Search

In recent decades, the demand for naked barley grain on the market has increased, that is why the breeders of the leading institutions of the Russian Federation resumed breeding research to develop new naked barley varieties. In this regard, the purpose of the current study was to develop a breeding material for naked spring barley, adapted to the conditions of the Rostov region and, on its basis, new varieties. There have been developed the new naked barley variety with high productivity and grain quality, adapted to the conditions of the North Caucasus. The study was carried out in the scientific crop rotation of the department of barley breeding and seed production of the FSBSI Agricultural Research Center “Donskoy” (Zernograd) in 2014–2020. Based on the best local spring barley varieties (‘Leon’ and ‘Shchedry’) and naked varieties of the VIR collection (‘Mancuria’ (Sweden), ‘NB-OWA’ (Nepal), ‘CDC-Dawn’ (Canada), ‘K-3780’ (Tajikistan) and ‘Golozerny’ (RF)) there have been developed and studied the new naked barley lines. There was found that according to the trait ‘plant height’ the lines ‘Leon x Golozerny’ and ‘Leon x Mancuria’ belonged to the middle-height group (90.2 cm and 83.4 cm, respectively). The rest of the lines belonged to the tall group, their height varied from 96.5 cm to 100.3 cm. According to the trait ‘earing date’ there has been identified the line ‘Leon x Golozerny’ (24V), which is more early-ripening (5 days less than that of the standard). The same line turned out to be the only one that significantly exceeded the standard (the standard variety ‘Ratnik’ on 4.5 t/ha) according to productivity +0.3 t / ha, and taking into account the absence of hulls, this difference significantly increased to +0.7 t/ha. There was established that according to the trait ‘1000-grain weight’, the lines ‘Leon x Mancuria’, ‘Leon x CDC-Dawn’, ‘Leon x K-3780’ and ‘Leon x Golozerny’ significantly exceeded the standard variety. The best indicators of grain quality were identified for the lines ‘Leon x K-3780’ and ‘Leon x Golozerny’. The lines ‘Leon x Mancuria’ and ‘Leon x Golozerny’ were the most resistant to powdery mildew. The current paper has presented the characteristics of the new naked spring barley variety ‘Zernogradsky 1717’, which has a great productivity and a set of positive economically valuable traits.

Relevance. To meet the needs of the population and commodity producers in the south of Russia in tomatoes, it is necessary to develop varieties and hybrids of various directions that have high productivity, increased fruit quality and resistance to fungal and bacterial diseases, that meet modern production technologies and market requirements, as well as have an acceptable price for seeds.Aim of study – creation of new maternal tomato lines with functional male sterility for open ground for various purposes (for fresh consumption, whole-fruit canning,), which will make it possible to create highly productive hybrids that meet the agro-climatic conditions of the south of Russia in the future.Materials and methods. The studies were carried out in a breeding film greenhouse and in field conditions on the territory of the experimental plot of FSBSI "Federal Scientific Rice Centre" of the central soil-climatic zone of Krasnodar region. The object of research were mid-ripening tomato varieties included in the competitive testing, sterile tomato lines, new F1 hybrids based on FMS, collection and breeding material. The types of crosses used in the work: analyzing, saturating, top-crosses. To speed up the process of developing new sterile tomato lines in winter, artificial climate chambers were used.Results. As a result of the research, promising new functionally sterile lines of tomato were identified, differing in the length of the vegetation period, weight, color, fruit shape, biochemical parameters, with a number of useful economically valuable traits.

Development of heterotic hybrids is the most efficient approach to solve the problem of increasing the yield of rapeseed ( Brassica napus L.), a leading oilseed crop. The cytoplasmic male sterility (CMS), widely used in rapeseed hybrid seed production, makes it possible to control hybridization between female and male lines. A review of publications on the nature of CMS systems in rapeseed and their utilization in breeding is presented. In rapeseed there are more than 10 known CMS systems of alloplasmic and homoplasmic origin. The male sterility character proved to be determined by chimeric mitochondrial genes, characterized by the presence of novel transcribed open reading frames ( orf ). Mitochondrial CMS genes associated with nap , pol , ogu and Nsa sterility types as well as nuclear Rf genes for pollen fertility restoration were identified. Molecular marker systems for identification of CMS-inducing and male fertility restoring genes were developed. The ogu , pol , MSL and inap CMS systems are commonly used for producing industrial rapeseed hybrids. The State Register of the Russian Federation for 2019 contains rapeseed hybrids of only foreign origin. Main achievements in domestic rapeseed hybrid production are highlighted. Research and breeding institutions developed new source material for rapeseed hete rotic hybrid breeding in various regions of the country. The sterility and fertility restoration sources were received from Canadian and French institutions as well as from domestic working collections. The yield structure traits did not deteriorate after transferring hybrid maternal lines to the sterile cytoplasm, while the glucosinolate content increased when pollen fertility restoring genes were transferred into paternal lines. Dihaploid (androclinium) lines and in vitro culture of unfertilized ovules were used to accelerate the breeding process. Experimental hybrids were developed using pol and ogu CMS.

Relevance. High lycopene fruit content has been regarded as a very important genetic trait in tomato breeding. Use lycopene molecular markers in combination with conventional breeding techniques allowed us to create hybrids with high lycopene accumulation, excellent organoleptic qualities, high yield production and resistance to pathogens, and to effectively optimize our breeding programmes for commercial greehouses production.Material and Methods. In this study tomato samples including selected lines and hybrids with various allelic combinations of genes determining carotene accumulation, and other genetic traits, such as disease resistance and yield production were tested. Introgression of spontaneous and induced mutations was used to increase carotenoid levels (og and hp) and improve fruit technological qualities (nor, alc, rin). The research material was tomato collection, mutants, breeding lines and hybrids listed in the State Register Russian Federation tomato hybrids of breeding SS Agrofirm "Ilyinichna" VNIIO branch of the All-Russian Scientific Research Institute of Vegetable Growing – Branch of the FSBSI Federal Scientific Vegetable Center. DNA typing of fruit quality genes was performed at the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus.Results. New domestic hybrids for industrial greenhouses, which characterised by improved organoleptic qualities and technological traits were developed with the help of phasedcross-breeding that allowed to combine the genes nor, rin, alc, leading to an extension of the shelf life with the genes B, og, hp1, etc., contributing to an increase in carotenoid content in fruits. It was established that for targeted selection and hybridization, despite the negative influence of the nor, rin, alc genes it is possible to raise the level of carotenoids to average values. Correlation between lycopene concentration in fruits and high temperature and level of insolation was confirmed. It was shown that pink-fruited forms contain significantly more lycopenethanred-fruitedones. Different all eliccombinations of structural genes involved in carotenoids biosynthesis and regulatory genes that provided maximal accumulation of lycopene in hybrid swithred and pink fruits were revealed. Hybrids with the combination of high concentrations of sugar (° Brix), dry matter and maximal lycopene values, combined defining excellent taste were selected: Prekrasnaiya lady, Olya, Quadrille, Victoria. New F1 hybrids one for industrial greenhouses: G950, G956, G960, Magistral and pink fruited G12897, surpassed the Dutch standard in productivity up to 21%, and in tastes/organoleptic qualities for 1-1.8 points.

Seven female lines were crossed with three males in a line × tester design. The resulting 21 hybrids along with 10 parents were evaluated for the combining ability and gene effects during 2012 Kharif season. The results revealed that there were significant differences among genotypes (parents and crosses) for yield and its component traits. The combining ability variance and the ratio of GCA to total genetic variance showed predominance of non-additive gene action for all the characters except plant height and days to maturity. The lines, Natu nunepalli and Viswanadha were found to possess significantly high GCA effects for yield and its component traits while the crosses Viswanadha x II-1872 and Natu (DWFC) x II-1871 exhibited significant desired SCA effects for first grade leaf yield and spangle score, respectively. From this study it was found that the parents Natu nunepalli and Viswanadha could be used for diallel selective mating for fostering greater recombination, especially in the yield traits. The results suggested that adoption of reciprocal recurrent selection would be the best method for improving the yield potential in natu tobacco.

The present study was accomplished to assess the general combining ability effects of parents and specific combining ability effects of hybrids for yield and yield related traits in maize. Sixty three F1 hybrids were developed and by crossing nine productive maize inbred lines with seven quality protein maize inbred testers in Line x Tester mating design and evaluated for twelve yield and yield related characters at CCSHAU, Regional Research Station, Karnal, Haryana. The combining ability analysis revealed the presence of higher magnitude of SCA than GCA variance for all characters under study except for number of grains per cob. The preponderance ratio of sca/gca variance revealed presence of non additive gene action in the expression of all the characters under study. Among the sixteen parents inbred lines viz., HKI 1040-4, HKI 323, HKI 161 and HKI 163 were found to be the best parent for grain yield and among the hybrids, HKI 1128 x HKI 163, HKI 659-3 x HKI 194-6, HKI 1126 x HKI 161, HKI 536YN x HKI 193-1 and HKI 488 x HKI 170(1+2) exhibited highest significant and favourable sca effects for yield and yield attributing characters.

Background The advent of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology marked the beginning of a new era in the field of molecular biology, allowing the efficient and precise creation of targeted mutations in the genome of every living cell. Since its discovery, different gene editing approaches based on the CRISPR/Cas9 technology have been widely established in mammalian cell lines, while limited knowledge is available on genetic manipulation in fish cell lines. In this work, we developed a strategy to CRISPR/Cas9 gene edit rainbow trout (Oncorhynchus mykiss) cell lines and to generate single cell clone-derived knock-out cell lines, focusing on the phase I biotransformation enzyme encoding gene, cyp1a1, and on the intestinal cell line, RTgutGC, as example. Results Ribonucleoprotein (RNP) complexes, consisting of the Cas9 protein and a fluorescently labeled crRNA/tracrRNA duplex targeting the cyp1a1 gene, were delivered via electroporation. A T7 endonuclease I (T7EI) assay was performed on flow cytometry enriched transfected cells in order to detect CRISPR-mediated targeted mutations in the cyp1a1 locus, revealing an overall gene editing efficiency of 39%. Sanger sequencing coupled with bioinformatic analysis led to the detection of multiple insertions and deletions of variable lengths in the cyp1a1 region directed by CRISPR/Cas9 machinery. Clonal isolation based on the use of cloning cylinders was applied, allowing to overcome the genetic heterogeneity created by the CRISPR/Cas9 gene editing. Using this method, two monoclonal CRISPR edited rainbow trout cell lines were established for the first time. Sequencing analysis of the mutant clones confirmed the disruption of the cyp1a1 gene open reading frame through the insertion of 101 or 1 base pair, respectively. Conclusions The designed RNP-based CRISPR/Cas9 approach, starting from overcoming limitations of transfection to achieving a clonal cell line, sets the stage for exploiting permanent gene editing in rainbow trout, and potentially other fish cells, for unprecedented exploration of gene function., Cell & Bioscience, 11 (1), ISSN:2045-3701

Dinoflagellates in the family Symbiodiniaceae are obligate endosymbionts of diverse marine invertebrates, including corals, and impact the capacity of their hosts to respond to climate change-driven ocean warming. Understanding the conditions under which increased genetic variation in Symbiodiniaceae arises via sexual recombination can support efforts to evolve thermal tolerance in these symbionts and ultimately mitigate coral bleaching, the breakdown of the coral-Symbiodiniaceae partnership under stress. However, direct observations of meiosis in Symbiodiniaceae have not been reported, despite various lines of indirect evidence that it occurs. We present the first cytological evidence of sex in Symbiodiniaceae based on nuclear DNA content and morphology using Image Flow Cytometry, Cell Sorting and Confocal Microscopy. We show the Symbiodiniaceae species, Cladocopium latusorum, undergoes gamete conjugation, zygote formation, and meiosis within a dominant reef-building coral in situ. On average, sex was detected in 1.5% of the cells analyzed (N = 10,000–40,000 cells observed per sample in a total of 20 samples obtained from 3 Pocillopora colonies). We hypothesize that meiosis follows a two-step process described in other dinoflagellates, in which diploid zygotes form dyads during meiosis I, and triads and tetrads as final products of meiosis II. This study sets the stage for investigating environmental triggers of Symbiodiniaceae sexuality and can accelerate the assisted evolution of a key coral symbiont in order to combat reef degradation., DIANAS, DETECCIÓN INNOVADORA DE PROLIFERACIONES ALGALES TÓXICAS: UNA NECESIDAD FRENTE AL CALENTAMIENTO GLOBAL

Mesenchymal cells of diverse origins differ in gene and protein expression besides producing varying effects on their organ-matched epithelial cells’ maintenance and differentiation capacity. Co-culture with rodent’s tissue-specific pancreatic mesenchyme accelerates proliferation, self-renewal, and differentiation of pancreatic epithelial progenitors. Therefore, in our study, the impact of three-dimensional (3D) co-culture of human fetal pancreatic-derived mesenchymal cells (hFP-MCs) with human embryonic stem cell-derived pancreatic progenitors (hESC-PPs) development towards endocrine and beta cells was assessed. Besides, the ability to maintain scalable cultures combining hFP-MCs and hESC-PPs was investigated. hFP-MCs expressed many markers in common with bone marrow-derived mesenchymal stem cells (BM-MSCs). However, they showed higher expression of DESMIN compared to BM-MSCs. After co-culture of hESC-PPs with hFP-MCs, the pancreatic progenitor (PP) spheroids generated in Matrigel had higher expression of NGN3 and INSULIN than BM-MSCs co-culture group, which shows an inductive impact of pancreatic mesenchyme on hESC-PPs beta-cells maturation. Pancreatic aggregates generated by forced aggregation through scalable AggreWell system showed similar features compared to the spheroids. These aggregates, a combination of hFP-MCs and hESC-PPs can be applied as an appropriate tool for assessing endocrine-niche interactions and developmental processes by mimicking the pancreatic tissue.

Simple Summary Head and neck cancers can be divided in two major groups according to their risk factors, being high-risk human papillomavirus related (HPV-positive) and alcohol and tobacco related (HPV-negative) head and neck cancers. The majority of the locally advanced patients are treated with radiotherapy. However, up to 50% of these patients show local recurrences. The majority of these recurrences are linked to resistance to radiotherapy treatment. It is known that the response to DNA damage, also a process called the DNA damage response, is an important factor that determines the effectivity of radiotherapy. Here, we assessed the role of the DNA damage response in the resistance process to radiotherapy of head and neck cancers, by generating head and neck cancer cells resistant to radiotherapy. We show that the DNA damage response is differentially involved in the resistance process of HPV-positive and HPV-negative head and neck cancer cells. More specifically, HPV-positive radiotherapy-resistant cells showed increased ability to repair the DNA damage induced by radiotherapy. HPV-negative radiotherapy-resistant cells showed increased capacity to replicate after radiotherapy treatment. Despite this difference, inhibition of the DNA damage response enhanced the effect of radiotherapy in both groups. Abstract Radioresistance is a major cause of recurrences and radiotherapy (RT) failure in head and neck squamous cell carcinoma (HNSCC). DNA damage response (DDR) is known to be important for RT response, but its role in radioresistance is not fully understood. Here, we assessed the role of DDR in the radioresistance process of HNSCC by generating radioresistant clones from both HPV-positive SCC154 and HPV-negative SCC61 cells. We show that fractionated RT decreased RT response of HPV-positive and HPV-negative radioresistant clones in vitro and in vivo. Moreover, HPV-positive and HPV-negative radioresistant clones were characterized by differential DDR response. HPV-positive radioresistant clones showed less residual double-strand break damage and increased G2/M arrest recovery after RT, indicating an acquisition of increased DDR kinetics. In contrast, HPV-negative radioresistant clones showed less micronucleated cells after RT and increased survival upon checkpoint inhibition, indicating an increased replicative capacity. Inhibiting key factors of DDR in combination with RT rescued the radioresistant phenotype of both HPV-positive and HPV-negative radioresistant clones. Altogether, our results not only highlight the importance of DDR response in the radioresistance process of HPV-positive and HPV-negative HNSCC, but also provide possibilities for new therapies for HNSCC patients in recurrent settings.

Seeds of maize hybrids in the Russian Federation are obtained using CMS, therefore it is necessary to transfer the developed hybrids to a sterile basis to introduce them into production. The purpose of the work is to study the reaction of self-pollinated maize lines to various types of CMS. There was estimated the response of 26 self-pollinated maize lines to the “M” type of CMS and the response of 23 lines to the “C” type of CMS. The simple sterile hybrids “Milena M”, “Madonna M”, “Krucha M”, “Istok C”, “Nimfa C”, “Malvina C”, which are the mother forms of the new three-linear maize hybrids have been used as the testers of the sources of sterility. The self-pollinated lines have been classified according to the response to the “M” type of CMS as sterility fixers (11), incomplete sterility fixers (7), fertility reducing agents (6), non-uniform in reconstructing ability (2). In relation to the “C” type of CMS there were identified 10 sterility fixers, 3 incomplete sterility fixers, 7 fertility reducing agents, 3 non-uniform in reconstructing ability. The reducing agents of Moldavian type of CMS “FC 18 A”, “T 22 A”, “DS 768/83-3”, “DS 768/85-4”, “DS 768/85”, “DS 768/85-5” are of the greatest practical interest as they can be used as natural reducing agents (paternal forms) in the developed hybrid combinations. The constant natural reducing agents of the “C” type of CMS (“T 22 A”, “PLS 61”, “DK 22/325”, as paternal forms of hybrids grown on sterile cytoplasm of the “C” type) are of practical value. The use of sterile testers in hybridization allows simultaneously with the development of new hybrids to start the first stage of their transfer to a sterile basis, namely to evaluate the response of the lines to the sterile cytoplasm.

Responses of an individual to food deprivation, such as a 16-h fast, are complex, and are influenced by environmental and genetic factors. Domestication is an ongoing process during which adaptations to changing environments occur over generations. Food deprivation by their caretakers is less for domestic chickens than for their junglefowl ancestors. Unlike domestic chicken, the junglefowl adapted over generations to periods of food deprivation, which may be reflected in differences in metabolic responses to brief periods without food. Here, we compared the blood glucose and plasma levels of non-esterified fatty acids (NEFA) among four populations when deprived of feed for 16 h. The four populations included a domestic White Rock experimental line (LWS) maintained for generations under ad libitum feeding, adult red junglefowl (RJF), and a reciprocal cross of the lines. Although there were significant differences in adult (31-week) body weight between the RJF (683 g) and LWS (1282 g), with the weight of F1 crosses being intermediate, the amount of abdominal fat relative to body weight was similar for all populations. Patterns for blood glucose responses to a glucose bolus after a 16-h fast were similar for the initial and final points in the parental and cross populations. However, RJF reached their peak faster than LWS, with the reciprocal cross intermediate to the parental populations. Plasma NEFA concentrations were higher after the 16-h fast than in fed states, with no population differences for the fasting state. However, in the fed state, NEFA levels were lesser for LWS than for others, which was reflected further in percentage change from fed to fasted. This larger change in LWS suggests differences in mobilization of energy substrates and implies that during domestication or development of the LWS line, thresholds for responses to acute stressors may have increased. Published version

Background. Sunflower as an oilseed crop originated from Russia. It was in this country that single-headed highly productive forms were obtained, industrial oil production was invented, first local varieties were developed, accessions with increased oil content in seeds were identified by V. S. Pustovoit and his associates, and the first high-yielding cultivars ('Peredovik', 'VNIIMK 8883') with high oil content in seeds (50-60%) were produced. Mutants with modified fatty acid composition were obtained, and high oleic cultivar 'Pervenets' was created. Soviet varieties formed the basis for the world's sunflower breeding, and further for breeding commercial hybrids using the effect of heterosis. In the article, the genetic diversity of lines originated from domestic varieties is demonstrated. The genealogy of many domestic lines and also of the American line HA89 which served as the reference in breeding trials and genetic experiments is discussed. The information on the lines developed by the researchers of VIR at Kuban Experiment Station in the years 1970-2015 as well as the available data of foreign researchers on the lines obtained from domestic varieties are presented. Materials and methods . The lines were developed using repeated self pollination of the varieties and selection by morphological characters, CMS, pollen fertility restoration ability, duration of the growing season, and downy mildew resistance in each inbred generation. As a rule, lines were homogeneous after 7-8 generations. Results and discussion . The information on the genealogy of 38 lines developed by the researches of VIR, two VNIIMK lines, 16 foreign lines, and also the data on the origin of the CMS PET1 source are analyzed. The genealogical overview of the lines has allowed us to draw a conclusion on the origin of the high oil content and high oleic content traits. It is these characters that in the first place distinguish cultivated sunflower from the wild species of the genus Helianthus L. Lines originated from one and the same variety can differ by the presence of diagnostic markers of the Rf1 gene controlling pollen fertility restoration in forms with cytoplasmic male sterility, and also by allelic variants of storage protein genes and morphological characters.

Clubroot disease (causative organism Plasmodiophora brassicae Wor.) is among the most economically important and harmful diseases of the cole crops, and the damage due to this disease may reach up to 50-75% of the yield and even 100% in epiphytotics years. Even resistant varieties become susceptible over the years, because of appearance of the new pathogen races and change of climatic conditions in the main growing areas of the crop. In this context the Laboratory of Plant Immunity and Protection, of Federal State Budgetary Scientific Institution “Federal Scientific Vegetable Center” implements continuous phytoimmunological evaluation of collection and selection specimens and also directional material rather than just annual monitoring of causative organism dissemination in order to find new resistance sources. For this purpose an artificial infection background is used: compost obtained from decomposed nodules on the cabbage roots affected by clubroot disease (infection load 105-106 spores/cm3). The resistance of white cabbage varieties was evaluated during the harvesting period using five-point score of the root system damage, which formed the basis for categorization into resistance groups. For the analysis of artificial background intensity and specimen ranking the individual plants of the white cabbage variety Slava 1305, which is a susceptibility standard, were randomly planted in the entire area of the infection background. The impact of atmospheric conditions in the study year on the results of phytopathological evaluation of cabbage selection specimens against the infection background is demonstrated. Under unfavorable conditions for pathogen development (2014) the most specimens (74%) were categorized as relatively resistant, while in favourable for pathogen year 2015 relatively resistant specimens comprised only 5% of the total number of studied specimens. Since the same specimen may show different level of resistance depending on the year conditions, the stability of character manifestation is the important criterion for identification of the resistance resources. Phytopathological evaluation aimed on selection of clubroot-resistant forms in the Moscow region should last for at least three years even with the use of infection background. Long-lasting evaluation showed that the strains No 234/15,140/14,216/17 exhibiting high resistance to clubroot against artificial infection background regardless of the year conditions are the most valuable for selection. The resistance of white cabbage selection varieties to clubroot disease was studied against the infection background.

Mutant genotypes of biomass sorghum for low lignin - bmr (brown midrib) and sensitive to photoperiod are interesting for the production of second-generation ethanol. This research aimed to estimate the combining ability between lines A and R of biomass sorghum, both carrying the bmr-6 allele, for bioenergy potential. Three male-sterile lines (A) and ten fertility restorative lines (R) were used, generating 30 hybrids bmr and sensitive to photoperiod. The 30 hybrids, their parents, and six controls were evaluated in a triple lattice design (7 × 7), in two experimental areas of Embrapa Maize and Sorghum, in Sete Lagoas and Nova Porteirinha (Minas Gerais). These lines were crossed in a partial diallel design, based on the Griffing IV model (1956) adapted for partial diallel by Miranda Filho and Geraldi (1984). The evaluated traits were: days to flowering (DTF, days after sowing), plant height (PH, m), fresh biomass yield (FBY, ton.ha−1), dry matter (DM, %), and dry matter yield (DMY, t. ha−1). In addition, the FAI-BLUP selection Index was used to assist the selection of hybrids that simultaneously meet favorable traits. Results revealed that treatments x environment interaction had a significant effect (p

Onion (A. cepa L.) is an outcrossing biennial species with a very large genome. Development of genetically uniform (inbred) lines highly desired by onion breeders is a difficult process due to high level of heterozygosity. Inbred onion development may take up to five generations (similar to 10 years) by classical selfing technique. Onion shows severe inbreeding depression, which further complicates production of lines with stabilized important agronomic traits. When applied successfully, haploidization technology can be useful in the development of fully homozygous onion lines in 2 years. Although production of haploid and doubled haploid (DH) onions via gynogenesis was reported more than three decades ago, successful production and utilization of DHs in onion breeding is still far behind of expectations of breeders. The main obstacles in front of the success include high variation in the response of donor materials to gynogenesis induction and difficulties faced in the process of obtaining DHs from haploid plants. We use a DH production procedure enabling us to develop DH plants from a wide range of onion donor materials. This procedure is based on production of haploid plants via single step culture of unopened flower buds, detection of haploid plants among gynogenic regenerants, and converting these plants to fecund DHs using a combination of ploidy manipulation techniques. The bulbs of DHs are produced in about 1 year after the initiation of induction cultures and selfed seeds are produced from fecund DH plants when they flower in the second year.

This article provides an overview of some problems of the breeding and reproduction of laboratory minipigs. The most obvious of these are the lack of centralized accounting of breeding groups, uniform selection standards for reproduction and evaluation of breeding animals, as well as minimizing the accumulation of fitness-reducing mutations and maintaining genetic diversity. According to the latest estimates, there are at least 30 breeding groups of mini-pigs systematically used as laboratory animals in the world. Among them, there are both breed formations represented by several colonies, and breeding groups consisting of a single herd. It was shown that the main selection strategy is selection for the live weight of adults of 50-80 kg and the adaptation of animals to a specific type of biomedical experiments. For its implementation in the breeding of foreign mini-pigs, selection by live weight is practiced at 140- and 154-day-old age. It was indicated that different herds of mini-pigs have their own breeding methods to counteract inbred depression and maintain genetic diversity. Examples are the maximization of coat color phenotypes, the cyclical system of matching parent pairs, and the structuring of herds into subpopulations. In addition, in the breeding of foreign mini-pigs, molecular genetic methods are used to monitor heterozygosity. Every effort is made to keep the number of inbred crosses in the breeding of laboratory mini-pigs to a minimum, which is not always possible due to their small number. It is estimated that to avoid close inbreeding, the number of breeding groups should be at least 28 individuals, including boars of at least 4 genealogical lines and at least 4 families of sows. The accumulation of genetic cargo in herds of mini-pigs takes place, but the harmful effect is rather the result of erroneous decisions of breeders. Despite the fact that when breeding a number of mini-pigs, the goal was to complete the herds with exclusively white animals, in most breeding groups there is a polymorphism in the phenotype of the coat color.В статье представлен обзор проблем разведения и селекции лабораторных мини-свиней. Наиболее очевидные из них – отсутствие централизованного учета селекционных групп, единых стандартов отбора для воспроизводства и оценки племенных животных, а также минимизация накопления снижающих приспособленность мутаций и поддержание генетического разнообразия. По последним данным, в мире насчитывают не менее 30 селекционных групп мини-свиней, систематически используемых в качестве лабораторных животных. Среди них существуют как породные образования, представленные несколькими колониями, так и селекционные группы, состоящие из одного стада. Показано, что основная стратегия отбора включает селекцию на живую массу взрослых особей 50–80 кг и приспособленность животных к конкретному типу биомедицинских экспериментов. Для ее реализации в разведении зарубежных мини-свиней практикуют отбор по живой массе в 140- и 154-дневном возрасте. Указано, что в стадах мини-свиней представлены разные селекционные методы противодействия инбредной депрессии и поддержания генетического разнообразия. Примерами служат максимизация фенотипов масти, цикличная система подбора родительских пар и структурирование стад на субпопуляции. Кроме того, в разведении зарубежных мини-свиней для мониторинга гетерозиготности используют молекулярно-генетические методы. Количество инбредных скрещиваний в разведении лабораторных мини-свиней стараются минимизировать, что не всегда возможно из-за их малочисленности. Подсчитано, что во избежание тесного инбридинга численность селекционной группы должна быть не менее 28 особей, включающих хряков как минимум четырех генеалогических линий и свиноматок из не менее четырех семейств. Накопление генетического груза в стадах мини-свиней возможно, но вредоносный эффект является скорее следствием ошибочных решений селекционеров. Несмотря на то что при выведении ряда мини-свиней стояла цель укомплектовать стада исключительно белыми животными, в большинстве селекционных групп наблюдается полиморфизм по фенотипу масти.

Breast cancer is one of the most prevalent types of cancers worldwide and yet, its pathophysiology is poorly understood. Single-cell electrophysiological studies have provided evidence that membrane depolarization is implicated in the proliferation and metastasis of breast cancer. However, metastatic breast cancer cells are highly dynamic microscopic systems with complexities beyond a single-cell level. There is an urgent need for electrophysiological studies and technologies capable of decoding the intercellular signaling pathways and networks that control proliferation and metastasis, particularly at a population level. Hence, we present for the first time non-invasive in vitro electrical recordings of strongly metastatic MDA-MB-231 and weakly/non-metastatic MCF-7 breast cancer cell lines. To accomplish this, we fabricated an ultra-low noise sensor that exploits large-area electrodes, of 2 mm2, which maximizes the double-layer capacitance and concomitant detection sensitivity. We show that the current recorded after adherence of the cells is dominated by the opening of voltage-gated sodium channels (VGSCs), confirmed by application of the highly specific inhibitor, tetrodotoxin (TTX). The electrical activity of MDA-MB-231 cells surpasses that of the MCF-7 cells, suggesting a link between the cells’ bioelectricity and invasiveness. We also recorded an activity pattern with characteristics similar to that of Random Telegraph Signal (RTS) noise. RTS patterns were less frequent than the asynchronous VGSC signals. The RTS noise power spectral density showed a Lorentzian shape, which revealed the presence of a low-frequency signal across MDA-MB-231 cell populations with propagation speeds of the same order as those reported for intercellular Ca2+ waves. Our recording platform paves the way for real-time investigations of the bioelectricity of cancer cells, their ionic/pharmacological properties and relationship to metastatic potential.

La arveja arbustiva tiene características potenciales para la agroindustria y comercio en fresco y puede ser una alternativa para la zona cerealista de Nariño. La presente investigación, se realizó en dos municipios (Yacuanquer y Guaitarilla), pertenecientes a la zona cerealista de Nariño, durante tres épocas de siembra, en el 2017. La combinación de localidades por épocas constituyeron los seis ambientes estudiados. Se evaluaron ocho líneas de arveja arbustiva por número de vainas por planta, adaptabilidad y estabilidad fenotípica, para rendimiento en vaina verde y reacción a enfermedades foliares. En número de vainas sobresalieron GRICAND404 y GRICAND402. El análisis de adaptabilidad y de estabilidad fenotípica permitió identificar, como ambientes favorables, a Yacuanquer abril, Guaitarilla abril y Yacuanquer marzo; GRICAND405, con un rendimiento promedio de 9,27t.ha-1, se consideró una línea adaptable y predecible en los seis ambientes evaluados y GRICAND406, con 9,07t.ha-1, fue predecible y mostró mejor adaptación a los ambientes más favorables. En reacción a las enfermedades evaluadas, todas las líneas fueron susceptibles al ataque del complejo Ascochyta y el porcentaje de infestación de la enfermedad, se evidenció mayor en épocas muy lluviosas, como la época 1 (siembra de marzo) y disminuyó en épocas secas, como la época 3 (siembra de mayo); el ataque de Oídio, se evidenció mayormente en la época de menor precipitación (época 3), siendo resistente la línea GRICAND402. The shrub pea has potential characteristics for agroindustry and fresh trade and can be an alternative for the cereal zone of Nariño. The present investigation was carried out in two municipalities (Yacuanquer and Guaitarilla) belonging to the Nariño cereal zone during three sowing seasons in 2.017; the combination of the localities by the times was the six environments studied. Eight shrub pea lines were evaluated by number of pods per plant, adaptability and phenotypic stability to yield in green pod and reaction to foliar diseases. The number of pods stood out GRICAND404 and GRICAND402. The analysis of adaptability and phenotypic stability made it possible to identify the favorable environments for Yacuanquer april, Guaitarilla april and Yacuanquer march; GRICAND405 with an average yield of 9.27t.ha-1 was considered the adaptable and predictable line in the six environments evaluated and GRICAND406 with 9.07t.ha-1 was predictable and showed better adaptation to the most favorable environments. In reaction to diseases, all the lines were susceptible to attack by the Ascochyta complex and the percentage of infestation of the disease was greater in very rainy seasons such as season 1 (march sowing) and decreased in dry seasons such as season 3 (planting of may), the attack of Oídio was evidenced mainly in the period of least precipitation (season 3), being resistant the GRICAND402 line. Incluye referencias bibliográficas

نفذت تجربة حقلية في المحطة الارشادية في المهناوية سدة الهندية/ بابل في الموسم الربيعي لعام 2015والخريفي2015 بهدف زراعة (6) سلالات نقية من الذرة الصفراء (OH40،ZP707،ZP670،IK8،R153 وDK) في الموسم الربيعي وتهجينها وفي الموسم الخريفي تقييم السلالات والهجن التبادلية الناتجة منها وذلك بزراعتها بمسافات مختلفة 30,25,20)) سم بين النباتات لدراسة تأثير المسافات بين النباتات في نمو وحاصل السلالات المذكورة وهجنها التبادلية وفق تصميم القطاعات العشوائية الكاملة RCBD)) اشارت النتائج الى وجود فروق معنوية بين التراكيب الوراثية والمسافات ببن النباتات والتداخل بينهما أذ تميزت السلالةR153) ) بإعطائها اعلى القيم للصفات التالية : المساحة الورقية, عدد العرانيص, عدد الحبوب بالعرنوص ,حاصل النبات الواحد،الحاصل الكلي (طن.ه-1) اذ اعطت القيم (3466 سم2،1.10 عرنوص. نبات-1،393.36 حبة. عرنوص غم. نبات-1، 6.3 طن.هـ-1) بالتتابع ، عند المسافة 30سم متفوقتا على المسافتين الاخريتين 25.20) ) سم الصفات المدروسة كافة . اما الهجن فقد تفوق الهجين R153)× IK8) لجميع الصفات المذكورة معطيا اعلى المعدلات وقدرها (5639سم2 (1.25 عرنوص.نبات-1, 635.30 حبة. عرنوص-1 ,185.39 غم. نبات-1 10.6 طن .هكتار-1. واعطت السلالة (R153) والهجن R153)× IK8) عند المسافة (25) سم افضل تداخل بينهما.

Ninety five lines of sunflower genetic collection differing by their ability to suppress the CMS phenotype were molecularly marked with the use of 7 primer pairs. Using the STS marker orfH522, a sterile (PET1) cytoplasmon was identified in 79 lines, which confirmed indirectly the presence of fertility restoration genes in their genotypes. The majority of these lines also have a complex of molecular markers linked to the Rf1 gene. The HRG01, HRG02 and STS115 markers showed the best diagnostic value in revealing the Rf1 gene in the examined material. The data on allelic variation of the microsatellite loci ORS224, ORS511 and ORS799 were obtained for the first time.

In multicellular organisms, such as vertebrates and flowering plants, horizontal transfer (HT) of genetic information is thought to be a rare event. However, recent findings unveiled unexpectedly frequent HT of RTE-clade LINEs. To elucidate the molecular footprints of the genomic integration machinery of RTE-related retroposons, the sequence patterns surrounding the insertion sites of plant Au-like SINE families were analyzed in the genomes of a wide variety of flowering plants. A novel and remarkable finding regarding target site duplications (TSDs) for SINEs was they start with thymine approximately one helical pitch (ten nucleotides) downstream of a thymine stretch. This TSD pattern was found in RTE-clade LINEs, which share the 3′-end sequence of these SINEs, in the genome of leguminous plants. These results demonstrably show that Au-like SINEs were mobilized by the enzymatic machinery of RTE-clade LINEs. Further, we discovered the same TSD pattern in animal SINEs from lizard and mammals, in which the RTE-clade LINEs sharing the 3′-end sequence with these animal SINEs showed a distinct TSD pattern. Moreover, a significant correlation was observed between the first nucleotide of TSDs and microsatellite-like sequences found at the 3′-ends of SINEs and LINEs. We propose that RTE-encoded protein could preferentially bind to a DNA region that contains a thymine stretch to cleave a phosphodiester bond downstream of the stretch. Further, determination of cleavage sites and/or efficiency of primer sites for reverse transcription may depend on microsatellite-like repeats in the RNA template. Such a unique mechanism may have enabled retroposons to successfully expand in frontier genomes after HT.

Pellucid punctuations and lines in the leaf blade are common in Casearia, but for some of its species these structures are mentioned as inconspicuous, not visible or obscure. Cavities and ducts have been anatomically described and correlated with pellucid punctuations and lines, respectively, in only six species from the genus; however, the distinction between these types of secretory structures remains unclear. Therefore, we aimed to anatomically describe the pellucid punctuations and lines in leaves of 47 species of Casearia. Leaf blade samples were obtained from herbarium and field collected material and processed according to standard methods for light microscopy analysis. Cavities and ducts were absent in only four of the 47 species (C. commersoniana, C. javitensis and C. spruceana of section Piparea, and C. eicheleriana of section Casearia). In the remaining species, cavities corresponded to pellucid punctuations, while ducts corresponded with lines. The majority of species have both structures randomly distributed throughout the leaf blade, however, in C. bahiensis, these structures are concentrated along the leaf margin, while in C. aquifolia and C. crassinervis they were predominant along the midrib. Our data show that anatomical analyses are essential for distinguishing between cavities and ducts. We also demonstrate that the distribution of cavities and ducts provides useful information for the taxonomy of Casearia at both the species and section levels.

A total of 300 eggs from 45 female quails from three different lines (Dark brown, light brown and white) aged 10 weeks old were equally divided into three experimental groups having 15 birds of each and caged separately, to estimate the external and internal egg quality traits. Half eggs were measured 24 hrs. post lay and others were measured after one week of storage at room temperature. The results showed that there were significant differences among lines for egg length, yolk height, yolk index, albumen diameter, albumen index and Haugh unit. The light brown line resulted in the best quality. All internal egg quality characters and egg length were differed significantly between fresh and storage eggs. The fresh eggs resulted in better quality than stored eggs. Phenotypic correlation appeared that there were significant correlation coefficients between some egg quality characteristics in dark brown line more than light brown and white lines.

Tree-shaped plants of tomato have many useful traits, which gain them advantages over common tomato forms. In world plant collection there are over 200 accessions, cultivars and hybrids of tree-shaped tomato. The aim of the study was to develop new initial tomato accessions for breeding program for tree-shaped cultivars and hybrids that are distinguished by a shape, fruit weight and color, improved by interspecific hybridization chemical composition and resistance to the major diseases. The tree-shaped breeding lines: ‘196/12’, ‘374/08’, ‘909/14’, ‘911/14’ and other were observed with the use of interspecific hybridization and selection in the population F2-F4. The selected lines passed the trial in artificially infected condition with Alternaria, viral diseases (MToV, TSWV) and phytoplasmas. Among breeding lines assessed the ‘911/14’, ‘374/08’ and‘40/11’ were less affected by complex of pathogens. The lines ‘196/12’ and ‘909/14’ passed an assessment in the nursery for variety trial in naturally and provocatively infected conditions with major pathogens. Both breeding lines had an advantage over standard accession ‘Laguna’ and ‘Maraphon’ for total yield capacity and standard fruit harvest; it was more by 9.6% and 52.2% and 9.5% and 53.4% respectively. Fruits of lines 196/12 and 909/14 had good taste quality with high content of dry matter (5.8% and 6.8%), sugars (3.3% and 3.1%), vitamin C (22.2 and 24.8 mg/100 grams). The selected tree-shaped accessions have been used to develop heterotic hybrids and to be sources of economically valuable traits.

IF 5.992; International audience; The inhibition of heat shock protein 90 (Hsp90) is a promising strategy to increase the radiosensitivity of tumor cells. However, Hsp90 inhibition induces the expression of Hsp70 which is a prominent cytoprotective protein. Therefore, dual targeting of Hsp70 and Hsp90 might be beneficial to increase the radiosensitivity of tumor cells. Hsp70 inhibiting peptide aptamers have been shown to increase the sensitivity of tumor cells to apoptosis induced by different anticancer drugs. Herein, we studied the radiosensitizing activity of the Hsp70 inhibiting peptide aptamer A17 in combination with the Hsp90 inhibitor NVP-AUY922. Whereas A17 significantly increased apoptosis induction by NVP-AUY922 it did not significantly affect the radiosensitivity of human lung and breast cancer cells. However, Hsp70 inhibition by the aptamer A17 potentiated the radiosensitizing effects of the Hsp90 inhibitor NVP-AUY922. Mechanistically we speculate that an increased number of DNA double strand breaks and an enhanced G(2)/M arrest might be responsible for the increased radiosensitization in A17 expressing tumor cells. Therefore, the simultaneous inhibition of Hsp90 and Hsp70 combined with radiotherapy might provide a promising anti-cancer strategy. (C) 2017 Elsevier B.V. All rights reserved.

The important stage of breeding program for F 1 hybrids with high yield capacity is to estimate the combining ability of parental lines for productivity characteristics. The information on combining ability may predict the productivity traits to exhibit and helps produce heterotic hybrids and use the initial breeding accessions in sweet pepper. The combining ability of breeding accessions for yield traits was studied with the use of complete diallel crossing. The research was carried out at experimental plots of State Institution ‘Transnistrian Institute of Agriculture’ in 2015-2016. In 2015, seven fertile lines were crossed through complete diallel cross to study combining ability. In 2016 the parental lines and 42 hybrids F 1 forms produced from crossing were tested in open field condition. Analysis of General Combining Ability and Specific Combining Ability showed that none of all parental components had any essential expected effect for all tested traits of productivity. The line 134 showed the high effects of General Combining Ability as maternal and parental components for four traits, such as early and general yield, average weight and thickness of fruit pericarp. Out of 42 hybrid combinations studied, some accessions have been selected. They had the high effects of General Combining Ability with high constants of Specific Combining Ability for early yield – 4; general – 12; average fruit weight – 9; thickness of fruit pericarp – 4; dry matter content – 10; ascorbic acid – 13.

Results of estimation of general and specific combining ability of male and female parental tomato forms were given for complex of traits. The estimation was carried out according to incomplete diallel crosses 15x15. Such varities as ‘Trapeza’, Rosovaya kapelka’, ‘Seniorita’, ‘Ocharovanie’, ‘Tigris’ ‘Vishnya Zheltaya’ (Gavrish), ‘Denezhnoye Derevo’ (national breeding), and lines: ‘46/06’, ‘49/09’, ‘295/09’, ‘336/11’, ‘354/11’, ‘357/11’, ‘388/09’ (nor), ‘498’ (selection of TARI) were used as intial breeding accessions, differing in bush type (determinate and indeterminate), duration of vegetative phase (ultraearly, early, medium early, middle-ripening), fruit shape (rounded, oval), fruit color (red, pink, black, orange, tiger and with the gene nor), fruit weight (10 g. and more), brush structure (dense, friable). As a results, ‘Trapeza’, ‘Vishnya Zheltaya’, ‘Ocharovaniye’, ‘Seniorita’, and lines: ‘295/10’, ‘49/09’, ‘498’, ‘357/11’, ‘354/11’, ‘388/09’ (nor) were selected and recommended to be used in breeding program for development of heterotic hybrids with high fruit setting and generative bush type. The promising hybrids F 1 with high constants of specific combining ability for a complex of economically valuable traits have been observed. Because of different lines and accessions were used in crossings, these hybrids varied in internode length of cluster type, classical or shorten; rounded or oval fruit shape; fruit color, red (F 1 combinations ‘354/11’ х ‘Seniorita’, ‘Trapeza’ х ‘L.49/09’, L. ‘49/09’ х L.354/11), pink (F 1 combination ‘Rosovaya Kapelka’ х ‘L.354/11’), yellow (F 1 combination ‘Ocharovaniye’ х ‘Vishiya Zheltaya’), deep brown (F 1 combination ‘L.357/11’ х ‘L.354/11’).

Recent progress has enabled the conversion of primed human embryonic stem cells (hESCs) to the naive state of pluripotency, resembling the well-characterized naive mouse ESCs (mESCs). However, a thorough histone epigenetic characterization of this conversion process is currently lacking, while its likeness to the mouse model has not been clearly established. Here, we profile the histone epigenome of hESCs during conversion in a time-resolved experimental design, using an untargeted mass spectrometry-based approach. In total, 23 histone post-translational modifications (hPTMs) changed significantly over time. H3K27Me3 was the most prominently increasing marker hPTM in naive hESCs. This is in line with previous reports in mouse, prompting us to compare all the shared hPTM fold changes between mouse and human, revealing a set of conserved hPTM markers for the naive state. Principally, we present the first roadmap of the changing human histone epigenome during the conversion of hESCs from the primed to the naive state. This further revealed similarities with mouse, which hint at a conserved mammalian epigenetic signature of the ground state of pluripotency.

Fungal diseases cause a lot of economic damage in castor and in some instances; it is a limiting factor to commercial cultivation of the crop. In the present study, survey on castor germplasm for sources of resistant genes to Cercospora leaf spot was carried out at National Cereals Research Institute Badeggi, Nigeria. The genotypes were grown in a resolvable incomplete block design with three replications. The results obtained revealed high variability of host resistance among the genotypes. Percentage disease incidence ranged between 16.67% and 100%. A range between 1.43 score and 4.17 scores with average score of 2.89 were recorded for disease severity among the genotypes. Highest (95.92%) and lowest (13.03%) damage index were observed in the genotypes Acc. 059 and Acc. 022 respectively. The Principal component (PC) 1 to 4 explained 100% of the variability in the germplasm. PC1 explained 69.72% of the variability with the major contributory parameters being the disease incidence, severity and damage index. Significant negative correlations were recorded between the seed yield and all the three disease indices. The result of cluster analysis revealed six cluster groups among the germplasm with cluster membership ranging between 4 and 40 members. The cluster I contained members (Acc. 002, Acc. 017, Acc. 022, Acc. 026, Acc. 027, Acc. 048 and Acc. 061) with low average disease incidence, severity and damage index. The results reported here could serve as a basis for further screening of the potential resistant genotypes under controlled condition to develop resistant lines.

Summary Gain of 20q11.21 is one of the most common recurrent genomic aberrations in human pluripotent stem cells. Although it is known that overexpression of the antiapoptotic gene Bcl-xL confers a survival advantage to the abnormal cells, their differentiation capacity has not been fully investigated. RNA sequencing of mutant and control hESC lines, and a line transgenically overexpressing Bcl-xL, shows that overexpression of Bcl-xL is sufficient to cause most transcriptional changes induced by the gain of 20q11.21. Moreover, the differentially expressed genes in mutant and Bcl-xL overexpressing lines are enriched for genes involved in TGF-β- and SMAD-mediated signaling, and neuron differentiation. Finally, we show that this altered signaling has a dramatic negative effect on neuroectodermal differentiation, while the cells maintain their ability to differentiate to mesendoderm derivatives. These findings stress the importance of thorough genetic testing of the lines before their use in research or the clinic., Highlights • Bcl-xL overexpression drives the transcriptomic profile of 20q11.21 mutant lines • 20q11.21 mutant lines downregulate CHCHD2, a known TGF-β pathway modulator • Mutant lines differentially express genes involved in TGF-β and SMAD signaling • Mutant lines show impaired ectoderm commitment due to TGF-β signaling deregulation, In this article Spits and colleagues show that Bcl-xL overexpression is the main driver of the distinct transcriptomic profile of human embryonic stem cells with a gain of 20q11.21. The cells have an abnormal TGF-β- and SMAD-mediated signaling, which impairs commitment to neuroectoderm while differentiation to the other lineages remains unaffected.

Tumor-associated cell-free DNAs (cfDNAs) are found to play some important roles at different stages of tumor progression; they are involved in the transformation of normal cells and contribute to tumor migration and invasion. DNase I is considered a promising cancer cure, due to its ability to degrade cfDNAs. Previous studies using murine tumor models have proved the high anti-metastatic potential of DNase I. Later circulating cfDNAs, especially tandem repeats associated with short-interspersed nuclear elements (SINEs) and long-interspersed nuclear elements (LINEs), have been found to be the enzyme's main molecular targets. Here, using Lewis lung carcinoma, melanoma B16, and lymphosarcoma RLS40 murine tumor models, we reveal that tumor progression is accompanied by an increase in the level of SINE and LINEs in the pool of circulating cfDNAs. Treatment with DNase I decreased in the number and area of metastases by factor 3-10, and the size of the primary tumor node by factor 1.5-2, which correlated with 5- to 10-fold decreasing SINEs and LINEs. We demonstrated that SINEs and LINEs from cfDNA of tumor-bearing mice are able to penetrate human cells. The results show that SINEs and LINEs could be important players in metastasis, and this allows them to be considered as attractive new targets for anticancer therapy.

[EN] The aim of the experiment was to compare 3 genetic groups, slaughtered at similar weights, to examine their productive and carcass traits and economic value. Three lines of the Pannon Breeding Programme, selected for different criteria, were examined in the experiment. Pannon Ka (PKa, maternal line) does were inseminated with semen of PKa, Pannon White (PWhite) or Pannon Large (PLarge, terminal line) bucks. The kits (PKa×PKa, PWhite×PKa, PLarge×PKa; n=60 in each genetic group) were weaned at 35 d of age and reared until 88, 83 and 79, respectively, when they reached similar body weights for slaughtering (2.8 kg). The weight gain of PLarge×PKa was the largest (51.0 g/d) and that of PKa×PKa was the smallest (47.2 g/d), while PWhite×PKa (41.8 g/d) was intermediate (P, This paper was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences, and by the GOP-1.1.1-11-2012-0132 project.

Summary Reports on the retention of somatic cell memory in induced pluripotent stem cells (iPSCs) have complicated the selection of the optimal cell type for the generation of iPSC biobanks. To address this issue we compared transcriptomic, epigenetic, and differentiation propensities of genetically matched human iPSCs derived from fibroblasts and blood, two tissues of the most practical relevance for biobanking. Our results show that iPSC lines derived from the same donor are highly similar to each other. However, genetic variation imparts a donor-specific expression and methylation profile in reprogrammed cells that leads to variable functional capacities of iPSC lines. Our results suggest that integration-free, bona fide iPSC lines from fibroblasts and blood can be combined in repositories to form biobanks. Due to the impact of genetic variation on iPSC differentiation, biobanks should contain cells from large numbers of donors., Graphical Abstract, Highlights • Isogenic iPSC from fibroblasts and blood have similar differentiation propensities • Donor-dependent variability affects molecular and differentiation propensities of iPSCs • Impact of donor variability exceeds source-cell-specific differences in iPSC lines • Bona fide iPSC lines from different tissues can be combined in the repositories, Trokovic and colleagues include epigenetic, transcriptional, and functional analyses in the same study, and show that the individual donor-dependent differences largely determine variability between iPSC lines. They show that fibroblast- and blood-derived iPSCs from a given donor are highly similar for their differentiation propensities while differences are most dramatic across donors. Thus bona fide iPSC lines from different tissues can be combined in biobanks.

Mouse embryonic stem cells (mESCs) exist in a naive, primed and ground state of pluripotency. While comparative analyses of these pluripotency states have been reported, the mESCs utilized originated from various genetic backgrounds and were derived in different laboratories. mESC derivation in conventional LIF + serum culture conditions is strain dependent, with different genetic backgrounds potentially affecting subsequent stem cell characteristics. In the present study, we performed a comprehensive characterization of naive, primed and ground state mESCs originating from the same genetic background within our laboratory, by comparing their transcriptional profiles. We showed unique transcriptional profiles for naive, primed and ground state mESCs. While naive and ground state mESCs have more similar but not identical profiles, primed state mESCs show a very distinct profile. We further demonstrate that the differentiation propensity of mESCs to specific germ layers is highly dependent on their respective state of pluripotency.

This study was conducted to determine the goodness of fit of Gompertz, Logistic, Von Bertalanffy, Richards, Levakovich and Janoschek growth models in Japanese quail. Therefore, weekly live-weight data obtained from 372 females and 339 males were fitted. Females' live weights were found to be higher than that of males, and the first divergence in the growth of female and male birds occurred in 21-28 days, and it survived until the experiment (P

Background: Lettuce is a globally important leafy vegetable and a model plant for biotechnology due to its adaptability to tissue culture and stable genetic transformation. Lettuce is also crucial for functional genomics research in the Asteraceae which includes species of great agronomical importance. The development of transgenic events implies the production of a large number of shoots that must be differentiated between transgenic and non-transgenic through the activity of the selective agent, being kanamycin the most popular. Results: In this work we adjusted the selection conditions of transgenic seedlings to avoid any escapes, finding that threshold concentration of kanamycin was 75 mg/L. To monitor the selection system, we studied the morphological response of transgenic and non-transgenic seedlings in presence of kanamycin to look for a visual morphological marker. Several traits like shoot length, primary root length, number of leaves, fresh weight, and appearance of the aerial part and development of lateral roots were affected in non-transgenic seedlings after 30 d of culture in selective media. However, only lateral root development showed an early, qualitative and reliable association with nptII presence, as corroborated by PCR detection. Applied in successive transgenic progenies, this method of selection combined with morphological follow-up allowed selecting the homozygous presence of nptII gene in 100% of the analyzed plants from T2 to T5. Conclusions: This protocol allows a simplified scaling-up of the production of multiple homozygous transgenic progeny lines in the early generations avoiding expensive and time-consuming molecular assays. Inst. de Biotecnología Fil: Darqui, Flavia Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Radonic, Laura Mabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Lopez, Nilda Ester. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Hopp, Horacio Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina Fil: López Bilbao, Marisa Gisela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina

The MTT assay is one of the often used cell viability/cytotoxicity assays. However, when the methanol extracts of plants are used to test their cytotoxic potential, interference may occur, resulting in false-positive viability results. Therefore, in this study, the reliability of the MTT assay was investigated in the case of plant use. The methanol extracts of three different plants (Hypericum adenotrichum, Salvia kronenburgii, and Pelargonium quercetorum) were tested in breast cancer cell lines (MCF-7 and MDA-MB-231) using the MTT assay and the results were compared to the ATP assay, which is a much more sensitive and reliable assay due to its interference-free feature. Additionally, decreased cell density was confirmed with phase-contrast microscopy and fluorescence staining (Hoechst 33342 dye). Although both of the viability/cytotoxicity assays are considered as metabolic assays, viabilities (in %) in the MTT assay were found to be strikingly higher when compared to the results with the ATP assay. Even in the case of total death, the MTT assay still produced artificial/false increases in viability. The morphology-based evaluation of viability/cytotoxicity by phase-contrast microscopy and Hoechst 33342 staining were greatly compatible with the ATP assay results. Overestimated (false) viabilities in the MTT assay suggests a serious interference between the MTT assay itself and the extracts used. Some ingredients of plants may have reducing activity (like the dehydrogenase activity of the cells) that converts the MTT compound into the colored formazan that is the principle of the assay. Therefore, the MTT assay may not be a suitable assay for some plant extracts, urging great caution when plants are used. Uludag University Scientific ResearchUludag University [BUAP(F)-2014/3] This study was supported by the Uludag University Scientific Research Projects Unit (BUAP(F)-2014/3 coded project). WOS:000418253100007 30814856 Q4

BACKGROUND We previously established parthenogenetic mouse embryonic stem cells (ESCs) and this study was subsequently conducted for elucidating the influence of oocyte parthenogenesis on gene expression profile of ESCs. METHODS Gene expression of parthenogenetic ESC (pESC)-1 or pESC-2 was separately compared with that of two normally fertilized ESC (nfESC) lines (B6D2F1 and R1 strains), and quantification of mRNA expression was conducted for validating microarray data. RESULTS In two sets of comparison, reaction of 11 347 and 15 454 gene probes were altered by parthenogenesis, while strain difference changed the expression of 15 750 and 14 944 probes. Level of correlation coefficient was higher in the comparisons between normal fertilization and parthenogenesis (0.974-0.985) than in the comparisons between strains of nfESCs (0.97-0.971). Overall, the expression of 3276-3329 genes was changed after parthenogenesis, and 88% (96/109) of major functional genes differentially (P < 0.01) expressed in one comparison set showed the same change in the other. When we monitored imprinted genes, expression of nine paternal and eight maternal genes were altered after parthenogenesis and 88% (14/16) of these was confirmed by mRNA quantification. CONCLUSIONS The change in gene expression after parthenogenesis was similar to, or less than, the change induced by a strain difference under a certain genetic background. These results may suggest the clinical feasibility of parthenogenesis-derived, pluripotent cells

34 págs., 5 tabl. The definitive version is available at: https://www.crops.org/publications/cs, Maize (Zea mays L.) for northern growing areas requires cold tolerance for extending the vegetative period. Our objectives were to evaluate two large panels of maize inbred lines adapted to Europe for cold tolerance and to estimate the effects of cold-related traits on biomass production. Two inbred panels were evaluated for cold tolerance per se and in testcrosses under cold and control conditions in a growth chamber and under field conditions. Comparisons of inbreds and groups of inbreds were made taking into account the single nucleotide polymorphisms (SNP)-based genetic structure of the panels, and the factors affecting biomass production were studied. Eight flint and one dent inbred with diverse origins were the most cold tolerant. The most cold-tolerant dent and flint groups were the Iodent Ph207 and the Northern Flint D171 groups, respectively. The relationships between inbred per se and testcross performance and between controlled and field conditions were low. Regressions with dry matter yield in the field as dependent variable identified plant height (R2 = 0.285) as the main independent variable, followed by quantum efficiency of photosystem II (R2 = 0.034) and other traits with minor contributions. Cold-tolerance–related traits had low and negative effects on dry matter yield. Models intending the prediction of final performance from traits scored in early developmental stages are not expected to be precise enough for breeding. For improving cold tolerance, inbreds released from crosses among the No Iodent group and the Northern Flint group may show high combining ability, as well as between both groups and the Northern Flint D171 group., This research was supported in the framework of the Plant-KBBE program (project acronym “Cornfed”) by the Spanish Ministry of Science and Innovation (proj. EUI2008-03642 and EUI2008-03635), the French National Agency for Research (ANR, Ministry of High Education and Research), and the German Federal Ministry of Education and Research (grant numbers 0315461AD).

The mechanism for generating double minutes chromosomes (dmin) and homogeneously staining regions (hsr) in cancer is still poorly understood. Through an integrated approach combining nextgeneration sequencing, single nucleotide polymorphism array, fluorescent in situ hybridization and polymerase chain reaction-based techniques, we inferred the fine structure ofMYC-containing dmin/hsr amplicons harboring sequences from several different chromosomes in seven tumor cell lines, and characterized an unprecedented number of hsr insertion sites. Local chromosome shattering involving a single-step catastrophic event (chromothripsis) was recently proposed to explain clustered chromosomal rearrangements and genomic amplifications in cancer. Our bioinformatics analyses based on the listed criteria to define chromothripsis led us to exclude it as the driving force underlying amplicon genesis in our samples. Instead, the finding of coexisting heterogeneous amplicons, differing in their complexity and chromosome content, in cell lines derived from the same tumor indicated the occurrence of a multistep evolutionary process in the genesis of dmin/hsr. Our integrated approach allowed us to gather a complete view of the complex chromosome rearrangements occurring within MYC amplicons, suggesting that more than one model may be invoked to explain the origin of dmin/hsr in cancer. Finally, we identified PVT1 as a target of fusion events, confirming its role as breakpoint hotspot in MYC amplification. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

RESUMO Estudos precoces com relação à seleção de linhagens de milho, com base em características de qualidade de sementes, podem ser explorados em programas de melhoramento nas primeiras etapas da obtenção de híbridos. O objetivo deste estudo foi analisar a divergência genética e a heterose associadas à qualidade fisiológica de sementes, em função do vigor, para a composição bioquímica na obtenção de milho híbrido. No experimento, foram utilizadas sementes de 8 linhagens e 3 híbridos de milho (híbrido simples, triplo e triplo modificado). Avaliaram-se os componentes bioquímicos nas sementes por meio dos teores de amido, proteína total, fósforo total, ferro e zinco. Com base no método de Singh, observou-se que a variável que mais contribuiu para a divergência genética quanto à composição bioquímica das sementes foi o teor de amido, com 46,47%. Constatou-se divergência genética e heterose para a composição bioquímica das sementes, indicando que essa característica pode ser explorada em programas de melhoramento visando à qualidade das sementes. O híbrido simples, composto pelas linhagens 2 e 4, foi indicado como a melhor combinação para a composição bioquímica nas sementes, com as maiores heteroses para os teores de amido, fósforo, ferro e zinco. ABSTRACT Early studies with regard to the selection of maize lines based on seed quality characteristics can be exploited in breeding programs in the early stages of hybrids achievement. The objective of this study was to analyze the genetic diversity and the heterosis associated with physiological quality of seeds, in function of the force, for biochemical composition in the achievement of hybrid corn. In the experiment, seeds from 8 lines and 3 maize hybrids (single hybrid, triple and modified triple) were used. Biochemical components in the seeds were evaluated by means of starch content, total protein, total phosphorus, iron and zinc. Based on Singh’s method, it was observed that the variable which most contributed to genetic divergence on the biochemical composition of the seeds was the starch content, with 46.47%. Genetic diversity and heterosis were found for the biochemical composition of the seeds, indicating that this feature can be exploited in breeding programs aiming at the quality of the seeds. The simple hybrid, composed of the strains 2 and 4, was indicated as the best match for the biochemical composition in seeds, with the highest heterosis for starch content, phosphorus, iron and zinc.

The present study was undertaken to assess the extent of heterosis for seed cotton yield and its attributing traits in 24 interspecific hybrids of cotton developed by crossing 8 lines with 3 testers in line × tester mating design during kharif, 2017- 18 at Agricultural Research Station Dharwad Farm. Significant heterosis was observed for the characters under study indicating the presence of genetic diversity among the parental lines. Except for number of sympodia per plant and boll weight, the mean sum of squares of parents vs crosses was significant for all the characters indicating presence of heterosis for these traits. None of the crosses was superior for all the traits studied. However, the cross CPD-462 x SBYF-425 was the best as it had the highest mean performance for economically important characters. Majority of the crosses exhibited significantly positive mid parent, better parent heterosis for all important yield contributing characters except ginning outturn. Most of the hybrids expressed significant standard heterosis for all the characters except boll weight and ginning outturn over the checks.

Yield and sugar content genetic analyzes for hybrids developed engaging pollinating lines of the first and the second inbred generations throughout two cycles of recurrent breeding has underpinned the determination of regularities for combinability expression in crossing components and the change of their gene interaction types. This unveiled the lack of correlation between the parents descriptions as selected from groups that displayed contrast combining of productivity components and the lines combinability derived thereof. This provided a ground for correcting the breeding technology of the development of MS hybrids through the exclusion of individual polarization as phenotype based breeding method with replacing this by the assessment of economical value of the topcrossing components. The genetic value was determined for pollinator lines of І1 first hybrid generation by productivity components derived from multiple seed populations by Ulad breeding. 7 economically valuable lines were derived from U752 population by yield and 17 by sugar content, whereas 6 and 10 limes were derived from КМ2, respectively . This provided a ground for the development of synthetic varieties that displayed yield and sugar content characteristics improved, respectively, by 9,4 and 2,4% (U752 initial population ) and by 4,4 and 8,3% ( КМ2 initial population) compared to group standard. The structure of genetic variability of MS hybrids productivity as developed engaging І2 .first and second inbred generation. In the genotype variation of characteristics of hybrids derived from І1 pollinators of recurrent breeding first cycle the additive effects of both parental forms prevailed, and as regards the hybrids developed engaging І2 pollinators of the second cycle of recurrent breeding non-additive effects of the components interaction prevailed. In the second cycle of recurrent breeding 7 І2 lines valuable for combining were singled out, which had high general combinability by yield, and 8 were singled out by sugar content. 5 lines were singled out: U752/5/7, U752/84/3, КМ2/33/4, КМ2/34/7, КМ2/75/4, which combined high general combinability by yield and sugar content at once. Heterosis effect of hybrid combinations was subject to total fusion of additive and non-additive effects of the genes controlling quantitative characteristics of the productivity. The best hybrids yield engaging І2, pollinator lines selected out of U752 and КМ2 multi-seed populations ranged within 104...117,3%, sugar content – 102,4...106,0%, while that of synthetic varieties – 113,4 and 103,6% (U752) and 106,7 and 109,3% (КМ2), respectively, against the group standard.

Purpose: Induction of viral lytic cycle, which induces death of host cells, may constitute a useful adjunct to current therapeutic regimens for Epstein–Barr virus (EBV)-driven malignancies. Human telomerase reverse transcriptase (hTERT), essential for the oncogenic process, may modulate the switch from latent to lytic infection. The possible therapeutic role of hTERT inhibition combined with antiviral drugs was investigated. Experimental Design: EBV-negative BL41 and convertant EBV-positive BL41/B95.8 Burkitt's lymphoma cell lines and lymphoblastoid cell lines (LCL) were infected with retroviral vector encoding short hairpin RNA (shRNA) anti-hTERT and cultured with or without the prodrug ganciclovir. The effects on EBV lytic replication, cell proliferation, and apoptosis were characterized. Results: hTERT silencing by shRNA induced the expression of BZLF1, EA-D, and gp350 EBV lytic proteins and triggered a complete lytic cycle. This effect was associated with downregulation of BATF, a negative regulator of BZLF1 transcription. hTERT silencing also resulted in antiproliferative and proapoptotic effects. In particular, hTERT inhibition induced an accumulation of cells in the S-phase, an effect likely due to the dephosphorylation of 4E-BP1, an AKT1-dependent substrate, which results in a decreased availability of proteins needed for cell-cycle progression. Besides inducing cell death through activation of complete EBV lytic replication, hTERT inhibition triggered AKT1/FOXO3/NOXA–dependent apoptosis in EBV-positive and -negative Burkitt's lymphoma cells. Finally, ganciclovir enhanced the apoptotic effect induced by hTERT inhibition in EBV-positive Burkitt's lymphomas and LCLs. Conclusions: These results suggest that combination of antiviral drugs with strategies able to inhibit hTERT expression may result in therapeutically relevant effects in patients with EBV-related malignancies. Clin Cancer Res; 19(8); 2036–47. ©2013 AACR.

Uniparental parthenotes are considered an unwanted byproduct of in vitro fertilization. In utero parthenote development is severely compromised by defective organogenesis and in particular by defective cardiogenesis. Although developmentally compromised, apparently pluripotent stem cells can be derived from parthenogenetic blastocysts. Here we hypothesized that nonembryonic parthenogenetic stem cells (PSCs) can be directed toward the cardiac lineage and applied to tissue-engineered heart repair. We first confirmed similar fundamental properties in murine PSCs and embryonic stem cells (ESCs), despite notable differences in genetic (allelic variability) and epigenetic (differential imprinting) characteristics. Haploidentity of major histocompatibility complexes (MHCs) in PSCs is particularly attractive for allogeneic cell-based therapies. Accordingly, we confirmed acceptance of PSCs in MHC-matched allotransplantation. Cardiomyocyte derivation from PSCs and ESCs was equally effective. The use of cardiomyocyte-restricted GFP enabled cell sorting and documentation of advanced structural and functional maturation in vitro and in vivo. This included seamless electrical integration of PSC-derived cardiomyocytes into recipient myocardium. Finally, we enriched cardiomyocytes to facilitate engineering of force-generating myocardium and demonstrated the utility of this technique in enhancing regional myocardial function after myocardial infarction. Collectively, our data demonstrate pluripotency, with unrestricted cardiogenicity in PSCs, and introduce this unique cell type as an attractive source for tissue-engineered heart repair.

Pluripotent stem cells offer unprecedented potential not only for human medicine but also for veterinary medicine, particularly in relation to the horse. Induced pluripotent stem cells (iPSCs) are particularly promising, as they are functionally similar to embryonic stem cells and can be generated in vitro in a patient-specific manner. In this study, we report the generation of equine iPSCs from skin fibroblasts obtained from a foal and reprogrammed using viral vectors coding for murine Oct4, Sox2, c-Myc, and Klf4 sequences. The reprogrammed cell lines were morphologically similar to iPSCs reported from other species and could be stably maintained over more than 30 passages. Immunostaining and polymerase chain reaction analyses revealed that these cell lines expressed an array of endogenous markers associated with pluripotency, including OCT4, SOX2, NANOG, REX1, LIN28, SSEA1, SSEA4, and TRA1-60. Furthermore, under the appropriate conditions, the equine iPSCs readily formed embryoid bodies and differentiated in vitro into cells expressing markers of ectoderm, mesoderm, and endoderm, and when injected into immunodeficient mice, gave raise to tumors containing differentiated derivatives of the 3 germ layers. Finally, we also reprogrammed fibroblasts from a 2-year-old horse. The reprogrammed cells were similar to iPSCs derived from neonatal fibroblasts in terms of morphology, expression of pluripotency markers, and differentiation ability. The generation of these novel cell lines constitutes an important step toward the understanding of pluripotency in the horse, and paves the way for iPSC technology to potentially become a powerful research and clinical tool in veterinary biomedicine.

Drug-induced hepatotoxicity is a leading cause of attrition for candidate pharmaceuticals in development. New preclinical screening methods are crucial to predict drug toxicity prior to human studies. Of all in vitro hepatotoxicity models, primary human hepatocytes are considered as 'the gold standard.' However, their use is hindered by limited availability and inter-individual variation. These barriers may be overcome by using primary mouse hepatocytes. We used differential in gel electrophoresis (DIGE) to study large-scale protein expression of primary mouse hepatocytes. These hepatocytes were exposed to three well-defined hepatotoxicants: acetaminophen, amiodarone, and cyclosporin A. Each hepatotoxicant induces a different hepatotoxic phenotype. Based on the DIGE results, the mRNA expression levels of deregulated proteins from cyclosporin A-treated cells were also analyzed. We were able to distinguish cyclosporin A from controls, as well as acetaminophen and amiodarone-treated samples. Cyclosporin A induced endoplasmic reticulum (ER) stress and altered the ER-Golgi transport. Moreover, liver carboxylesterase and bile salt sulfotransferase were differentially expressed. These proteins were associated with a protective adaptive response against cyclosporin A-induced cholestasis. The results of this study are comparable with effects in HepG2 cells. Therefore, we suggest both models can be used to analyze the cholestatic properties of cyclosporin A. Furthermore, this study showed a conserved response between primary mouse hepatocytes and HepG2 cells. These findings collectively lend support for use of omics strategies in preclinical toxicology, and might inform future efforts to better link preclinical and clinical research in rational drug development.

Imaging techniques such as immunofluorescence (IF) and the expression of fluorescent protein (FP) fusions are widely used to investigate the subcellular distribution of proteins. Here we report a systematic analysis of >500 human proteins comparing the localizations obtained in live versus fixed cells using FPs and IF, respectively. We identify systematic discrepancies between IF and FPs as well as between FP tagging at the N and C termini. The analysis shows that for 80% of the proteins, IF and FPs yield the same subcellular distribution, and the locations of 250 previously unlocalized proteins were determined by the overlap between the two methods. Approximately 60% of proteins localize to multiple organelles for both methods, indicating a complex subcellular protein organization. These results show that both IF and FP tagging are reliable techniques and demonstrate the usefulness of an integrative approach for a complete investigation of the subcellular human proteome. QC 20130521