Brachypodium distachyon seedling growth visualization under osmotic stress and overexpression of MIR7757 to increase drought tolerance

Brachypodium distachyon a monocot model plant has facilitated the downscaling for studying the most important cereal crops of the world both genetically and phenetically. This owes to its dwarf stature, small genome size and rapid life cycle which was utilized in our research for analysing its morphological features under osmotic stress. The purpose of this study was to visualize Brachypodium seedlings under osmotic pressure to observe morphological adaptation under drought-like conditions. It was found that Brachypodium displays the typical adaptive mechanisms of cereal plants mainly root apical meristem showing lateral hair growth and stunted growth. The root cells also displayed change in single cell morphology by swelling into compartment like structures as compared to non-stressed cells. This observation was made in the elongation and maturation zones of the root. Lateral hair growth was observed from the root apical meristem after 18 hours of PEG-mediated osmotic stress. Brachypodium not only manifests physiological adaptations to drought stress but also elicits molecular adaptation to counter it. To explore the genetic basis of drought tolerance the microRNAs involved in water deficit were traced out through a reverse genetics approach. The T-DNA mutant library of Brachypodium distachyon allowed for the investigation of a newly discovered microRNA miR7757 involved in water deficit to be overexpressed in Brachypodium to rapidly produce drought tolerant varieties bypassing conventional breeding techniques

A short overview on the latest updates on cereal crop plant genome sequencing with an emphasis on cereal crops and their wild relatives

The advent of next generation sequencing has brought a revolution in the sequencing and availability of whole genome data for numerous plant species. However the genome sequencing of major staple food crops has been noticeably obscure and till relatively recently majorly unaccomplished. The obstacles for sequencing of genomes of the Poaceae grasses including sugarcane and the Triticeae wheat, barley and rye has been largely ascribed to the complex polyploid nature of their genomes, having undergone numerous evolutionary changes duplications and additions resulting in their huge modern genomes of today. Undertaking their sequencing has been a daunting task however due to the sequencing of wild grass relatives such as Brachypodium and Aegilops has been an encouraging step providing an essential framework and reference for deciphering the complex genomes particularly Triticum aestivum. This paper discusses the major challenges involved, the approaches taken and the up to date accomplished tasks for sequencing a few of the major large grass crop genome

Barriers and Promoters of Retention of Direct Care Workers in Community Mental Health Agencies

Demand for behavioral health direct-care providers is increasing due to shortage of licensed behavioral health providers. However, high turnover has been reported among them with limited exploratory research. The present study aimed to identify a wide variety of barriers and promoters of retention and strategies to retain direct care workers. An online, self-administered survey designed to measure demographics, job satisfaction, perceived importance of various job aspects, intention to leave, perceived stress and sources of stress was administered among 179 direct care workers from four agencies. Multiple logistic regression exhibited higher odds of intending to leave for those who had higher general perceived stress (OR=1.3, CI=1.1-1.7) and those who experienced stress from supervisor (OR=5.0, CI=1.7-14.4) and organizational culture (OR=4.2, CI=1.1-18.4). Work-related stress is a prevalent issue among direct-care providers and may be associated with turnover. Policy formulation and implementation directed at strategies to reduce stress may be warranted to improve retention.https://digitalcommons.unmc.edu/bhecn_report/1000/thumbnail.jp

History and current status of wheat miRNAs using next-generation sequencing and their roles in development and stress

As small molecules that aid in posttranscriptional silencing, microRNA (miRNA) discovery and characterization have vastly benefited from the recent development and widespread application of next-generation sequencing (NGS) technologies. Several miRNAs were identified through sequencing of constructed small RNA libraries, whereas others were predicted by in silico methods using the recently accumulating sequence data. NGS was a major breakthrough in efforts to sequence and dissect the genomes of plants, including bread wheat and its progenitors, which have large, repetitive and complex genomes. Availability of survey sequences of wheat whole genome and its individual chromosomes enabled researchers to predict and assess wheat miRNAs both in the subgenomic and whole genome levels. Moreover, small RNA construction and sequencing-based studies identified several putative development- and stress-related wheat miRNAs, revealing their differential expression patterns in specific developmental stages and/or in response to stress conditions. With the vast amount of wheat miRNAs identified in recent years, we are approaching to an overall knowledge on the wheat miRNA repertoire. In the following years, more comprehensive research in relation to miRNA conservation or divergence across wheat and its close relatives or progenitors should be performed. Results may serve valuable in understanding both the significant roles of species-specific miRNAs and also provide us information in relation to the dynamics between miRNAs and evolution in wheat. Furthermore, putative development- or stress-related miRNAs identified should be subjected to further functional analysis, which may be valuable in efforts to develop wheat with better resistance and/or yield

THROUGH THE LOOKING GLASS: Real-Time Imaging in Brachypodium Roots and Osmotic Stress Analysis

To elucidate dynamic developmental processes in plants, live tissues and organs must be visualised frequently and for extended periods. The development of roots is studied at a cellular resolution not only to comprehend the basic processes fundamental to maintenance and pattern formation but also study stress tolerance adaptation in plants. Despite technological advancements, maintaining continuous access to samples and simultaneously preserving their morphological structures and physiological conditions without causing damage presents hindrances in the measurement, visualisation and analyses of growing organs including plant roots. We propose a preliminary system which integrates the optical real-time visualisation through light microscopy with a liquid culture which enables us to image at the tissue and cellular level horizontally growing Brachypodium roots every few minutes and up to 24 h. We describe a simple setup which can be used to track the growth of the root as it grows including the root tip growth and osmotic stress dynamics. We demonstrate the system’s capability to scale down the PEG-mediated osmotic stress analysis and collected data on gene expression under osmotic stress

Inactivation of Nosema spp. with zinc phthalocyanine

Most honey bee pathogens, such as Vairimorpha (Nosema), cannot be rapidly and definitively diagnosed in a natural setting, consequently there is typically the spread of these diseases through shared and re-use of beekeeping equipment. Furthermore, there are no viable treatment options available for Nosema spores to aid in managing the spread of this bee disease. We therefore aimed to develop a new method using novel Zinc Phthalocyanine (ZnPc) as a photosensitizer for the photodynamic inactivation of Nosema spores that could be used for the decontamination of beekeeping equipment. Nosema spores were propagated for in vitro testing using four caged Apis mellifera honey bees. The ZnPc treatment was characterized, encapsulated with a liposome, and then used as either a 10 or 100 µM treatment for the freshly harvested Nosema spores, for either a 30 and or 60-minute time period, under either light or dark conditions, in-vitro, in 96-well plates. In the dark treatment, after 30-min, the ZnPc 100 µM treatment, caused a 30 % Nosema mortality, while this increased to 80 % at the same concentration after the light treatment. The high rate of anti-spore effects, in a short period of time, supports the notion that this could be an effective treatment for managing honey bee Nosema infections in the future. Our results also suggest that the photo activation of the treatment could be applied in the field setting and this would increase the sterilization of beekeeping equipment against Nosema

Regulation of forager honey bee appetite independent of the glucose-insulin signaling pathway

Introduction: To maintain energetic homeostasis the energetic state of the individual needs to communicate with appetite regulatory mechanisms on a regular basis. Although hunger levels indicated by the energetic state and appetite levels, the desire for food intake, tend to be correlated, and on their own are well studied, how the two cross-talk and regulate one another is less known. Insects, in contrast to vertebrates, tend to have trehalose as the primary sugar found in the hemolymph, which could possibly serve as an alternative monitor of the energetic state in comparison to the glucose-insulin signaling pathway, found in vertebrates. Methods: We investigate how manipulating hemolymph sugar levels alter the biogenic amines in the honey bee brain, appetite levels, and insulin like peptide gene expression, across three age classes, to determine how the energetic state of the honey bee might be connected to appetite regulation. Results: We found that only in the forager bees, with a lowering of hemolymph trehalose levels, there was an increase in octopamine and a decrease in tyramine levels in the honey bee brain that corresponded with increased appetite levels, while there was no significant changes in Insulin Like Peptide-1 or 2 gene expression. Discussion: Our findings suggest that hemolymph trehalose levels aid in regulating appetite levels, in forager bees, via octopamine and tyramine, and this regulation appears to be functioning independent of the glucose insulin signaling pathway. Whether this potentially more direct and rapid appetite regulatory pathway can be generalized to other insects, which also undergo energy demanding activities, remains to be investigated

3D bioprinting of tyramine modified hydrogels under visible light for osteochondral interface

Recent advancements in tissue engineering have demonstrated a great potential for the fabrication of three-dimensional (3D) tissue structures such as cartilage and bone. However, achieving structural integrity between different tissues and fabricating tissue interfaces are still great challenges. In this study, an in situ crosslinked hybrid, multi-material 3D bioprinting approach was used for the fabrication of hydrogel structures based on an aspiration-extrusion microcapillary method. Different cell-laden hydrogels were aspirated in the same microcapillary glass and deposited in the desired geometrical and volumetric arrangement directly from a computer model. Alginate and carboxymethyl cellulose were modified with tyramine to enhance cell bioactivity and mechanical properties of human bone marrow mesenchymal stem cells-laden bioinks. Hydrogels were prepared for extrusion by gelling in microcapillary glass utilizing an in situ crosslink approach with ruthenium (Ru) and sodium persulfate photo-initiating mechanisms under visible light. The developed bioinks were then bioprinted in precise gradient composition for cartilage-bone tissue interface using microcapillary bioprinting technique. The biofabricated constructs were co-cultured in chondrogenic/osteogenic culture media for three weeks. After cell viability and morphology evaluations of the bioprinted structures, biochemical and histological analyses, and a gene expression analysis for the bioprinted structure were carried out. Analysis of cartilage and bone formation based on cell alignment and histological evaluation indicated that mechanical cues in conjunction with chemical cues successfully induced MSC differentiation into chondrogenic and osteogenic tissues with a controlled interface