Search results for: genotypic
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 74

Search results for: genotypic

74 Study of Phenotypic Polymorphism and Detection of Genotypic Polymorphism in Menochilus sexmaculatus (Coleoptera: Insecta) Using RAPD PCR

Authors: Huma Balouch

Abstract:

Menochilus sexmaculatus commonly known as six spotted zig zag ladybird, is an aphidophagus and the most misidentified Coccinellids due to the occurrence of numerous color variants. The correct identification of Menochilus sexmaculatus and its strains is necessary to implement the use of biological control. In the present study phenotypic and genotypic polymorphism was investigated in Menochilus sexmaculatus collected from Punjab, NWFP and Sindh provinces of Pakistan. Six different morphs of the species were distinguished by analyzing its Elytral color and spot pattern and then Polymerase Chain Reaction was used to generate random amplification of polymorphic DNA (RAPD) from six different types of Menochilus sexmaculatus. Forty primers (OPA & OPC Kit) were used to perform RAPD PCR on six different types of Menochilus sexmaculatus of which, seven primers revealed different patterns related to the Menochilus sexmaculatus types. These seven primers (OPA-04, OPA-09, OPA-18, OPC-04, OPC-12, OPC-15 and OPC-18) produced 111 clear polymorphic bands and 6 scorable strain specific markers. The cluster analysis applied to RAPD data showed high polymorphism among six types and it can be concluded that these six types are six polymorphic strains of the same species.

Keywords: Menochilus sexmaculatus, aphidophagus, coccinellids, phenotypic and genotypic polymorphism, RAPD-PCR, strain specific markers

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73 Genotypic Response Differences among Faba Bean Accessions under Regular Deficit Irrigation (RDI)

Authors: M. Afzal, Salem Safer Alghamdi, Awais Ahmad

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Limited amount of irrigation water is an alarming threat to arid and semiarid agriculture. However, genotypic response differences to water deficit conditions within species have been reported frequently. Present study was conducted in order to measure the genotypic differences among faba bean accessions under Regular Deficit Irrigation (RDI). Five seeds from each accession were sown in 135 silt filled pots (30 x 24 cm). Experiment was planned under split plot arrangement and replicated thrice. Treatments consisted of three RDI levels (100% (control), 60% and 40% of the field capacity) and fifteen faba bean accessions (two local accessions as reference while thirteen from different sources around the world). Irrigation treatment was started from the very first day of sowing. Plant height, shoot dry weight, stomatal conductance and total chlorophyll contents (SPAD reading) were measured one month after germination. Irrigation, faba bean accessions and the all possible interactions has stood significantly high for all studied parameters. Regular deficient irrigation has hampered the plant growth and associated parameters in decreasing order (100% < 60% < 40%). Accessions have responded differently under regular deficient irrigation and some of them are even better than local accession. A highly significant correlation among all parameters has also been observed. It was concluded from results that above parameters could be used as markers to identify the genotypic differences for water deficit stress response. This outcome encouraged the use of superior faba bean genotypes in breeding programs for improved varieties to enhance water use efficiency under stress conditions.

Keywords: accessions, stomatal conductance, total chlorophyll contents, RDI, regular deficient irrigation

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72 Phenotypic and Genotypic Diagnosis of Gaucher Disease in Algeria

Authors: S. Hallal, Z. Chami, A. Hadji-Lehtihet, S. Sokhal-Boudella, A. Berhoune, L. Yargui

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Gaucher disease is the most common lysosomal storage in our population, it is due to a deficiency of β –glucosidase acid. The enzyme deficiency causes a pathological accumulation of undegraded substrate in lysosomes. This metabolic overload is responsible for a multisystemic disease with hepatosplenomegaly, anemia, thrombocytopenia, and bone involvement. Neurological involvement is rare. The laboratory diagnosis of Gaucher disease consists of phenotypic diagnosis by determining the enzymatic activity of β - glucosidase by fluorimetric method, a study by genotypic diagnosis in the GBA gene, limiting the search recurrent mutations (N370S, L444P, 84 GG); PCR followed by an enzymatic digestion. Abnormal profiles were verified by sequencing. Monitoring of treated patients is provided by the determination of chitotriosidase. Our experience spaning a period of 6 years (2007-2014) has enabled us to diagnose 78 patients out of a total of 328 requests from the various departments of pediatrics, internal medicine, neurology. Genotypic diagnosis focused on the entire family of 9 children treated at pediatric CHU Mustapha, which help define the clinical form; or 5 of them had type III disease, carrying the L444P mutation in the homozygous state. Three others were composite (N370/L444P) (N370S/other unintended mutation in our study), and only in one family no recurrent mutation has been found. This molecular study permits screening of heterozygous essential for genetic counseling.

Keywords: Gaucher disease, mutations, N370S, L444P

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71 Genotypic Variation in the Germination Performance and Seed Vigor of Safflower (Carthamus tinctorius L.)

Authors: Mehmet Demir Kaya, Engin Gökhan Kulan, Onur İleri, Süleyman Avcı

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Due to variation in seed size, shape and oil content of safflower cultivars, germination and emergence performance have been severely influenced by seed characteristics. This study aimed to determine genotypic variation among safflower genotypes for one thousand seed weight, oil content, germination and seed vigor using electrical conductivity (EC) and cold test. In the study, safflower lines ES37-5, ES38-4, ES43-11, ES55-14 and ES58-11 which were developed by single seed selection method, and Dinçer and Remzibey-05 were used as standard varieties. The genotypes were grown under rainfed conditions in Eskişehir, Turkey with four replications. The seeds of each genotype were subjected to standard germination and emergence test at 25°C for 10 days with four replications and 50 seeds per replicate. Electrical conductivity test was performed at 25°C for 24 h to assess the seed vigor. Also, cold test were applied to each safflower genotype at 10°C for 4 days and 25°C for 6 days. Results showed that oil content of the safflower genotypes were different. The highest oil content was determined in ES43-11 with 36.6% while the lowest was 25.9% in ES38-4. Higher germination and emergence rate were obtained from ES55-14 with 96.5% and 73.0%, respectively. There was no significant difference among the safflower genotypes for EC values. Cold test showed that ES43-11 and ES55-14 gave the maximum germination percentages. It was concluded that genotypic factors except for soil and climatic conditions play an important role for determining seed vigor because safflower genotypes grown at the same condition produced various seed vigor values.

Keywords: Carthamus tinctorius L., germination, emergence, cold test, electrical conductivity

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70 Genetic Variability and Heritability Among Indigenous Pearl Millet (Pennisetum Glaucum L. R. BR.) in Striga Infested Fields of Sudan Savanna, Nigeria

Authors: Adamu Usman, Grace Stanley Balami

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Pearl millet (Pennisetum glaucum L. R. Br.) is a cereal cultivated in arid and semi-arid areas of the world. It supports more than 100 million people around the world. Parasitic weed (Striga hermonthica Del. Benth) is a major constraint to its production. Estimated yield losses are put at 10 - 95% depending on variety, ecology and cultural practices. Potentials in selection of traits in pearl millets for grain yield have been reported and it depends on genotypic variability and heritability among landraces. Variability and heritability among cultivars could offer opportunities for improvement. The study was conducted to determine the genetic variability among cultivars and estimate broad sense heritability among grain yield and related traits. F1 breeding populations were generated with 9 parental cultivars, viz; Ex-Gubio, Ex-Monguno, Ex-Baga as males and PEO 5984, Super-SOSAT, SOSAT-C88, Ex-Borno and LCIC9702 as females through Line × Tester mating during 2017 dry season at Lushi Irrigation Station, Bauchi Metropolitan in Bauchi State, Nigeria. The F1 population and the parents were evaluated during cropping season of 2018 at Bauchi and Maiduguri. Data collected were subjected to analysis of variance. Results showed significant difference among cultivars and among traits indicating variability. Number of plants at emergence, days to 50% flowering, days to 100% flowering, plant height, panicle length, number of plants at harvest, Striga count at 90 days after sowing, panicle weight and grain yield were significantly different. Significant variability offer opportunity for improvement as superior individuals can be isolated. Genotypic variance estimates of traits were largely greater than environmental variances except in plant height and 1000 seed weight. Environmental variances were low and in some cases negligible. The phenotypic variances of all traits were higher than genotypic variances. Similarly phenotypic coefficient of variation (PCV) was higher than genotypic coefficient of variation (GCV). High heritability was found in days to 50% flowering (90.27%), Striga count at 90 days after sowing (90.07%), number of plants at harvest (87.97%), days to 100% flowering (83.89%), number of plants at emergence (82.19%) and plant height (73.18%). Greater heritability estimates could be due to presence of additive gene. The result revealed wider variability among genotypes and traits. Traits having high heritability could easily respond to selection. High value of GCV, PCV and heritability estimates indicate that selection for these traits are possible and could be effective.

Keywords: variability, heritability, phenotypic, genotypic, striga

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69 Pharmacogenetic Analysis of Inter-Ethnic Variability in the Uptake Transporter SLCO1B1 Gene in Colombian, Mozambican, and Portuguese Populations

Authors: Mulata Haile Nega, Derebew Fikadu Berhe, Vera Ribeiro Marques

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There is no epidemiologic data on this gene polymorphism in several countries. Therefore, this study aimed to assess the genotype and allele frequencies of the gene variant in three countries. This study involved healthy individuals from Colombia, Mozambique, and Portugal. Genomic DNA was isolated from blood samples using the Qiamp DNA Extraction Kit (Qiagen). The isolated DNA was genotyped using Polymerase Chain Reaction (PCR) - Restriction Fragment Length Polymorphism. Microstat and GraphPad quick cal software were used for the Chi-square test and evaluation of Hardy-Weinberg equilibrium, respectively. A total of 181 individuals’ blood sample was analyzed. Overall, TT (74.0%) genotype was the highest, and CC (7.8%) was the lowest. Country wise genotypic frequencies were Colombia 47(70.2%) TT, 12(17.9%) TC and 8(11.9%) CC; Mozambique 47(88.7%) TT, 5(9.4%) TC, and 1(1.9%) CC; and Portugal 40(65.6%) TT, 16(26.2%) TC, and 5(8.2%) CC. The reference (T) allele was highest among Mozambicans (93.4%) compared to Colombians (79.1%) and Portuguese (78.7%). Mozambicans showed statistically significant genotypic and allelic frequency differences compared to Colombians (p<0.01) and Portuguese (p <0.01). Overall and country-wise, the CC genotype was less frequent and relatively high for Colombians and Portuguese populations. This finding may imply statins risk-benefit variability associated with CC genotype among these populations that needs further understanding.

Keywords: c.521T>C, polymorphism, SLCO1B1, SNP, statins

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68 Investigation of Genetic Variation for Agronomic Traits among the Recombinant Inbred Lines of Wheat from the Norstar × Zagross Cross under Water Stress Condition

Authors: Mohammad Reza Farzami Pour

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Determination of genetic variation is useful for plant breeding and hence production of more efficient plant species under different conditions, like drought stress. In this study, a sample of 28 recombinant inbred lines (RILs) of wheat developed from the cross of Norstar and Zagross varieties, together with their parents, were evaluated for two years (2010-2012) under normal and water stress conditions using split plot design with three replications. Main plots included two irrigation treatments of 70 and 140 mm evaporation from Class A pan and sub-plots consisted of 30 genotypes. The effect of genotypes and interaction of genotypes with years and water regimes were significant for all characters. Significant genotypic effect implies the existence of genetic variation among the lines under study. Heritability estimates were high for 1000 grain weight (0.87). Biomass and grain yield showed the lowest heritability values (0.42 and 0.50, respectively). Highest genotypic and phenotypic coefficients of variation (GCV and PCV) belonged to harvest index. Moderate genetic advance for most of the traits suggested the feasibility of selection among the RILs under investigation. Some RILs were higher yielding than either parent at both environments.

Keywords: wheat, genetic gain, heritability, recombinant inbred lines

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67 Agro-Morphological Traits Based Genetic Diversity Analysis of ‘Ethiopian Dinich’ Plectranthus edulis (Vatke) Agnew Populations Collected from Diverse Agro-Ecologies in Ethiopia

Authors: Fekadu Gadissa, Kassahun Tesfaye, Kifle Dagne, Mulatu Geleta

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‘Ethiopian dinich’ also called ‘Ethiopian potato’ is one of the economically important ‘orphan’ edible tuber crops indigenous to Ethiopia. We evaluated the morphological and agronomic traits performances of 174 samples from Ethiopia at multiple locations using 12 qualitative and 16 quantitative traits, recorded at the correct growth stages. We observed several morphotypes and phenotypic variations for qualitative traits along with a wide range of mean performance values for all quantitative traits. Analysis of variance for each quantitative trait showed a highly significant (p<0.001) variation among the collections with eventually non-significant variation for environment-traits interaction for all but flower length. A comparatively high phenotypic and genotypic coefficient of variation was observed for plant height, days to flower initiation, days to 50% flowering and tuber number per hill. Moreover, the variability and coefficients of variation due to genotype-environment interaction was nearly zero for all the traits except flower length. High genotypic coefficients of variation coupled with a high estimate of broad sense heritability and high genetic advance as a percent of collection mean were obtained for tuber weight per hill, number of primary branches per plant, tuber number per hill and number of plants per hill. Association of tuber yield per hectare of land showed a large magnitude of positive phenotypic and genotypic correlation with those traits. Principal components analysis revealed 76% of the total variation for the first six principal axes with high factor loadings again from tuber number per hill, number of primary branches per plant and tuber weight. The collections were grouped into four clusters with the weak region (zone) of origin based pattern. In general, there is high genetic-based variability for ‘Ethiopian dinich’ improvement and conservation. DNA based markers are recommended for further genetic diversity estimation for use in breeding and conservation.

Keywords: agro-morphological traits, Ethiopian dinich, genetic diversity, variance components

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66 Bayesian Estimation of Hierarchical Models for Genotypic Differentiation of Arabidopsis thaliana

Authors: Gautier Viaud, Paul-Henry Cournède

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Plant growth models have been used extensively for the prediction of the phenotypic performance of plants. However, they remain most often calibrated for a given genotype and therefore do not take into account genotype by environment interactions. One way of achieving such an objective is to consider Bayesian hierarchical models. Three levels can be identified in such models: The first level describes how a given growth model describes the phenotype of the plant as a function of individual parameters, the second level describes how these individual parameters are distributed within a plant population, the third level corresponds to the attribution of priors on population parameters. Thanks to the Bayesian framework, choosing appropriate priors for the population parameters permits to derive analytical expressions for the full conditional distributions of these population parameters. As plant growth models are of a nonlinear nature, individual parameters cannot be sampled explicitly, and a Metropolis step must be performed. This allows for the use of a hybrid Gibbs--Metropolis sampler. A generic approach was devised for the implementation of both general state space models and estimation algorithms within a programming platform. It was designed using the Julia language, which combines an elegant syntax, metaprogramming capabilities and exhibits high efficiency. Results were obtained for Arabidopsis thaliana on both simulated and real data. An organ-scale Greenlab model for the latter is thus presented, where the surface areas of each individual leaf can be simulated. It is assumed that the error made on the measurement of leaf areas is proportional to the leaf area itself; multiplicative normal noises for the observations are therefore used. Real data were obtained via image analysis of zenithal images of Arabidopsis thaliana over a period of 21 days using a two-step segmentation and tracking algorithm which notably takes advantage of the Arabidopsis thaliana phyllotaxy. Since the model formulation is rather flexible, there is no need that the data for a single individual be available at all times, nor that the times at which data is available be the same for all the different individuals. This allows to discard data from image analysis when it is not considered reliable enough, thereby providing low-biased data in large quantity for leaf areas. The proposed model precisely reproduces the dynamics of Arabidopsis thaliana’s growth while accounting for the variability between genotypes. In addition to the estimation of the population parameters, the level of variability is an interesting indicator of the genotypic stability of model parameters. A promising perspective is to test whether some of the latter should be considered as fixed effects.

Keywords: bayesian, genotypic differentiation, hierarchical models, plant growth models

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65 Variability Parameters for Growth and Yield Characters in Fenugreek, Trigonella spp. Genotypes

Authors: Anita Singh, Richa Naula, Manoj Raghav

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India is a leading producer and consumer of fenugreek for its culinary uses and medicinal application. In India, most of the people are of vegetarian class. In such a situation, a leafy vegetable, such as fenugreek is of chief concern due to its high nutritional property, medicinal values and industrial uses. One of the most important factors restricting their large scale production and development of superior varieties is that very scanty knowledge about their genetic diversity, inter and intraspecific variability and genetic relationship among the species. Improvement of the crop depends upon the magnitude of genetic variability for economic characters. Therefore, the present research work was carried out to analyse the variability parameters for growth and yield character in twenty-eight fenugreek genotypes along with two standard checks Pant Ragini and Pusa Early Bunching. The experiment was laid out in Randomized Block Design with three replication during rabi season 2015-2016 at Pantnagar Centre for Plant Genetic Resources, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand. The analysis of variance revealed highly significant differences among all the genotypes for all traits. High genotypic and phenotypic coefficient variation were observed for characters, namely the number of primary branches per plant, number of leaves at 30, 45 and 60 DAS, green leaf yield per plant, green leaf yield q/ha . The genetic advance recorded highest in green leaf yield q/ha (33.93) followed by green leaf yield per plant (21.20g). Highest percent of heritability were shown by 1000 seed weight (99.12%) followed by the number of primary branches per plant (97.18%). Green leaf yield q/ha showed high heritability and high genetic advance. These superior genotypes can be further used in crop improvement programs of fenugreek.

Keywords: genetic advance, genotypic coefficient variation, heritability, phenotypic coefficient variation

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64 Genotypic Characterization of Gram-Positive Bacteria Isolated on Ornamental Animals Feed

Authors: C. Miranda, R. Soares, S. Cunha, L. Ferreira, G. Igrejas, P. Poeta

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Different animal species, including ornamental animals, are reported as potential reservoirs of antibiotic resistance genes. Consequently, these resistances can be disseminated in the environment and transferred to humans. Moreover, multidrug-resistant bacteria reduce the efficacy of antibiotics, as the case of vancomycin-resistant enterococci. Enterococcus faecalis and E. faecium are described as the main nosocomial pathogens. In this line, the aim of this study was to characterize resistance and virulence genes of enterococci species isolated from samples of food supplied to ornamental animals during 2020. The 29 enterococci isolates (10 E. faecalis and 19 E. faecium) were tested for the presence of the resistance genes for the following antibiotics: erythromicyn (ermA, ermB and ermC), tetracycline (tetL, tetM, tetK and tetO), quinupristin/dalfopristin (vatD and vatE), gentamicin (aac(6’)-aph(2’’)-Ia), chloramphenicol (catA), streptomycin (ant(6)-Ia) and vancomycin (vanA and vanB). The same isolates were also tested for 10 virulence factors genes (esp, ace, gelE, agg, fsr, cpd, cylA, cylB, cylM and cylLL). The resistance and virulence genes were performed by PCR, using specific primers and conditions. Negative and positive controls were used in all PCR assays. The most prevalent resistance genes detected in both enterococci species were ermB (n=15, 52%), ermC (n=7, 24%), tetK (n=8, 28%) and vatE (n=4, 14%). Resistance genes for vancomycin were found in ten (34%) E. faecalis and ten (34%) E. faecium isolates. Only E. faecium isolates showed the presence of ermA (n=2, 7%), tetL (n=13, 45%) and ant(6)-Ia gene (n=4, 14%). A total of nine (31%) enterococci isolates were classified as multidrug-resistant bacteria (3 E. faecalis and 6 E. faecium). In three E. faecalis and one E. faecium were not detected resistance genes. The virulence genes detected in both species were agg (n=6, 21%) and cylLL (n=11, 38%). In general, each isolate showed only one of these virulence genes. Five E. faecalis and eleven E. faecium isolates were negative for all analyzed virulence genes. These preliminary results showed the presence of multidrug-resistant enterococci in food supplied to ornamental animals, in particular vancomycin-resistant enterococci. This genotypic characterization reinforces the relevance to public health in the control of antibiotic-resistant bacteria.

Keywords: antibiotic resistance, enterococci, feed, ornamental animals

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63 Genotypic Identification of Oral Bacteria Using 16S rRNA in Children with and without Early Childhood Caries in Kelantan, Malaysia

Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

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Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

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62 Association of a Genetic Polymorphism in Cytochrome P450, Family 1 with Risk of Developing Esophagus Squamous Cell Carcinoma

Authors: Soodabeh Shahid Sales, Azam Rastgar Moghadam, Mehrane Mehramiz, Malihe Entezari, Kazem Anvari, Mohammad Sadegh Khorrami, Saeideh Ahmadi Simab, Ali Moradi, Seyed Mahdi Hassanian, Majid Ghayour-Mobarhan, Gordon A. Ferns, Amir Avan

Abstract:

Background Esophageal cancer has been reported as the eighth most common cancer universal and the seventh cause of cancer-related death in men .recent studies have revealed that cytochrome P450, family 1, subfamily B, polypeptide 1, which plays a role in metabolizing xenobiotics, is associated with different cancers. Therefore in the present study, we investigated the impact of CYP1B1-rs1056836 on esophagus squamous cell carcinoma (ESCC) patients. Method: 317 subjects, with and without ESCC were recruited. DNA was extracted and genotyped via Real-time PCR-Based Taq Man. Kaplan Meier curves were utilized to assess overall and progression-free survival. To evaluate the relationship between patients clinicopathological data, genotypic frequencies, disease prognosis, and patients survival, Pearson chi-square and t-test were used. Logistic regression was utilized to assess the association between the risk of ESCC and genotypes. Results: the genotypic frequency for GG, GC, and CC are respectively 58.6% , 29.8%, 11.5% in the healthy group and 51.8%, 36.14% and 12% in ESCC group. With respect to the recessive genetic inheritance model, an association between the GG genotype and stage of ESCC were found. Also, statistically significant results were not found for this variation and risk of ESCC. Patients with GG genotype had a decreased risk of nodal metastasis in comparison with patients with CC/CG genotype, although this link was not statistically significant. Conclusion: Our findings illustrated the correlation of CYP1B1-rs1056836 as a potential biomarker for ESCC patients, supporting further studies in larger populations in different ethnic groups. Moreover, further investigations are warranted to evaluate the association of emerging marker with dietary intake and lifestyle.

Keywords: Cytochrome P450, esophagus squamous cell carcinoma, dietary intake, lifestyle

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61 Genetic Divergence Study of Rice on the Basis of Various Morphological Traits

Authors: Muhammad Ashfaq, Muhammad Saleem Haider, Muhammad Ali, Muhammad Sajjad, Amna Ali, Urooj Mubashar

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Phenotypic diversity was confirmed by measuring different morphological traits i.e. seed traits (seed length, seed width, seed thickness, seed length-width ratio, 1000 grain weight) and root-shoot traits (shoot length, root length, shoot fresh weight, root fresh weight, root-shoot ratio, root numbers and root thickness). Variance and association study of desirable traits determine the genotypic differences among the rice germplasm. All the traits showed significant differences among the genotypes. The traits were studied in Randomized complete block design (RCBD) at different water levels. Some traits showed positive correlation with each other and beneficial for increasing the yield and production of the crop. Seed thickness has positive correlation with seed length and seed width (r= 0.104**, r=0.246**). On the other hand, various root shoot traits showed positive highly significant association at different water levels i.e. root length, fresh root weight, root thickness, shoot thickness and root numbers. Our main focus to study the performance/correlation of root shoots traits under stress condition. Fresh root weight, shoot thickness and root numbers showed positive significant association with shoot length, root length, fresh root and shoot weight (r=0.2530**, r=0.2891**, r=0.4626**, r=0.4515**, r=0.5781**, r=0.7164**, r=0.0603**, r= 0.5570**, r=0.5824**). Long root length genotypes favors and suitable for drought stress conditions and screening of diverse genotypes for the further development of new plant material that performing well under different environmental conditions. After screening genetic diversity of potential rice, lines were studied to check the polymorphism by using some SSR markers. DNA was extracted, and PCR analyses were done to study PIC values and allelic diversity of the genotypes. The main objective of this study is to screen out the genotypes on the basis of various genotypic and phenotypic traits.

Keywords: rice, morphological traits, association, germplasm, genetic diversity, water levels, variation

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60 Asparagus racemosus Willd for Enhanced Medicinal Properties

Authors: Ashok Kumar, Parveen Parveen

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India is bestowed with an extremely high population of plant species with medicinal value and even has two biodiversity hotspots. Indian systems of medicine including Ayurveda, Siddha and Unani have historically been serving humankind across the world since time immemorial. About 1500 plant species have well been documented in Ayurvedic Nighantus as official medicinal plants. Additionally, several hundred species of plants are being routinely used as medicines by local people especially tribes living in and around forests. The natural resources for medicinal plants have unscientifically been over-exploited forcing rapid depletion in their genetic diversity. Moreover, renewed global interest in herbal medicines may even lead to additional depletion of medicinal plant wealth of the country, as about 95% collection of medicinal plants for pharmaceutical preparation is being carried out from natural forests. On the other hand, huge export market of medicinal and aromatic plants needs to be seriously tapped for enhancing inflow of foreign currency. Asparagus racemosus Willd., a member of family Liliaceae, is one of thirty-two plant species that have been identified as priority species for cultivation and conservation by the National Medicinal Plant Board (NMPB), Government of India. Though attention is being focused on standardization of agro-techniques and extraction methods, little has been designed on genetic improvement and selection of desired types with higher root production and saponin content, a basic ingredient of medicinal value. The saponin not only improves defense mechanisms and controls diabetes but the roots of this species promote secretion of breast milk, improved lost body weight and considered as an aphrodisiac. There is ample scope for genetic improvement of this species for enhancing productivity substantially, qualitatively and quantitatively. It is emphasized to select desired genotypes with sufficient genetic diversity for important economic traits. Hybridization between two genetically divergent genotypes could result in the synthesis of new F1 hybrids consisting of useful traits of both the parents. The evaluation of twenty seed sources of Asparagus racemosus assembled different geographical locations of India revelled high degree of variability for traits of economic importance. The maximum genotypic and phenotypic variance was observed for shoot height among shoot related traits and for root length among root related traits. The shoot height, genotypic variance, phenotypic variance, genotypic coefficient of variance, the phenotypic coefficient of variance was recorded to be 231.80, 3924.80, 61.26 and 1037.32, respectively, where those of the root length were 9.55, 16.80, 23.46 and 41.27, respectively. The maximum genetic advance and genetic gain were obtained for shoot height among shoot-related traits and root length among root-related traits. Index values were developed for all seed sources based on the four most important traits, and Panthnagar (Uttrakhand), Jodhpur (Rajasthan), Dehradun (Uttarakhand), Chandigarh (Punjab), Jammu (Jammu & Kashmir) and Solan (Himachal Pradesh) were found to be promising seed sources.

Keywords: asparagus, genetic, genotypes, variance

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59 Determination of Genotypic Relationship among 12 Sugarcane (Saccharum officinarum) Varieties

Authors: Faith Eweluegim Enahoro-Ofagbe, Alika Eke Joseph

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Information on genetic variation within a population is crucial for utilizing heterozygosity for breeding programs that aim to improve crop species. The study was conducted to ascertain the genotypic similarities among twelve sugarcane (Saccharum officinarum) varieties to group them for purposes of hybridizations for cane yield improvement. The experiment was conducted at the University of Benin, Faculty of Agriculture Teaching and Research Farm, Benin City. Twelve sugarcane varieties obtained from National Cereals Research Institute, Badeggi, Niger State, Nigeria, were planted in three replications in a randomized complete block design. Each variety was planted on a five-row plot of 5.0 m in length. Data were collected on 12 agronomic traits, including; the number of millable cane, cane girth, internode length, number of male and female flowers (fuss), days to flag leaf, days to flowering, brix%, cane yield, and others. There were significant differences, according to the findings among the twelve genotypes for the number of days to flag leaf, number of male and female flowers (fuss), and cane yield. The relationship between the twelve sugarcane varieties was expressed using hierarchical cluster analysis. The twelve genotypes were grouped into three major clusters based on hierarchical classification. Cluster I had five genotypes, cluster II had four, and cluster III had three. Cluster III was dominated by varieties characterized by higher cane yield, number of leaves, internode length, brix%, number of millable stalks, stalk/stool, cane girth, and cane length. Cluster II contained genotypes with early maturity characteristics, such as early flowering, early flag leaf development, growth rate, and the number of female and male flowers (fuss). The maximum inter-cluster distance between clusters III and I indicated higher genetic diversity between the two groups. Hybridization between the two groups could result in transgressive recombinants for agronomically important traits.

Keywords: sugarcane, Saccharum officinarum, genotype, cluster analysis, principal components analysis

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58 A Diagnostic Accuracy Study: Comparison of Two Different Molecular-Based Tests (Genotype HelicoDR and Seeplex Clar-H. pylori ACE Detection), in the Diagnosis of Helicobacter pylori Infections

Authors: Recep Kesli, Huseyin Bilgin, Yasar Unlu, Gokhan Gungor

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Aim: The aim of this study was to compare diagnostic values of two different molecular-based tests (GenoType® HelicoDR ve Seeplex® H. pylori-ClaR- ACE Detection) in detection presence of the H. pylori from gastric biopsy specimens. In addition to this also was aimed to determine resistance ratios of H. pylori strains against to clarytromycine and quinolone isolated from gastric biopsy material cultures by using both the genotypic (GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection) and phenotypic (gradient strip, E-test) methods. Material and methods: A total of 266 patients who admitted to Konya Education and Research Hospital Department of Gastroenterology with dyspeptic complaints, between January 2011-June 2013, were included in the study. Microbiological and histopathological examinations of biopsy specimens taken from antrum and corpus regions were performed. The presence of H. pylori in all the biopsy samples was investigated by five differnt dignostic methods together: culture (C) (Portagerm pylori-PORT PYL, Pylori agar-PYL, GENbox microaer, bioMerieux, France), histology (H) (Giemsa, Hematoxylin and Eosin staining), rapid urease test (RUT) (CLOtest, Cimberly-Clark, USA), and two different molecular tests; GenoType® HelicoDR, Hain, Germany, based on DNA strip assay, and Seeplex ® H. pylori -ClaR- ACE Detection, Seegene, South Korea, based on multiplex PCR. Antimicrobial resistance of H. pylori isolates against clarithromycin and levofloxacin was determined by GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection, and gradient strip (E-test, bioMerieux, France) methods. Culture positivity alone or positivities of both histology and RUT together was accepted as the gold standard for H. pylori positivity. Sensitivity and specificity rates of two molecular methods used in the study were calculated by taking the two gold standards previously mentioned. Results: A total of 266 patients between 16-83 years old who 144 (54.1 %) were female, 122 (45.9 %) were male were included in the study. 144 patients were found as culture positive, and 157 were H and RUT were positive together. 179 patients were found as positive with GenoType® HelicoDR and Seeplex ® H. pylori -ClaR- ACE Detection together. Sensitivity and specificity rates of studied five different methods were found as follows: C were 80.9 % and 84.4 %, H + RUT were 88.2 % and 75.4 %, GenoType® HelicoDR were 100 % and 71.3 %, and Seeplex ® H. pylori -ClaR- ACE Detection were, 100 % and 71.3 %. A strong correlation was found between C and H+RUT, C and GenoType® HelicoDR, and C and Seeplex ® H. pylori -ClaR- ACE Detection (r:0.644 and p:0.000, r:0.757 and p:0.000, r:0.757 and p:0.000, respectively). Of all the isolated 144 H. pylori strains 24 (16.6 %) were detected as resistant to claritromycine, and 18 (12.5 %) were levofloxacin. Genotypic claritromycine resistance was detected only in 15 cases with GenoType® HelicoDR, and 6 cases with Seeplex ® H. pylori -ClaR- ACE Detection. Conclusion: In our study, it was concluded that; GenoType® HelicoDR and Seeplex ® H. pylori -ClaR- ACE Detection was found as the most sensitive diagnostic methods when comparing all the investigated other ones (C, H, and RUT).

Keywords: Helicobacter pylori, GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection, antimicrobial resistance

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57 Detection of Tetracycline Resistance Genes in Lactococcus garvieae Strains Isolated from Rainbow Trout

Authors: M. Raissy, M. Shahrani

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The present study was done to evaluate the presence of tetracycline resistance genes in Lactococcus garvieae isolated from cultured rainbow trout, West Iran. The isolates were examined for antimicrobial resistance using disc diffusion method. Of the 49 strains tested, 19 were resistant to tetracycline (38.7%), 32 to enrofloxacin (65.3%), 21 to erythromycin (42.8%), 20 to chloramphenicol and trimetoprim-sulfamethoxazole (40.8%). The strains were then characterized for their genotypic resistance profiles. The results revealed that all 49 isolates contained at least one of the tetracycline resistance genes. Tet (A) was found in 89.4% of tetracycline resistant isolates and the frequency of other gene were as follow: tet (E) 42.1%, tet (B) 47.3%, tet (D) 15.7%, tet (L) 26.3%, tet (K) 52.6%, tet (G) 36.8%, tet (34) 21%, tet (S) 63.1%, tet (C) 57.8%, tet (M) 73.6%, tet (O) 42.1%. The results revealed high levels of antibiotic resistance in L. garvieae strains which is a potential danger for trout culture as well as for public health.

Keywords: Lactococcus garvieae, tetracycline resistance genes, rainbow trout, antimicrobial resistance

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56 Improving the Efficiency of Wheat and Triticale Androgenesis: Ultrastructural and Transcriptomic Study

Authors: M. Szechynska-Hebda, M. Sobczak, E. Rozanska, J. Troczynska, Z. Banaszak, N. Hordyńska, M. Dyda, M. Wedzony

Abstract:

Chloroplasts, as essential organelles for photosynthesis, play a critical role in plant development. However, disturbances in the proper functioning of chloroplasts, in the extreme case manifesting as albinism of tissues and whole plants, are a phenomenon often occurring in conditions deviating from natural (e.g., in vitro cultures applied in breeding programs). Using whole-transcriptome analysis (RNA-Seq) together with light, fluorescent and electron microscopy, it was shown, that development of chloroplasts and formation of green or albino plants in the androgenesis process are genotype-dependent; however, they could be modulated by sub-optimal temperature treatment. The reprogramming of the microspore development from gametophytic to sporophytic, and then regeneration of green plant can be positively regulated by cold stress (4 ⁰C). A high temperature stress (32 ⁰C) can induce androgenesis, but it is a factor negatively influencing green plant regeneration (promoting albinism). A similar effect on microspores, androgenesis, and subsequent chloroplast formation, is elicited as a result of postponing the date of spike collection from spring to summer in field conditions (natural temperature rise). It is determined in both environmental or genotypic manner. The delay of the sowing date (environmental effect) or growing of late genotypes (genotypic effect) result in spike maturation at higher temperatures and significantly enhance albino plant formation in androgenesis process. Such a temperature system (4 ⁰C vs. 32 ⁰C) was used to study the chloroplast biogenesis process in wheat and triticale. It was shown, that efficiency of physiological processes differentiates microspore development during cold reprograming in genotypes susceptible and resistant to androgenesis. Moreover, a great variation in developmental stages of the microspores in one anther is observed for susceptible genotypes. Microspores that are more physiologically active under cold conditions can activate signaling pathways and processes, which provide an appropriate supply of metabolites to cell compartments. This, in turn, fully correlates with the genotype-dependent efficiency of chloroplast formation (or different types of plastid) at particular steps of androgenesis. The effect obtained after applying a high temperature stress is different. High temperature causes a significant acceleration of microspore development and less variation in developmental stages at the end of the treatment. Therefore, the developmental diversity of the microspores in one anther seems to be a critical factor for subsequent cell and chloroplast differentiation. The work was financed by Ministry of Agriculture and Rural Development within Program: 'Biological Progress in Plant Production', project no HOR.hn.802.15.2018

Keywords: androgenesis, chloroplast biogenesis, temperature stress, wheat

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55 Some Probiotic Traits of Lactobacillus Strains Isolated from Pollen

Authors: Hani Belhadj, Daoud Harzallah, Seddik Khennouf, Saliha Dahamna, Mouloud Ghadbane

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In this study, Lactobacillus strains isolated from pollen were identified by means of phenotypic and genotypic methods, At pH 2, most strains proved to be acid resistants, with losses in cell viability ranging from 0.77 to 4.04 Log orders. In addition, at pH 3 all strains could grew and resist the acidic conditions, with losses in cell viability ranging from 0.40 to 3.61 Log orders. It seems that, 0.3% and 0.5% of bile salts does not affect greatly the survival of most strains, excluding Lactobacillus sp. BH1398. Survival ranged from 81.0±3.5 to 93.5±3.9%. In contrast, in the presence of 1.0% bile salts, survival of five strains was decreased by more than 50%. Lactobacillus fermentum BH1509 was considered the most tolerant strain (77.5% for 1% bile) followed by Lactobacillus plantarum BH1541 (59.9% for 1% bile). Furthermore, all strains were resistant to colistine, clindamycine, chloramphenicol, and ciprofloxacine, but most of the strains were susceptible to Peniciline, Oxacillin, Oxytetracyclin, and Amoxicillin. Functionally interesting Lactobacillus isolates may be used in the future as probiotic cultures for manufacturing fermented foods and as bioactive delivery systems.

Keywords: probiotics, lactobacillus, pollen, bile, acid tolerance

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54 Glycine Betaine Affects Antioxidant Response and Lipid Peroxidation in Wheat Genotypes under Water-Deficit Conditions

Authors: S. K. Thind, Neha Gupta

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Glycine betaine (N, N’, N’’– trimethyl glycine), (GB) as aqueous solution (100 mM) containing 0.1% TWEEN-20 (Ploythylene glycol sorbitan monolaurate) was sprayed on selected nineteen wheat genotypes at maximum tillering and anthesis stages. Water-deficit conditions resulted in lipid peroxidation. GB applications reduced lipid peroxidation in all wheat genotypes at both the stages. Catalase (CAT) activity was recorded more in control than under stressed conditions in selected wheat genotypes at both the stages; GB had no effect. The ascorbic acid content in leaves of selected genotypes increased under water deficit. A genotypic variability in Ascorbate peroxidase (APx) activity was recorded and GB treatment decreased it. Superoxide dismutase (SOD) activity was increased significantly under water-deficit at both stages in all genotypes. In present study, prolonged water-deficit conditions caused CAT deficiency/suppression which was compensated by APX and SOD; and GB exogenous application mitigated negative effect of water-deficit stress on lipid peroxidation.

Keywords: glycine-betaine, lipid peroxidation, ROS, water deficit stress

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53 How Does Vicia faba-rhizobia Symbiosis Improve Its Performance under Low Phosphorus Availability?

Authors: B. Makoudi, R. Ghanimi, M. Mouradi, A. Kabbadj, M. Farissi, J. J. Drevon, C. Ghoulam

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This work focuses on the responses of Vicia fabarhizobia symbiosis to phosphorus deficiency and their contribution to tolerate this constraint. The study was carried out on four faba bean varieties, Aguadulce, Alfia, Luz Otono, and Reina Mora submitted to two phosphorus treatments, deficient and sufficient and cultivated under field and greenhouse hydroaeroponic culture. Plants were harvested at flowering stage for growth, nodulation and phosphorus content assessment. Phosphatases in nodules and rhizospheric soil were analyzed. The impact of phosphorus deficiency on yield component was assessed at maturity stage. Under field conditions, phosphorus deficiency affected negatively nodule biomass and nodule phosphorus content with Alfia and Reina Mora showing the highest biomass reduction. The phosphatase activities in nodules and rhizospheric soil were increased under phosphorus deficiency. At maturity stage, under soil low available phosphorus, the pods number and 100 seeds weight were reduced. The genotypic variation was evident for almost all tested parameters.

Keywords: faba bean, phosphorus, rhizobia, yield

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52 Genetic Variability and Principal Component Analysis in Eggplant (Solanum melongena)

Authors: M. R. Naroui Rad, A. Ghalandarzehi, J. A. Koohpayegani

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Nine advanced cultivars and lines were planted in transplant trays on March, 2013. In mid-April 2014, nine cultivars and lines were taken from the seedling trays and were evaluated and compared in an experiment in form of a completely randomized block design with three replications at the Agricultural Research Station, Zahak. The results of the analysis of variance showed that there was a significant difference between the studied cultivars in terms of average fruit weight, fruit length, fruit diameter, ratio of fruit length to its diameter, the relative number of seeds per fruit, and each plant yield. The total yield of Sohrab and Y6 line with and an average of 41.9 and 36.7 t/ ha allocated the highest yield respectively to themselves. The results of simple correlation between the analyzed traits showed the final yield was affected by the average fruit weight due to direct and indirect effects of fruit weight and plant yield on the final yield. The genotypic and heritability values were high for fruit weight, fruit length and number of seed per fruit. The first two principal components accounted for 81.6% of the total variation among the characters describing genotypes.

Keywords: eggplant, principal component, variation, path analysis

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51 Extraction of Saponins and Cyclopeptides from Cow Cockle (Vaccaria hispanica (Mill.) Rauschert) Seeds Grown in Turkey

Authors: Ihsan Burak Cam, Ferhan Balci-Torun, Ayhan Topuz, Esin Ari, Ismail Gokhan Deniz, Ilker Genc

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The seeds of Vaccaria hispanica have been used in food and pharmaceutical industry. It is an important product due to its superior starch granules, triterpenic saponins, and cyclopeptides suitable for drug delivery. V. hispanica naturally grows in different climatic regions and has genotypes that differ in terms of seed content and composition. Sixty-six V. hispanica seed specimens were collected based on the representation of the distribution in all regions of Turkey and the determination of possible genotypic differences between regions. The seeds, collected from each of the 66 locations, were grown in greenhouse conditions in Akdeniz University, Antalya. Saponin and cyclopeptide contents of the V. hispanica seeds were determined after harvest. Accelerated solvent extraction (ASE) was applied for the extraction of saponins and cyclopeptides. Cyclopeptide (segetalin A) and saponin content of V. hispanica seeds were found in the range of 0.165-0.654 g/100 g and 0.15-1.14 g/100 g, respectively. The results were found to be promising for the seeds from Turkey in terms of saponin content and quality. Acknowledgment: This study was supported by the Scientific and Research Council of Turkey (TUBITAK) (project no 112 O 136).

Keywords: Vaccaria hispanica, saponin, cyclopeptid, cow cockle seeds

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50 The Use of SD Bioline TB AgMPT64® Detection Assay for Rapid Characterization of Mycobacteria in Nigeria

Authors: S. Ibrahim, U. B. Abubakar, S. Danbirni, A. Usman, F. M. Ballah, C. A. Kudi, L. Lawson, G. H. Abdulrazak, I. A. Abdulkadir

Abstract:

Performing culture and characterization of mycobacteria in low resource settings like Nigeria is a very difficult task to undertake because of the very few and limited laboratories carrying out such an experiment; this is a largely due to stringent and laborious nature of the tests. Hence, a rapid, simple and accurate test for characterization is needed. The “SD BIOLINE TB Ag MPT 64 Rapid ®” is a simple and rapid immunochromatographic test used in differentiating Mycobacteria into Mycobacterium tuberculosis (NTM). The 100 sputa were obtained from patients suspected to be infected with tuberculosis and presented themselves to hospitals for check-up and treatment were involved in the study. The samples were cultured in a class III Biosafety cabinet and level III biosafety practices were followed. Forty isolates were obtained from the cultured sputa, and there were identified as Acid-fast bacilli (AFB) using Zeihl-Neelsen acid-fast stain. All the isolates (AFB positive) were then subjected to the SD BIOLINE Analyses. A total of 31 (77.5%) were characterized as MTBC, while nine (22.5%) were NTM. The total turnaround time for the rapid assay was just 30 minutes as compared to a few days of phenotypic and genotypic method. It was simple, rapid and reliable test to differentiate MTBC from NTM.

Keywords: culture, mycobacteria, non tuberculous mycobacterium, SD Bioline

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49 Doubled Haploid Production in Wheat Using Imperata cylindrica Mediated Chromosome Elimination Technique

Authors: Madhu Patial, Dharam Pal, Jagdish Kumar, H. K. Chaudhary

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Doubled haploid breeding serves as a useful technique in wheat improvement by providing instant and complete homozygosity. Of the various techniques employed for haploid production chromosome elimination has a large scale practical application in wheat improvement. Barclay (1975) initiated the technique in wheat by crossing wheat variety Chinese spring with Hordeum bulbosum, but due to presence of the dominant crossability inhibitor genes Kr7 and Kr2 in many wheat varieties, the technique was however genotypic specific. The discovery of wheat X maize system of haploid production being genotype non-specific is quite successful but still maize needs to be grown in greenhouse to coincide flowering with wheat crop. Recently, wheat X Imperate cylindrica has been identified as a new chromosome mediated DH approach for efficient haploid induction. An experiment to use this technique in wheat was set up by crossing six F1s and two three way F1s with Imperata cylindrica. The data was recorded for the three component traits of haploid induction viz., seed formation, embryo formation and regeneration frequency. Variation among wheat F1s was observed and higher frequency for all the traits were recorded in cross HD 2997/2*FL-8/DONSK-POLL and KLE/BER/2*FL-8/DONSK-POLL.

Keywords: wheat, haploid, imperata cylindrica, chromosome elimination technique

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48 Association of MIR146A rs2910164 Variation with a Predisposition to Sporadic Breast Cancer in a Pakistani Cohort

Authors: Mushtaq Ahmad, Bashir Rahman, Taqweem-ul-Haq, Fazal Jalil, Aftab Ali Shah

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Single nucleotide polymorphisms (SNPs) in genes coding for microRNAs (miRNAs) play a pivotal role in the progression of breast cancer (BC). We investigated the association of miR-146a rs2910164 G/C polymorphism with the risk of BC in the Pakistani population. The miR-146a rs2910164 polymorphism was genotyped in 300 BC-cases and 300 age- and gender-matched healthy controls using T-ARMS-PCR. Genotype and allele frequencies were calculated, and the association between genotypes and the risk of BC was calculated by odds ratios (OR) and confidence intervals (95%). A significant difference in genotypic frequencies (χ2=63.10; p ≤ 0.0001) and allelic frequencies (OR=0.3955 (0.3132-0.4993); p ≤ 0.0001) was observed between cases and controls. Furthermore, we also found that miR-146 rs2910164 CC homozygote increased the risk of breast cancer in the dominant (OR=0.2397 (0.1629-0.3526); p=0.0001; GG vs GC+CC) and recessive (OR=2.803 (1.865- 4.213); P ≤ 0.0001; CC vs GC+GG) inheritance models. In summary, miR-146a rs2910164 G/C is significantly associated with BC in the Pakistani population. To our knowledge, this is the first study that assessed MIR146a rs2910164 G > C SNP in Pakistani population. By analyzing the secondary structure of MIR146A variant, a significant structural modification was noted. Study with a larger sample size is needed to further confirm these findings.

Keywords: breast cancer, MIR146A, microRNA, SNP

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47 Isolation and Screening of Fungal Strains for β-Galactosidase Production

Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

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Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: beta-galactosidase, enzyme, fungal, isolation

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46 Functional Profiling of a Circular RNA from the Huntingtin (HTT) Gene

Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

Abstract:

Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

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45 Genomic and Proteomic Variation in Glycine Max Genotypes towards Salinity

Authors: Faheema Khan

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In order to investigate the influence of genetic background on salt tolerance in Soybean (Glycine max) ten soybean genotypes released/notified in India were selected. (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712). The 10-day-old seedlings were subjected to 0, 25, 50, 75, 100, 125, and 150 mM NaCl for 15 days. Plant growth, leaf osmotic adjustment, and RAPD analysis were studied. In comparison to control plants, the plant growth in all genotypes was decreased by salt stress, respectively. Salt stress decreased leaf osmotic potential in all genotypes however the maximum reduction was observed in genotype Pusa-24 followed by PK-416 and Pusa-20. The difference in osmotic adjustment between all the genotypes was correlated with the concentrations of ion examined such as Na+ and the leaf proline concentration. These results suggest that the genotypic variation for salt tolerance can be partially accounted for by plant physiological measures. The genetic polymorphisms between soybean genotypes differing in response to salt stress were characterized using 25 RAPD primers. These primers generated a total of 1640 amplification products, among which 1615 were found to be polymorphic. A very high degree of polymorphism (98.30%) was observed. UPGMA cluster analysis of genetic similarity indices grouped all the genotypes into two major clusters. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings. Our results show that RAPD technique is a sensitive, precise and efficient tool for genomic analysis in soybean genotypes.

Keywords: glycine max, NaCl, RAPD, proteomics

Procedia PDF Downloads 551