A Combined Reach Out and Read and Imagination Library Program on Kindergarten Readiness.

OBJECTIVES: Sharing books with preschoolers is known to improve kindergarten readiness. Both Reach Out and Read (ROR) and Dolly Parton's Imagination Library (DPIL) have shown positive effects on book sharing at home. We developed a novel combined ROR/DPIL program and examined the effect on kindergarten readiness assessment (KRA) scores. METHODS: At urban ROR primary care sites, patients <5 years living in the city school district were enrolled from July 2015 through January 2019 in the ROR/DPIL program when seen for a clinic visit. The literacy subtest of the KRA was examined for participants entering kindergarten in the fall of 2016, 2017, and 2018. The "on-track" rate of participants was compared with nonparticipant groups. RESULTS: A total of 797 kindergarten-aged ROR/DPIL participants were matched to Ohio KRA scores for 2016, 2017, and 2018 school years. The percentages of students "on-track" on KRA literacy subtests increased significantly by cohort (2016, 42.9% [95% confidence interval (CI): 34.9%-50.9%] versus 2017, 50.9% [95% CI: 44.9%-56.9%] versus 2018, 58.3% [95% CI: 53.3%-63.3%], P = .004). ROR/DPIL participants were compared with a proportionate stratified random sample of 1580 non-ROR/DPIL peers. On-track in literacy did not significantly differ between groups (2016 [P = .262], 2017 [P = .653], 2018 [P = .656]), nor did they differ after restricting analysis to economically disadvantaged children (2016 [P = .191], 2017 [P = .721], 2018 [P = .191]). CONCLUSIONS: With these results, we suggest that a program combining literacy anticipatory guidance at clinic visits and more books in the home can potentially improve kindergarten readiness. Pediatric health care providers can play an important role in promoting kindergarten readiness through literacy promotion.

Modulation of nonessential amino acid biosynthetic pathways in virulent Hessian fly larvae (Mayetiola destructor), feeding on susceptible host wheat (Triticum aestivum).

Compatible interactions between wheat (Triticum aestivum), and its dipteran pest Hessian fly (Hf, Mayetiola destructor) result in successful establishment of larval feeding sites rendering the host plant susceptible. Virulent larvae employ an effector-based feeding strategy to reprogram the host physiology resulting in formation of a protein- and sugar-rich nutritive tissue beneficial to developing larvae. Previous studies documented increased levels of nonessential amino acids (NAA; that need not be received through insect diet) in the susceptible wheat in response to larval feeding, suggesting importance of plant-derived NAA in larval nutrition. Here, we investigated the modulation of genes from NAA biosynthetic pathways (NAABP) in virulent Hf larvae. Transcript profiling for 16 NAABP genes, annotated from the recently assembled Hf genome, was carried out in the feeding first-, and second-instars and compared with that of the first-instar neonate (newly hatched, migrating, assumed to be non-feeding) larvae. While Tyr, Gln, Glu, and Pro NAABP genes transcript abundance declined in the feeding instars as compared to the neonates, those for Ala, and Ser increased in the feeding larval instars, despite higher levels of these NAA in the susceptible host plant. Asp, Asn, Gly and Cys NAABP genes exhibited variable expression profiles in the feeding first- and second-instars. Our results indicate that while Hf larvae utilize the plant-derived NAA, de novo synthesis of several NAA may be necessary to: (i) provide larvae with the requisite amount for sustaining growth before nutritive tissue formation and, (ii) overcome any inadequate amounts in the host plant, post-nutritive tissue formation.

A Novel, Economical Way to Assess Virulence in Field Populations of Hessian Fly (Diptera: Cecidomyiidae) Utilizing Wheat Resistance Gene H13 as a Model.

Mayetiola destructor (Say) is a serious pest of wheat, Triticum aestivum L., in North America, North Africa, and Central Asia. Singly deployed resistance genes in wheat cultivars have provided effective management of Hessian fly populations for >50 yr. Thirty-five H genes have been documented. Defense mediated by the H gene constitutes strong selection on the Hessian fly population, killing 100% of larvae. A mutation in a matching Hessian fly avirulence gene confers virulence to the H gene, leading to survival on the resistant plant. As the frequency of virulence rises in the population, the H gene loses its effectiveness for pest management. Knowing the frequency of virulence in the population is not only important for monitoring but also for decisions about which H gene should be deployed in regional wheat breeding programs. Here, we present a novel assay for detecting virulence in the field. Hessian fly males were collected in Alabama, Georgia, North Carolina, and South Carolina using sticky traps baited with Hessian fly sex pheromone. Utilizing two PCR reactions, diagnostic molecular markers for the six alleles controlling avirulence and virulence to H13 can be scored based on band size. Throughout the southeast, all three avirulence and three virulence alleles can be identified. In South Carolina, the PCR assay was sensitive enough to detect the spread of virulence into two counties previously documented as 100% susceptible to H13. The new assay also indicates that the previous methods overestimated virulence in the field owing to scoring of the plant instead of the insect.

A massive expansion of effector genes underlies gall-formation in the wheat pest Mayetiola destructor.

Gall-forming arthropods are highly specialized herbivores that, in combination with their hosts, produce extended phenotypes with unique morphologies [1]. Many are economically important, and others have improved our understanding of ecology and adaptive radiation [2]. However, the mechanisms that these arthropods use to induce plant galls are poorly understood. We sequenced the genome of the Hessian fly (Mayetiola destructor; Diptera: Cecidomyiidae), a plant parasitic gall midge and a pest of wheat (Triticum spp.), with the aim of identifying genic modifications that contribute to its plant-parasitic lifestyle. Among several adaptive modifications, we discovered an expansive reservoir of potential effector proteins. Nearly 5% of the 20,163 predicted gene models matched putative effector gene transcripts present in the M. destructor larval salivary gland. Another 466 putative effectors were discovered among the genes that have no sequence similarities in other organisms. The largest known arthropod gene family (family SSGP-71) was also discovered within the effector reservoir. SSGP-71 proteins lack sequence homologies to other proteins, but their structures resemble both ubiquitin E3 ligases in plants and E3-ligase-mimicking effectors in plant pathogenic bacteria. SSGP-71 proteins and wheat Skp proteins interact in vivo. Mutations in different SSGP-71 genes avoid the effector-triggered immunity that is directed by the wheat resistance genes H6 and H9. Results point to effectors as the agents responsible for arthropod-induced plant gall formation.

A genome-wide survey of small interfering RNA and microRNA pathway genes in a galling insect.

Deployment of resistance (R) genes is the most effective control for Hessian fly, Mayetiola destructor (Say); however, deployment of R genes results in an increased frequency of pest genotypes that display virulence to them. RNA interference (RNAi) is a useful reverse genetics tool for studying such insect virulence pathways, but requires a systemic phenotype, which is not found in all species. In an effort to correlate our observed weak RNAi phenotype in M. destructor with a genetic basis, we have aggregated and compared RNAi related genes across M. destructor, three other insect species, and the nematode Caenorhabditis elegans. We report here the annotation of the core genes in the small interfering RNA (siRNA) and microRNA (miRNA) pathways in M. destructor. While most of the miRNA pathway genes were highly conserved across the species studied, the siRNA pathway genes showed increased relative variability in comparison to the miRNA pathway. In particular, the Piwi/Argonaute/Zwille (PAZ) domain of Dicer-2 (DCR-2) had the least amount of sequence similarity of any domain among species surveyed, with a trend of increased conservation in those species with amenable systemic RNAi. A homolog of the systemic interference defective-1 (Sid-1) gene of C. elegans was also not annotated in the M. destructor genome. Indeed, it is of interest that a Sid-1 homolog has not been detected in any dipteran species to date. We hypothesize the sequence architecture of the PAZ domain in the M. destructor DCR-2 protein is related to reduced efficacy of this enzyme and this taken together with the lack of a Sid-1 homolog may account for the weak RNAi response observed to date in this species as well as other dipteran species.

Tissue and developmental expression of a gene from Hessian fly encoding an ABC-active-transporter protein: implications for Malpighian tubule function during interactions with wheat.

We report on the transcriptional patterns of a putative white (w) gene encoding an ABC-active-transporter protein during development in Hessian fly, Mayetiola destructor. The deduced amino acid sequence for the Hessian fly white showed 74-77% similarities to white/ATP-binding-cassette proteins and 52-57% similarities to scarlet/ATP-binding-cassette proteins from other dipterans. Conserved ATP-binding motifs and transmembrane alpha-helix segments were identified in the Hessian fly white protein further supporting its function as an ABC-active-transporter similar to the Drosophila white protein. Spatial analysis of transcript levels for white in larval Hessian fly tissues by quantitative real-time PCR revealed the greatest level of transcript in the Malpighian tubules, while analysis of temporal expression during development revealed the highest transcript levels in late 2nd- and early 3rd-instar larvae. Analysis of transcript levels for white in Hessian fly larvae feeding on susceptible and resistant wheat showed greater levels of the transcript in larvae feeding on resistant plants. We speculate the increased transcript level for white in larvae feeding on resistant wheat could be correlated with stress and increased Malpighian tubule activity associated with the metabolism and detoxification of toxic substrates generated either endogenously or encountered exogenously from the host plant.