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Thirty-one varieties of wheat cultivated in Romania were analyzed regarding the genetic diversity and physicochemical properties, including the following determinations: moisture, ash, protein, wet gluten, lipid, starch content, falling number and damaged starch, considering the particularity of each species, its biological status and origin. The physicochemical data showed that the wheat samples presented large variability. The physicochemical properties of wheat flour were assessed by multivariable data analysis, using principal component analysis (PCA). All wheat samples clustered together according to their physicochemical data showed an association between all wheat species. The protein and ash contents were more related to the ancient wheat species, while the amounts of starch and damaged starch were associated with the modern ones. Positive correlations were obtained between protein and wet gluten content and between lipid and ash content. ISSR markers were used to analyze and compare genetic diversity among selected wheat cultivars. The obtained data were analyzed using NTSYSpc software considering the coefficients of similarity (Jaccard) and dissimilarity (Neighbor joining). The Jaccard coefficients varied from 0.53 to 1, reflecting the high genetic diversity characteristic of all wheat varieties.

Amaranthus retroflexus L. and Chenopodium album L. (Amaranthaceae) are weedy plants that cause severe ecological and economic damage. In this study, we collected DNA from three different countries and assessed genetic diversity using inter-simple sequence repeat (ISSR) markers. Our analysis shows both weed species have low genetic diversity within a population and high genetic diversity among populations, as well as a low value of gene flow among the populations. UPGMA clustering and principal coordinate analysis indicate four distinct groups for A. retroflexus L. and C. album L. exist. We detected significant isolation-by-distance for A. retroflexus L. and no significant correlation for C.album L. These conclusions are based data from 13 ISSR primers where the average percentage of polymorphism produced was 98.46% for A. retroflexus L. and 74.81% for C. album L.These data suggest that each population was independently introduced to the location from which it was sampled and these noxious weeds come armed with considerable genetic variability giving them the opportunity to manifest myriad traits that could be used to avoid management practices. Our results, albeit not definitive about this issue, do not support the native status of C. album L. in Iran.

Pine marshy forests Vaccinio uliginosi−Pinetum Kleist 1929 are associations included in Natura 2000 areas. They are at the highest risk of dismemberment due to global climate change. The study assessed the possibility of regeneration and genetic variability of Scots pine in a marshy forest habitat. The research was carried out in the Polesie National Park on the Durne Bagno and Lake Moszne peatbog. In the examined locations the phytocenosis of pine marshy forest was characterized. Subsequently the density of pine seedlings was estimated and their height measured. The genetic variability of the pine regeneration was estimated using the ISSR technique. Based on the research results, it was concluded that the regeneration potential of pine was low. On average, 200 regeneration plants were inventoried per hectare, and their average height was 26.34 cm. Significantly more pine regeneration was recorded (123 plants) in Lake Moszne, with an average height of 6.99 cm. By contrast, only 36 pine trees were inventoried in Durne Bagno, but their height was 45.69 cm. Scots pine genetic variability was similar in both locations. The percentage of polymorphic loci was 74.24. The average number of alleles per locus reached 1.685 and the effective number of alleles per locus was 1.405. The expected heterozygosity and Shannon index were 0.239 and 0.362, respectively. The phytocenosis was scarce, but characteristic of the Vaccinio uliginosi−Pinetum habitat. In total, 17 species were inventoried, of which 11 occurred in both locations. Light, humidity and habitat fertility indices were characteristic of this type of habitat and amounted to 6.98, 7.70 and 1.53, respectively. No relationship between undergrowth vegetation and the Scots pine regeneration was found. A negative effect of bog blueberry Vaccinium uliginosum L. on the number of pine regeneration was demonstrated only in the Durne Bagno location. Shading of the tree and shrub layer did not suppress the growth of Scots pine regeneration or undergrowth vegetation. Based on the study results, we concluded that the current state of preservation of pine marshy forests in the Polesie National Park was satisfactory. The presence of pine in the shrub layer allowed to conclude that the survival of seedlings in pine marshy forests was possible and could lead to their transition to the next developmental phases. However, as a result of the ongoing climate change and those predicted in the near future, we recommend constant monitoring of the habitat in order to maintain its sustainability., Sylwan 167 (1): 37-48

Nitraria schoberi L. (Nitrariaceae) is a halophytic plant with a continuous range in Central Asia and with only two populations in the westernmost distribution limit of species, in Romania. Currently, there is no documented explanation for the species’ presence in Europe, outside the main distribution area. Considering that marginal populations genetics are important in establishing range limits and species adaptative potential, genetic diversity was assessed using Inter-simple sequence repeat markers (ISSR). Both the Shannon’s Information Index (I) and Expected Heterozygosity (He) suggested a relatively low level of genetic diversity within the two populations. However, the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) dendrogram and Principal Coordinates Analysis clearly distinguished the two populations. Our presumptions, based on current results, are that the marginal westernmost population of N. schoberi was established due to the unique conditions from the “islands of desert” developed in a temperate continental climate. The European establishment of this species was likely accidental and probably due to ornithochory. Genetic relatedness between populations could be a consequence of their common origin, presumably from proximal Asian N. schoberi populations, while the separation can be explained by the lack of genetic material exchange between the two populations.

In vitro mutagenesis is an alternative to induce genetic variation in vanilla (Vanilla planifolia Jacks. ex Andrews), which is characterized by low genetic diversity. The objective of this study was to induce somaclonal variation in V. planifolia by gamma radiation and detect it using inter-simple sequence repeat (ISSR) molecular markers. Shoots previously established in vitro were multiplied in Murashige and Skoog culture medium supplemented with 2 mg·L−1 BAP (6-benzylaminopurine). Explants were irradiated with different doses (0, 20, 40, 60, 80 and 100 Gy) of 60Co gamma rays. Survival percentage, number of shoots per explant, shoot length, number of leaves per shoot, and lethal dose (LD50) were recorded after 60 d of culture. For molecular analysis, ten shoots were used for each dose and the donor plant as a control. Eight ISSR primers were selected, and 43 fragments were obtained. The percentage of polymorphism (% P) was estimated. A dendrogram based on Jaccard’s coefficient and the neighbor joining clustering method was obtained. Results showed a hormetic effect on the explants, promoting development at low dose (20 Gy) and inhibition and death at high doses (60–100 Gy). The LD50 was observed at the 60 Gy. Primers UBC-808, UBC-836 and UBC-840 showed the highest % P, with 42.6%, 34.7% and 28.7%, respectively. Genetic distance analysis showed that treatments without irradiation and with irradiation presented somaclonal variation. The use of gamma rays during in vitro culture is an alternative to broaden genetic diversity for vanilla breeding.

Background.Studying genetic polymorphism of the Prunus pumilaL. representatives with a wide range of variability in morphological characters will make it possible to identify promising genotypes with high combining ability for hybridization.Material and methods.The study covered 39 genotypes from the Chelyabinsk cultivated population and 6 genotypes from Khakassia, with extreme or intermediate expressions of such characters as fruit size, color and shape; stone shape and surface structure; leaf blade serration and shape; plant habitus, and some others. Genetic distances were calculated using the data matrix for binary states of presence (1) or absence (0) of amplified fragments with the Past Program (Gower). The UPGMA dendrograms were obtained in TREECON (version 1.3b).Results and discussion. The selected 5 ISSR and 2 RAPD primers demonstrated their effectiveness in detecting genetic polymorphism in the studied accessions. In individual cases, a weak connection of separate markers with leaf blade edge serration, stone surface structure, plant habitus, and geographic origin of the genotype was detected. Concurrent inclusion of all primers and genotypes in the cluster analysis failed to group the studied P. pumilasamples into clades according to the presence/absence of any traits, which may attest to their close genetic affinity. An independent clade with reliable bootstrap support (100%) was formed only for the genotypes of the taxonomically close sp. P. tomentosaThunb., analyzed as an external group. Moreover, this clade manifested (with the 86% reliability of bootstrap support) differences between the hybrid genotypes of P. pumila× P. tomentosa(F3) and the genotype of P. tomentosa. Parent pairs promising for fruit quality and shrub shape improvement through breeding were identified using the produced indices of genetic distinction.

Self-fertilization (also termed selfing) is a mode of reproduction that occurs in hermaphrodites and has evolved several times in various plant and animal species. A transition from outbreeding to selfing in hermaphroditic flowers is typically associated with changes in flower morphology and functionality. This study aimed to identify genetic effects of selfing in the F2 progeny of F1 hybrid developed by crossing Lilium lancifolium with the Asiatic Lilium hybrid ‘Dreamland.’ Fluorescence in situ hybridization (FISH) and inter-simple sequence repeats (ISSR) techniques were used to detect genetic variations in plants produced by selfing. The FISH results showed that F1 hybrid were similar to the female parent (L. lancifolium) regarding the 45S loci, but F2 individuals showed variation in the number and location of the respective loci. In F2 progeny, F2-2, F2-3, F2-4, F2-5, and F2-8 hybrids expressed two strong and one weak 5S signal on chromosome 3, whereas F2-7 and F2-9 individuals expressed one strong and two weak signals. Only two strong 5S signals were detected in an F2-1 plant. The ISSR results showed a maximum similarity value of 0.6269 between the female parent and the F2-2 hybrid. Regarding similarity to the male parent, a maximum value of 0.6119 was found in the F2-1 and F2-2 hybrids. The highest genetic distance from L. lancifolium and the Asiatic Lilium hybrid ‘Dreamland’ was observed in the F2-4 progeny (0.6352 and 0.7547, respectively). Phylogenetic relationships showed that the F2 progeny were closer to the male parent than to the female parent. Self-fertilization showed effects on variation among the F2 progeny, and effects on the genome were confirmed using FISH and ISSR analyses.

Syzygium cumini L. (ver Jamun; BlackBerry) is a native, evergreen multipurpose tree species of India. Besides being a fruit tree and for agroforestry in different regions, it is medicinally important too. This study aimed to determine genetic diversity using molecular and phytochemical markers in sixteen genotypes of Indian S. cumini from different agro-ecological zones. The present study used a combination of ISSR markers and the HPLC technique to explore these genotypes. The results showed a wide genetic diversity range based on the similarity coefficient values observed in S. cumini sixteen accessions from different sites. Four primary phenolic acids were discovered in all the accessions; caffeic acid (CA) was found in high concentrations. The intraspecific association between molecular and phytochemical characteristics was the primary goal of this investigation. By employing gene-specific markers for the route of secondary metabolites (polyphenols) production, it further investigated the progressive research of diversity analysis of polyphenol content in S. cumini accessions, which may also expand its nutraceutical and pharmaceutical utilization.

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.

An important area of crop development is the introduction of new vegetable plants, one of which is the sweet potato Ipomoea batatas (L.) Lam. The aim of our research was genetic, biological and economic assessment of sweet potato accessions held in the collection of the Plant Introduction and Acclimatization Dept., Udmurt Federal Research Center, Ural Branch of the RAS (UdmFRC UB RAS).Within this experiment, DNA polymorphism of 16 accessions of I. batatas was tested using ISSR markers. All accessions were found to represent different genotypes; the VM 17 accession obtained as a result of a bud mutation differed significantly from the original accession in its genotype.Evaluation of 16 accessions of sweet potato cultivars was carried out in a small-plot field experiment. The results of biological and economic assessment witnessed to the prospects of sweet potato cultivation within the territory of the Udmurt Republic for marketable tuber production. The productivity of the accessions was 140–2700 g of tubers per plant. According to their productivity, the accessions were divided into clusters: the low-productivity group (up to 500 g), with ‘Betty’, Fioletovy, ‘Beauregard’, ‘Covington’ and Bezhevy; medium-productivity group (500–1000 g), with ‘Jewel’, Fioletovy Sochi, Brazilsky, Afgansky, Vinnitsky rozovy, Lyubitelsky and ‘Bayou Bell’; and high-productivity group (more than 1000 g), with Pobeda 100, ВM 17, Druzhkovsky and Bely NBS. On the basis of the evaluation, 11 sweet potato accessions were recommended for cultivation in the Middle Urals: Afgansky, ‘Bayou Bell’, Bely NBS, ВM 17, Brazilsky, Druzhkovsky, Fioletovy Sochi, ‘Jewel’, Lyubitelsky, Pobeda 100, and Vinnitsky rozovy.

PURPOSE: The genus Nicotiana is a well-defined group of species of which tobacco (Nicotiana tabacum L.) is the most important crop plant and plays a significant role in the economies of many countries. Recent advances in molecular genetics of the crop allowed the identification of many important genes and their location on chromosomes. However, the genetic analysis of this inbreeding, highly homozygous plant that serves as a model in many other studies is still very much work in progress. METHODS: A large proportion of the tobacco genome is represented by highly repeated DNA sequences, which makes molecular markers, based on them, an obvious first choice in diversity studies. Here we present an assessment with Inter-Simple Sequence Repeat (ISSR) markers of the intra- and inter-cultivar variability in a set of Bulgarian tobacco varieties. RESULTS: The screening of the 4 primers revealed a varying number of bands generated by different primers. The total number of bands varied between 4 and 13. No correlation between the total number of bands and the number of polymorphic bands was observed. Testing with ISSR primers revealed that plants with different DNA profiles can be found in varieties used. The highly similar profiles when E7 primer was used in two of the tested varieties from different production types indicate that the DNA fingerprinting technique may present a particular challenge in this crop. CONCLUSIONS: The present study demonstrates the power of the ISSR technique as a tool for diversity identification in tobacco.

Artificial selection related with important agronomic characteristics of Stevia rebaudiana may cause genetic divergence and formation of genetically structured populations with genetic uniformity or diversity within cultivars. Current study employed inter simple sequence repeats of DNA (ISSR markers) to assess genetic diversity within and among a single cultivated population maintained through sexual propagation (SR1) and four cultivated populations generated by artificial selection and maintained by vegetative propagation (SR2–SR5). Highest polymorphism rate was reported in SR1 (89.24%), whilst the lowest rate of polymorphism occurred in SR2 (60.13%). ISSR markers revealed that selection of plants with traits of vegetative-propagated interest may lead towards the generation of genetically more uniform DNA-level populations, while plants maintained by sexual propagation have high genetic variability. High estimated genetic divergence level indicated that the five areas of stevia form genetically structured populations. SR2 and SR4 are constituted by plants more homogeneous at DNA level for the selected characteristics than plants of SR3 and SR5 populations. Predominant and homogeneous genotypes selected at SR2 and SR4 areas could be valuable for tracing strategies to obtain stevia plants with the desirable agronomic characteristics through crosses between contrasting individuals in future breeding programs.

Background Coleus (family Lamiaceae) is a popular garden ornamental plant known for its bright-colored foliage. The effective role of different plant growth regulators have been widely reported in plant cultivars (Singh A and Prasad SM Int. J. Sci. Eng. Appl. Sci. 2 (1): 1-10, (2016)). Kinetin has received great interest due to its richness in bioactive phytochemicals including polyphenolic compounds with its antioxidant activity. Molecular markers have been very helpful in enhancing our understanding of genome structure and function. In our study, we investigated the effect of three treatments of kinetin on the phytochemical contents of five Coleus cultivars and molecular characterization by ISSR markers. Results The studied cultivars of Coleus revealed adequate amounts of pigments chlorophyll a (Cha), chlorophyll b (Chb), and total carotenoids (Carot) as well as phytochemical ingredients including total carbohydrates (Carb), total phenolic (TP), total flavonoids (TF), and total tannins (TT). Finger paint cultivar (C5) revealed the highest values (0.266, 10.853, 11.648, and 6.722 mg/g) of Carb, TP, and TT, respectively, while the highest contents (11.814 and 11.648 mg/g) of TF were identified in Beckwhites gem species (C2) and finger paint cultivar (C5), respectively. Inter-simple sequence repeat (ISSR) marker analysis was performed on the samples with 5 primer pairs. These bands were identified as 21 polymorphic and 18 monomorphic ones with 53.8% polymorphism. Ten unique bands were identified in the resulted ISSR profile. Conclusion These results suggested that the effect of kinetin treatments stimulates secondary metabolites production. Also, ISSR marker is helpful for possible distinguishing, identifying, characterizing, and selection processing of Coleus cultivars.

Sodium azide (NaN3) is widely used as a chemical mutagen in mutation studies, especially to induce point mutations. This study was carried out to determine genetic diversity of six-rowed barley cultivar Finola and 14 mutant lines induced by sodium azide mutagenesis agent and identified by using 5 Inter Simple Sequence Repeat (ISSR) markers. According to the results, 24 bands were produced by five ISSR markers, 18 of them were found polymorphic, and the average polymorphism rate was 76.33%. The average allele number was determined as 4.8, and UBC-808 marker had the highest allele number with 6 alleles, while UBC-820 and UBC-825 markers had the lowest allele numbers with 4 alleles. The average polymorphism information content of the markers used in the study was 0.83, and ranked from 0.78 to 0.86. A dendogram was created using marker data, and according to the dendogram, genotypes were divided into two main groups. Fnl-4, Fnl-8, Fnl-10, Fnl-11 and Fnl-12 mutants existed in the first group, while the others including Finola cultivar were in the second group. Based on the principal component analysis (PCA), Finola cultivar and mutant lines Fnl-5, Fnl-13 and Fnl-14 were evaluated as they are similar to Finola although, differed for some alleles., Sodyum azid (NaN3) özellikle nokta mutasyon çalışmalarında yoğun olarak kullanılan kimyasal bir mutajendir. Bu çalışmada, Finola altı sıralı arpa çeşidi, kimyasal bir mutajen olan sodyum azid ile muamele edilerek 14 adet mutant arpa hattı elde edilmiştir. Elde edilen hatlar arasındaki genetik farklılık, 5 adet basit tekrarlı diziler arası polimorfizm (Inter Simple Sequence Repeat, ISSR) markörü yardımıyla belirlenmiştir. Elde edilen sonuçlara göre 5 ISSR marköründen 24 adet bant elde edilmiş ve bu bantlardan 18 adet polimorfik allel belirlenmiş ve ortalama polimorfizm oranı %76.33 olarak saptanmıştır. Ortalama allel sayısı 4.8 olup, 6 bant ile UBC-808 en fazla bant üreten primer olurken, en az bant üreten primerler ise 4’er bant ile UBC-820 ve UBC-825 primerleri olmuştur. Kullanılan markörlere ait ortalama polimorfizm bilgi içeriği (PIC) 0.83 olarak hesaplanmış ve PIC değeri 0.78 ile 0.86 aralığında değişmiştir. Markör verileri dendrogramda 2 ana grup oluşturmuştur. Birinci kümede Fnl-4, Fnl-8, Fnl-10, Fnl-11 ve Fnl-12 mutantları yer alırken, ikinci kümede ise Finola çeşidi ile birlikte diğer mutant hatlar yer almaktadır. Temel bileşenler analizi, Finola çeşidi ve mutant hatlardan Fnl-5, Fnl-13 ve Fnl-14 hatlarının birbirine benzemekle birlikte, bazı alleller bakımından farklı olduklarını ortaya koymuştur.

Мета. Виявити генетичний поліморфізм та дивергенцію сортів і гібридів кабачка (C. pepo L.) різного географічного походження за ISSR маркерами. Методи. Для оцінки генетичного поліморфізму 29 сортів і гібридів C. pepo L. різного походження використовували ISSR аналіз. Коефіцієнт подібності між досліджуваними зразками кабачка визначали за формулою Nei–Li. Розрахунок коефіцієнтів подібності та побудування філогенетичного дерева здійснювали за допомогою пакету програм Phylip-3.69. Кластерний аналіз проводили методом приєднання сусідів (Neighbor-joining). Достовірність групування зразків у кластери перевіряли методом бутстреп-аналізу. Результати. Використання 13 праймерів до міжмікросателітних ділянок дозволило виявити 129 локусів ДНК кабачка, серед яких 109 були поліморфними, 20 – мономорфними, 11 – унікальними для певних зразків. Електрофореграми продуктів ампліфікації дослідних зразків різнилися кількістю і розміром ампліконів. Виявлено високий поліморфізм ДНК дослідних зразків кабачка, який варіював від 62,5 % (праймер UBC 810) до 100 % (праймери UBC 2, UBC 3 і UBC 846). Рівень молекулярно-генетичного поліморфізму зразків кабачка варіював від 55,8 до 63,6 % у гібридів ‘Rimini’ і ‘Eight Ball’, відповідно. Встановлено низьку генетичну дивергенцію між дослідними зразками C. pepo L., коефіцієнт подібності за Nei–Li становив 0,0005–0,0092. За допомогою кластерного аналізу зразки кабачка були згруповані у два кластери. Основним критерієм кластеризації був рівень генетичної дивергенції. Географічне походження зразків не впливало на особливості їх групування. Висновки. За результатами вивчення зразків кабачка різного географічного походження за допомогою ISSR аналізу встановлено високий генетичний поліморфізм і незначна генетична дивергенція між дослідними зразками. Виявлено унікальні фрагменти ДНК, які можуть бути використані для паспортизації відповідних зразків, а також для розробки інших молекулярно-генетичних маркерів. Отримана інформація може бути корисною для оптимізації селекційного процесу кабачка і подальших досліджень в області молекулярної генетики цієї культури.

Anise (Pimpinella anisum L.) is an annual spice and medicinal plant belonging to the family Apiaceae. Only a limited number of anise breeding studies are available. Therefore, genetic variation among anise landraces has not been extensively examined with molecular markers. in this study, the molecular diversity of 15 Turkish anise landraces collected from different regions of Turkey and four foreign landraces obtained from Cyprus, Syria and France were analyzed using randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers. the discriminatory power of these markers was assessed by calculating various marker parameters, namely percent polymorphism, polymorphism information content (PIC), resolving power (RP), and marker index (MI). the genomic DNA of 19 anise landraces was amplified with nine RAPD primers that generated 71 polymorphic bands and five ISSRs primers that produced 45 polymorphic bands. the percentage of polymorphic loci was 59.8% in RAPD profiling and 51.5% in ISSR profiling. the average PIC values obtained using RAPD and ISSR markers were 0.40 and 0.38, respectively. the mean RP (2.90 and 2.67) and MI values (1.23 and 1.05) of the RAPD assay were found to be somewhat higher than those of the ISSR assay, but ISSR was more favorable than the RAPD techniques in terms of presenting the phylogenetic relationships in anise landraces. A UPGMA cluster grouped the 19 anise landraces into two major clusters based on both markers. the RAPD and ISSR marker systems were found to be useful for determining genetic diversity in P. anisum L. and identifying variation among landraces., Commission of Scientific Research Projects of Ege University [2013/OMYO/001], I thank the Commission of Scientific Research Projects of Ege University for financial support to carry out this study (Project No: 2013/OMYO/001). I am grateful to Dr. Tansel Kaygisiz Asciog. ul and Dr. Dilsat Yegenoglu for their assistance in statistical calculations of the data presented in this manuscript. I also thank Elif Denizaslani and Christine Taylor for their suggestions to improve the language of the text. Finally, I would like to thank the reviewers for providing useful comments on the previous draft of this article.

Cichorium intybus L. (chicory) is an important medicinal plant with significant economic potential and has recently gained rapid momentum in the functional food sector. In the present study, soil chemistry, phytochemical, and molecular diversity were assessed for 50 accessions of chicory collected from diverse agro-climatic zones. In total, 64 common metabolites were identified from the leaves of 7 chicory accessions collected from different altitudes and among them, the predominant metabolites included methyl commate B (6.3–10.14%), gamma sitosterol (2.79–9.3%), and 9, 12, 15-octadecatrienoic-acid (2.55–8.42%). Three terpenoid compounds, viz., betulin, kolavelool and betulinaldehyde, were observed at high altitudes (1790, 1901, and 2172 m) and not observed at low altitudes. Among these compounds, betulin had the highest concentration with an average value of 23.53% followed by kolavelool with 7.37% and betulinaldehyde with 7.21%. For molecular diversity analysis, 12 ISSR primers were selected for PCR amplification and 86 bands were generated with an overall polymorphism percentage of 67.44%. The observed Nei’s genetic diversity (H) and Shannon’s information indices (I) were highest for the Pulwama (CIN-PU) group of accessions (H = 0.222 ± 0.018; I = 0.343 ± 0.027) and lowest for the Baramulla (CIN-BM) group of accessions (H = 0.115 ± 0.019; I = 0.173 ± 0.028). The Analysis of Molecular Variance (AMOVA) analysis revealed 56% variation existing within the groups and 44% among the groups of chicory accessions. This study shows that chicory populations vary considerably in terms of their molecular and phytochemical composition as a function of their geographic location. Furthermore, this study demonstrates that chicory phytochemical and molecular diversity are significantly influenced by altitude, soil chemistry, and growing conditions. Using metabolomics and altitudinal variation, cluster analysis showed that geographic origin was correlated with diversity patterns.

Background Sugar beet is considered a very important strategic economic crop, as it comes at the second place for white sugar production in Egypt after sugar cane and the sugar percentage in its roots is 20%. This work was conducted in Egypt as a direct response to bridge the gap which began to expand significantly between the production and consumption of white sugar and the desire for the genetic improvement process to increase the productivity of this crop. Further, as serious attempt to understand the nature of the genetic diversity among a group of sugar beet accessions with different origins, it constitutes an important source for establishing a program of breeding and improving sugar beet crop under Egyptian conditions. Results The process of verifying the existence of significant genetic variation between the eight sugar beet varieties with various origins was carried out through a set of genetic parameters through two growing seasons besides, the data of ISSR markers profile. Further, all the sugar beet varieties were superior in all the studied traits during their evaluation over two agricultural years, where the two sugar beet accessions Oscar Poly and Rass Poly are superior in TSS % and sucrose % traits, Mont Bianco and Sultan in apparent purity % trait, Blino and Sultan in root yield and top yield traits and Rass Poly and Blino in sugar yield trait, respectively. In addition, the values of heritability in broad sense were high in all studied traits except the two traits; sucrose % and sugar yield where their results were appeared medium during both growing seasons. In the same regard, expected genetic advance values showed significant genetic progress in all studied traits during the two growing seasons. As well, highly genetic diversity was obtained between the eight sugar beet accessions through generated 169 amplified fragments from the twelve ISSR primers, 122 of them were polymorphic with 72.18% polymorphism. Conclusions The eight sugar beet varieties achieved remarkable and distinctive results for all traits under study during two agricultural years. However, they were differentiated and various from each other. As they differed in their superiority in all estimated traits and they proved to be fertile material for studying in this investigation and very suitable for enriching the breeding and genetic improvement program to promote the sugar beet yield in order to increase the productivity of white sugar in Egypt.

Barley production is essential in Egypt. In the present study, 15 different six-rowed Egyptian barley cultivars were studied. To differentiate between the different cultivars under study in terms of morphological characteristics and ISSR, molecular characterization reactions were carried out. Moreover, four cultivars (Giza 123, Giza 126, Giza 136, and Giza 138) were selected for further studies using scanning electron microscopy (SEM). Computational analysis of the DNA barcoding sequences of the two plastid markers rbcL and matK was executed, and the results were deposited in the NCBI database. The morphological traits showed low statistical significance among the different cultivars under study via the data collected from two seasons, suggesting that the mean field performance of these Egyptian cultivars may be equal under these conditions. The results showed that the phylogenetic tree was divided into four groups, one of which contained the most closely related genotypes in the genetic distance, including Giza 124, Giza 130, Giza 138, Giza 136, and Giza 137, which converge in the indicative uses of farmers. The seed coat of the studied cultivars was “rugose”. The elevation folding of the rugose pattern ranged from 11 ± 1.73 µm (Giza 126) to 14.67 ± 2.43 µm (Giza 123), suggesting variation in seed quality and its uses in feed and the food industry. According to the similarity matrix of ISSR analysis, the highest similarity value (93%) was recorded between Giza 133 and Giza 132, as well as between Giza 2000 and Giza 126. On the other hand, the lowest similarity value (80%) was recorded between Giza 130 and (Giza 133 and Giza 132), indicating that these cultivars were distantly related. Polymorphism information content (PIC) ranged from 0.26 for the primer ISSR UBC 835 to 0.37 for the primers ISSR UBC 814 and ISSR UBC 840. The current study showed that the matK gene is more mutable than the rbcL gene among the tested cultivars.

Orientadores: Renato Goldenberg, Eric de Camargo Smidt Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: Myrtaceae é conhecida como uma das mais complexas famílias de angiospermas. Myrcia teve sua última revisão taxonômica realizada há 153 anos e desde então inúmeras espécies novas foram descritas, deixando Myrcia como o segundo maior gênero da tribo Myrteae. O elevado número de espécies no gênero e a falta de uma revisão taxonômica completa e atualizada fez com que vários complexos de espécies mal delimitadas fossem formados em Myrcia. Um destes complexos é abordado no presente trabalho e inclui quatro espécies (M. laruotteana, M. lajeana, M. selloi e M. tomentosa) de ampla distribuição e grande variação morfológica, tornando difícil a separação entre os táxons. Neste estudo foram usados marcadores ISSR para investigar a correlação entre a variabilidade genética de populações destas espécies com aspectos morfológicos, geográficos, fitogeográficos e taxonômicos, a fim de esclarecer a delimitação das espécies. Levando em conta os resultados obtidos nestas análises, uma nova proposta de classificação específica é apresentada, com descrições dos táxons aceitos, dados de distribuição geográfica, floração e frutificação, e chave de identificação. No estudo de genética de populações com ISSR, o valor de diversidade genética encontrado (He = 0,215) foi mais baixo que o valor para espécies perenes, de distribuição ampla e fecundação cruzada. Isto pode ser explicado em partes pela auto-compatibilidade que as espécies do complexo M. laruotteana podem apresentar, como já relatado na literatura. Nenhum dos grupos que refletem aspectos morfológicos, geográficos e fitogeográficos testados apresentou boa estruturação genética, ou seja, nenhum deles mostrou alguma descontinuidade genética que pudesse representar os táxons envolvidos no complexo. Os grupos testados que representam a circunscrição taxonômica atual do complexo também não apresentaram boa estruturação genética. Os grupos formados a partir da análise Bayesiana mostraram melhor estruturação genética a partir da AMOVA (12% de variação entre as populações dentro dos grupos e 27% de variação entre os grupos). A análise Bayesiana juntamente com o dendrograma formado demonstra uma separação em dois grandes grupos, o primeiro formado por todas as populações de M. tomentosa, e o segundo contendo as populações das outras espécies misturadas. A partir destes resultados e levando em consideração toda a variação existente entre os extremos morfológicos das espécies, é proposta uma nova classificação para o complexo, com sinonimização de M. lajeana sob M. laruotteana. Myrcia tomentosa e M. laruotteana podem ser reconhecidas principalmente pelo indumento, geralmente denso na primeira e quase ausente na segunda. Myrcia selloi pode ser reconhecida pela queda dos remanescentes do hipanto e do cálice, deixando os frutos com uma cicatriz circular apical Abstract: Myrtaceae is known as one of the most complex families of angiosperms. The last taxonomic revision of Myrcia was made 153 years ago and since then many new species were described, leaving Myrcia as the second largest genus of the tribe Myrteae. The high number of species in the genus and the lack of a complete and updated taxonomic revision resulted in many species complexes in Myrcia. One of these complexes is investigated in this work and includes four species (M. laruotteana, M. lajeana, M. selloi and M. tomentosa) with wide distribution and a lot of morphological variation, making it difficult to distinguish these taxa. This study used ISSR markers to investigate the correlation between the genetic variability of populations of these species with morphological, geographical, phytogeographic and taxonomic aspects, in order to clarify the delimitation of species. Taking into account the results obtained in these analyzes, a new proposal of specific classification is presented with descriptions of accepted taxa, geographical, flowering and fruiting data, and identification key. In the study of population genetics with ISSR, the value of genetic diversity (He = 0.215) was lower than the value for perennial, widely distributed and cross-fertilization species. This can be explained in part by self-compatibility that species within M. laruotteana complex apparently present, as previously reported in the literature. None of the groups that reflect morphological, geographical and phytogeographic aspects showed good genetic structure, i.e., none of them showed any genetic discontinuity that could represent the taxa involved in the complex. The groups represented by the current taxonomic circumscription of the complex also did not show good genetic structure. The groups formed from the Bayesian analysis showed better genetic structure from the AMOVA (12% variation among populations within groups and 27% variation between groups). The Bayesian analysis with the dendrogram formed showed two major groups, the first with all populations of M. tomentosa, and the second containing mixed populations from other species. From these results and taking into account the large amount of variation between the morphological extremes of the species, we propose a new classification for the complex, with the synonymization of M. lajeana under M. laruotteana. Myrcia tomentosa and M. laruotteana differ from each other mainly by the indumenta, which is dense in the former and almost absent in the latter. Myrcia selloi differs from the others by the caduceus remnants of the hypanthium and calyx, leaving circular apical scar on the fruit Mestrado Biologia Vegetal Mestra em Biologia Vegetal

The runner bean is the third most economically important Phaseolus species, which is cultivated on small-scale agriculture for the production of immature pods or to obtain dry seeds. However, despite the economic importance and agronomic potential of this species, the runner bean has been little studied from the genetic standpoint. Therefore, the main objective of this study was to characterize ten selected lines of runner bean obtained from Central (Santiago) and Southern (Valdivia and Villarica) Chile based on morphological and agronomic traits. In addition, the genetic variability of these lines was determined using 12 Inter-Simple Sequence Repeat (ISSR) markers to evaluate the potential of this germplasm for breeding and commercial purposes. As a result, the lines from Central Chile were characterized, and had a higher number of pods per plant compared to the Southern lines, although the size and weight of their seeds were lower. Moreover, a low level of genetic diversity (He = 0.251) was encountered in this population. Finally, this is one of the first studies that generate relevant and novel information on the morphological, agronomic and genetic characterization of the P. coccineus germplasm present in Chile.

Landraces are sources of genetic variability, especially with respect to alleles that confer tolerance to biotic and abiotic factors and, therefore, can aid breeding programs in the development of promising cultivars. However, it is necessary to know this genetic patrimony at a level that allows its alleged use in breeding programs. In this sense, the objective of this study was: (i) to identify cowpea landraces that present agronomic potential and (ii) to evaluate the genetic diversity for future cross-breeding. For this, two trials (coastal and sertão of the state of Ceará) were carried out in distinct periods for the morphological and agronomic characterization of eight landraces of the state of Ceará plus two control cultivars. Both assays were conducted in a randomized complete block design with four replicates. Additionally, the molecular characterization by ISSR markers was done. Due to the presence of interaction genotypes by environments, it was observed for the conditions of the coast (Fortaleza), the traditional variety Boi Deitado and the conditions of the sertão (Madalena) to Vinagre Barrigudo de Caldo, as the most indicated to be superior in grain yield and in other agronomic characters. In order to increase the genetic base of the cowpea, we suggest crosses between genotypes Boi deitado and Cojó for the generation of segregating populations of future breeding programs. RESUMO Variedades tradicionais são fontes de variabilidade genética, principalmente no que se refere a alelos que conferem tolerância a fatores bióticos e abióticos e, portanto, podem auxiliar os programas de melhoramento no desenvolvimento de cultivares promissoras. Contudo, torna-se necessário conhecer esse patrimônio genético em nível que permita sua pretensa utilização em programas de melhoramento genético. Assim, objetivou-se com esse estudo: (i) identificar variedades tradicionais de feijão-caupi que apresentem potencial agronômico e (ii) avaliar a diversidade genética das mesmas para orientação de futuros cruzamentos. Para tal, dois ensaios (litoral e sertão do estado do Ceará) foram realizados em períodos distintos para a caracterização morfoagronômica de oito variedades tradicionais do estado do Ceará mais duas cultivares testemunhas. Ambos os ensaios foram realizados no delineamento em blocos ao acaso com quatro repetições. Adicionalmente, foi feita a caracterização molecular por marcadores ISSR. Devido a presença de interação genótipos por ambientes, observou-se que para as condições do litoral (Fortaleza), a variedade tradicional Boi Deitado foi a mais indicada, e para as condições do sertão (Madalena) foi a Vinagre Barrigudo de Caldo, por serem superiores quanto ao rendimento de grãos e quanto a outros caracteres agronômicos. Buscando ampliar a base genética do feijão-caupi, sugerem-se cruzamentos entre os genótipos Boi deitado e Cojó para a geração de populações segregantes em futuros programas de melhoramento.

Cucumber (Cucumis sativus L.) damping-off disease caused by Phytophthora melonis is a limiting factor affecting cucurbit production. This study assessed a collection of 38 domestic and exotic commercial cucumber genotypes for damping-off resistance at transplanting and 50% flowering. Plants were classified using a four-score scale, the average damping-off percentage ranging from 7.92% (resistant) to 88.01% [highly susceptible (HS)]. Most of the genotypes were susceptible and HS to P. melonis. These genotypes were analyzed with 15 inter-simple sequence repeat (ISSR) markers. A total of 317 bands were produced, of which 297 bands (93.69%) were polymorphic. The primer ISSR29 possessed the highest resolving power, polymorphic information content and marker index value, which could be the most informative primer for distinguishing cucumber genotypes. Cluster analysis and principal component analysis were carried out and there was a probable connection between cucumber genotypes and resistance level against P. melonis, indicating possible application for ISSR in studying disease resistance of cucumber and producing cultivars that resist damping-off. The expression of three candidate genes probably involved in the defence against P. melonis was quantified by quantitative real-time polymerase chain reaction in five cucumber genotypes showing different susceptibility to the pathogen. CsWRKY20, CsLecRK6.1 and LOX1 genes were differentially expressed depending on genotype in early stages of infection. LOX1 was the most expressed gene across experiments and the one that best discriminated between susceptible and resistant genotypes. These results are a valuable resource for future functional genomics studies to unravel the molecular mechanisms of C. sativus/P. melonis interaction.

The objective of this study was to establish an efficient—direct or indirect—regeneration system for pineapple (Ananas comosus L.) plants, with a high rate of multiplication and that would preserve the genetic identity of the donor genotype (Hybrid ‘MD2’) in the regenerated plants. Ten treatments, with different concentrations of 2,4-Dichlorophenoxyacetic (2,4-D) and Picloram (P), in the absence or presence of 6-Benzylaminopurine (BAP), were used for in vitro morphogenesis induction, as well as histological and molecular techniques, in order to characterize the morphogenic responses induced. Significant differences between treatments tested, to induce callus and buds, were assessed by the Kruskal Wallis method and the Mann–Whitney U-tests. Different pineapple regeneration routes were identified, showing the high regeneration potential of this species. The medium containing 2 mg L−1 2,4-D and 2 mg L−1 BAP, where indirect somatic embryogenesis occurred, was selected as the most efficient treatment, with an average of 120 somatic embryos per explant, differing significantly from the rest of the treatments. It was also demonstrated that the pineapple plants regenerated in vitro preserved the genetic identity of the donor genotype, which represents a high degree of confidence for the application of indirect somatic embryogenesis for A. comusus clonal propagation.

Eleven populations of the endangered plant Moehringia jankaе and twenty-eight of the rare plant Moehringia grisebachii were collected across its natural range from Bulgaria. Their genetic diversity was investigated through fifteen selected Inter Simple Sequence Repeats (ISSR) primers. The ISSR primers produced a total of 285 bands, of which 275 were polymorphic and 10 – monomorphic. Capability of the primers was assessed through the high mean values for the polymorphic information content (0.78), effective multiplex ratio (14.73), resolving power (27.90) and marker index (11.36). Based on the obtained mean values of the molecular data the species M. grisebachii (effective number of alleles = 1.39, Shannon’s information index = 0.38, expected heterozygosity = 0.24, Nei's genetic diversity = 0.25, gene flow= 0.65) demonstrated higher genetic diversity than the species M. jankaе (effective number of alleles = 1.28, Shannon’s information index = 0.26, expected heterozygosity = 0.17, Nei's genetic diversity = 0.23, gene flow = 0.52). These results were supported by Analysis of molecular variance (AMOVA), showing higher variability within populations of M. jankaе (90%) and M. grisebachii (62%), than among populations - 10% and 38%, respectively, and 25% among both species. Neighbor joining and principal coordinate analysis (PCoA) grouped the thirty-nine studied populations by species and region of spread. The data are applicable in conservation programs for protecting and keeping of both species. The registered genetic similarity between the populations of the two species (from Eastern Balkan Range) does not exclude the possibility of hybridization between their natural populations.

The investigation of genetic diversity in natural populations of species that show potential for use in reforestation programs is a key step in making management decisions. However, reforestation programs with native species in Brazil are still rarely based on a genetic understanding of the seed matrices used for seedling production. This is also the case for Myrsine umbellata, a dioecious shrub within the family Primulaceae that has been used in reforestation programs in Brazil, mainly due to its high production capacity of fruits attractive to the avifauna. The goal of this study was to measure intra- and interpopulational genetic diversity in natural populations of M. umbellata in six forest remnants of the Atlantic Forest using ISSR markers. The results revealed that the intrapopulational genetic diversity was greater than the genetic diversity among the studied populations. For this reason, the cultivation of seedlings from seeds obtained in more than one population seems the most appropriate strategy for reforestation purposes. Even though the most isolated populations are also the ones with highest genetic structure, all populations of M. umbellata included in this study revealed to be an important germplasm bank conserved in situ. Resumo Investigar a diversidade genética em populações naturais de espécies com potencial para uso em programas de reflorestamento é um passo importante para a tomada de decisões relacionadas ao planejamento e manejo dos plantios. No entanto, programas de reflorestamento com espécies nativas no Brasil raramente são realizados com base no conhecimento do genótipo das matrizes doadoras das sementes utilizadas para produção de mudas. Este é o caso de Myrsine umbellata, um arbusto dioico pertencente à família Primulaceae, que vem sendo utilizado em programas de reflorestamento no Brasil principalmente por apresentar elevada capacidade de produção de frutos atrativos à avifauna. O objetivo deste estudo foi mensurar a diversidade genética intra e interpopulacional em populações naturais de M. umbellata em seis remanescentes florestais da Floresta Atlântica, utilizando marcadores ISSR. Os resultados revelaram que a diversidade genética intrapopulacional foi maior que a diversidade genética entre as populações estudadas. Por essa razão, o cultivo de mudas a partir de sementes obtidas em mais de uma população parece ser a estratégia mais apropriada para reflorestamento. Mesmo considerando que as populações mais isoladas são aquelas mais estruturadas, todas as populações de M. umbellata incluídas neste estudo revelaram ser importantes bancos de germoplasma conservados in situ.

Abstract The paper presents a technique for micropropagation of endangered in Europe and extinct in Poland Pulsatilla vulgaris for ex situ conservation of the genetic resources. Genotype-dependent induction of somatic embryogenesis and rooting was revealed in series of two experiments (I and II) performed under the same experimental conditions. Shoot tips of seedlings were the best explants in both experiments and Murashige and Skoog (MS) medium supplemented with 0.25 or 0.5 mg L−1 BAP was suitable for induction of somatic embryos (SE) and adventitious shoots. Mass SE was obtained in experiment I after explants transfer on ½ MS (2% sucrose) + 0.45 mg L−1 B1 and extending culture to 2–3 months without passages. Rooting of adventitious shoots was a critical point. Out of seven rooting media used in experiment I, only two, ½ MS hormone free (2% sucrose) + 0.45 mg L−1 B1 or MS + 5 mg L−1 NAA + 3.76 mg L−1 B2 resulted in altogether 36.4% rooted shoots. In experiment II, somatic embryogenesis, rooting and acclimatization of adventitious shoots failed. Regenerated plantlets and seedlings converted from SE from experiment I were acclimatized to ex vitro conditions. Both genome size, determined by flow cytometry, and genetic diversity analyzed by ISSR markers, confirmed the compatibility of regenerants from experiment I with P. vulgaris initial seedlings and commercial cultivar. Regenerants obtained in experiment II differed genetically from the regenerants of experiment I and cultivar. Propagated in vitro tissues/organs (SE, adventitious shoots) of P. vulgaris could be a source of material for cryopreservation, artificial seed production and/or for acclimatization of regenerated plantlets and could be used for restoration of the extinct populations. Key Message The micropropagation technique via organogenesis and somatic embryogenesis of endangered in Europe pasqueflower was developed as a tool for species recovery. The critical point is that somatic embryogenesis is genotype-dependent, which affects the repeatability of the experiments and also imposes applying molecular techniques to confirm the genetic fidelity of the regenerants with the initial material and other genotypes.

Habitat disturbance and excessive collection of wild orchids from their natural habitat have threatened many orchids species at risk of extinction. In this study, the in vitro regeneration protocol for Macodes limii, a jewel orchid endemic to Sabah was established. The effects of explant source and plant growth regulators (PGRs) including naphthaleneacetic acid, picloram, 2,4-dichlorophenoxyacetic acid, 6-benzylaminopurine, kinetin, and thidiazuron on the in vitro regeneration capacity of M. limii plantlets were examined. Both factors showed a significant interaction in promoting axillary shoot formation. Nodal explants from the third and fourth positions cultured with 1.0 mg/L TDZ, induced 95% of shoot regeneration, with an average of three shoots/explant (1.6–1.8 cm of shoot length) after 90 days of culture. The well-developed plantlets went through an acclimatization phase for 60 days with a 60% of survival rate. An inter simple sequence repeat (ISSR) marker analysis confirmed the genetic stability of the in vitro regenerated plants to the mother plant. The successfully acclimatized plantlets were finally transferred to Poring Orchid Conservation Centre for reintroduction. The established protocol provides the means for large-scale production of this endemic jewel orchid, as well as a basis for further research aimed at the conservation and genetic improvement of this plant.

The mangabeira is a native fruit tree from Brazil with fruits that present significant potential for exploitation. This species is experiencing genetic erosion, which increases the importance of elucidating the genetic diversity that exists in mangabeira populations to support conservation programs. Thus, this study aimed to evaluate the diversity and genetic structure of mangabeira populations from the Germplasm Bank of Embrapa Meio-Norte using inter simple sequence repeat (ISSR) markers. A total of 29 accessions from Brazil were characterized, including one from Sergipe, one from Bahia, three from the Distrito Federal, 11 from Piauí and 13 from Paraíba. The 11 ISSR primers provided 166 loci, among which 120 were polymorphic. The analysis of molecular variance (AMOVA) indicated that 69.66% of the observed genetic variability occurred within populations and that the populations showed high genetic differentiation. The results obtained from the STRUCTURE analysis indicated the existence of two genetic groups. The Nei and Shannon indices of genetic diversity varied from 0.15 to 0.24 and from 0.22 to 0.34, respectively. The coefficient of similarity ranged from 0.57 to 0.94, with a mean of 0.76. The mean was used as the cut-off point in the dendrogram, and seven groups were identified. In conclusion, this study demonstrates the presence of low or moderate genetic diversity within the studied mangabeira populations and high genetic differentiation between the populations. The results indicate a need to increase the number of mangabeira population samples from different collection sites as a strategy to achieve more significant results for the conservation and genetic improvement of this species. The mangabeira is a native fruit tree from Brazil with fruits that present significant potential for exploitation. This species is experiencing genetic erosion, which increases the importance of elucidating the genetic diversity that exists in mangabeira populations to support conservation programs. Thus, this study aimed to evaluate the diversity and genetic structure of mangabeira populations from the Germplasm Bank of Embrapa Meio-Norte using inter simple sequence repeat (ISSR) markers. A total of 29 accessions from Brazil were characterized, including one from Sergipe, one from Bahia, three from the Distrito Federal, 11 from Piauí and 13 from Paraíba. The 11 ISSR primers provided 166 loci, among which 120 were polymorphic. The analysis of molecular variance (AMOVA) indicated that 69.66% of the observed genetic variability occurred within populations and that the populations showed high genetic differentiation. The results obtained from the STRUCTURE analysis indicated the existence of two genetic groups. The Nei and Shannon indices of genetic diversity varied from 0.15 to 0.24 and from 0.22 to 0.34, respectively. The coefficient of similarity ranged from 0.57 to 0.94, with a mean of 0.76. The mean was used as the cut-off point in the dendrogram, and seven groups were identified. In conclusion, this study demonstrates the presence of low or moderate genetic diversity within the studied mangabeira populations and high genetic differentiation between the populations. The results indicate a need to increase the number of mangabeira population samples from different collection sites as a strategy to achieve more significant results for the conservation and genetic improvement of this species.

The Viola epipsila-V. palustris complex is a highly taxonomically complicated group of species in its entire circumboreal range of distribution. Habitat loss, forest flooding, and hybridization could lead to the extinction of V. epipsila. A hybrid index and principal component analysis (PCA) were used to select qualitative and quantitative morphological features to distinguish parent species and hybrids, inter simple sequence repeat (ISSR) markers to determine the genetic diversity of the populations, flow cytometry to estimate the genome size (GS), and non-coding chloroplast DNA (cpDNA) regions to indicate the directions of crosses. All taxa are very morphologically variable, and their features can change within a season. The most stable feature is the distance of the bracts on the pedicel from the rhizome. The genetic diversity of all taxa populations is low and highly influenced by selfing and vegetative propagation. The population structure is differentiated: populations of V. epipsila or V. palustris, mixed populations with both parent species, F1 hybrids and populations with introgressive forms occur in different regions. The interspecific GS variation corresponds to the ploidy level (4x = 2.52 pg, 8x = 4.26 pg, 6x = 3.42 pg). Viola epipsila is the mother plant of the hybrids. Research has shown the risk of V. epipsila extinction in Central Europe and the importance of local populations in studying the role of hybridization in reducing/maintaining/increasing biodiversity.

The paper presents the results of the analysis of relationship between genetic distance and geographical distance in different broomrape populations (Orobanche cumana) - a rhizopathogen specific for sunflower that essentially affects the production of this important oil crop. Thirty-nine broomrape populations collected from different regions of the Republic of Moldova and belonging to different races (≤ E, F, G and H) have been included in the study. Correlative analysis of the data with reference to the genetic distance calculated based on the results of fingerprinting with SSR (Simple Sequence Repeats) and ISSR (Inter Simple Sequence Repeats) markers and to the geographical distance suggested that no correlation exists between these two parameters. The matrix of genetic distances was used as input for a principal coordinate analysis (PCoA). The PCoA revealed a more clear grouping of populations from the same region in the case of ISSR analysis, the main part of populations from central and southern part of the country being grouped into two clusters, regardless of their race.

Fusarium head blight (FHB) is a devastating disease of small grains cereal in Argentina. FHB is mainly associated with species within the Fusarium graminearum species complex, potential trichothecene producers. The control strategy success will be determined by the interaction among genetic variability of the pathogen, the host susceptibility and the environmental factors. The aim of this study was to evaluate the genetic variability of F. graminearum ss populations defined as temporal and spatial populations. Symptomatic spikes were collected during an FHB outbreak (2012/13) and a non-outbreak (2014/15) wheat-harvest season from wheat fields in Argentina. Strains were morphological and molecular analysed, of which four strains were Fusarium meridionale and 71 were Fusarium graminearum ss. The fungal populations assessed by ISSR molecular markers showed a genetic diversity (GD = 0.999–1.00) and migration rate Nm = 5 and structure population determined K = 2. Subpopulation 1 (K1) was established by 29 strains isolated from Marcos Juárez (MJ) and Pergamino (PER) during the 2012/13 and 2014/15 harvest seasons and subpopulation 2 (K2) was established by 46 strains isolated from Corral de Bustos (CBU) and Carlos Pellegrini (CPE) during the 2012/13 and 2014/15 harvest seasons. High genotypic and genetic diversity together with great differentiation within subpopulations and high gene flow would indicate that the strains isolated in Argentina are part of the same, unique and widespread population, as was already shown in a previous study. Understanding fungal genetic structure and continue monitoring of F. graminearum ss populations through different harvest seasons will assist not only breeders but also the development of improved management control strategies. Fil: Yerkovich, Nadia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales, Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Fumero, María Verónica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales, Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Cantoro, Renata. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales, Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Palazzini, Juan Manuel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales, Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales, Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigación en Micología y Micotoxicología; Argentina

We report the first comprehensive multilocus molecular characterization of cultivated Ilex paraguariensis plants belonging to a breeding program. Using nuclear and homologous chloroplastidic molecular markers, we have genotyped 158 plants from four plantation sites. Analyses of the nuclear data (187 variable dominant loci) allowed detecting high diversity (0.569), the occurrence of four distinct genetic clusters, and a low but significant differentiation among sites. Additionally, 20 chloroplastidic alleles were identified applying five microsatellite polymorphic markers, and a high chloroplastidic diversity was recognized (0.505); two haplogroups were distinguished amongst the 63 haplotypes detected. Our results from both nuclear and plastidic markers indicate that most genetic variation reside within plantations sites (≥ 95%), and that these plantations were established on highly variable materials (either as seeds or plantlets) derived from, at least, 63 maternal lineages. Moreover, our study suggests that the genetic structure within each plantation site was most likely shaped by past admixture favored by farmers´ practices during the establishment of each plantation. Also, subsequent constraints in gene flow and/or a low level of shared polymorphism among plantations could have contributed to current structure.

In vitro regeneration of Viola stagnina Kit., endangered in most part of its European distribution range, was successfully obtained based on the newly developed protocol. Adventitious shoots via direct and indirect organogenesis were induced on leaf blade and petiole fragments on solidified Murashige and Skoog (MS) medium supplemented with 0.5 or 1 mg l−1 thidiazuron, respectively. Shoots were rooted on half-strength MS medium with 2% sucrose and 0.5 mg l−1 indole-3-acetic acid (IAA), and plantlets were successfully acclimatized. Sixty-five of the regenerated plants (72% of isolated shoots cultured on rooting medium) survived at the experimental plot conditions. Among recovered via organogenesis plants, individuals ‘true-to-type’ derived from initial plant G1 and also plants genetically distant from initial plants were detected by ISSR markers. In all groups of clones genetic indices (number of genotypes, polymorphic markers, gene diversity, total gene diversity, mean gene diversity) were lower than in natural populations. Regenerated plantlets had the same genome size estimated by flow cytometry as initial material and plants from natural populations. They developed chasmogamous flowers with highly viable pollen grains (over 90%), cleistogamous flowers, and set seeds from both flower types in the first and second seasons cultivated at experimental plots. This is the first report of a successfully developed micropropagation protocol of V. stagnina, and the first detailed genetic analysis of recovered plants with the use of ISSR markers and genome size measurements allowing to discuss the advantageous role of somaclonal variation in ex situ plant conservation with the use of in vitro micropropagation.

Мета. Виявити генетичний поліморфізм та дивергенцію сортів і гібридів кабачка (C. pepo L.) різного географічного походження за ISSR маркерами.Методи. Для оцінки генетичного поліморфізму 29 сортів і гібридів C. pepo L. різного походження використовували ISSR аналіз. Коефіцієнт подібності між досліджуваними зразками кабачка визначали за формулою Nei–Li. Розрахунок коефіцієнтів подібності та побудування філогенетичного дерева здійснювали за допомогою пакету програм Phylip-3.69. Кластерний аналіз проводили методом приєднання сусідів (Neighbor-joining). Достовірність групування зразків у кластери перевіряли методом бутстреп-аналізу.Результати. Використання 13 праймерів до міжмікросателітних ділянок дозволило виявити 129 локусів ДНК кабачка, серед яких 109 були поліморфними, 20 – мономорфними, 11 – унікальними для певних зразків. Електрофореграми продуктів ампліфікації дослідних зразків різнилися кількістю і розміром ампліконів. Виявлено високий поліморфізм ДНК дослідних зразків кабачка, який варіював від 62,5 % (праймер UBC 810) до 100 % (праймери UBC 2, UBC 3 і UBC 846). Рівень молекулярно-генетичного поліморфізму зразків кабачка варіював від 55,8 до 63,6 % у гібридів ‘Rimini’ і ‘Eight Ball’, відповідно. Встановлено низьку генетичну дивергенцію між дослідними зразками C. pepo L., коефіцієнт подібності за Nei–Li становив 0,0005–0,0092. За допомогою кластерного аналізу зразки кабачка були згруповані у два кластери. Основним критерієм кластеризації був рівень генетичної дивергенції. Географічне походження зразків не впливало на особливості їх групування.Висновки. За результатами вивчення зразків кабачка різного географічного походження за допомогою ISSR аналізу встановлено високий генетичний поліморфізм і незначна генетична дивергенція між дослідними зразками. Виявлено унікальні фрагменти ДНК, які можуть бути використані для паспортизації відповідних зразків, а також для розробки інших молекулярно-генетичних маркерів. Отримана інформація може бути корисною для оптимізації селекційного процесу кабачка і подальших досліджень в області молекулярної генетики цієї культури., Цель. Выявить генетический полиморфизм и дивергенцию сортов и гибридов кабачка (C. pepo L.) разного географического происхождения по ISSR маркерами.Методы. Для оценки генетического полиморфизма 29 сортов и гибридов C. pepo L. различного происхождения использовали ISSR анализ. Коэффициент сходства между исследуемыми образцами кабачка определяли по формуле Nei-Li. Расчет коэффициентов сходства и построение филогенетического дерева осуществляли с помощью пакета программ Phylip-3.69. Кластерный анализ проводили методом присоединения соседей (Neighbor-joining). Достоверность группировки образцов в кластеры проверяли методом бутстреп-анализа.Результаты. Использование 13 праймеров к межмикросателлитным участкам позволило выявить 129 локусов ДНК кабачка, среди которых 109 были полиморфные, 20 – мономорфные, 11 – уникальные для определенных образцов. Электрофореграммы продуктов амплификации опытных образцов отличались количеством и размером ампликонов. Выявлен высокий полиморфизм ДНК опытных образцов кабачка, который варьировал от 62,5% (праймер UBC 810) до 100% (праймеры UBC 2, UBC 3 и UBC 846). Уровень молекулярно-генетического полиморфизма образцов кабачка варьировал от 55,8 до 63,6% у гибридов "Rimini" и "Eight Ball", соответственно. Установлено низкую генетическую дивергенцию между опытными образцами C. pepo L., коэффициент сходства по Nei-Li составил 0,0005-0,0092. С помощью кластерного анализа образцы кабачка были сгруппированы в два кластера. Основным критерием кластеризации был уровень генетической дивергенции. Географическое происхождение образцов не влияло на особенности их группировки.Выводы. По результатам изучения образцов кабачка разного географического происхождения с помощью ISSR анализа установлены высокий генетический полиморфизм и незначительная генетическая дивергенция между опытными образцами. Обнаружены уникальные фрагменты ДНК, которые могут быть использованы для паспортизации соответствующих образцов, а также для разработки других молекулярно-генетических маркеров. Полученная информация может быть полезной для оптимизации селекционного процесса кабачка и последующих исследований в области молекулярной генетики этой культуры., Purpose. Identify genetic polymorphism and divergence of C. pepo L. varieties and hybrids of different geographical origin using ISSR markers.Methods. ISSR analysis was used to evaluate the genetic polymorphism of 29 C. pepo L. varieties and hybrids of different origin. The similarity coefficient between the investigated courgetti accessions was calculated by the Nei – Li’s formula. Calculating coefficients of similarity and phylogenetic tree construction was performed with the Phylip-3.69 software package. The cluster analysis was performed by the Neighbor-joining method. The validity of the accessions grouping into clusters was tested by bootstrap analysis.Results. The use of 13 primers in the intermicrosatellite regions revealed 129 loci of courgetti DNA, among them 109 were polymorphic, 20 were monomorphic, 11 were unique to certain accessions. Electrophoregrams of the amplification products of the investigated accessions differed in the number and size of the amplicons. High DNA polymorphism of the investigated courgetti accessions was found, which ranged from 62,5% (primer UBC 810) to 100% (primers UBC 2, UBC 3, and UBC 846). The level of molecular genetic polymorphism of courgetti accessions varied from 55,8 to 63,6% in the 'Rimini' and 'Eight Ball' hybrids correspondingly. Low genetic divergence was determined between the C. pepo L. specimens, the Nei – Li similarity coefficient was 0,0005–0,0092. Using the cluster analysis, courgetti accessions were grouped into two clusters. The main criterion for clustering was the level of genetic divergence. The geographical origin of the accessions did not affect the peculiarities of their grouping.Conclusions. The results of the study of courgetti accessions of different geographical origin using ISSR analysis revealed high genetic polymorphism and little genetic divergence between the experimental accessions. Unique DNA fragments have been identified and can be used for the certification of relevant samples, as well as for the development of other molecular genetic markers. The obtained information may be useful for optimizing the courgetti breeding process and for further studies in the molecular genetics of this culture.

Purpose. Identify genetic polymorphism and divergence ofC. pepoL. varieties and hybrids of different geographical origin using ISSR markers. Methods. ISSR analysis was used to evaluate the genetic polymorphism of 29C. pepoL. varieties and hybrids of different origin. The similarity coefficient between the investigated courgetti accessions was calculated by the Nei – Li’s formula. Calculating coefficients of similarity and phylogenetic tree construction was performed with the Phylip-3.69 software package. The cluster analysis was performed by the Neighbor-joining method. The validity of the accessions grouping into clusters was tested by bootstrap analysis. Results. The use of 13 primers in the intermicrosatellite regions revealed 129 loci of courgetti DNA, among them 109 were polymorphic, 20 were monomorphic, 11 were unique to certain accessions. Electrophoregrams of the amplification products of the investigated accessions differed in the number and size of the amplicons. High DNA polymorphism of the investigated courgetti accessions was found, which ranged from 62,5% (primer UBC 810) to 100% (primers UBC 2, UBC 3, and UBC 846). The level of molecular genetic polymorphism of courgetti accessions varied from 55,8 to 63,6% in the 'Rimini' and 'Eight Ball' hybrids correspondingly. Low genetic divergence was determined between theC. pepoL. specimens, the Nei – Li similarity coefficient was 0,0005–0,0092. Using the cluster analysis, courgetti accessions were grouped into two clusters. The main criterion for clustering was the level of genetic divergence. The geographical origin of the accessions did not affect the peculiarities of their grouping. Conclusions. The results of the study of courgetti accessions of different geographical origin using ISSR analysis revealed high genetic polymorphism and little genetic divergence between the experimental accessions. Unique DNA fragments have been identified and can be used for the certification of relevant samples, as well as for the development of other molecular genetic markers. The obtained information may be useful for optimizing the courgetti breeding process and for further studies in the molecular genetics of this culture.

Genetic diversity of two local populations of the Ural endemic and the Pleistocene relict Scorzonera glabra Rupr. (Asteraceae) from the surroundings of Karabashskiy Copper-Smelting Plant (impact) and mountain Egosa (background) was studied using ISSR markers. The results show that the level of polymorphism is high in both populations. Analysis of molecular variance (AMOVA) show that the large portion of the variability (85 %) determines by intrapopulation one. There are no significant differences in genetic diversity among local populations. We suppose that S. glabra being a plastic species has pre-adaptations that allow it to grow under impact conditions

Any restoration and improvement programs for Persian oak stands require assessment of the structure of genetic variation within and among the natural populations and realize the genetic distance patterns. Vulnerability against biotic and abiotic stresses is the consequence of the reduction in genetic diversity. Genetic variation is crucial for natural populations of organisms to evolve in response to changing environmental variables. Low level of diversity in the gene pool of plant spices may lead to increased risk of extinction in the face to abiotic and biotic environmental stresses. Genetic diversity of 180 Persian oak trees from central Zagros region including Kermanshah, Ilam and Lorestan provinces was investigated. Fifteen ISSR loci amplified and 157 alleles produced which 99.17 percent (156 alleles) were polymorph. Allele number per locus was in a range from 4 (UBC814) to 17 (ISSR16) with average 10.47; polymorphism information content was different from 0.052 (UBC895) to 0.425 (UBC807). Amplified fragments were in a range from 150bp to 1800bp. AMOVA shows 20 percent of variation among and 80 percent of variation within populations. The minimum genetic distance was manifested in Lorestan populations and the maximum distance was in Kermanshah. Results showed the higher levels of differentiation among Kermanshah and Lorestan genotypes. Heterogeny of evaluated loci in populations in Kermanshah was more than others in Lorestan and Ilam. Cluster analysis led to groping of genotypes in five distinct clusters. Amplification and suitable polymorphism of ISSR markers in Persian oak and also a high level of molecular variance between populations are promising for the capacity of this molecular marker system to detect the informative alleles for phenotypic traits.

Fagopyrum spp. (buckwheat) is a dicotyledonous pseudocereal crop mainly cultivated in the north-western Himalayan regions for its highly nutritional, antioxidant and therapeutic values. In the present investigation, molecular characterization was performed by using ISSR (inter simple sequence repeat) markers on 42 accessions of four buckwheat species (Fagopyrum esculentum, F. sagittatum, F. tataricum and F. kashmirianum). The 12 pre-screened ISSR primers amplified 102 bands, and amongst them 85 bands exhibited polymorphism with an average polymorphism of 82.73%. The results revealed that Shannon’s information indices (I) and Nei’s genetic diversity (H) were low for F. tataricum (I = 0.1028 ± 0.2307, H = 0.0707 ± 0.1617) and high for F. esculentum (I = 0.1715 ± 0.2622, H = 0.1164 ± 0.1796). It was estimated that within the accessions of Fagopyrum species, the species diversity (HT) and mean diversity (HS) were 0.3200 and 0.1041, respectively. Molecular variance partitioning by AMOVA also indicated a significant genetic differentiation accounting for 73% among and 27% within the accessions of Fagopyrum species. Overall, accessions of F. esculentum had the greatest distance from the other accessions of buckwheat species, which includes F. sagittatum, F. tataricum and F. kashmirianum as revealed by FST distance and Nei’s unbiased genetic distance. The dendograms based on UPGMA and PCoA segregated 42 accessions of four buckwheat species into three major groups. This study clearly reveals a considerable amount of genetic diversity at the intra-specific level in F. esculentum, F. sagittatum and F. kashmirianum accessions. The factors responsible for it are diverse geographical conditions, pollinating behavior and cultivation practices adapted in these regions. The study also indicated a close phylogenetic relationship between F. tataricum and F. kashmirianum.

The runner bean is the third most economically important Phaseolus species, which is cultivated on small-scale agriculture for the production of immature pods or to obtain dry seeds. However, despite the economic importance and agronomic potential of this species, the runner bean has been little studied from the genetic standpoint. Therefore, the main objective of this study was to characterize ten selected lines of runner bean obtained from Central (Santiago) and Southern (Valdivia and Villarica) Chile based on morphological and agronomic traits. In addition, the genetic variability of these lines was determined using 12 Inter-Simple Sequence Repeat (ISSR) markers to evaluate the potential of this germplasm for breeding and commercial purposes. As a result, the lines from Central Chile were characterized, and had a higher number of pods per plant compared to the Southern lines, although the size and weight of their seeds were lower. Moreover, a low level of genetic diversity (He = 0.251) was encountered in this population. Finally, this is one of the first studies that generate relevant and novel information on the morphological, agronomic and genetic characterization of the P. coccineus germplasm present in Chile.

Banana and plantains are generally classified based on morphological characteristics namely AA, AAA, AB, AAB and ABB. Further, there are four sub-groups in AAB genomic group. Presently we analyzed diversity using ISSR markers in 18 cultivars of AAB genomic group along with two each of AA types and BB wild accessions for comparison. The results have shown that AAB cultivars form a separate group. Dendrogram analysis showed that the subgroups ‘Plantain’ ‘Silk’ and ‘Mysore’ were placed in between AA and BB type. Whereas ten cultivars of ‘Pome’ sub group of the cultivars were unique and was placed in a separate cluster. In this study using ISSR markers, we are able to identify the subgroups clearly and their genetic relationships within the AAB group. The cultivars Rasthali and Nendran were clearly separated. The Pome sub-group cultivars found to be in a group which may be based on their geographical origin.

Currently three species are recognized in Russia within the genus Elymus subsection Pendulini, namely, E. pendulinus, E. brachypodioides and E. vernicosus. The degree and quality of lemma and stem node vestitures and ecology are considered important characters for species delineation. In Primorsky krai and Altai Republic we have found a number of mixed populations in which there is visually continuous variation in lemmas vestitures from being completely glabrous to densely puberulent or pilose. Additionally, there are a number of populations in Primorsky krai in which individuals having either smooth or pilous stem nodes grow together. A study of polymorphism of ISSR markers among selective accessions of three species from different locations was carried out together with the reference species E. ciliaris and E. gmelinii. The consensus dendrogramm constructed by results of ISSR markers has shown no taxon specific patterns in accessions of the subsection Pendulini. Hybrids between the individuals of different species of the subsection in six cross combinations have been created. All F1 plants had normally developed open anthers, seed fertility (SF) ranged from 8 % to 89 % in different combinations. In F2 populations no sterile individuals or increase of SF up to normal values were observed. Hence, three taxa form a unified recombination gene pool. The diagnostic characters were found to be controlled by one pair of alternative alleles for glabrous vs. hairy stem nodes, and by one or two pairs of alleles in different cross combinations for glabrous vs. hairy lemmas. These characters can be considered as normal inter- and/or intra-population variation. Therefore, E. brachypodioides and E. vernicosus need to be relegated to infraspecific rank within Elymus pendulinus s. l. as varieties.

Information about the genetic variation among cultivars of vegetable crops is of vital importance for improvement of plant breeding programmes worldwide. The objectives of this study were to group 19 pepper (Capsicum annuum L.) cultivars from the collection of Maritsa Vegetable Crops Research Institute, Plovdiv, Bulgariainto clusters according to their distances as estimated by agronomic traits and 9 di-and tri -nucleotide inter simple sequence repeat polymorphism (ISSR) markers and to assess the relationships between them. The phenotypic characterization during 3 consecutive years revealed significant differences among Bulgarian cultivars for the studied 13 phenotypic traits. The biplot analysis of quantitative traits showed that the most strongly correlating traits with the first axis (55.6% of variance) were fruit width, fruit weight and pericarp thickness (in the negative direction of the axis), and plant height (PH) (in the positive direction). The most discriminative traits, considering the second axis (22.6% of variance) were fruit length (FL) and to a lesser extent the stem height (StH). The correspondence analysis of the qualitative traits showed that the intensity of the colour of the fruit (before and at maturity), fruit colourbefore maturity and fruit shape in longitudinal section were the most discriminative characteristics for the first two dimensions. The agronomic traits data and 7 dinucleotide ISSR primers were used to estimate the pairwise genetic distances. Higher mean phenotypic distance (0.414) in comparison to the genotypic ones (0.214) among the cultivars was observed, indicating higher phenotypic diversity among them. A highly signi?cant, positive correlation between the agronomic data and ISSR marker-based matrices (r=0.41, p=0.001)was detected. This indicates that ISSR distance tended to re?ect that of the agronomics ones. However, additional molecular studies and large collection of highly diverse genotypes are needed to reveal associations between the marker loci and specific QTLs. The research initiated is a base for more precise estimation of genetic distances between pepper genotypes from the available large collection of landraces, local and modern cultivars including large number of highly polymorphic markers.

Genotypes of the chrysanthemum cultivars Harman and Indianapolis were transformed using Agrobacterium tumefaciens strain LBA4404, which carries the binary plasmid pBIN19::Rd29A::ScTPS1TPS2::nos and pBIN19::35S::ScTPS1TPS2::nos, respectively. ISSR markers were used to discriminate two cultivars from each other, and independent transgenic plants generated in each cultivar. Firstly, the nptII gene and Tnos sequence in the transformed genotypes was confirmed by PCR. For genotyping, 10 ISSR profile markers produced 131 DNA bands. The percentage of polymorphism ranged from 20 to 82.4 %. Primer UBC 872 provided the highest percentage of polymorphic bands (PBP), polymorphic information content (PIC), marker index (MI) and resolving power (Rp). A positive correlation was found between the Rp value of each primer and the number of identified genotypes (r=0.822***). The UPGMA analysis generated two groups with a similarity coefficient of 0.67. The genotype grouping was confirmed by PCoA. The ISSR technique was efficient to discriminate transgenic cultivars from non-transgenic plants.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2022-06-08T15:15:31Z No. of bitstreams: 1 2019 - Gustavo Torres Dos Santos Amorim.pdf: 4496184 bytes, checksum: 97c5d949fe3471296b9348cded51297a (MD5) Made available in DSpace on 2022-06-08T15:15:31Z (GMT). No. of bitstreams: 1 2019 - Gustavo Torres Dos Santos Amorim.pdf: 4496184 bytes, checksum: 97c5d949fe3471296b9348cded51297a (MD5) Previous issue date: 2019-07-30 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior The periwinkle (Catharanthus roseus [L.] G. Don) is distinguished because it is exclusive source of vincristine and vinblastine alkaloids, however they occur in low concentrations in the species. Such alkaloids are used in chemotherapy for the treatment of tumors, leukemias and lymphomas. Therefore, it is essential to develop a breeding program in search of superior genotypes for vincristine and vinblastine production. For this, it is necessary, initially, the study of the preferential reproduction mode of the species for the correct application of the breeding methods. Given the above, this research aimed to study the preferential mode of reproduction in 10 genotypes of periwinkle from the germplasm collection of the Federal Rural University of Rio de Janeiro, based on the pollen:ovule (P:O) ratio, histochemical pollen, emission and development of pollen tubes in vivo, pollen viability and receptivity of stigma. As well as to estimate genetic diversity using ISSR molecular markers (Inter Simple Sequence Repeats) among the analyzed genotypes. To estimate the ratio (P:O), anthers of each flower were excised, and pollen grains were counted under a microscope; ovaries of the same flowers were dissected, and the ovules were counted under stereomicroscope. In the histochemical analysis of pollen grains were used the dyes Sudan IV and the solution I2KI for the detection of lipid and starch, respectively. The emission and development of pollen tubes in vivo were evaluated in flower buds and flowers in anthesis, under fluorescence microscopy, using 0.1% aniline blue fluorochrome. For the pollen viability, in flower buds, the triple solution of Alexander was used and for the receptivity of the stigma was adopted the solution of alpha-naphthyl acetate, in closed flowers and anthesis. In the genotypic analysis, 15 ISSR primers were used. The phenetic matrix was estimated by dissimilarity index of Jaccard and the grouping of the genotypes was performed by UPGMA (Unweighted Pair Groups Method with Arithmetic Mean). The consistency of the clustering pattern was verified through the cophenetic correlation. The results of the ratio (P:O) indicated that eight of the 10 genotypes presented both facultative self-pollinated and facultative cross pollinated (mixed mating). However, two genotypes presented only one mode of reproduction, facultative cross pollinated, in UFRRJ VIN007, and facultative self-pollinated in UFRRJ VIN004. The receptive stigmas and the high pollen viability in flower buds established favorable conditions for the occurrence of cleistogamy. Based on the emission and development of pollen tubes in vivo, it was verified that of the eight genotypres with mixed reproduction mode, six of them had a strong tendency to allogamy and all were self-compatible. It was also verified the occurrence of cleistogamy in 50% of the genotypes. In the case of UFRRJ VIN004 such occurrence was accentuated, which reinforces its tendency to autogamy. However, it was observed that seven genotypes, including UFRRJ VIN007, showed a strong tendency to allogamy. In view of this, a great diversity of reproductive strategies can be observed in periwinkle, since both self-pollinated and allogamy may be present. The high genetic diversity obtained can be explained by the strong tendency to allogamy found in 70% of the analyzed genotypes A vinca (Catharanthus roseus [L.] G. Don) distingue-se por ser fonte exclusiva dos alcaloides vincristina e vimblastina, no entanto ocorrem em baixas concentra??es na esp?cie. Tais alcaloides s?o utilizados em quimioter?picos para o tratamento de tumores, leucemias e linfomas. Sendo assim, ? fundamental desenvolver um programa de melhoramento gen?tico na busca de gen?tipos superiores quanto a produ??o de vincristina e vimblastina. Para isso, ? necess?rio, inicialmente, o estudo do modo de reprodu??o preferencial da esp?cie para a aplica??o correta dos m?todos de melhoramento. Diante do exposto, esta pesquisa teve como objetivo estudar o modo de reprodu??o preferencial em 10 gen?tipos de vinca da cole??o de germoplasma da Universidade Federal Rural do Rio de Janeiro, com base na rela??o p?len:?vulo (P:O), histoqu?mica dos gr?os de p?len, emiss?o e desenvolvimento de tubos pol?nicos in vivo, viabilidade pol?nica e receptividade do estigma. Bem como estimar a diversidade gen?tica via marcadores moleculares ISSR (Inter Simple Sequence Repeats) entre os gen?tipos analisados. Para estimar a raz?o (P:O), anteras de cada flor foram excisadas, e os gr?os de p?len foram contados sob microsc?pio; ov?rios das mesmas flores foram dissecados, e os ?vulos foram contados sob estereomicrosc?pio. Na an?lise histoqu?mica dos gr?os de p?len utilizou-se os corantes Sudan IV e a solu??o de I2KI para detec??o de lip?deo e amido, respectivamente. A emiss?o e desenvolvimento de tubos pol?nicos in vivo foram avaliados em bot?es florais e flores na antese, em microscopia de fluoresc?ncia, utilizando-se o fluorocromo azul de anilina a 0,1%. Para a viabilidade pol?nica, em bot?es florais, empregou-se a solu??o tripla de Alexander e para a receptividade do estigma adotou-se a solu??o de acetato de alfa-naftil, em flores fechadas e na antese. Na an?lise genot?pica foram utilizados 15 primers ISSR. A matriz fen?tica foi estimada pelo ?ndice de dissimilaridade de Jaccard e o agrupamento dos gen?tipos foi realizado via UPGMA (Unweighted Pair Groups Method with Arithmetic Mean). Verificou-se a consist?ncia do padr?o de agrupamento atrav?s da correla??o cofen?tica. Os resultados da rela??o (P:O) indicaram que oito, dos 10 gen?tipos, apresentaram, simultaneamente, autogamia facultativa e alogamia facultativa (modo de reprodu??o misto). Por?m, dois gen?tipos apresentaram somente um modo de reprodu??o, al?gamo facultativo, no UFRRJ VIN007, e aut?gamo facultativo no UFRRJ VIN004. Os estigmas receptivos e a elevada viabilidade pol?nica em bot?es florais estabeleceram condi??es favor?veis para a ocorr?ncia da cleistogamia. Com base na emiss?o e desenvolvimento de tubos pol?nicos in vivo, constatou-se que dos oito gen?tipos com modo de reprodu??o misto, seis deles tiveram forte tend?ncia a alogamia e todos foram autocompat?veis. Verificou-se tamb?m a ocorr?ncia da cleistogamia em 50% dos gen?tipos. No caso, do UFRRJ VIN004 tal ocorr?ncia foi acentuada, o que refor?a a sua tend?ncia a autogamia. Entretanto, observou-se que sete gen?tipos, incluindo o UFRRJ VIN007, exibiram forte tend?ncia a alogamia. Diante disso, percebe-se grande diversidade de estrat?gias reprodutivas em vinca, j? que tanto a autofecunda??o quanto a alogamia podem estar presentes. A alta diversidade gen?tica obtida pode ser explicada pela forte tend?ncia a alogamia constatada em 70% dos gen?tipos analisados

Intra-species genetic variability assessment is an effective tool in formulating genetic improvement and germplasm conservation strategies. Houttuynia cordata Thunb. is a semidomesticated medicinal herb consumed widely in traditional diet in North-Eastern India. In the present study, an effort has been made to assess the genetic diversity of H. cordata Thunb. from Brahmaputra valley of North-East India. A total of 545 genotypes from 18 populations of H. cordata Thunb. from four different regions, i.e. North-East, North-West, South-East and South-West, with respect to river Brahmaputra were collected and population genetic diversity and structure were analysed using ISSR molecular markers. Population genetic structure analysis using unweighted pair group method with averages (UPGMA)-based hierarchical cluster analysis, principal coordinate analysis (PCoA) and model-based clustering in STRUCTURE program revealed that the population of H. cordata Thunb. grouped according to regional distribution and forms four genetically distinct clusters. The analysis of molecular variance showed that differentiation among regions was significant with 60% genetic variation among region, 3% genetic variation among population within region and 37% genetic variation within population. We found wide variation in Nei’s gene diversity (Hj) ranging from 0.07782 in Margherita population to 0.13634 in Barapani population. Furthermore, Nei’s gene diversity within population (Nei’s Hs) and total gene diversity (Ht) were found to be 0.1081 and 0.1769 respectively. The genetic differentiation among 18 population was high (Fst = 0.3894; p

The Collared Kingfisher (Todiramphus chloris) is widely distributed across the Indian and western Pacific Oceans and consists of about 50 subspecies. Two different subspecies of T. chloris occur in the Arabian Peninsula: T. c. abyssinicus from the Red Sea coast and T. c. kalbaensis from the Arabian Sea coast in the United Arab Emirates and Oman. The aim of this study was to determine the molecular relationship between the two Arabian subspecies and to establish the first DNA barcodes from the Arabian Peninsula for this species. Three different mitochondrial genes were used: (i) cytochrome c oxidase subunit I (COI), (ii) 12S rRNA (12S) and (iii) NADH dehydrogenase-1 (ND1). The COI gene sequences of the two subspecies were 100% identical, while the 12S and ND1 gene sequences revealed a unique single nucleotide variation between the two subspecies. Thus, this single nucleotide variation can be used as a DNA barcode to discriminate between two subspecies. Furthermore, the genetic profile or fingerprint for both subspecies were compared using ten primers of the highly polymorphic nuclear markers (Inter Simple Sequence Repeat, ISSR). As expected, the DNA analysis of the ISSR markers was able to distinguish between the specimens of the two subspecies. These results suggest that T. c. abyssinicus and T. c. kalbaensis are not identical and thus belong to different subspecies. Besides, the sequences of the COI gene for T. c. abyssinicus and T. c. kalbaensis differs by only 1.28% from T. sanctus suggesting that the Arabian subspecies are closely related to the Sacred Kingfisher (T. sanctus).

The objective of this work was to evaluate the use of RAPD and ISSR molecular markers to determine the genetic variability among cashew (Anacardium spp.) genotypes, and to indicate possible promising crosses based on cashew genetic variability and phenotypic performance. Ten hybrids - derived from the crosses CCP 76 x BGC 589, CCP 76 x BRS 226, CCP 76 x HAC 276-1, CCP 76 x Embrapa 51, CCP 76 x BRS 253, CCP 76 x HAC222-4, and BRS226 x Embrapa 51 - and their parents were assessed at the molecular level. The hybrids were evaluated for nut yield, mean nut weight, bored nuts, and powdery mildew on nuts (scale 0-4). The RAPD and ISSR markers were efficient in the determinaton of the genetic variability among cashew genotypes, allowing of the grouping of 21 clusters. Associated with the phenotypic characterization of cashew nut for yield, weight, and health, the used markers can efficiently identify possible combinations with higher genetic variability and higher probability of developing transgressive genotypes in segregating populations. Resumo: O objetivo deste trabalho foi avaliar o uso de marcadores moleculares RAPD e ISSR para determinar a variabilidade genética entre genótipos de cajueiro (Anacardium spp.) e indicar possíveis cruzamentos promissores com base na variabilidade genética e no desempenho fenotípico do cajueiro. Foram avaliados, em nível molecular, dez híbridos - originados dos cruzamentos CCP 76 x BGC 589, CCP 76 x BRS 226, CCP 76 x HAC 276-1, CCP 76 x Embrapa 51, CCP 76 x BRS 253, CCP 76 x HAC222-4 e BRS226 x Embrapa 51 - e seus respectivos genitores. Os híbridos foram avaliados quanto à produção de castanha, à massa média da castanha, à incidência de castanhas furadas e à incidência de oídio na castanha (escala 0-4). Os marcadores RAPD e ISSR foram eficientes em determinar a variabilidade genética entre esses genótipos de cajueiro, tendo permitido o agrupamento de 21 grupos. Associados à caracterização fenotípica da castanha de caju quanto à produtividade, à massa e à sanidade da castanha, os marcadores utilizados são eficientes para identificar possíveis combinações capazes de proporcionar maior variabilidade genética e maior probabilidade de obtenção de genótipos transgressivos em populações segregantes.

One of the hypotheses about the causes of invasive transmission of Phalaris arundinacea in the US is, that this species have repeatively introducted from Europe or by hybridization of the introduced genotypes with the native species. This is the same situation as Phragmites australis. Part of this intercontinental gene flow is enhanced by the production of Phalaris cultivars in Minnesota. The consequences of these exchanges have a significant impact on the management of invasive crops from both continents. Previous studies confirm only minimal morphological heterogeneity between cultivated and native (wild) forms of Phalaris arundinacea. In Phalaris arundinacea, we analyzed genetic similarities and differences between the US populations (Minnesota) and population of the Czech Republic. The extent of genetic variation in native and invasive populations was compared using ISSR markers. The occurence of commercial genotypes was observed across the full spectrum of invasive genotypes, which suggests a relatively frequent exchange of genes among forage, ornamental, and native US populations. The genus Myriophyllum is mainly significant because of its invasive species, which have always been in a special position as the course of ecological and evolutionary changes may be observed in real time. In Central Europe it is represented by three native species (M. spicatum L., M. alterniflorum and M. verticillatum) and two non-native species of North American M. heterophyllum and South American M. aquaticum. Species M. spicatum, together with M. heterophyllum and M. aquaticum, is one of the most important invasive plants that, due to their aggressive growth, cause many problems, especially in North America. The main theme of this Ph.D. is the evaluation of the role of polyploidization in processes leading to invasive character of some species of the genus Myriophyllum in the USA, cytological and population-genetic comparison of native (Eurasia) and invasive populations (US) of Myriophyllum spicatum. Moreover, possible "Pools" of genetic and cytotype variability of invasive populations and evaluation of usability of simple genome size were determinated in taxonomic determination by classical morphological procedure of unclassifiable plants, which might reduce the cost and efficiency of invasive stool biomonitoring and thus the management of its invasive populations

Viola banksii, the type species of section Erpetion, is endemic in eastern mainland Australia. In this paper we characterise morphological and anatomical features and assess genome size and genetic diversity in combination with the breeding system. V. banksii develops exclusively chasmogamous flowers. Ovules are anatropous, crassinucellate and bitegmic, the female gametophyte is of the Polygonum type, and the embryo is of Asterad type surrounded by nuclear endosperm. Pollen is non-heteromorphic, 3-aperturate, and highly viable. V. banksii grows in shade on moist, well drained, often sandy soils, and this is reflected in the anatomy of its organs, which includes a lack of subepidermal collenchyma in aerial parts, large leaf epidermal cells with thin cell walls, a narrow cuticle layer, and vascular bundles with xylem that are not rich in vessels. V. banksii is tolerant to zinc and lead based on phytotoxicity test. The high chromosome number (2n = 10x = 50) does not correspond to a small genome size (2C DNA = 1.27 pg). Low mean intra-populational gene diversity (HS = 0.077) detected by ISSR markers confirms the strong influence of selfing and clonal propagation by pseudostolons. Unique morphological traits of V. banksii include nyctinastic petal movement, the lack of a floral spur, the presence of gland-like protuberances on two stamens, and the presence of pseudostolons, which could be a synapomorphy for the whole section.