Content of zinc, iron and their absorption inhibitors in Nicaraguan common beans (Phaseolus vulgaris L.).

Protein-energy malnutrition and mineral deficiencies are two of the three forms of nutritional deficiencies that affect most developing countries due to inadequate access to food and diets based on a sole crop. Common bean (Phaseolus vulgaris L.) is the staple crop of Nicaragua and it has the potential to improve the nutritional status of the poorest group of the nation. Its high content of both protein and nonhaem iron provides many nutrients, but inhibitors also may prevent absorption of iron and zinc by the human consumer. A proper production chain must be followed to ensure the best grain quality for the consumer. To achieve food security, both production and high nutritional content must be maintained. Four nationally important accessions of common bean, with different harvesting dates, were selected to be submitted to two treatments: to evaluate the impact of storage conditions on the end quality of the grain. The duration of the study was six months with sampling every six weeks, and the two treatments were controlled one stored at 40°C and 75 RH %, and the other was stored in in-situ conditions. Proximate and mineral composition was evaluated as well as tannin, phytate and bioavailability. Significant differences among different accessions were found, being the most significant in protein, Fe and Zn content, tannins and phytate. Protein values ranged from 21-23%. Iron content was 61-81 mg/kg but only 3-4% was bioavailable. Zinc content was 21-25 mg/kg and 10-12% was bioavailable. The concentration of phytate ranged from 8.6-9.6 mg/g while tannin values ranged within 37.7-43.8 mg/g. Storage at high temperatures was demonstrated to have an impact on certain nutritional compounds and proved detrimental to final grain quality. Soluble sugar content and tannin content decreased after six months in both storage conditions, IDF decreased in the in-situ and SDF in the stress. The iron content and bioavailability in INTA Biofortificado were not as outstanding as expected, so experiments should be conducted to compare its iron uptake and delivery with other cultivars.

Potato virus A as a heterologous protein expression tool in plants

Viruksien käyttö tuotekehityksen ja tutkimuksen vaatimien proteiinien tuottamiseen, syötävien rokotteiden kehittämiseen ja geeniterapiaan edustavat kasvavia biotekniikan sovellusalueita. Perunan A-virus (PVA) kuuluu potyviruksiin, joiden proteiinit tuotetaan aluksi yhtenä suurena molekyylinä, joka pilkotaan yksittäisiksi proteiineiksi viruksen itsensä tuottamilla entsyymeillä. Siten virusgenomiin lisätty vieras geeni käännetään proteiiniksi virusproteiinien mukana. Lopputuloksena kaikkia proteiineja tuotetaan kasvisoluissa samansuuruinen määrä. Lisäksi, viruksen proteiinikuoren koontimekanismi sallii perintöaineksen merkittävän lisäyksen ilman että viruksen tartutuskyky merkittävästi heikkenee. Koska virus monistuu ja leviää koko kasviin, jo melko pieni määrä kasveja riittää huomattavan proteiinimäärän tuottamiseen esimerkiksi säännösten mukaisessa kasvihuoneessa. Tämän työn tarkoituksena oli muuntaa PVA:n genomia siten, että virus soveltuisi yhden vieraan proteiinin tai useiden erilaisten proteiinien samanaikaiseen tuottamiseen kasveissa. Aluksi kokeiltiin viruksen replikaasia ja kuoriproteiinia koodaavien genomialueiden välistä kohtaa ja ihmisestä peräisi olevaa geeniä, joka tuotti S-COMT-entsyymiä (katekoli-O-metyylitransferaasi). Sen aktiivisuuden rajoittaminen auttaa Parkinsonintaudin hoidossa. Kasvissa tuotettua S-COMT:ia voitaisiin käyttää lääkekehityksessä estolääkkeiden testaukseen. Kahden viikon kuluttua tartutuksesta tupakan lehdissä oli entsymaattisesti aktiivista S-COMT:ia n. 1 % lehden liukoisista proteiineista. PVA:n P1-proteiinia koodaavalta alueelta oli paikannettu kohta, johon ehkä voitaisiin siirtää vieras geeni. Asia varmistettiin siirtämällä tähän kohtaan meduusan geeni, joka tuottaa UV-valossa vihreänä fluoresoivaa proteiinia (GFP). GFP-geeniä kantava PVA levisi kasvissa ja lisääntyi n. 30-50 %:iin viruksen normaalista pitoisuudesta. Koko kasvi fluoresoi vihreänä UV-valossa. Vieras geeni voidaan sijoittaa myös potyviruksen P1- ja HCpro-proteiineja koodaavien alueiden väliin. Samaan PVA-genomiin siirrettiin kolme geeniä, yksi kuhunkin kolmesta kloonauskohdasta: GFP-geeni P1:n sisälle, merivuokon lusiferaasigeeni P1/HCpro-kohtaan ja bakteerin beta-glukuronidaasigeeni (GUS) replikaasi/kuoriproteiini-kohtaan. Virusgenomin ja itse viruksen pituudet kasvoivat 38 %, mutta virus säilytti tartutuskykynsä. Se levisi kasveissa saavuttaen n. 15 % viruksen normaalista pitoisuudesta. Kaikki kolme vierasta proteiinia esiintyivät lehdissä aktiivisina.

Production of F4 Fimbrial Adhesin in Plants: a Model for Oral Porcine Vaccine against Enterotoxigenic Escherichia coli

F4 fimbriae of enterotoxigenic Escherichia coli (ETEC) are highly stable multimeric structures with a capacity to evoke mucosal immune responses. With these characters F4 offer a unique model system to study oral vaccination against ETEC-induced porcine postweaning diarrhea. Postweaning diarrhea is a major problem in piggeries worldwide and results in significant economic losses. No vaccine is currently available to protect weaned piglets against ETEC infections. Transgenic plants provide an economically feasible platform for large-scale production of vaccine antigens for animal health. In this study, the capacity of transgenic plants to produce FaeG protein, the major structural subunit and adhesin of F4 fimbria, was evaluated. Using the model plant tobacco, the optimal subcellular location for FaeG accumulation was examined. Targeting of FaeG into chloroplasts offered a superior accumulation level of 1% of total soluble proteins (TSP) over the other investigated subcellular locations, namely, the endoplasmic reticulum and the apoplast. Moreover, we determined whether the FaeG protein, when isolated from its fimbrial background and produced in a plant cell, would retain the key properties of an oral vaccine, i.e. stability in gastrointestinal conditions, binding to porcine intestinal F4 receptors (F4R), and inhibition of the F4-possessing (F4+) ETEC attachment to F4R. The chloroplast-derived FaeG protein did show resistance against low pH and proteolysis in the simulated gastrointestinal conditions and was able to bind to the F4R, subsequently inhibiting the F4+ ETEC binding in a dose-dependent manner. To investigate the oral immunogenicity of FaeG protein, the edible crop plant alfalfa was transformed with the chloroplast-targeting construct and equally to tobacco plants, a high-yield FaeG accumulation of 1% of TSP was obtained. A similar yield was also obtained in the seeds of barley, a valuable crop plant, when the FaeG-encoding gene was expressed under an endosperm-specific promoter and subcellularly targeted into the endoplasmic reticulum. Furthermore, desiccated alfalfa plants and barley grains were shown to have a capacity to store FaeG protein in a stable form for years. When the transgenic alfalfa plants were administred orally to weaned piglets, slight F4-specific systemic and mucosal immune responses were induced. Co-administration of the transgenic alfalfa and the mucosal adjuvant cholera toxin enhanced the F4-specific immune response; the duration and number of F4+ E. coli excretion following F4+ ETEC challenge were significantly reduced as compared with pigs that had received nontransgenic plant material. In conclusion, the results suggest that transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against porcine F4+ ETEC infections. The findings here thus present new approaches to develop the vaccination strategy against porcine postweaning diarrhea.

Distribution of non-biting midges (Diptera, Chironomidae) in subarctic lakes of Finnish Lapland applications in lake classification and palaeolimnology

Climate change contributes directly or indirectly to changes in species distributions, and there is very high confidence that recent climate warming is already affecting ecosystems. The Arctic has already experienced the greatest regional warming in recent decades, and the trend is continuing. However, studies on the northern ecosystems are scarce compared to more southerly regions. Better understanding of the past and present environmental change is needed to be able to forecast the future. Multivariate methods were used to explore the distributional patterns of chironomids in 50 shallow (≤ 10m) lakes in relation to 24 variables determined in northern Fennoscandia at the ecotonal area from the boreal forest in the south to the orohemiarctic zone in the north. Highest taxon richness was noted at middle elevations around 400 m a.s.l. Significantly lower values were observed from cold lakes situated in the tundra zone. Lake water alkalinity had the strongest positive correlation with the taxon richness. Many taxa had preference for lakes either on tundra area or forested area. The variation in the chironomid abundance data was best correlated with sediment organic content (LOI), lake water total organic carbon content, pH and air temperature, with LOI being the strongest variable. Three major lake groups were separated on the basis of their chironomid assemblages: (i) small and shallow organic-rich lakes, (ii) large and base-rich lakes, and (iii) cold and clear oligotrophic tundra lakes. Environmental variables best discriminating the lake groups were LOI, taxon richness, and Mg. When repeated, this kind of an approach could be useful and efficient in monitoring the effects of global change on species ranges. Many species of fast spreading insects, including chironomids, show a remarkable ability to track environmental changes. Based on this ability, past environmental conditions have been reconstructed using their chitinous remains in the lake sediment profiles. In order to study the Holocene environmental history of subarctic aquatic systems, and quantitatively reconstruct the past temperatures at or near the treeline, long sediment cores covering the last 10000 years (the Holocene) were collected from three lakes. Lower temperature values than expected based on the presence of pine in the catchment during the mid-Holocene were reconstructed from a lake with great water volume and depth. The lake provided thermal refuge for profundal, cold adapted taxa during the warm period. In a shallow lake, the decrease in the reconstructed temperatures during the late Holocene may reflect the indirect response of the midges to climate change through, e.g., pH change. The results from three lakes indicated that the response of chironomids to climate have been more or less indirect. However, concurrent shifts in assemblages of chironomids and vegetation in two lakes during the Holocene time period indicated that the midges together with the terrestrial vegetation had responded to the same ultimate cause, which most likely was the Holocene climate change. This was also supported by the similarity in the long-term trends in faunal succession for the chironomid assemblages in several lakes in the area. In northern Finnish Lapland the distribution of chironomids were significantly correlated with physical and limnological factors that are most likely to change as a result of future climate change. The indirect and individualistic response of aquatic systems, as reconstructed using the chironomid assemblages, to the climate change in the past suggests that in the future, the lake ecosystems in the north do not respond in one predictable way to the global climate change. Lakes in the north may respond to global climate change in various ways that are dependent on the initial characters of the catchment area and the lake.

Turvemaan männyn (Pinus sylvestris L.) ja kuusen (Picea abies [L.] Karst.) kanto- ja juuripuun ominaisuudet ja asetoniliukoisten uuteaineiden määrä

Tutkimus on osa Metsäklusteri Oy:n Future Biorefinery –tutkimusohjelmaa, jossa kartoitetaan mahdollisuuksia hyödyntää metsäteollisuuden raaka-aineita aiempaa tarkemmin ja uusissa tuotteissa. Tutkimuksen tavoitteena on selvittää männyn (Pinus sylvestris L.) ja kuusen (Picea abies [L.] Karst.) juurten ja kantopuun rakenne ja ominaisuudet. Tutkimuksessa selvitetään löytyykö männyn ja kuusen juurista reaktiopuuta ja määritetään asetoniliukoisten uuteaineiden osuus kantoja juuripuussa. Tutkimusaineistona oli viisi eri-ikäistä mäntyä ja viisi eri-ikäistä kuusta. Juuri- ja kantoaineisto kerättiin Metsäntutkimuslaitoksen toimesta Parkanon seudulta (62.017°N, 23.017°E) hakkuun jälkeen. Maanalaisista juurista otettiin näytteet kolmelta eri etäisyydeltä juurenniskaan nähden. Kummankaan lajin juurista ei löytynyt varsinaista reaktiopuuta, mutta joissakin näytteissä havaittiin lievää reaktiopuuta. Lievää reaktiopuuta löytyi enemmän männyn kuin kuusen juurista, eikä sitä löytynyt lainkaan kaikkein ohuimmista, noin 2 cm paksuisista juurenosista. Männyn kannoissa uuteaineprosentti oli korkeampi kuin kuusen. Männyn juurissa uuteaineprosentti kasvoi edettäessä kohti juuren kärkeä. Kuusella uuteaineprosentti laski aluksi, mutta lähellä juuren kärkeä taas kasvoi. Kuoren uuteainepitoisuus oli molemmilla puulajeilla korkeampi kuin puuaineen. Tutkimusaineisto oli suppea, eikä tutkimuksessa pyritty tilastolliseen yleistettävyyteen. Laajemmasta aineistosta tehdylle tutkimukselle on tarvetta, sillä turvekankailta saatavan puun tarjonta on Suomessa kasvussa, mutta juurten uuteaine- ja reaktiopuututkimuksia on tehty vain kivennäismailta kerätyistä aineistoista.

Helsingin Östersundomin pienvesien kartoitus

The area of Östersundom (29,1 square kilometers) was attached to Helsinki in the beginning of the year 2009. Östersundom is formed mostly from the municipality of Sipoo, and partly from the city of Vantaa. Nowadays Östersundom is still quite rural, but city planning has already started, and there are plans to develop Östersundom into a district with 45 000 inhabitants. In this study, the headwaters, streams and small lakes of Östersundom were studied to produce information as a basis for city planning. There are six main streams and five small lakes in Östersundom. The main methodology used in this study was the examination of the physical and the chemical quality of the water. The hygienic quality of the water was also studied. It was also examined whether the waters are in their natural state, or have they been treated and transformed by man. In addition, other factors affecting the waters were examined. Geographical information data was produced as a result of this work. Östersundom is the main area looked at in this study, some factors are examined in the scope of the catchment areas. Water samples were collected in three sampling periods: 31.8 4.9.2009, 3. 4.2.2010, and 10. 14.4.2010. There were 20 sampling points in Östersundom (5 in small lakes, 15 in streams). In the winter sampling period, only six samples were collected, from which one was taken from a small lake. Field measurements associated with water sampling included water temperature, oxygen concentration, pH and electoral conductivity. Water samples were analyzed in the Laboratories of Physical Geography in the University of Helsinki for the following properties: total suspended solids (TSS), total dissolved substances (TDS), organic matter, alkalinity, colour, principal anions and cations and trace elements. Metropolilab analyzed the amount of faecal coliform bacteria in the samples. The waters in Östersundom can be divided to three classes according to water quality and other characteristics: the upper course of the streams, the lower course of the streams and the small lakes. The streams in their upper course are in general acidic, and their acid neutralization capacity is low. The proportion of the organic matter is high. Also the concentrations of aluminium and iron tend to be high. The streams in the lower course have acidity closer to neutral, and the buffering capacity is good. The amounts of TSS and TDS are high, and as a result, the concentrations of many ions and trace elements are high as well. Bacteria were detected at times in the streams of the lower course. Four of the five small lakes in Östersundom are humic and acidic. TSS and TDS concentrations tend to be low, but the proportion of organic matter is often high. There were no bacteria in the small lakes. The fifth small lake (Landbonlampi) differs from the others by its water colour, which is very clear. This lake is very acidic, and its buffering capacity is extremely low. Compared to the headwaters in Finland in general, the concentrations of many ions and trace elements are higher in Östersundom. On the other hand, the characteristics of water were different according to the classification upper course streams, lower course streams, and small lakes. Generally, the best water quality was observed in the stream of Gumbölenpuro and in the lakes Storträsk, Genaträsk, Hältingträsk and Landbonlampi. Several valuable waters in their natural state were discovered from the area. The most representative example is the stream of Östersundominpuro in its lower course, where the stream flows through a broad-leaf forest area. The small lakes of Östersundom, and the biggest stream Krapuoja, with its meandering channel, are also valuable in their natural state.