ENABLING MARKER-ASSISTED BREEDING (MAB) FOR BLUSH IN PEACH [Prunus persica (L.) Batsch]

ABSTRACT Phenotyping is a crucial component for using DNA-based tools in gene discovery and marker development. Phenotypic and genotypic data are essential for linking genetic variation with biological function, thus documenting gene function. However, phenotypic data gathering is not keeping pace with the immensely increasing amount of available genomic information, brought forth by current Next Generation Sequencing technologies. Standardized phenotyping protocols for peach [Prunus persica (L.) Batsch] have been developed for 6 productivity traits (on the tree) and 16 fruit quality traits. Documentation of fruit quality phenotypes has been performed applying developed standardized phenotyping protocol in two seasons, at nine locations, on 513 peach and almond accessions, cultivars, advanced selections, lines, and or populations. In this study blush (i.e. red skin pigmentation) inheritance and associated genes were investigated extensively. Blush is an important trait for marketing peaches. The red skin pigmentation develops through the flavonoid and anthocyanin pathways. Both genetic and environmental stimuli and their interaction (genotype x environment) control the regulation of this pathway. Sunlight induces the progression of red skin development. To study the genetic control of blush in peach a controlled cross between two cultivars with contrasting phenotypes, Zin Dai (~30% red) and Crimson Lady (~100% red), was made. One F1 hybrid, BY02p4019, with intermediate levels of blush (~65% red) was selfed to generate a segregating F2 blush population (ZC2). The segregating population was phenotyped for blush for four years (2007, 2008, 2010, 2011) using a visual rating scale (0-5) and in 2011 using a colorimeter (L*, a*, b*). Twenty-five individuals, exhibiting a blush range from 0 (0% red) to 5 (100% red) and a normal distribution for this trait, were genotyped with an IPSC 9K peach SNP array v1. A ZC2 genetic linkage map was constructed with 1,335 SNP markers, comprising 14 linkage groups. This map covers a genetic distance of ~452.51 cM with an average marker spacing of 2.38 cM/marker and an average number of 95 markers per LG. A major QTL for blush has been located on LG3, denoted Blush.Pp.ZC-3.1. This QTL spanned 21-41cM on LG3 and explained on average 72% of phenotypic variation for the trait. QTL analysis for four different seasons confirms the identification of this major QTL for blush in peach, and supports its stability. Three minor QTL\u27s were located on LG\u27s 4, and 7 indicating the presence of minor genes involved with blush development. Candidate genes involved in skin and flesh coloration of cherry (PavMYB10) and apple (MdMYB10) are located within the interval of the major QTL on LG3 suggesting the same genetic control for color development in the Rosaceae family. A standardized protocol for collecting phenotypic data in peach will facilitate discovery of genes associated with fruit quality and other agronomically important traits

From Establishment to Final Independence: A Study of the National Archives of the United States of America from 1934–1985

This thesis is a study of the National Archives of the United States from the institution’s establishment in 1934 under the presidency of Franklin Roosevelt to becoming the National Archives and Record Administration in 1985. The Archives during the 1930’s and 1940’s functioned as an independent agency, until the Archives lost their independence under the Hoover Commission. In 1949 the Archives became part of the newly formed General Services Administration. During the 1950’s and 1960’s National Archives helped change the archival profession. Furthermore, we see how the two independence movements in the 1960’s and 1980’s that were ultimately successful in the restoration of the Archives status as an independent agency

Roles and reflections : using team teaching to improve literacy teaching

This article describes the development of a team teaching model by a first grade team consisting of classroom and Title 1 teachers. This project grew out of a study group designed to look at best practice in literacy education and evolved in response to student achievement data. Four factors influenced the progress of the project including peer collaboration and demonstration, professional reading, reflection by teachers and the facilitator, and leadership roles. The role of the facilitator receives special attention, as – unlike most efforts at classroom change involving a grade level team – this effort began within the team, and without an external facilitator

Genetic Determinism of Xanthomonas arboricola pv. pruni (Xap) Resistance, Fruit Quality, and Phenological Traits in Peach and Incorporation of Marker-Assisted Selection (MAS) in the University of Arkansas Peach and Nectarine Breeding Program

For over two decades the incorporation of marker-assisted selection (MAS) has been discussed as an innovative approach to increase peach breeding efficiency. Although hundreds of quantitative trait loci (QTLs) have been identified, only a few have been converted into usable MAS tools. This highlights a disconnect between genomic discoveries and practical application in breeding programs, which has delayed use of MAS. In this dissertation, three sequential steps were performed with the objective to bridge this gap and develop breeding-relevant DNA tests for MAS of fruit bacterial spot resistance (Xap1 and Xap6), fruit quality [blush (Rf), acidity (D), and acidity and soluble solid content (G7Flav)], and phenological traits [maturity date (G4mat), fruit type (G), and flesh color (Y)] across four RosBREED peach breeding programs. First, previously identified QTLs were further investigated and 32 SNPs were grouped into haplotypes and validated for association with breeding-relevant trait differences across RosBREED germplasm. The SNPs were divided into two mini-arrays and advanced breeding material from all programs were outsourced for design and testing. The validated SNP loci were used in marker-assisted parent selection (MAPS) in 2013-2015 at the University of Arkansas (UA) program to combine horizontal Xap resistance with high fruit quality spanning the season. Secondly, four of the SNP-based tests (Xap1, Xap6, G, and Y loci) were converted into sequence length polymorphism-based (SLP-based) tests (Ppe-XapF1-SSR, Ppe-XapF6-SSR, indelG, and PpCCD4b-SSR) and screened on the UA RosBREED germplasm and 22 additional F1 populations. Results were compared across both DNA tests to identify the most efficient genotyping approach for each trait. In 2015, two of the SLP-based DNA tests, the indelG (pubescent vs. glabrous) and PpCCD4b-SSR (white vs. yellow flesh) were advanced to test in marker-assisted seedling selection (MASS). In the final step, QTL analysis was conducted for Xap fruit, Xap leaf, and Xap leaf-assay resistance along with seven fruit quality and phenological traits using the Pedigree-Based Analysis (PBA) approach and the UA RosBREED pedigree. Overall 20 QTLs were identified, 10 for Xap resistance and 10 for quality and phenological traits. These 20 QTLs are optimal targets for future DNA test development, validation, and use in MAS

Rational Design, Synthesis and Biological Evaluation of Pyrimidine-4,6-diamine derivatives as Type-II inhibitors of FLT3 Selective Against c-KIT.

FMS-like Tyrosine Kinase 3 (FLT3) is a clinically validated target for acute myeloid leukemia (AML). Inhibitors targeting FLT3 have been evaluated in clinical studies and have exhibited potential to treat FLT3-driven AML. A frequent, clinical limitation is FLT3 selectivity, as concomitant inhibition of FLT3 and c-KIT is thought to cause dose-limiting myelosuppression. Through a rational design approach, novel FLT3 inhibitors were synthesized employing a pyridine/pyrimidine warhead. The most potent compound identified from the studies is compound 13a, which exhibited an IC50 value of 13.9 ± 6.5 nM against the FLT3 kinase with high selectivity over c-KIT. Mechanism of action studies suggested that 13a is a Type-II kinase inhibitor, which was also supported through computer aided drug discovery (CADD) efforts. Cell-based assays identified that 13a was potent on a variety of FLT3-driven cell lines with clinical relevance. We report herein the discovery and therapeutic evaluation of 4,6-diamino pyrimidine-based Type-II FLT3 inhibitors, which can serve as a FLT3-selective scaffold for further clinical development

Effect of trunk exercise upon lumbar IVD height and vertebral compliance when performed supine with 1 g at the CoM compared to upright in 1 g

Background: Spinal unloading in microgravity is associated with stature increments, back pain, intervertebral disc (IVD) swelling and impaired spinal kinematics. The aim of this study was to determine the effect of lateral stabilization, trunk rotation and isometric abdominal exercise upon lumbar IVD height, and both passive and active vertebral compliance when performed supine on a short-arm human centrifuge (SAHC)-a candidate microgravity countermeasure-with 1 g at the CoM, compared to that generated with equivalent upright exercise in 1 g. Methods: 12 (8 male) healthy subjects (33.8 ± 7 years, 178.4 ± 8.2 cm, 72.1 ± 9.6 kg) gave written informed consent. Subjects performed three sets of upper body trunk exercises either when standing upright (UPRIGHT), or when being spun on the SAHC. Lumbar IVD height and vertebral compliance (active and passive) were evaluated prior to SAHC (PRE SAHC) and following the first SAHC (POST SPIN 1) and second Spin (POST SPIN 2), in addition to before (PRE UPRIGHT), and after upright trunk exercises (POST UPRIGHT). Results: No significant effect upon IVD height (L2-S1) when performed UPRIGHT or on the SAHC was observed. Trunk muscle exercise induced significant (p < 0.05) reduction of active thoracic vertebral compliance when performed on the SAHC, but not UPRIGHT. However, no effect was observed in the cervical, lumbar or across the entire vertebral column. On passive or active vertebral compliance. Conclusion: This study, the first of its kind demonstrates that trunk exercise were feasible and tolerable. Whilst trunk muscle exercise appears to have minor effect upon IVD height, it may be a candidate approach to mitigate-particularly active-vertebral stability on Earth, and in μg via concurrent SAHC. However, significant variability suggests larger studies including optimization of trunk exercise and SAHC prescription with MRI are warranted. Trial registration: North Rhine ethical committee (Number: 6000223393) and registered on 29/09/2020 in the German Clinical Trials Register (DRKS00021750). Keywords: Artificial gravity; Countermeasures; Exercise; IVD; Spine; Stiffnes

Decreased Proliferation Kinetics of Mouse Myoblasts Overexpressing FRG1

Although recent publications have linked the molecular events driving facioscapulohumeral muscular dystrophy (FSHD) to expression of the double homeobox transcription factor DUX4, overexpression of FRG1 has been proposed as one alternative causal agent as mice overexpressing FRG1 present with muscular dystrophy. Here, we characterize proliferative defects in two independent myoblast lines overexpressing FRG1. Myoblasts isolated from thigh muscle of FRG1 transgenic mice, an affected dystrophic muscle, exhibit delayed proliferation as measured by decreased clone size, whereas myoblasts isolated from the unaffected diaphragm muscle proliferated normally. To confirm the observation that overexpression of FRG1 could impair myoblast proliferation, we examined C2C12 myoblasts with inducible overexpression of FRG1, finding increased doubling time and G1-phase cells in mass culture after induction of FRG1 and decreased levels of pRb phosphorylation. We propose that depressed myoblast proliferation may contribute to the pathology of mice overexpressing FRG1 and may play a part in FSHD