination of ISSR and SCoT molecular markers to study population genetic variability in H. persicum geographical populations for future genetic conservation and germplasm collection of this medicinal plant.

Centaurea is a value-ultimate genus of medicinal plants showing high diversification levels, especially within the Mediterranean basin, and is still traditionally recognized as a complicated taxon. So far, few studies utilizing molecular markers have been done on Centaurea spp. towards a better dissection of its phylogeny and accurate assessment of genetic diversity. Here, two functional marker systems, start codon targeted (SCoT) polymorphism and CAAT box-derived polymorphism (CBDP), were implemented to assess the genetic diversity between eight wild Centaurea species in Egypt. Seventeen SCoT and 19 CBDP primers generated 197 and 179 bands, respectively. These primers generated 158 (80.2%) and 131 (73.18%) polymorphic amplicons with an average of 9.29 and 6.89 amplicons per primer, respectively. SCoT primers exhibited higher levels in % of polymorphism, % of heterozygosity, effective multiplex ratio, discriminating power, resolving power parameters compared to the CBDP. In contrast, no significant differences were observed between SCoT and CBDP in polymorphism information content, marker index and mean heterozygosity parameters. The UPGMA (unweighted pair group method with arithmetic mean) dendrogram of the combined data classified the Centaurea species under two major clades; the first comprised two species, whereas the second grouped five species. C. pallescens was kept separated as the most diverged among the eight Centaurea species. Principal component analysis (PCA) topology revealed highly similar results to those obtained from the cluster analysis. Ultimately, our results represent the first report utilising two gene-targeting marker systems as powerful techniques for assessing the genetic variability and relatedness among these eight valuable Centaurea species.

The assessment of the genetic diversity of cultivated varieties is a very important element of breeding programs. This allows the determination of the level of genetic differentiation of cultivated varieties, their genetic distinctiveness, and is also of great importance in the selection of parental components for crossbreeding. The aim of the present study was to determine the level of genetic diversity of oat varieties currently grown in Central Europe based on two marker systems: ISSR and SCoT. The research conducted showed that both these types of markers were suitable for conducting analyses relating to the assessment of genetic diversity. The calculated coefficients showed that the analyzed cultivars were characterized by a high genetic similarity. However, the UPGMA and PCoA analyses clearly indicated the distinctiveness of the breeding programs conducted in Central European countries. The high genetic similarity of the analyzed forms allow us to conclude that it is necessary to expand the genetic pool of oat varieties. Numerous studies show that landraces may be the donor of genetic variation.

Characterization of genetic diversity in germplasm collections requires an efficient set of molecular markers. We assessed the efficiency of 36 new SCoT markers, 10 new ISSR markers, and 5 microsatellites for the characterization of genetic diversity in chrysanthemum core collection of 95 accessions (Russian and foreign cultivars). Seven new SCoT (SCoT12, 20, 21, 23, 29, 31, 34) and six new ISSR markers ((GA)8T, (CT)8G, (CTTCA)3, (GGAGA)3, (TC)8C, (CT)8TG) were efficient for the genetic diversity analysis in Chrysanthemum × morifolium collection. After STRUCTURE analysis, most Russian cultivars showed 20–50% of genetic admixtures of the foreign cultivars. Neighbor joining analysis based on the combination of SSR, ISSR, and SCoT data showed the best accordance with phenotype and origin compared to the separate analysis by each marker type. The position of the accessions within the phylogenetic tree corresponded with the origin and with some important traits, namely, plant height, stem and peduncle thickness, inflorescence type, composite flower and floret types, flower color, and disc color. In addition, several SCoT markers were suitable to separate the groups distinctly by the phenotypical traits such as plant height (SCoT29, SCoT34), thickness of the stem and peduncle (SCoT31, SCoT34), and leaf size and the floret type (SCoT31). These results provide new findings for the selection of markers associated with important traits in Chrysanthemum for trait-oriented breeding and germplasm characterization.

Pomegranate is one of the oldest known fruit crops, well adapted to hot and dry areas and successfully grown in Mediterranean countries. During the last two decades, numerous publications have revealed the traditionally known associations between pomegranate consumption and health benefits, which led to increased demand by consumers and expansion in cultivation areas. Pomegranate is well adapted to areas with diverse pedoclimatic conditions and local cultivars could provide an essential source of genes for breeding. In this study, fruit phenotypic and genetic variability, and relationships, were studied in 26 Greek and foreign pomegranate cultivars/accessions grown in an ex situ collection located in Naoussa Greece, using Inter Simple Sequence Repeat (ISSR) and Start Codon Targeted (SCoT) molecular markers. Results from the principal component analysis made on fruit phenotypic characters revealed five components that accounted for 74.8% of the total variance, the first being related to skin color parameters and the second to juice antioxidant contents and aril color. Clustering from phenotypic data allocated individuals into four clusters. A total of 184 bands were generated for all markers applied across the 26 pomegranate cultivars/accessions, with an average of 77 bands per ISSR markers and 82 bands for SCoT markers. Low variability in the phenotypic and genotypic level was indicated, nevertheless, results from the association study between phenotypic traits and molecular markers that were obtained using Multiple Regression Analysis (MRA) could prove valuable for marker-assisted breeding programs, especially from SCoT markers that were found to be strongly or averagely associated with the morphological traits and chemical components.

The stems of Dendrobium officinale have been used as a rare and valuable Chinese tonic medicine, known as “Tiepi Fengdou”, since the Qing dynasty. Because of the increased market demand and continued exploitation of this plant, the reserves of wild D. officinale resources have been depleted, and D. officinale products on the market are being increasingly adulterated. Such changes have strongly affected the sustainable utilization of this valuable medicinal plant resource and the development of related industries. In this study, a species-specific DNA marker was developed for the rapid and accurate authentication of D. officinale. In total, 36 start codon-targeted (SCoT) polymorphism primers were screened in 36 definite Dendrobium species, and a distinct species-specific DNA amplicon (SCoT13-215) for D. officinale was obtained. After the sequence was cloned and sequenced, a sequence-characterized amplified region marker was developed (named SHF/SHR) and validated through PCR amplification of all 38 Dendrobium samples. The marker’s specificity for D. officinale was confirmed through the consistent amplification of a clear 197-bp band. This SCAR marker can be used to rapidly, effectively, and reliably identify D. officinale among various Dendrobium species and may play an important role in ensuring the quality of medicinal preparations and protecting the germplasm of this important medicinal species.

The genus Mentha is taxonomically and phylogenetically challenging due to complex genomes, polyploidization and an extensive historical nomenclature, potentially hiding cryptic taxa. A straightforward interpretation of phylogenetic relationships within the section Mentha is further hindered by dominant but outdated concepts on historically identified hybrid taxa. Mentha spicata is traditionally considered to be of hybrid origin, but the evidence for this is weak. Here, we aim to understand the phylogenetic relationships within the section Mentha using large sample sizes and to revisit the hybrid status and identity of M. spicata. We show that two of three traditional species in the subsection Spicatae are polyphyletic, as is the subsection as a whole, while the real number of cryptic species was underestimated. Compared to previous studies we present a fundamentally different phylogeny, with a basal split between M. spicata s.s. and M. longifolia s.s. Cluster analyses of morphological and genotypic data demonstrate that there is a dissociation between morphologically and genotypically defined groups of samples. We did not find any evidence that M. spicata is of hybrid origin, and we conclude its taxonomic status should be revised. The combination of genetic and phenotypic information is essential when evaluating hyperdiverse taxonomic groups.

Tomatoes are usually consumed daily in the human diet. High temperatures reduce the number of tomato yields per year. Heat stress has been considered one of the most prominent causes of alterations in morphological and molecular characteristics in crops that decrease normal growth, production, and yield in diverse plants, including tomatoes (Solanum lycopersicum L.). In this study, we evaluated six tomato lines, namely G1, G2, G3, G4, G5, and G6, at morphological, molecular, and cytological levels under heat stress. The average results of two seasons (2018 and 2019) clarified that the G6, G1, and G2 lines recorded the highest flowering values, as well as some fruit and vegetative growth traits. Furthermore, G6 and G2 had the maximum number of fruits/plant, whereas G2 and G1 produced the highest yield/plant under high temperatures. The number of chromosomes in all lines was 2n = 24, except for G5, in which the number was 2n = 26, whereas chromosome sizes were small, ranging from 323.08 to 464.48 µm. The G1 cultivar was a symmetrical cultivar (primitive), having the highest total form percentage (TF%) and symmetry index (Syi) values and the minimum karyotype asymmetry index (ASK) value, whereas G4 was asymmetrical (advanced). Molecular marker analysis demonstrated that intersimple sequence repeat (ISSR) primers 49A, HB-14, 49A, 49B, and 89B presented the highest values for polymorphism percentage P%, marker index (MI), effective multiplex ratio (EMR), and polymorphism information content (PIC), respectively. In contrast, OP-A3, OP-B3, SCoT 2, and SCoT 12 primers showed the highest PIC, EMR, MI, P%, and resolving power (Rp) values across the studied random amplified polymorphic DNA (RAPD) and start codon-targeted (SCoT) primers. Moreover, ISSR revealed the highest number of unique specific markers (6), followed by RAPD (4) and SCoT (3) markers. Cluster analysis of combined cytological data and data relating to molecular marker attributes separated the G1, G2, and G3 lines into one group, whereas the other lines were clustered in another group. On the whole, the application of combined analysis using morphological, cytological, and molecular genetics techniques could be considered to provide suitable parameters for studying the evolution of the genetic divergence between the studied tomato lines.

This study aims to determine grafting&rsquo, s efficiency to improve tomato growth and production under salinity stress conditions. A commercial tomato hybrid (cv. Bark) and eight wild tomato accessions were evaluated at molecular, physiological and agronomic levels. At the molecular level, two robust gene-targeting marker systems (Conserved DNA-Derived Polymorphism, CDDP and Start Codon Targeted Polymorphism, SCoT) were employed. Bark cv. was grafted as a scion onto the four tomato genotypes&rsquo, roots as stocks. The rootstocks effect was evaluated by growing plants at 0, 100 and 200 mM NaCl. Our results showed that grafting enhanced plant shoots and roots growth (plant height, number of branches, plant fresh weight, root length, and root fresh and dry weight), fruit yield (total yield, number and weight of fruits) and fruit quality (Vitamin C, firmness and total soluble solids) in Bark on most tested rootstocks. A significant interaction between salinity levels and rootstocks for all measured hormones, antioxidants and proline was observed. In conclusion, our consistent results from the three approaches (molecular, physiological and agronomical) revealed that the four genotypes (LA1995, LA2711, LA2485 and LA3845) were found to be grouped and exhibit better performance under salinity stress conditions. Furthermore, grafting could be a low-cost alternative method to improve salt tolerance in sensitive tomato genotypes.

The aim of this study is to optimize and evaluate the effectiveness of vitrification, droplet-vitrification, and encapsulation-vitrification techniques in the cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara &lsquo, Gold Heart&rsquo, a popular medicinal and ornamental plant species. In vitro-derived shoot tips were used in the experiments. All three techniques were based on explant dehydration with plant vitrification solution 3 (PVS3, 50% glycerol and 50% sucrose) for 0, 30, 60, 90, 120, 150, or 180 min. The recovered microshoots were subjected to morphometric, biochemical, and molecular analyses (RAPD, ISSR, SCoT). The highest recovery level was reported with the encapsulation-vitrification protocol based on 150 min dehydration (73.1%), while the vitrification technique was the least effective (maximum 25.8% recovery). Explants cryopreserved with the encapsulation-vitrification technique produced the highest mean number of shoots (4.9), moreover, this technique was optimal in terms of rooting efficiency. The highest fresh weight of shoots, on the other hand, was found with the vitrification protocol based on a 30-min PVS3 treatment. The concentrations of chlorophyll a and b were lower in all cryopreservation-derived plants, compared to the untreated control. On the other hand, short dehydration and cryopreservation of non-encapsulated explants stimulated the synthesis of anthocyanins. A small genetic variation in 5% of all samples analyzed was detected by RAPD and ISSR marker systems. Only plants recovered from the encapsulation-vitrification protocol had no DNA sequence alternations.

Karyotype characters, mitotic metaphase chromosomes, and haploid ideograms of six species of Plantago L. species. Analysis of somatic metaphases showed that the chromosome numbers of these species were 2n = 8 for P. ovata, 2n = 10 for P. albicans, P. amplexicaulis, P. ciliata and P. psammophilia, and 2n = 12 for P. lanceolata. The karyotype formulae were nsm (+), nsm (–) and nm. According to TF% and Syi index, and according to A and A1, P. ciliata may be considered the most symmetrical and P. lanceolata may be had the most asymmetrical karyotype. There are 63 total bands and 11unique bands resulting from RAPD; 23 total bands and 3unique bands resulting from both ISSR and 23 total bands in addition to 2 unique bands generated from SCoT markers. Perceptual mapping (Bilpot) and cluster analysis based on cytological and molecular attributes gave the same results that distinguished the studied taxa into two main groups; P. ovata and the other taxa also classified into two subgroups.

Genetic diversity and variation of four naturally distributed Huperzia serrata populations in Vietnam were analyzed based on the DNA fingerprint data obtained by ISSR and SCoT techniques separately and combined together. Both of ISSR and SCoT markers showed that the genetic diversity in Vietnam was relatively high at population and species level, except for the Hoang Lien population. The genetic diversity parameters of the Hoang Lien population were Nei’s gene diversity index (He) = 0.1436, Shannon index (I) = 0.2161, percentage of polymorphic bands (PPB) = 45.45%; of the Bach Ma population, He = 0.21, I = 0.33, PPB = 71.97%; of the Ngoc Linh population, He = 0.20, I = 0.31, PPB = 74.24%; of the Bidoup population, He = 0.19, I = 0.30, PPB = 74.24%; and at species level in Vietnam, He = 0.23, I = 0.36, PPB = 100%. The genetic differentiation was high with a value of GST = 0.19 and the number of migrants was estimated through gene flow as Nm = 2.16 individuals per generation among populations. The results of analysis of molecular variance (AMOVA) indicated that 28 and 72% of the total variation were among and within populations, respectively. The genetic diversity and genetic variation parameters of the investigated populations depended on the type of techniques used for DNA fingerprinting and also on the nature of each population. However, there was a high correlation and general trends among the obtained data sets when using ISSR, SCoT and the combination of these techniques.

The genetic diversity and relationships among 48 landrace and improved safflower (Carthamus tinctorius L.) genotypes were analyzed using three gene-targeted markers, start codon-targeted (SCoT) polymorphism, conserved DNA-derived polymorphism (CDDP) and CAAT box-derived polymorphism (CBDP). A total of 30 primers (10 primers from each marker) detected genetic polymorphism among the safflower genotypes. Three different marker types showed high level of polymorphism and the CDDP markers produced a higher number of polymorphic bands (74) in comparisons with SCoT and CBDP. The average PIC values for SCoT, CDDP, and CBDP were 0.39, 0.43, and 0.41, respectively. The marker index (MI) in CDDP markers was higher than that in SCoT and CBDP markers. Cluster analysis using SCoT and CDDP markers grouped the safflower genotypes into three distinct groups, whereas the CBDP markers divided the genotypes into two clusters. There were positive correlations between the similarity matrices obtained by each marker type. The results from the cluster analysis and STRUCTURE analysis using combined data grouped the genotypes into two clusters, generally in agreement with their origins. To our knowledge, this is the first detailed report of using gene-targeted molecular markers for genetic diversity analysis in safflower. Our results showed the efficiency of these markers for genetic diversity analysis in safflower and their potential for genome diversity and germplasm conservation.

In this study, inter-simple sequence repeats (ISSR) and start codon targeted (SCoT) markers were used for genetic diversity and relationship analysis of nine Salvia species. Twenty-one and twenty selected ISSR and SCoT primers amplified 350 and 329 loci, respectively, of which all were polymorphic. The obtained average polymorphism information content (ISSR, 0.38; SCoT, 0.40), average band informativeness (ISSR, 16.67; SCoT, 16.45) and resolving power (ISSR, 9.75; SCoT, 12.52) revealed high genetic diversity prevailing among Salvia accessions. Considering the ISSR and SCoT data, the species with a basic chromosome number of x = 8 showed higher values of the percentage polymorphism loci (PPL), the number of observed alleles (Na) and Shannon index (I) than the other species. The partition of clusters in the neighbour-joining dendrogram based on ISSR, SCoT and combined data was similar and grouped all individuals into four clusters. However, the dendrogram generated based on SCoT separated the individuals into sub-clusters in accordance with their species and section. The Mantel test revealed a similar polymorphism distribution pattern between ISSR and SCoT techniques, the correlation coefficient (r) was 0.83, and the results showed that both techniques were effective to assess the genetic diversity. Our results indicated that SCoT markers can be used as a reliable and informative technique for evaluation of genetic diversity and relationships among Salvia species.

In the present investigation 40 genotypes of maize from Czechoslovakia, Hungary, Poland, Union of Soviet Socialist Republics, Slovakia and Yugoslavia were analysed using 20 start codon targeted (SCoT) markers, 10 simple sequence repeat (SSR) markers, 13 random amplified polymorphic (RAPD) markers and using SDS-PAGE markers. Twenty SCoT primers produced 114 DNA fragments with an average of 5.7 bands per primer. Out of the total of 114 amplified fragments, 86 (76.43 %) were polymorphic, with an average of 4.30 polymorphic bands per primer. Ten SSR primers revealed a total of 65 alleles ranging from 4 (UMC1060) to 8 (UMC2002 and UMC1155) alleles per locus with a mean value of 6.50 alleles per locus. 20 SCoT primers produced total 114 fragments across 40 maize genotypes, of which 86 (76.43 %) were polymorphic with an average of 4.30 polymorphic fragments per primer and number of amplified fragments ranged from 2 (SCoT 45) to 8 (SCoT 28 and SCoT 63). The number of total scorable protein bands was twentythree as a result of SDS-PAGE technique but those that were not cosistent in reproducibility and showed occasional variation in sharpness and density were not considered. Based on these bands forty accessions of maize were screened. Out of twentythree polypeptide bands, 6 (31%) were commonly present in all accessions and considered as monomorphic, while 17 (65%) showed variations and considered as polymorphic. The dendrogram of 40 old maize genotypes based on SSR, SCoT, RAPD and SDS-PAGE markers using UGMA algorithm was constructed.

Greenhouse experiment is carried out to explore the outcome of γ-radiation on physiological and genetic variation in red radish (Raphanus sativus) for two generations. Gamma rays from 60Co were used to penetrate red radish seeds with different dose levels (0.0, 10, 20, 40 and 80 Gy). Plants generated from irradiated seeds and from self-pollination of these plants, called M1 and M2 generations, respectively. Some morphological and physiological traits were then determined, and the genetic diversity of both generations was studied using Start Codon Targeted (SCoT) and Conserved DNA-Derived Polymorphism (CDDP) molecular markers. All studied morphological traits (number of leaves/plants, leave height, root diameter, and root weight) were steadily improved by raising irradiation dose rate, reaching a cumulative raise at the irradiation doe level 40 Gy and decreased at dose level 80 Gy. Photosynthetic pigments of red radish plants released a notable increase by increasing gamma rays dose level for chlorophyll (a), chlorophyll (b) and carotenoids for 40 Gy dose rate. Proline content was elevated proportionally to the irradiation dose level, with the greatest increase seen at dose level of 80 Gy. Moreover, phytochemical screening was detected for the both two generations. Fourteen SCoT primers generated a total number of banding patterns of 194 with average 13.86 and the primer SCoT-33 released the highest number banding patterns (21). The percentage mean of polymorphism for all the SCoT primers was 74.66% and was 66.49 and 63.74% for M1 and M2 respectively. Furthermore, fifteen CDDP primers generated a total number of banding patterns of 186 and the primer CDDP-5 relieved the highest number of banding patterns (20). The percentage mean of polymorphism for all the CDDP primers was 73.41% and was 64.38 and 65.91% for M1 and M2 respectively. It could be concluded that gamma irradiation exhibited an appropriate variation in red radish M1 and M2 which was detected by SCoT and CDDP molecular markers.

In this study, the molecular mechanism underlying ethephon-induced shortened internodes in maize was investigated using Zhengdan958 maize variety. The ethephon treatment was performed by spraying 225 mL/ha Ethephon 40% SL onto the foliage at the eight-expanded-leaves stage. The differentially expressed genes in the young internodes under ethephon treatment were identified through methylation-sensitive amplified fragment length polymorphism (MSAP), start codon targeted polymorphism (SCoT) and cDNA-amplified fragment length polymorphism (AFLP) analyses. MSAP results showed the methylation sites were widely distributed in both the ethephon-treated (at 27.8%) and control plants (30.1%). This suggested that ethephon treatment modified the methylation patterns; with 3.0% of the sites being hyper-methylated and 7.3% demethylated compared with the control. Based on SCoT analysis, 148 transcript derived fragments (TDFs) were obtained in the ethephon-treated plants. Among them, 38 were up-regulated (25.7%) and 47 down-regulated (31.8%). cDNA-AFLP analysis using 70 primer pairs identified 1635 TDFs in the ethephon-treated and the control plants. Of these, 600 and 564 TDFs were up- and down-regulated by the ethephon treatment, respectively. BLASTX analysis on 50 (randomly selected) differentially expressed TDFs divided them into several categories based on their putative biological functions: signal transduction (6%), resistance-related (14%), energy and metabolism (22%), transcription (4%), cell apoptosis (2%), unknown functional proteins (42%) and unknown genes (10%). Our results revealed that ethephon treatment could induce DNA methylation variation principally by increasing the demethylation tendency. This is suggested to play roles in stress-defence genes expression regulation and the differentially expressed genes could be associated with shortened internodes in maize.

Cactaceae plant family comprises over 130 genera and 2000 species of succulent flowering plants. The genera Mammillaria and Notocactus (Parodia), which have medicinal and nutritional applications as well as aesthetic appeal, are considered to be among the major genera of the family. Several species of both genera show morphological and chemical similarities and diversities according to environmental conditions and genotypes. Here, we assessed the genetic relationships of nine species belonging to two major genera Mammillaria and Notocactus under the family Cactaceae, using two modern gene-targeting marker techniques, the Start Codon Targeted (SCoT) Polymorphism and the Conserved DNA-Derived Polymorphism (CDDP). Besides, we screened the various phytochemicals and evaluated the antioxidant activities of the nine species of cacti. Five out of the 10 SCoT and eight CDDP primers used to screen genetic variations within the nine species yielded species-specific reproducible bands. The entire 156 loci were detected, of which 107 were polymorphic, 26 were monomorphic, and 23 were unique loci. The nine species were categorized into two groups based on the dendrogram and similarity matrix. Phytochemical profiling revealed that sterols, triterpenes, flavonoids, and tannins were found in all the tested species. Additionally, two Notocactus species (N. shlosserii and N. roseoluteus) and one Mammillaria species (M. spinosissima) revealed a considerable antioxidant activity. Our results demonstrated that gene-targeting marker techniques were highly powerful tools for the classification and characterization of the nine investigated species, despite displaying high similarities at both morphological and phytochemical levels.

Background Bletilla striata is a well-known traditional Chinese herb with varieties of functions. In China, the natural resources of Bletilla striata have been severely damaged because of the excessive exploitation and destruction of natural habitats. The aim of present study was to provide a reference for fully exploiting and utilizing the germplasm resources of Bletilla striata. Results The genetic diversity of 50 varieties of Bletilla striata from different area in China was analyzed by SCoT and IRAP molecular marker technique. A total of 209 bands were amplified by 20 groups of SCoT primers, of which 201 (96.17%) were polymorphic, and 47 polymorphic bands (94%) were observed in 50 bands amplified by 8 groups of IRAP primers. The 50 populations of Bletilla striata were divided into two major groups by SCoT and IRAP at the genetic similarity coefficient value of 0.60 and 0.68 individually. The partition of clusters in the unweighted pair group method with arithmetic mean dendrogram and principal coordinate analysis plot based on the SCoT and IRAP markers was similar. Conclusions Results indicated the abundant genetic diversity of Bletilla striata among different areas. Our results will provide useful information for resource protection.

Coconut (Cocos nucifera L.) is one of the important palms grown both as a homestead and plantation crop in countries and most island territories of tropical regions. Different DNA-based marker systems have been utilized to assess the extent of genetic diversity in coconut. Advances in genomics research have resulted in the development of novel gene-targeted markers. In the present study, we have used a simple and novel marker system, start codon targeted polymorphism (SCoT), for its evaluation as a potential marker system in coconut. SCoT markers were utilized for assessment of genetic diversity in 23 coconut accessions (10 talls and 13 dwarfs), representing different geographical regions. Out of 25 SCoT primers screened, 15 primers were selected for this study based on their consistent amplification patterns. A total of 102 scorable bands were produced by the 15 primers, 88 % of which were polymorphic. The scored data were used to construct a similarity matrix. The similarity coefficient values ranged between 0.37 and 0.91. These coefficients were utilized to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA). The extent of genetic diversity observed based on SCoT analysis of coconut accessions was comparable to earlier findings using other marker systems. Tall and dwarf coconut accessions were clearly demarcated, and in general, coconut accessions from the same geographical region clustered together. The results indicate the potential of SCoT markers to be utilized as molecular markers to detect DNA polymorphism in coconut accessions.

Twenty-four start codon targeted (SCoT) markers were used to assess genetic diversity and population structure of indigenous, introduced and domesticated ramie (Boehmeria nivea L. Gaudich.). A total of 155 genotypes from five populations were investigated for SCoT polymorphism, which produced 136 amplicons with 87.5% polymorphism. Polymorphism information content and resolving power of the SCoT markers were 0.69 and 3.22, respectively. The Indian ramie populations exhibited high SCoT polymorphism (> 50%), high genetic differentiation (GST = 0.27) and moderate gene flow (Nm = 1.34). Analysis of molecular variance identified significant differences for genetic polymorphism among the populations explaining 13.1% of the total variation. The domesticated population exhibited higher genetic polymorphism and heterozygosity compared to natural populations. Cluster analysis supported population genetic analysis and suggested close association between introduced and domesticated genotypes. The present study shows effectiveness of employing SCoT markers in a cross pollinated heterozygous species like Boehmeria, and would be useful for further studies in population genetics, conservation genetics and cultivar improvement., Highlights • Genetic diversity and population structure of ramie (Boehmeria nivea) were assessed by SCoT markers. • SCoT markers revealed high polymorphism and resolving ability in Boehmeria nivea. • The wild ramie showed moderate heterozygosity and gene flow with high genetic variation, suggesting unidirectional gene flow. • The domesticated population exhibited highest heterozygosity and close association with introduced population.

An efficient genetically stable regeneration protocol with increased phytochemical production has been established for Dendrobium nobile, a highly prized orchid for its economic and medicinal importance. Protocorm like bodies (PLBs) were induced from the pseudostem segments using thidiazuron (TDZ; 1.5 mg/l), by-passing the conventional auxin–cytokinin complement approach for plant regeneration. Although, PLB induction was observed at higher concentrations of TDZ, plantlet regeneration from those PLBs was affected adversely. The best rooting (5.41 roots/shoot) was achieved in MS medium with 1.5 mg/l TDZ and 0.25% activated charcoal. Plantlets were successfully transferred to a greenhouse with a survival rate of 84.3%, exhibiting normal development. Genetic stability of the regenerated plants was investigated using randomly amplified polymorphic DNA (RAPD) and start codon targeted (SCoT) polymorphism markers which detected 97% of genetic fidelity among the regenerants. The PIC values of RAPD and SCoT primers were recorded to be 0.92 and 0.76 and their Rp values ranged between 3.66 and 10, and 4 and 12 respectively. The amplification products of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. A comparative phytochemical analysis among the mother and the micropropagated plants showed a higher yield of secondary metabolites. The regeneration protocol developed in this study provides a basis for ex-situ germplasm conservation and also harnesses the various secondary metabolite compounds of medicinal importance present in D. nobile., Highlights • An efficient micropropagation protocol for Dendrobium nobile, has been developed. • Genetic stability evaluated by ScoT and RAPD markers. • Higher antioxidant activity within the micropropagated plants over the mother plant. • Efficiency of TDZ as a potent growth regulator.

International audience; Succinyl-CoA:3-ketoacid CoA transferase (SCOT) deficiency is an inborn error of ketone body metabolism and causes episodic ketoacidosis. We report clinical and molecular analyses of 5 patients with SCOT deficiency. Patients GS07, GS13, and GS14 are homozygotes of S405P, L327P, R468C, respectively. GS17 and GS18 are compound heterozygotes for S226N and A215V, and V404F and E273X, respectively. These mutations have not been reported previously. Missense mutations were further characterized by transient expression analysis of mutant cDNAs. Among 6 missense mutations, mutants L327P, R468C, and A215V retained some residual activities and their mutant proteins were detected in immunoblot analysis following expression at 37°C. They were more stable at 30°C than 37°C indicating their temperature sensitive character. The R468C mutant is a distinct temperature sensitive mutant which retained 12% and 51% of wild-type residual activities at 37°C and 30°C, respectively. The S226N mutant protein was detected but retained no residual activity. Effects of missense mutations were predicted from the tertiary structure of the SCOT molecule. Main effects of these mutations were destabilization of SCOT molecules, and some of them also affected catalytic activity. Among 5 patients, GS07 and GS18 had null mutations in both alleles and the other three patients retained some residual SCOT activities. All 5 developed a first severe ketoacidotic crisis with blood gas pH

This article belongs to the Section Molecular Diversity., Orchardgrass (Dactylis glomerata L.), is a well-known perennial forage species; however, rust diseases have caused a noticeable reduction in the quality and production of orchardgrass. In this study, genetic diversity was assessed and the marker-trait associations for rust were examined using 18 EST-SSR and 21 SCoT markers in 75 orchardgrass accessions. A high level of genetic diversity was detected in orchardgrass with an average genetic diversity index of 0.369. For the EST-SSR and SCoT markers, 164 and 289 total bands were obtained, of which 148 (90.24%) and 272 (94.12%) were polymorphic, respectively. Results from an AMOVA analysis showed that more genetic variance existed within populations (87.57%) than among populations (12.43%). Using a parameter marker index, the efficiencies of the EST-SSR and SCoT markers were compared to show that SCoTs have higher marker efficiency (8.07) than EST-SSRs (4.82). The results of a UPGMA cluster analysis and a STRUCTURE analysis were both correlated with the geographic distribution of the orchardgrass accessions. Linkage disequilibrium analysis revealed an average r2 of 0.1627 across all band pairs, indicating a high extent of linkage disequilibrium in the material. An association analysis between the rust trait and 410 bands from the EST-SSR and SCoT markers using TASSEL software revealed 20 band panels were associated with the rust trait in both 2011 and 2012. The 20 bands obtained from association analysis could be used in breeding programs for lineage selection to prevent great losses of orchardgrass caused by rust, and provide valuable information for further association mapping using this collection of orchardgrass., This work was supported by the National Basic Research Program of China (973 Program) (2014CB138705) and the National Natural Science Foundation of China, NSFC (31101760).