Indigenous Australian grass seeds as grains: macrostructure, microstructure and histochemistry

Utilization of grains of local grasses by Australia's First Nations people for food and connection to Country has largely been lost due to colonization. Native Australian grain production has the potential to deliver environmental, economic, nutritional and cultural benefits to First Nations people and the wider community. Revitalization of the native grain food system can only be achieved if relevant properties of the grains are elucidated. This study aimed to characterize the grain structure and histochemistry of four Australian native grasses: Dactyloctenium radulans (Button Grass), Astrebla lappacea (Curly Mitchell Grass), Panicum decompositum (Native Millet) and Microlaena stipoides (Weeping Grass). For these species, as well as wheat and sorghum, whole-grain images were obtained via stereo microscopy, starch and the embryo were visualized, and sections of fixed grains were imaged via bright-field and fluorescence microscopy. The shape, size and colour of the whole native grains varied between the species. The aleurone layer was one-cell thick in the native species, as in the domesticated grains, except for Weeping Grass, which had a two-cell-thick aleurone. In the native grains, endosperm cell walls appeared thinner than in wheat and sorghum. Starch granules in Button Grass, Curly Mitchell Grass and Native Millet were found mainly in the central region of the starchy endosperm, with very few granules in the sub-aleurone layer, whereas Weeping Grass had abundant starch in the sub-aleurone. Protein appeared most abundant in the aleurone and sub-aleurone layers of the native grains, although in Button Grass, the starchy endosperm was observed to be rich in protein, as in wheat and sorghum. As a proportion of the whole grain, the embryo was larger in the native species than in wheat. The differences found in the grain properties among the four native Australian species have important implications for the agri-food industry in a changing climate.Revitalization of native grain food systems in Australia requires knowledge of the structure and composition of grain components so that the grains can be utilized effectively in commercial food processing. We studied the grain morphology of four native Australian grass species that have been used as foods by Aboriginal communities for millenia: Button Grass (Dactyloctenium radulans ), Curly Mitchell Grass (Astrebla lappacea ), Native Millet (Panicum decompositum ) and Weeping Grass (Microlaena stipoides ). We characterized important macrostructural, microstructural and histochemical properties of the grains using stereo, bright-field and fluorescence microscopy. We found differences in the size, shape and colour of the whole grains, aleurone structure, spatial distribution of starch, beta-glucan/cellulose, protein and relative size and location of the embryo among the four native Australian species. Our results should stimulate further research into the connections between the structural, processing and nutritional properties of these grains

Ukulelen alkeita muksuille : Soitto-oppaan matka ideasta toteutukseen

Opinnäytetyönäni olen luonut ukulelen alkeisoppaan Muksulele – Ukulelen alkeita muksuille. Opas sisältää 18 uutta lastenlaulua sekä ohjeet ukulelen hallintaan ja neljän perussoinnun soittamiseen. Lisäksi laulujen yhteyteen olen kirjannut vinkkejä, kuinka soittimen kanssa kannattaa lapsiryhmässä edetä ja miten lauluja voi musiikkileikkikoulussa hyödyntää. Tässä opinnäytetyöni kirjallisessa osassa kuvaan soitto-oppaan syntyprosessin kaikkine vaiheineen. Kerron, kuinka suunnittelin opetuksen etenevän ja miten päädyin näihin ratkaisuihin. Mitä sointuja olisi hyvä opettaa ja missä järjestyksessä edetä soinnusta toiseen? Avaan myös laulujen sävellysprosessia, ja sitä, kuinka pedagoginen ajattelu vaikutti sävellystyöhöni. Muksulele-oppaan kohderyhminä ovat musiikkileikkikoulunopettajien ja –oppilaiden lisäksi myös varhaiskasvattajat ja muut lasten kanssa työskentelevät aikuiset. Tein yhteistyötä kolmen Riihimäkeläisen päiväkodin kanssa, joiden henkilökunta vastasi sähköiseen kyselyyn yhteistyömme aluksi. Kyselyllä kartoitin, olisivatko he kiinnostuneita käyttämään ukulelea työssään ja millaista ohjeistusta ja materiaaleja he tarvitsisivat, jotta se olisi mahdollista. Avaan työssäni heidän toiveidensa vaikutuksia oppaan sisältöön. Soitto-oppaan materiaalit kävivät valmistuttuaan läpi monta testaus- ja arviointitilannetta. Yhteistyöpäiväkodeissa pidin koulutustilaisuudet, joissa henkilökunta pääsi antamaan palautetta sekä lauluista että opetuksen etenemisestä. Lisäksi kantaa materiaaleihin ottivat myös musiikkileikkikoulun ukuleleryhmäläiset sekä työni asiantuntijaohjaaja. Kuvaan myös yksityiskohtaisesti palautteiden pohjalta tehtyjä muutoksia. Opinnäytetyöni toinen osa Muksulele –kirja julkaistaan heinäkuussa 2017.As my final project, I have created a kids’ guide to ukulele, Muksulele – Ukulelen alkeita muksuille. In the guidebook, there are 18 new songs to be accompanied by the ukulele and instructions on how to play four basic chords. Also, the book includes tips on how to use the instrument with children and what to do with the songs in class. In this written report, I describe the process of creating the guidebook. I describe decisions I made concerning the pedagogical process, i.e. what chords to teach and how to proceed from one chord to another. I also explain what I had to take into account when composing the pieces and how the pedagogical thinking was present through the whole experience. The main target group for the guidebook are early childhood music teachers, but it can also benefit early childhood educators and other adults working with children. I collaborated with three nurseries in Riihimäki and did an electrical survey among the staff. In the survey, I asked, for example, what kind of instructions they would need in order to use ukulele in their work. Some alterations were made to the guidebook on the basis of the survey results. The guidebook went through a series of tests and reviews. The staff of the collaborating nurseries had a chance to comment on my work during in-service training sessions I had with them. My music playschool ukulele students also gave me feedback, as well as a consulting expert I had the pleasure of working with. I describe the changes I made to the materials due to the feedback given in detail. The second part of my thesis, the Muksulele book, will be released in July 2017

Starch biosynthetic genes and enzymes are expressed and active in the absence of starch accumulation in sugar beet tap-root

BACKGROUND: Starch is the predominant storage compound in underground plant tissues like roots and tubers. An exception is sugar beet tap-root (Beta vulgaris ssp altissima) which exclusively stores sucrose. The underlying mechanism behind this divergent storage accumulation in sugar beet is currently not fully known. From the general presence of starch in roots and tubers it could be speculated that the lack in sugar beet tap-roots would originate from deficiency in pathways leading to starch. Therefore with emphasis on starch accumulation, we studied tap-roots of sugar beet using parsnip (Pastinaca sativa) as a comparator. RESULTS: Metabolic and structural analyses of sugar beet tap-root confirmed sucrose as the exclusive storage component. No starch granules could be detected in tap-roots of sugar beet or the wild ancestor sea beet (Beta vulgaris ssp. maritima). Analyses of parsnip showed that the main storage component was starch but tap-root tissue was also found to contain significant levels of sugars. Surprisingly, activities of four main starch biosynthetic enzymes, phosphoglucomutase, ADP-glucose pyrophosphorylase, starch synthase and starch branching enzyme, were similar in sugar beet and parsnip tap-roots. Transcriptional analysis confirmed expression of corresponding genes. Additionally, expression of genes involved in starch accumulation such as for plastidial hexose transportation and starch tuning functions could be determined in tap-roots of both plant species. Conclusion: Considering underground storage organs, sugar beet tap-root upholds a unique property in exclusively storing sucrose. Lack of starch also in the ancestor sea beet indicates an evolved trait of biological importance. Our findings in this study show that gene expression and enzymatic activity of main starch biosynthetic functions are present in sugar beet tap-root during storage accumulation. In view of this, the complete lack of starch in sugar beet tap-roots is enigmatic

CRISPR/Cas9 genome editing of potato StDMR6-1 results in plants less affected by different stress conditions

Potato is the third most important food crop, but cultivation is challenged by numerous diseases and adverse abiotic conditions. To combat diseases, frequent fungicide application is common. Knocking out susceptibility genes by genome editing could be a durable option to increase resistance. DMR6 has been described as a susceptibility gene in several crops, based on data that indicates increased resistance upon interruption of the gene function. In potato, Stdmr6-1 mutants have been described to have increased resistance against the late blight pathogen Phytophthora infestans in controlled conditions. Here, we present field evaluations of CRISPR/Cas9 mutants, in a location with a complex population of P. infestans, during four consecutive years that indicate increased resistance to late blight without any trade-off in terms of yield penalty or tuber quality. Furthermore, studies of potato tubers from the field trials indicated increased resistance to common scab, and the mutant lines exhibit increased resistance to early blight pathogen Alternaria solani in controlled conditions. Early blight and common scab are problematic targets in potato resistance breeding, as resistance genes are very scarce. The described broad-spectrum resistance of Stdmr6-1 mutants may further extend to some abiotic stress conditions. In controlled experiments of either drought simulation or salinity, Stdmr6-1 mutant plants are less affected than the background cultivar. Together, these results demonstrate the prospect of the Stdmr6-1 mutants as a useful tool in future sustainable potato cultivation without any apparent trade-offs

High-throughput method for detection and quantification of lesions on leaf scale based on trypan blue staining and digital image analysis

Background: Field-grown leafy vegetables can be damaged by biotic and abiotic factors, or mechanically damaged by farming practices. Available methods to evaluate leaf tissue damage mainly rely on colour diferentiation between healthy and damaged tissues. Alternatively, sophisticated equipment such as microscopy and hyperspectral cameras can be employed. Depending on the causal factor, colour change in the wounded area is not always induced and, by the time symptoms become visible, a plant can already be severely afected. To accurately detect and quantify damage on leaf scale, including microlesions, reliable diferentiation between healthy and damaged tissue is essential. We stained whole leaves with trypan blue dye, which traverses compromised cell membranes but is not absorbed in viable cells, followed by automated quantifcation of damage on leaf scale. Results: We present a robust, fast and sensitive method for leaf-scale visualisation, accurate automated extraction and measurement of damaged area on leaves of leafy vegetables. The image analysis pipeline we developed automatically identifes leaf area and individual stained (lesion) areas down to cell level. As proof of principle, we tested the methodology for damage detection and quantifcation on two feld-grown leafy vegetable species, spinach and Swiss chard. Conclusions: Our novel lesion quantifcation method can be used for detection of large (macro) or single-cell (micro) lesions on leaf scale, enabling quantifcation of lesions at any stage and without requiring symptoms to be in the visible spectrum. Quantifying the wounded area on leaf scale is necessary for generating prediction models for economic losses and produce shelf-life. In addition, risk assessments are based on accurate prediction of the relationship between leaf damage and infection rates by opportunistic pathogens and our method helps determine the sevn.erity of leaf damage at fne resoluti

Mobilization of lipid reserves during germination of oat (Avena sativa L.), a cereal rich in endosperm oil

Since the cereal endosperm is a dead tissue in the mature grain, β-oxidation is not possible there. This raises the question about the use of the endosperm oil in cereal grains during germination. In this study, mobilization of lipids in different tissues of germinating oat grains was analysed using thin-layer and gas chromatography. The data imply that the oat endosperm oil [triacylglycerol (TAG)] is not a dead-end product as it was absorbed by the scutellum, either as free fatty acids (FFAs) released from TAG or as intact TAG immediately degraded to FFAs. These data were supported by light and transmission electron microscopy (LM and TEM) studies where close contact between endosperm lipid droplets and the scutellum was observed. The appearance of the fused oil in the oat endosperm changed into oil droplets during germination in areas close to the aleurone and the scutellar epithelium. However, according to the data obtained by TEM these oil droplets are unlikely to be oil bodies surrounded by oleosins. Accumulation of FFA pools in the embryo suggested further transport of FFAs from the scutellum. Noticeably high levels of TAG were also accumulated in the embryo but were not synthesized by re-esterification from imported FFAs. Comparison between two oat cultivars with different amounts of oil and starch in the endosperm suggests that an increased oil to starch ratio in oat grains does not significantly impact the germination process

Asiakaslähtöisten työterveyspalveluiden kehittäminen yksityisessä työterveyshuollossa

Asiakaslähtöisyys on työterveyshuollon keskeinen toimintaperiaate. Asiakaslähtöinen toimintatapa on työterveyshuollon ja työpaikan yhteistyösuhde, joka perustuu luottamukseen ja sopimuksiin. Hyvin toimiva yhteistyö työpaikan ja työterveyshuollon välillä mahdollistaa asiakassuhteen kehittymisen ja lisää toiminnan arvoa sekä vaikuttavuutta. Asiakaslähtöinen toimintamalli huomioi työterveysyrityksen lähtökohdat ja ympäristön. Asiakaslähtöisten sosiaali- ja terveyspalveluiden kehittämisen suhteen on meneillään historiallinen murrosvaihe. Nykyisellään palveluilta edellytetään, että asiakkailla on palvelujärjestelmän toimijoiden kanssa tasavertainen vaikuttajan ja kehittäjän rooli. Keskeistä muutosprosessissa onkin asiakastarpeisiin vastaaminen, palvelutoiminnan asiakaslähtöisyys sekä asiakkaiden osallisuus. Asiakaslähtöisyys ja asiakaslähtöisten palveluiden kehittäminen on keskeinen tavoite myös Rinteen hallitusohjelmassa Osallistava ja osaava Suomi. Yksi hallitusohjelman strateginen kokonaisuus käsittää sosiaali- ja terveydenhuollon palveluiden parantamisen. Palveluiden kehittämisen tavoitteena on taata hoitoa ja huolenpitoa oikea-aikaisesti, yhdenvertaisesti, vaikuttavasti, kustannustehokkaasti, kokonaisvaltaisesti ja joustavasti. Tapaustutkimuksena toteutetussa kehittämistyössä tarkoituksena oli laatia asiakaslähtöinen toimintamalli yksityiselle palveluntarjoajalle työterveyspalveluiden kehittämiseksi. Tavoitteena oli kehittää työterveyshuollon palveluita asiakaslähtöisemmiksi. Tapaustutkimuksen aineistot kerättiin asiakaskyselyllä asiakasyrityksille (n=64) asiakasyrityksiä hoitavat työterveyshoitajat teemahaastateltiin (n=4) ja asiakasyrityksiin järjestettiin workshopit (n=3). Aineistojen avulla kuvattiin ensiksi asiakasyrityksien ja työterveyshuollon välistä yhteistyötä sekä asiakasyrityksien tarpeita ja toiveita työterveyshuollon palveluiden kehittämiseksi. Asiakaslähtöinen toimintamalli luotiin työterveyshoitajien teemahaastattelujen ja asiakasyrityksissä järjestettyjen workshopien pohjalta. Asiakaslähtöisessä toimintamallissa keskeisiksi asioiksi tulivat moniammatillisuuden hyödyntäminen, asiakas- ja tarvelähtöisyys, palveluiden saatavuus sekä työterveyshuollon sisällöstä tiedottaminen. Jatkossa työterveyshuollon kehittämistä olisikin hyvä kohdentaa näiden asioiden kehittämiseen asiakaslähtöisyyden takaamiseksi ja parantamiseksi.Customer orientation is a core principle of occupational health care. The customer-oriented approach can be defined as a partnership between occupational health care and the workplace based on trust and contracts. Well-functioning co-operation between the workplace and occupational health care enables the development of the client relationship and adds value and impact of the occupational health care measures. The customer-oriented approach considers the starting point and environment of the client company. When it comes to the development of customer oriented social and health care services, a historical change process is taking place. At present, clients are required to have an equal role as an influencer and developer with the service providers in the service system. Responding to customer needs, customer orientation in service operations and customer involvement are central to the change process. Customer orientation and the development of customer-oriented services is also a key objective of the program of the current Finnish Government (Inclusive and Competent Finland). One strategic component of the Government Program is the improvement of social and health care services. The goal of service development is to provide care in a timely, equitable, effective, cost-effective, holistic and flexible manner. The development work was carried out as a case study and its purpose was to develop a customer-oriented approach for a private service provider to develop occupational health care services. The aim was to make occupational health services more customer oriented. The research material of the case study was collected in three ways: a customer survey for client companies (n=64), occupational health care nurses managing client companies were interviewed (theme-oriented interviews) (n=4) and workshops were organized in client companies (n=3). Based on this material, the cooperation between client companies and occupational health care was described, as well as the needs and aspirations of client companies to develop occupational health services. The customer-oriented approach was created based on theme interviews with occupational health care nurses and workshops organized in client companies. In the customer oriented operating model, the following points emerged as key is-sues: utilization of multi-professionalism, customer- and need-based approach, availability of services and communication of the content of occupational health care. In the future, it would be a good idea to focus on these issues in the development of occupational health care services in order to ensure and improve customer orientation

Musiikkia maalaten : Kuinka 5-vuotiaat kuvaavat instrumentaalimusiikkia

Tässä opinnäytetyössä tutkin sitä, miten 5-vuotiaat lapset kuvaavat klassista instumentaalimusiikkia musiikkimaalauksen keinoin. Lisäksi tutkin, onko maalauksissa havaittavissa eroa niiden lasten kohdalla, jotka ovat kotonaan tottuneet kuuntelemaan klassista musiikkia. Soitin lapsille kolme erilaista orkesterikappaletta, joista he maalasivat vesivärimaalaukset. Selvitin lasten kotitaustat suhteessa musiikin harrastamiseen, haastattelin lapsia maalaustilanteiden jälkeen ja analysoin heidän maalauksiaan. Tutkimuskysymykseni olivat seuraavat: 1. Ottavatko 5-vuotiaat lapset maalatessaan maalauksiinsa ideoita tai vaikutteita soivasta instrumentaalimusiikista? 2. Onko instrumentaalimusiikin tulkitseminen lapselle helpompaa ja luontevampaa, jos hän on tottunut kuulemaan sitä kotona vanhempiensa kanssa? Tutkimuksesta käy ilmi, että osa lapsista ottaa musiikista vaikutteita maalauksiinsa ja osa taas ei kokenut musiikin ohjaavan maalausprosessiaan. Vaikutteita ja ideoita saaneiden lasten joukossa oli sekä niitä, joiden kotona kuunnellaan aktiivisesti klassista musiikkia, että myös lapsia, joilla ei ollut tällaista taustaa. Yhdenmukaisuutta samaan kappaleeseen maalatuissa maalauksissa oli lähinnä värimaailmoissa: voidaan siis päätellä kappaleen tunnelman välittyneen lasten värinkäyttöön. Tutkimusta tehdessäni totesin lasten nauttivan suuresti musiikkimaalauksesta. He ovat hyvin avoimia musiikin eri tyyleille, olivat ne sitten tuttuja entuudestaan tai eivät.In this thesis I study how 5-year-old children portray classical instrumental music in their paintings. I also study, whether there can be seen a difference in the paintings of children who are used to listening to classical music at home. I played the children three different orchestral pieces, from which they painted with water-colours. I researched the children’s backgrounds regarding their musical habits, interviewed them after the painting moments and analysed their paintings. My research questions were: 1. Do 5-year-old children take ideas or influence to their paintings from the instrumental music playing? 2. Is it easier and more natural for a child to interpret instrumental music if he or she is used to listening to it at home? Based on the research, it can be seen that some of the children take influence from the music and others didn't find it guiding their painting process. Among the children that took influence and ideas from the music were both those who had been listening classical music in their home and those who did not have that kind of background. The similarities in the paintings painted to the same music were mainly in their colouring so it can be concluded that the music's atmosphere influenced the childrens' use of colours. While conducting this research I noticed that children greatly enjoy music painting. They are very open to different types of music, whether they are familiar to them or not

Differential expression of aspartic and cysteine proteinases, glutamine synthetase, and a stress protein, HVA1, in germinating barley

Nitrogen is considered as a rate-limiting factor in plant growth. In barley (Hordeum vulgare, L.) seed, most of the nitrogen is deposited in storage proteins, which are remobilized for the growing seedling during germination. Proteolytic enzymes are of central importance in the hydrolysis of the reserve proteins. The main transport form of nitrogen in cereal seeds is glutamine, which is synthesized by glutamine synthetase. An ABA-responsive protein, HVAl, is induced during seed maturation but also due to desiccation stress during germination. In the present study, the expression of two different acid proteases, an aspartic and a cysteine proteinase, together with glutamine synthetase and a stress protein, HVAl, was studied by immuno-microscopy during barley germination. Aspartic proteinase is already present in the mature barley seed. During germination, it was gradually expressed in all tissues of the seed, excluding the dead starchy endosperm. Subcellulary, aspartic proteinase was localized in the protein storage vacuoles. Tissue distribution and the amount of the two molecular forms of the enzyme varied, suggesting different physiological roles for the two heterodimers. Contrary to the case of aspartic proteinase, the synthesis and secretion of the 30 kD cysteine proteinase was started in the scutellar epithelium after the beginning of germination, and continued in the aleurone layer. The enzyme was found to be secreted via the Golgi complex and the putative secretory vesicles to the cell exterior. Glutamine synthetase (GS) was expressed as a cytosolic isozyme in the scutellum and the aleurone layer of germinating barley seed. At the beginning of germination, GS was likely to be expressed as a homo-octamer, and later as different hetero-octamers. Under the stress conditions, a barley LEA3 protein, HVAl, was strongly induced, although it was present in the mature seed and declined gradually during germination. Although HVAl protein has no putative signal peptide, it was localized both in the cytoplasm and protein storage vacuoles, suggesting the presence of a direct uptake mechanism. HV Al might be degraded by the aspartic proteinase, as they were colocalized in the protein storage vacuoles. The exact function of HVAl remains an open question