Discrimination Of Genotypes Research Articles

Molecular markers useful to discriminate Coffea arabica cultivars with high genetic similarity

New cultivars are released every year to meet market demands. However, in species with a narrow genetic base, such as Coffea arabica, the cultivars are closely related and phenotypically similar. This hinders the accurate discrimination of genotypes using morphological descriptors in distinctness, uniformity, and stability (DUS) testing, which is required for the registration and protection of new cultivars. In this sense, molecular markers are an auxiliary tool for accurate and precise discrimination of cultivars. This study aimed to verify the informative capacity and effectiveness of a molecular marker set to discriminate among C. arabica varieties, create a database of DNA profiles and allele frequencies, analyze the genetic diversity in this collection, and explore genetic kinships. Thirty-four C. arabica cultivars/progenies, which belong to the Brazilian Cultivar Trial, were analyzed using 31 microsatellite markers. Markers with weak bands were removed, and of the remaining, 74.07% were polymorphic and revealed 47 alleles. The obtained molecular profiles revealed segregation between and within cultivars/progenies, and genetic variability was observed between all the cultivars/progenies. Sixteen markers were selected for dendrogram construction and for fingerprinting analysis of the cultivars. The ability of these markers to detect varietal mixture and analyze diversity between and within cultivars was also discussed in detail. The results demonstrated the effectiveness of markers in distinguishing related genotypes from those with similar phenotypic traits. This biotechnological tool will assist breeders in DUS testing of cultivars.

Strategy for selection of soybean genotypes tolerant to drought during germination.

Water deficit is the main reason for instability in the context of soybean culture. The development of strategies for the selection of more tolerant genotypes is necessary. These strategies include the use of polyethylene glycol 6000 solutions (PEG-6000) for conducting the germination test under conditions of water restriction. Thus, the objective of this study was to determine the osmotic potential and the main characteristics that promote the discrimination of soybean genotypes with regard to water stress tolerance during germination and the vigor test. Thirteen soybean cultivars were used. The seeds were allowed to germinate on sheets of germitest paper moistened in solution with PEG-6000, simulating different levels of water availability, which is expressed as osmotic potential (0.0, -0.2, -0.4, and -0.6 MPa). We assessed germination, length, and dry mass for seedlings and seeds, as well as reserve dynamics. Germination and variables related to the dynamics of reservation have great influence on the expression of variability in environments under stress. Among the different osmotic potentials, the -0.2 MPa was the most efficient for the expression of genetic variability among the cultivars. Conducting the germination test with PEG-6000 solution to -0.2 MPa was efficient for selecting soybean cultivars tolerant to water stress. This was accomplished by evaluating the percentage of germination, along with variables related to the dynamics of reservation.

Genotyping and Molecular Identification of Date Palm Cultivars Using Inter-Simple Sequence Repeat (ISSR) Markers.

Molecular markers are credible for the discrimination of genotypes and estimation of the extent of genetic diversity and relatedness in a set of genotypes. Inter-simple sequence repeat (ISSR) markers rapidly reveal high polymorphic fingerprints and have been used frequently to determine the genetic diversity among date palm cultivars. This chapter describes the application of ISSR markers for genotyping of date palm cultivars. The application involves extraction of genomic DNA from the target cultivars with reliable quality and quantity. Subsequently the extracted DNA serves as a template for amplification of genomic regions flanked by inverted simple sequence repeats using a single primer. The similarity of each pair of samples is measured by calculating the number of mono- and polymorphic bands revealed by gel electrophoresis. Matrices constructed for similarity and genetic distance are used to build a phylogenetic tree and cluster analysis, to determine the molecular relatedness of cultivars. The protocol describes 3 out of 9 tested primers consistently amplified 31 loci in 6 date palm cultivars, with 28 polymorphic loci.

Effect of the CEP72 Genotype and CYP3A5-Mediated Metabolism in Predicting Vincristine-Associated Peripheral Neuropathy

Introduction:Chemotherapy induced peripheral neuropathy (CIPN) is a recognized clinical outcome of vinca alkaloid based therapy that frequently results in dose reduction or therapy discontinuation, but the determinants of interpatient variability remain unpredictable.Previous pharmacogenetics studies have demonstrated that vincristine is metabolized through the CYP3A system with the CYP3A5*3 variant (GG allele at rs776746) having the least efficacy in metabolizing vincristine to its inactive metabolite M1 and conferring higher risk of neurotoxicity. Similarly, a single nucleotide polymorphism (SNP) in the promoter of the CEP72 gene (TT allele at rs924607) has been described as being significantly associated with increased risk and severity of vincristine-associated peripheral neuropathy in the pediatric population. Our recent studies have further studied the incidence of this risk allele and its correspondence with CIPN in the adult population treated with vincristine. This study aimed to investigate if an additive SNP analysis, both for CYP3A5*3 and CEP72, would determine increased predictability of CIPN in patients treated with vinca alkaloids.Methods:A retrospective case-control study of patients was completed in patients that were exposed to vinca alkaloids (vincristine, vinorelbine, and vinblastine) at Geisinger Medical Center from 2007-2015. Available samples of these patients were identified in the Geisinger MyCode bio banking project and the SNP of interest was genotyped using the TaqMan® SNP Genotyping Assay, with genotypic discrimination performed using SDS software (Applied Biosystems). Cases of CIPN were confirmed with retrospective chart review. Statistical analysis was conducted using SAS (SAS Institute Inc.) software, version 9.4. All genotyping and chart review was conducted by personnel blinded to sample identity.Results:A total of 121 patients had genotype mapping completed, of which 60 patients received vincristine, 33 received vinblastine, and 28 received vinorelbine. CIPN was detected in 35.5% of the study population. The CEP72 risk allele (TT) was detected in 14.9% (n=18) of the cohort and the CYP3A5*3 risk allele (GG) was detected in 89.2% (n=108) of the cohort, with similar incidence in each drug class. The median age of this population was 61 years of age with 52.9% (n=64) being males.Multivariate logistic regression analysis, controlling for age and sex, did not demonstrate a significant model for the recessive genetic model of both SNPs with all vinca alkaloids (p=0.33), vincristine alone (p=0.26), vinorelbine alone (p=0.98), or vinblastine alone (p=0.97). The incidence of the inactive G/G allele for CYP3A5*3 was seen in 87.5% (n=21) of the vincristine cohort with neuropathy,The incidence of neuropathy was higher in patients with the TT genotype (75%) in patients receiving vincristine therapy than with the CT or CC genotype (35.7%). As consistent with prior findings, a similar dominant effect was not detected in vinorelbine or vinblastine groups. In the vincristine population (n=60), diseases that were treated included diffuse large B-cell lymphoma (n=29), acute lymphoblastic leukemia (n=6), oligoastrocytoma (n=5), and Ewing's sarcoma (n=2). No disease specific variation in neuropathy incidence and association with TT genotype was detected. Patients received a median three doses of vincristine at 2 mg each (total 6 mg) before detection of CIPN. Four patients (6.67%) of the patients required dose adjustment of chemotherapy, including dose reduction (n=2) or early discontinuation (n=2).Conclusions:This study demonstrates that the CEP72 genotype confers predictive modeling for patients receiving vincristine-based therapy and that including CYP3A5*3 analysis in an additive model is not associated with an increased predictive model for CIPN in patients receiving vinca alkaloid based therapy. In our population, the high incidence of the inactive G/G allele of CYP3A5*3 made it difficult to discern a significant effect in patients that experienced neuropathy. Larger studies are needed to include a more diverse population to confirm this predictive association and to understand if this can help guide precision-based chemotherapy administration and favorably impact disease prognosis. DisclosuresNo relevant conflicts of interest to declare.

Identifying Barley Genotypes for Optimum Input Conditions in the WANA Regions

Abstract Barley (Hordeum vulgare L.) is an important cereal crop for its high demand for grain feed and forage/grazing for the animals as well as for human food in the WANA region under the low rainfall situations and thus creating an urgent need for developing high yielding barley genotypes. A total of 542 hulled and hulless genotypes with seven checks were evaluated at three locations with diverse agro-ecological conditions, Terbol in Lebanon and Marchouch and Allal Tazi in Morocco. The aim of this study was to understand the nature of genotype × environment interaction (GEI), quantify the genotypic variability and identify high yielding genotypes. The mixed models were fitted to evaluate heritability and predicted means to identify genotypes with specific adaptation to the locations using GGE-biplot. GEI across locations was significant for days to heading, days to maturity, plant height, spike length and grain yield. On a trial-wise analysis, days to maturity was most heritable (49 – 50% broad sense heritability on mean basis) while the grain yield was the least (5 – 13%). The genotype G427 (4812 kg/ha) at Marchouch, the check VMorales (4889 kg/ha) at Allal Tazi and G528 (6995 kg/ha) at Terbol were the highest yielding genotypes. Several hulled and hulless genotypes with higher grain yield and early flowering time in the three environments were identified for utilization by the national programs globally. The test locations, Marchouch and Allal Tazi, were found comprising one mega environment while Terbol showed maximum discrimination of genotypes for grain yield.

6 HCV genotyping 9G test and its comparison with VERSANT HCV genotype 2.0 assay (LiPA) for the hepatitis C virus genotyping

In this article, we describe the 6 HCV Genotyping 9G test and its evaluation by using clinical samples and plasmid DNA standards. In tests with 981 plasmid DNA standards, the 6 HCV Genotyping 9G test showed higher than 92.5% sensitivity and 99.4% specificity. The 6 HCV Genotyping 9G test was compared with the VERSANT HCV Genotype 2.0 assay (LiPA 2.0) for detection and discrimination of HCV genotypes in clinical samples. The results of both tests were verified by genomic sequencing. The 6 HCV Genotyping 9G test demonstrated a 100% agreement with the sequencing results, which was higher than LiPA 2.0. These results indicate that the 6 HCV Genotyping 9G test can be a reliable, sensitive, and accurate diagnostic tool for the correct identification of HCV genotypes in clinical specimens. 6 HCV Genotyping 9G test can genotype six HCV types in 1 PCR in 30min after PCR amplification. The 6 HCV Genotyping 9G test, thus provide critical information to physicians and assist them to apply accurate drug regimen for the effective hepatitis C treatment.

High‐resolution melt analysis without DNA extraction affords rapid genotype resolution and species identification

Extracting and sequencing DNA from specimens can impose major time and monetary costs to studies requiring genotyping, or identification to species, of large numbers of individuals. As such, so-called direct PCR methods have been developed enabling significant savings at the DNA extraction step. Similarly, real-time quantitative PCR techniques (qPCR) offer very cost-effective alternatives to sequencing. High-resolution melt analysis (HRM) is a qPCR method that incorporates an intercalating dye into a double-stranded PCR amplicon. The dye fluoresces brightly, but only when it is bound. Thus, after PCR, raising the temperature of the amplicon while measuring the fluorescence of the reaction results in the generation of a sequence-specific melt curve, allowing discrimination of genotypes. Methods combining HRM (or other qPCR methods) and direct PCR have not previously been reported, most likely due to concerns that any tissue in the reaction tube would interfere with detection of the fluorescent signal. Here, we couple direct PCR with HRM and, by way of three examples, demonstrate a very quick and cost-effective method for genotyping large numbers of specimens, using Rotor-Gene HRM instruments (QIAGEN). In contrast to the heated-block design of most qPCR/HRM instruments, the Rotor-Gene's centrifugal rotor and air-based temperature-regulation system facilitate our method by depositing tissues away from the pathway of the machine's fluorescence detection optics.

Investigating morpho-physiological traits and agro-meteorological indices in green gram [Vigna radiata L. Wilczek] for terminal heat tolerance

Forty genotypes of greengram were studied to ascertain the genetic variability and trait association among some important morpho-physiological traits and agro-meterological indices under heat stress condition. The results indicated that both GCV and PCV estimates were high for photo thermal index, heat use efficiency and seed yield. High heritability coupled with moderate genetic advance as per cent of mean was recorded for photo-thermal unit and relative temperature depression indicated that involvement of both additive and non-additive type of gene action and possibilities of effective selection for improvement of these traits. Seed yield showed significant and positive association with days to maturity, growing degree days, relative temperature depression and heat use efficiency. Based on variability, association and path analysis; heat use efficiency, maturity, photo thermal index and growing degree days were found most contributing indices/ traits should be considered as selection criteria for discrimination of outstanding greengram genotypes under heat stress condition.

High Resolution Melting (HRM) applied to wine authenticity

Wine authenticity methods are in increasing demand mainly in Denomination of Origin designations. The DNA-based methodologies are a reliable means of tracking food/wine varietal composition. The main aim of this work was the study of High Resolution Melting (HRM) application as a screening method for must and wine authenticity. Three sample types (leaf, must and wine) were used to validate the three developed HRM assays (Vv1–705bp; Vv2–375bp; and Vv3–119bp). The Vv1 HRM assay was only successful when applied to leaf and must samples. The Vv2 HRM assay successfully amplified all sample types, allowing genotype discrimination based on melting temperature values. The smallest amplicon, Vv3, produced a coincident melting curve shape in all sample types (leaf and wine) with corresponding genotypes. This study presents sensitive, rapid and efficient HRM assays applied for the first time to wine samples suitable for wine authenticity purposes.

Genotypic discrimination of <i>Aspergillus fumigatus</i> strain from related species within section fumigati

The aim was to make an exact diagnosis of 20 strains using molecular biological methods which were isolated from the atmosphere of the inpatient rooms in the Oncology and other departments of the Ege University Medical Faculty Hospital, and identified as Aspergillus fumigatus through phenotypic tests, and to determine their antibiotic susceptibility patterns. It was confirmed that the 20 phenotypically-identified A. fumigatus strains belonged to the section Fumigati after they were tested by the ITS-PCR method. Their sequence analysis was performed and the results sent to the NCBI GenBank, and their accession numbers were obtained. For their exact diagnosis at the species level, the β-tub (β-tubulin) and rodA (RodletA) genes were examined with the multiplex PCR. Anti-fungal susceptibility of the 20 strains was determined according to the M38-A2 standards. As a result of ITS-PCR, it was confirmed that 19 of the 20 strains identified as A. fumigatus through the phenotypic methods belonged to the section Fumigati. However, after the detection of β-tub and rodA genes, all 20 strains were identified as A. fumigatus. Although the results of the phenotypic and molecular biological tests applied to filamentous fungi do not often overlap, in this study, the results obtained from the molecular analysis confirmed the results of the phenotypic tests. However, 1 of the 20 strains phenotypically-identified as A. fumigatus was identified as Penicillium spp. as a result of ITS-PCR and sequence analysis. On the other hand, the profile obtained from β-tub and rodA tests indicated that the strain was A. fumigatus. Based on these results, this strain is thought to belong to the Aspergilloides genus which has the features of both genera.

Insight into Phenotypic and Genotypic Discrimination of Bacterial Pathogens: From Pre-Genomic to Post-Genomic Era

Insight into Phenotypic and Genotypic Discrimination of Bacterial Pathogens: From Pre-Genomic to Post-Genomic Era

Genotype Discrimination Using Facial and Body Anthropometry among Hausas

Individual's genotype can be classified as normal, carrier or sickle cell patient when it is AA, AS and SS respectively. Several researches attempted to discriminate the sickle cell patient with normal subject using anthropometry, however less attention is paid to the discrimination of an individual with normal genotype from carrier especially among Hausa population. The present study aimed at discriminating between normal and carrier from Hausa ethnic population using body and facial anthropometry. A total of 219 participants with mean age of 20.41 ± 2.94 years were involved. The genotype analyses revealed 197 and 22 participants as normal and carrier respectively. Direct anthropometry was used to measure the height, weight and 17 facial features. Independent sample t – test was used for the test of differences. Binary logistic regression (BLR) analysis was used to obtain a model for genotype discrimination. The predicted probabilities of BLR for the variables that contributed best to the prediction were analyzed using Receiver Operating Characteristic (ROC) curve. The results indicated that none of the facial features showed significant difference between normal and carrier. Of the three parameters of height, weight and BMI only weight exhibited significant differences between the groups. The model prediction accuracy with all variables in the equation was 90.9% with only 19.7% contribution to the prediction (Nagelkerke R Square = 0.197). Using stepwise BLR only weight was included in the equation (GENOTYPE= 0.058×WT −1.001, cut value is 0.5, Wald statistics= 5.022, p = 0.025) with overall accuracy of 90%. The ROC curve analyses for the weight only showed the area under curve (AUC) of 0.654 indicating 65.4% best prediction probability of the model. In conclusion, weight demonstrated a superior potential in discriminating the genotype over other variables considered in the study among Hausas of Nigeria.

Molecular analysis and MIRU-VNTR typing of Mycobacterium avium subsp. avium, ‘hominissuis’ and silvaticum strains of veterinary origin

Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and ‘M. avium subsp. hominissuis’ (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds.Strains were tested for the presence of large sequence polymorphism LSPA17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined.LSPA17 was present only in 19.9% of the strains. All LSPA17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific.Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains.

A novel exonuclease (TaqMan) assay for rapid haptoglobin genotyping.

Haptoglobin is an acute-phase binding protein that scavenges free hemoglobin. The human haptoglobin gene (HP) is polymorphic with two main alleles, haptoglobin allele 1 (Hp1) and haptoglobin allele 2 (Hp2). The smaller Hp1 allele features no duplication and consists of four exons, whereas the larger Hp2 allele, containing a 1.7 kb duplication, consists of six exons, with the fifth and sixth being highly homologous to exons 3 and 4 of Hp1. We designed an exonuclease (TaqMan) assay targeting single nucleotide differences between the homologous regions of Hp1 and Hp2. The assay contained one probe specifically binding to a site in intron 4 of Hp2, and another probe binding equally to intron 4 of Hp1 and intron 6 of Hp2. Measurement of post-PCR fluorescence allowed unambiguous discrimination of HP genotypes. Comparison with genotypes obtained by a method based upon allele-specific primers yielded fully corresponding results. The new HP genotyping method is fast, reliable, does not require real-time instruments and may be especially useful for high-throughput genotyping.

Screening for Abiotic Stress Tolerance in Rice: Salt, Cold, and Drought.

Rice (Oryza sativa) is the primary source of food for more than half of the world population. Most rice varieties are severely injured by abiotic stresses, with strong social and economic impact. Understanding rice responses to stress may help breeding for more tolerant varieties. However, papers dealing with stress experiments often describe very different experimental designs, thus making comparisons difficult. The use of identical setups is the only way to generate comparable data. This chapter is organized into three sections, describing the experimental conditions established at the Genomics of Plant Stress (GPlantS) unit of ITQB to assess the response of rice plants to three different abiotic stresses--high salinity, cold stress, and drought. All sections include a detailed description of the materials and methodology, as well as useful notes gathered from the GPlantS team's experience. We use rice seedlings as plants at this stage show high sensitivity to abiotic stresses. For the salt and cold stress assays we use hydroponic cultures, while for the drought assay plants are grown in soil and subjected to water withholding. All setups enable visual score determination and are suitable for sample collection along the imposition of stress. The proposed methodologies are simple and affordable to implement in most labs, allowing the discrimination of several rice genotypes at the molecular and phenotypic level.

A rapid test for detection of tribenuron-methyl resistance in sunflower

A rapid laboratory test for detection of tribenuron-methyl resistant sunflower genotypes was developed. Four homozygous and four heterozygous tribenuron-methyl resistant hybrids, as well as a susceptible one were grown on MS medium supplemented with different concentrations of herbicide (2.0 µM, 2.5 µM, 3.0 µM, 3.5 µM and 4.0 µM) and with pH either 7 or 8. The effect of medium pH and herbicide concentration on above-ground part and root mass of sunflower seedlings was observed. The test enabled visual discrimination between resistant and susceptible genotypes in 5 days, and discrimination between homozygous and heterozygous resistant genotypes in 12 days. All tested herbicide concentrations and morphological parameters were found to be suitable for discrimination of tribenuron-methyl susceptible genotypes. The best pH and herbicide concentration combination and morphological parameter for discrimination between homozygous and heterozygous resistant sunflower genotypes were 3.0 µM at pH 7 and root mass, respectively.

Biplot analysis of phenotypic stability in upland cotton genotypes in Mato Grosso.

Seed cotton yield is a trait governed by multiple genes that cause changes in the performance of genotypes depending on the cultivation environment. Breeding programs examine the genotype x environment interaction (GE) using precise statistical methods, such as AMMI (additive main effects and multiplicative interaction) and GGE biplot (genotype main effects + genotype x environment interaction). The AMMI method combines the analysis of variance and principal components, to adjust the main effects (genotypes and environments) and the effects of GE interaction, respectively. The GGE biplot groups the genotype additive effect together with the multiplicative effect of the GE interaction, and submits both of these to the principal components analysis. The aim of this study was to investigate the association between the AMMI and GGE biplot methods and select cotton genotypes that simultaneously showed high productivity of seed cotton and stability in Mato Grosso environments. Trials were conducted with cotton cultivars in eight environments across Mato Grosso State in the 2008/2009 crop season. The experiment used a randomized block design with 16 genotypes and four replicates per genotype x environment combination. Data for seeds cotton productivity were analyzed by AMMI and GGE biplot methods. Both methods were concordant in the discrimination of environments and genotypes for phenotypic stability. The genotypes BRS ARAÇÁ and LD 05 CV had high seed cotton productivity and phenotypic stability, and could be grown in all environments across Mato Grosso State.

GENETIC DIVERGENCE AMONG Passiflora cristalina Vanderpl & Zappi. GENOTYPES BASED ON FLOWER AND FRUIT CHARACTERISTICS

ABSTRACT This study aimed to evaluate the genetic divergence among Passiflora cristalina genotypes and quantify the relative contribution of 30 flower and fruit characteristics, seeking to support the preservation and characterization of genetic resources of the species for preservation and use in future breeding programs. We evaluated 150 fruit and 150 flowers collected in 15 genotypes with naturally occurring in the municipality of Alta Floresta, MT. The characterization of genotypes was performed through 30 morphological characteristics of flowers and fruits, 21 of these for flower and 9 for fruit. Data were evaluated using the principal components and cluster methods obtained by UPGMA method from the similarity matrix (Euclidian mean distance), using the Genes software. By principal component analysis, it has been found that the first three components have absorbed 52.11% of the accumulated variation. The characteristics that most contributed to the discrimination of genotypes were fresh fruit weight, stigma length, length of corona filaments, fruit width, petal width and pulp weight, which are more responsive for the selection of P.cristalina genotypes. Smaller contributions to diversity were obtained from anther width, bract width and fruit length. The smallest contributions for diversity were obtained from the following characteristics: anther width, bract width and fruit length. Through UPGMA clustering method, it was found that there is a large genetic divergence among genotypes analyzed because all genotypes were grouped with over 50% of dissimilarity. This study identified genotypes 4, 5 and 9 as the most divergent and therefore the most suitable for breeding in future breeding programs and genetic conservation of the species.

Number of repetitions for evaluating technological traits in cotton genotypes.

With the changes in spinning technology, technological cotton traits, such as fiber length, fiber uniformity, fiber strength, fineness, fiber maturity, percentage of fibers, and short fiber index, are of great importance for selecting cotton genotypes. However, for accurate discrimination of genotypes, it is important that these traits are evaluated with the best possible accuracy. The aim of this study was to determine the number of measurements (repetitions) needed to accurately assess technological traits of cotton genotypes. Seven experiments were conducted in four Brazilian States (Ceará, Rio Grande do Norte, Goiás, and Mato Grosso do Sul). We used nine brown and two white colored fiber lines in a randomized block design with four replications. After verifying the assumptions of residual normality and homogeneity of variances, analysis of variance was performed to estimate the repeatability coefficient and calculating the number of repetitions. Trials with four replications were found to be sufficient to identify superior cotton genotypes for all measured traits except short fiber index with a selective accuracy >90% and at least 81% accuracy in predicting their actual value. These results allow more accurate and reliable results in future researches with evaluating technological traits in cotton genotypes.

Inter-Specific Variation Studies among Nephrolepis using SDS-PAGE

The present study was aimed to reveal the biochemical similarity and varaition among the three taxonomically confused species Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret using SDS-PAGE analysis. To reveal the inter-speciifc variation among the selected Nephrolepis, SDS-PAGE was carried out to obtain protein bands following the method described by Anbalagan. Multiple regions (8) of activity were observed from protein electrophoretic system of Nephrolepis. A total of 19 bands with various Rf values and molecular weight were demonstrated in the SDS-PAGE gel system of Nephrolepis. Among the three species of Nephrolepis, N. exaltata showed maximum number of protein bands (9) followed by N. multiflora (6) and N. auriculata (4). Each region expressed different proteins which act as representative of the expression of a particular gene in the studied species of Nephrolepis. The similarity indices were calculated and cladogram was constructed based on the protein profiles of Nephrolepis and revealed the similarities and variation among the studied Nephrolepis species. The results obtained in this work also showed that SDS-PAGE analysis can provide an easy, low cost and quick way for the identification of inter-specific variation among the selected Nephrolepis species. SDS-PAGE analysis provided strong basis for the discrimination of genotypes on the basis of specific polypeptide fragments.