The 141-year period for Dr. Beal's seed viability experiment: A hybrid surprise.

PREMISE: In 1879, Dr. William Beal buried 20 glass bottles filled with seeds and sand at a single site at Michigan State University. The goal of the experiment was to understand seed longevity in the soil, a topic of general importance in ecology, restoration, conservation, and agriculture, by periodically assaying germinability of these seeds over 100 years. The interval between germination assays has been extended and the experiment will now end after 221 years, in 2100. METHODS: We dug up the 16th bottle in April 2021 and attempted to germinate the 141-year-old seeds it contained. We grew germinants to maturity and identified these to species by vegetative and reproductive phenotypes. For the first time in the history of this experiment, genomic DNA was sequenced to confirm species identities. RESULTS: Twenty seeds germinated over the 244-day assay. Eight germinated in the first 11 days. All 20 belonged to the Verbascum genus: Nineteen were V. blattaria according to phenotype and ITS2 genotype; and one had a hybrid V. blattaria × V. thapsus phenotype and ITS2 genotype. In total, 20/50 (40%) of the original Verbascum seeds in the bottle germinated in year 141. CONCLUSIONS: While most species in the Beal experiment lost all seed viability in the first 60 years, a high percentage of Verbascum seeds can still germinate after 141 years in the soil. Long-term experiments such as this one are rare and invaluable for studying seed viability in natural soil conditions.

The Influence of Youth Gender and Complex Trauma on the Relation Between Treatment Conditions and Outcomes in Therapeutic Residential Care.

Complex trauma (CT) is the experience, or witness, of prolonged abuse or neglect that negatively affects children's emotional and psychological health. Youth in residential care experience higher incidences of complex trauma than youth in community-based care, with notable gender differences and presentation of psychological symptoms. This study examined the effects of trauma-informed residential care and the relation between CT and gender. A sample (n = 206) from an evaluation of a youth psychiatric residential facility in the Midwest that transitioned from a traditional care model to a trauma-informed care model was used. A hierarchical regression was used to model the main effects of model of care, gender, CT, length of stay, and crisis response on treatment outcomes; and the moderating effects of gender and CT. The results support the high prevalence of CT in residential care populations. The final model explained 30.2% of the variance with a statistically significant interaction between gender and length of stay in treatment, indicating that longer lengths of stay in treatment are associated with less change in functional impairment for girls than boys. Youth gender and prior trauma are important factors to consider when monitoring experiences and treatment outcomes in youth residential care.

The Trauma Symptom Checklist for Young Children: A Psychometric Review.

The Trauma Symptom Checklist for Young Children (TSCYC) is a 90-item caregiver-report measure of childhood traumatic stress and abuse-related experiences in children, ages three to twelve years old. PURPOSE: The objective of this review is to examine the current empirical evidence (n = 22) regarding the psychometric properties of the TSCYC. METHOD: A variety of study designs were reviewed for psychometric evidence supporting the reliability and validity of the TSCYC. RESULTS: The psychometric evidence for the TSCYC indicates it is a reliable scale. However, evidence of validity is moderate and focuses on older children. DISCUSSION: Clinicians may utilize the TSCYC to support a PTSD diagnosis in children. Further psychometric exploration would strengthen the body of evidence for younger children (ages 3-5) who have had traumatic experiences.

Developmental Genetics of Corolla Tube Formation: Role of the tasiRNA-ARF Pathway and a Conceptual Model.

Over 80,000 angiosperm species produce flowers with petals fused into a corolla tube. The corolla tube contributes to the tremendous diversity of flower morphology and plays a critical role in plant reproduction, yet it remains one of the least understood plant structures from a developmental genetics perspective. Through mutant analyses and transgenic experiments, we show that the tasiRNA-ARF pathway is required for corolla tube formation in the monkeyflower species Mimulus lewisii Loss-of-function mutations in the M. lewisii orthologs of ARGONAUTE7 and SUPPRESSOR OF GENE SILENCING3 cause a dramatic decrease in abundance of TAS3-derived small RNAs and a moderate upregulation of AUXIN RESPONSE FACTOR3 (ARF3) and ARF4, which lead to inhibition of lateral expansion of the bases of petal primordia and complete arrest of the upward growth of the interprimordial regions, resulting in unfused corollas. Using the DR5 auxin-responsive promoter, we discovered that auxin signaling is continuous along the petal primordium base and the interprimordial region during the critical stage of corolla tube formation in the wild type, similar to the spatial pattern of MlARF4 expression. Auxin response is much weaker and more restricted in the mutant. Furthermore, exogenous application of a polar auxin transport inhibitor to wild-type floral apices disrupted petal fusion. Together, these results suggest a new conceptual model highlighting the central role of auxin-directed synchronized growth of the petal primordium base and the interprimordial region in corolla tube formation.

A Tetratricopeptide Repeat Protein Regulates Carotenoid Biosynthesis and Chromoplast Development in Monkeyflowers (Mimulus).

Little is known about the factors regulating carotenoid biosynthesis in flowers. Here, we characterized the REDUCED CAROTENOID PIGMENTATION2 (RCP2) locus from two monkeyflower (Mimulus) species, the bumblebee-pollinated species Mimulus lewisii and the hummingbird-pollinated species Mimulus verbenaceus We show that loss-of-function mutations of RCP2 cause drastic down-regulation of the entire carotenoid biosynthetic pathway. The causal gene underlying RCP2 encodes a tetratricopeptide repeat protein that is closely related to the Arabidopsis (Arabidopsis thaliana) REDUCED CHLOROPLAST COVERAGE proteins. RCP2 appears to regulate carotenoid biosynthesis independently of RCP1, a previously identified R2R3-MYB master regulator of carotenoid biosynthesis. We show that RCP2 is necessary and sufficient for chromoplast development and carotenoid accumulation in floral tissues. Simultaneous down-regulation of RCP2 and two closely related paralogs, RCP2-L1 and RCP2-L2, yielded plants with pale leaves deficient in chlorophyll and carotenoids and with reduced chloroplast compartment size. Finally, we demonstrate that M. verbenaceus is just as amenable to chemical mutagenesis and in planta transformation as the more extensively studied M. lewisii, making these two species an excellent platform for comparative developmental genetics studies of closely related species with dramatic phenotypic divergence.

Two MYB Proteins in a Self-Organizing Activator-Inhibitor System Produce Spotted Pigmentation Patterns.

Many organisms exhibit visually striking spotted or striped pigmentation patterns. Developmental models predict that such spatial patterns can form when a local autocatalytic feedback loop and a long-range inhibitory feedback loop interact. At its simplest, this self-organizing network only requires one self-activating activator that also activates a repressor, which inhibits the activator and diffuses to neighboring cells. However, the molecular activators and inhibitors fully fitting this versatile model remain elusive in pigmentation systems. Here, we characterize an R2R3-MYB activator and an R3-MYB repressor in monkeyflowers (Mimulus). Through experimental perturbation and mathematical modeling, we demonstrate that the properties of these two proteins correspond to an activator-inhibitor pair in a two-component, reaction-diffusion system, explaining the formation of dispersed anthocyanin spots in monkeyflower petals. Notably, disrupting this pattern impacts pollinator visitation. Thus, subtle changes in simple activator-inhibitor systems are likely essential contributors to the evolution of the remarkable diversity of pigmentation patterns in flowers.

Transcriptional Regulation of Carotenoid Biosynthesis in Plants: So Many Regulators, So Little Consensus.

In plants, the carotenoid biosynthesis pathway (CBP) is essential for the production of photosynthetic and protective pigments, plant hormones, and visual/olfactory attractants for animal pollinators and seed dispersers. The regulation of carotenoid biosynthesis at the transcriptional level is vitally important for all of these functions and has been the subject of intensive research. Many putative transcriptional regulators, both direct and indirect, have been identified through conventional mutant analysis, transcriptome profiling, yeast one-hybrid screening, and candidate gene approaches. Despite this progress, our understanding of the transcriptional regulation of carotenoid biosynthesis remains fragmented and incomplete. Frequently, a stimulus or regulator is known, but the mechanism by which it affects transcription has not been elucidated. In other cases, mechanisms have been proposed (such as direct binding of a CBP gene promoter by a transcription factor), but function was tested only in vitro or in heterologous systems, making it unclear whether these proteins actually play a role in carotenoid regulation in their endogenous environments. Even in cases where the mechanism is relatively well understood, regulators are often studied in isolation, either in a single plant species or outside the context of other known regulators. This presents a conundrum: why so many candidate regulators but so little consensus? Here we summarize current knowledge on transcriptional regulation of the CBP, lay out the challenges contributing to this conundrum, identify remaining knowledge gaps, and suggest future research directions to address these challenges and knowledge gaps.

Colorectal Cancer Screening: Role of Family Physicians.

The purpose of this study was to look at the differences in colorectal cancer screening awareness between two rural communities in Texas. In Clifton, patients have access to colonoscopies in their local hospital, while in Haskell, patients have to travel to a tertiary center. A 24-question survey pertaining to colon cancer from the Behavior Risk Factor Surveillance System (BRFSS) was given to patients at primary care clinics in Clifton and Haskell. To take the survey, participants had to have been patients for at least 1 year in either Clifton or Haskell clinic and be at least age 50 years or older. A total of 168 surveys were collected, 92 at Clifton and 76 at Haskell. A higher odds ratio (OR=3.61; CI = [1.11, 11.69]) was seen in Clifton compared with Haskell for patient colon cancer screening awareness. Also, a higher odds ratio (OR=2.50; CI = [1.13, 5.54]) was found of knowing at what age a person should be screened for colon cancer in Clifton compared with Haskell. A higher odds ratio (OR=3.61; CI = [1.42, 9.20]) was seen in Clifton compared with Haskell for patients ever having a colonoscopy or sigmoidoscopy. This study supports the idea that providing colorectal cancer screening and screening procedures locally in the rural community by family medicine physicians helps to contribute to an improved awareness of colorectal cancer screening guidelines as opposed to communities that do not offer screening locally.

Is the Preschool PATHS Curriculum Effective? A Review.

The preschool PATHS curriculum was designed as a universal preschool classroom curriculum to improve children's social-emotional learning and reduce problem behaviors. Purpose: This review examined the published outcome studies (n = 5) of the preschool PATHS curriculum examining social-emotional and problem behavior outcome measures. Method: One randomized control trial, two quasi-experimental designs and two pre-experimental designs met the inclusion criteria and were reviewed. Results: The evidence supporting the preschool PATHS curriculum positively impacting social-emotional competencies is mild to moderate. Evidence supporting the preschool PATHS curriculum as a universal curriculum to reduce problem behavior is sparse and equivocal. Discussion: Early childhood professionals should implement the preschool PATHS curriculum with the understanding that a child's acquisition of social competencies may not readily equate to a reduction in problem behaviors.

Competition between anthocyanin and flavonol biosynthesis produces spatial pattern variation of floral pigments between Mimulus species.

Flower color patterns have long served as a model for developmental genetics because pigment phenotypes are visually striking, yet generally not required for plant viability, facilitating the genetic analysis of color and pattern mutants. The evolution of novel flower colors and patterns has played a key role in the adaptive radiation of flowering plants via their specialized interactions with different pollinator guilds (e.g., bees, butterflies, birds), motivating the search for allelic differences affecting flower color pattern in closely related plant species with different pollinators. We have identified LIGHT AREAS1 (LAR1), encoding an R2R3-MYB transcription factor, as the causal gene underlying the spatial pattern variation of floral anthocyanin pigmentation between two sister species of monkeyflower: the bumblebee-pollinated Mimulus lewisii and the hummingbird-pollinated Mimulus cardinalis. We demonstrated that LAR1 positively regulates FLAVONOL SYNTHASE (FLS), essentially eliminating anthocyanin biosynthesis in the white region (i.e., light areas) around the corolla throat of M. lewisii flowers by diverting dihydroflavonol into flavonol biosynthesis from the anthocyanin pigment pathway. FLS is preferentially expressed in the light areas of the M. lewisii flower, thus prepatterning the corolla. LAR1 expression in M. cardinalis flowers is much lower than in M. lewisii, explaining the unpatterned phenotype and recessive inheritance of the M. cardinalis allele. Furthermore, our gene-expression analysis and genetic mapping results suggest that cis-regulatory change at the LAR1 gene played a critical role in the evolution of different pigmentation patterns between the two species.

An R2R3-MYB transcription factor regulates carotenoid pigmentation in Mimulus lewisii flowers.

Carotenoids are yellow, orange, and red pigments that contribute to the beautiful colors and nutritive value of many flowers and fruits. The structural genes in the highly conserved carotenoid biosynthetic pathway have been well characterized in multiple plant systems, but little is known about the transcription factors that control the expression of these structural genes. By analyzing a chemically induced mutant of Mimulus lewisii through bulk segregant analysis and transgenic experiments, we have identified an R2R3-MYB, Reduced Carotenoid Pigmentation 1 (RCP1), as the first transcription factor that positively regulates carotenoid biosynthesis during flower development. Loss-of-function mutations in RCP1 lead to down-regulation of all carotenoid biosynthetic genes and reduced carotenoid content in M. lewisii flowers, a phenotype recapitulated by RNA interference in the wild-type background. Overexpression of this gene in the rcp1 mutant background restores carotenoid production and, unexpectedly, results in simultaneous decrease of anthocyanin production in some transgenic lines by down-regulating the expression of an activator of anthocyanin biosynthesis. Identification of transcriptional regulators of carotenoid biosynthesis provides the 'toolbox' genes for understanding the molecular basis of flower color diversification in nature and for potential enhancement of carotenoid production in crop plants via genetic engineering.