PaLAR3 locus in Norway spruce genome and its link to resistance against Heterobasidion parviporum in drought stress

Heterobasidion –sukuun kuuluvien patogeenisten sienten aiheuttama juurikääpäinfektio on merkittävin hävikkipuuta aiheuttava patogeeni kuusella (Picea abies) Pohjoismaissa. Kuusen genomissa leucoantosyanidini reduktaasi -entsyymiä koodaavan PaLAR3 –geenin tiedetään vaikuttavan puuyksilön resistenssiin H. parviporum -juurikääpäinfektota vastaan. Geenin lokuksessa kuusella voi olla AA, AB tai BB alleelit. B-alleelin läsnäolon kuusen PaLAR3 lokuksessa on todettu vaikuttavan kuusen entsyymituotantoon siten, että (+)-catechiini pitoisuudet ovat korkeammat juurikääpäinokulaation jälkeen niillä yksilöillä, joiden PaLAR3 lokuksesta löytyy B-alleeli. Ilmastonmuutoksen seurauksena häiriöt ja muutokset hydrologisissa kierroissa lisääntyvät, ja sään ääri-ilmiöt yleistyvät. Tämän seurauksena kuivuuden uskotaan yleistyvän etenkin pohjoisilla alueilla, lisäten patogeenien ja tuholaisten leviämisen riskiä. Metsänjalostusohjelmilla pyritään jalostamaan metsiä siten, että ne olisivat taloudellisesti tuottavia ja kannattavia tulevaisuudessakin. Tutkimalla kuusen genetiikkaa jalostusohjelmaan voidaan lisätä sellaisia yksilöitä, jotka todennäköisimmin kestävät tulevaisuuden ilmastohaasteet paremmin. Tässä tutkielmassa tutkittiin kuivuuden vaikutusta juurikääpäinfektion leviämiseen kuusessa kuivuusstressin alaisena. Kuusten PaLAR3 geenin alleelit määritettiin ja tuloksissa tarkasteltiin inokuloidun juurikäävän aiheuttaman nekroosin pinta-alaa suhteessa alleeleihin, puiden kasteluolosuhteisiin, sekä inokuloinnissa käytettyyn juurikääpäkantaan. Tulosten valossa PaLAR3 geenin alleeleilla ei ole vaikutusta nekroosin pinta-alaan. Tilastollisen analyysin perusteella vaikutusta ei havaittu kuivuusstressistä kärsineessä, eikä normaaleissa kasteluoloissa olleessa ryhmässä. Ainoa tilastollisesti merkittävä havainto oli sen, että eri juurikääpäkannat (Hpa1 ja Hpa2) infektoivat eri tavoin. Hpa2 kannan infektiokyky vaikuttaa olevan korkeampi normaaleissa kasteluolosuhteissa. Näiden tulosten valossa metsänjalostusohjelmaan ei tulisi lisätä PaLAR3 geeniä. Tulevaisuudessa tutkimuksen tulisi pyrkiä tutkimaan eri geenien vuorovaikutuksia patogeeni-infektion alaisuudessa. Esimerkiksi kuusella myös toinen geeni (PaLAC5) on jo yhdistetty juurikääpäresistenssiin, ja sen tutkiminen yhdessä PaLAR3-geenin tuottaman entsyymitoiminnan kanssa voisi tuoda ratkaisuja tulevaisuudessa todennäköisesti pahenevaan Heterobasidion –suvun juurikääpäinfektioiden torjumiseksi.Heterobasidion genus fungi are the most significant pathogens in Finland causing root rot. These fungi infect Norway spruce (Picea abies), causing wood deficit in the forest industry. Leucoanthosyanidin reductase enzyme encoding gene, PaLAR3, has been linked in to the resistance of Norway spruce against H.parviporum induced root rot infections. The alleles in this locus can be AA, AB or BB, and the enzyme production and (+)-catechin concentrations have been proven to be higher in inoculation experiments in individuals with B allele present in their PaLAR3 locus. Climate change is predicted to increase disturbances in not only on the hydrological cycles, but also on the weather conditions. The drought is predicted to increase in the Northern hemisphere, increasing the risk of pathogens to spread into new areas. Forest breeding programs aim to make the forests more profitable and productive in the future as well. By studying the genetics of Norway spruce, it is possible to add more resilient individuals into the breeding program, to tackle the climate challenges the future might hold. In this Master’s thesis the spreading of the root rot infection under drought stress was studied. The PaLAR3 alleles were determined, and the area of necrosis caused by the fungal infection was compared against the alleles, watering treatments and the fungal strain used in the inoculations. Regarding the results the alleles in an individual’s PaLAR3 locus did not have any effect on the area of the necrosis. There were no statistically significant differences between the watering treatments. The only statistically significant result was that the different fungal strains’ (Hpa1 and Hpa2) infectiveness varied between different watering treatments. This is probably due to the different routes of pathogenesis. Hpa2 strain seems to be more infective in normal watering conditions. However, considering these results the PaLAR3 gene should not be added into the forest breeding program as itself. In the future, the research should focus more on gene interactions, since also other genes (e.g., PaLAC5) have been linked in the resistance against root rot infections in Norway spruce. By studying these genes together, the solutions for the deteriorating situation of the spreading of the fungal diseases could be discovered

Can a tympanic thermometer be used for measurement of body temperature in clinically healthy cats? : a comparative study between a tympanic thermometer and a rectal thermometer

På de flesta djursjukhus och kliniker används idag rektaltermometer för att mäta kroppstemperatur hos katter. Denna metod anses som tillförlitlig för att mäta kroppstemperatur men kan upplevas som en obehaglig och stressande metod för katten. Vid vissa sjukdomstillstånd såsom andnöd, smittsam sjukdom eller smärta vid ändtarmen kan en alternativ metod vara önskvärd. Syftet med denna valideringsstudie var att undersöka om temperaturmätning med en human örontermometer är en tillförlitlig metod för att mäta kroppstemperatur hos friska katter jämfört med en rektaltermometer, samt att undersöka metodens varians. Denna studie ämnade också undersöka om örontemperaturmätningen uppfattades som mindre stressande för katterna jämfört med rektaltemperaturmätningen. Studien inkluderade 26 friska katter i deras hemmiljö. Kroppstemperaturen mättes en gång med en rektaltermometer och en till tre gånger i varje öra med en örontermometer där mätningarna i öronen gjordes först i vänster sedan i höger öra. Vilken av metoderna som varje katt började med randomiserades. Katternas beteende bedömdes utifrån Fear free pets stresskala (FAS) vid varje temperaturmätning. En örontermometer designad för barn användes samt en digital rektaltermometer. Studien indikerade att temperaturen mätt med en human örontermometer i vänster öra var mer jämförbar med rektaltemperaturen än vad temperaturmätning i höger öra var. Differensen mellan medeltemperaturen uppmätt i vänster öra och rektalt för hela kohorten var endast 0,2 °C i denna studie. Vid användande av örontermometer för mätning av kroppstemperaturen hos katt bör två mätningar göras i vänster öra och medelvärdet av dessa användas. Detta för att få ett så tillförlitligt mätresultat som möjligt. Ingen statistisk signifikant skillnad i temperatur i parvisa jämförelser mellan höger och vänster öra samt rektalt observerades, även om hela modellen var signifikant, vilket troligen kan förklaras av den stressberoende temperaturförändring som höger öra uppvisade jämfört med rektaltemperaturen. En signifikant högre stressnivå kunde påvisas vid rektaltemperaturmätningen jämfört med örontemperaturmätningarna scorat med FAS. Detta visar att ur ett djuretiskt perspektiv så är en örontermometer ett lämpligt alternativ för temperaturmätning hos katt. Studien visar att örontermometer är en någorlunda tillförlitlig mätmetod för att mäta kroppstemperatur hos friska katter i en forskningssituation, med hänsyn till metodens egen varians och vid jämförelse med en rektaltermometer. För att kunna dra vidare slutsatser om validiteten av örontemperaturmätning och dess applicerbarhet för katt bör vidare studier av sjuka katter i en klinisk situation utföras.Most veterinary hospitals and clinics today use a rectal thermometer when measuring body temperature in cats. This method is considered to be a reliable method for that purpose, but it can be uncomfortable and stressful for the patient. During certain medical conditions such as dyspnea, contagious diseases and rectal pain, an alternative method would be desirable. The aim of this validation study was to determine whether temperature measurement with a tympanic membrane thermometer can be used as a reliable method for measuring body temperature in clinically healthy cats compared to a rectal thermometer and to explore the variance of the method. This study also aimed to examine if temperature measurement with a tympanic membrane thermometer was perceived as less stressful for the cats than temperature measurement with a rectal thermometer. The study included 26 healthy cats where temperature was measured in their respective home environments. The body temperature was measured once with the rectal thermometer and one to three times in each ear with the tympanic membrane thermometer, where the ear measurements always begun with the left ear followed by the right. The order in which the methods were used was randomized. The cats’ stress levels were assessed according to a fear, anxiety and stress scale (FAS) for each method. A tympanic membrane thermometer designed for children and a digital rectal thermometer were used in the trials. The study showed that measurement of body temperature with a tympanic membrane thermometer designed for humans in the left ear was best comparable with measurement with a rectal thermometer. The mean difference for the temperature measured in the left ear compared to the rectal temperature was 0.2 °C in this study cohort. When the body temperature is measured in cats two measurements should be performed in the left ear and a mean value should be calculated, this to achieve as reliable measurements as possible. No statistically significant difference in pairwise comparisons between temperature measured in the right ear and the left ear as well as rectally was detected even if the model in itself was significant. This can likely be explained by the stress-dependent temperature change that the right ear displayed compared to the rectal temperature. A significantly higher stress level could be detected when measurements with the rectal thermometer was performed compared to the tympanic thermometer, scored with FAS. This indicates that from an animal ethic point of view the tympanic membrane thermometer is a good alternative for temperature measurement in cats. The study shows that temperature measurements with a tympanic membrane thermometer is a fairly reliable method for measuring the body temperature in clinically healthy cats in a research situation considering the variance of the methods and compared to a rectal thermometer. To make any further conclusions about the validity of tympanic membrane temperature measurements and the applicability for cats, further studies of unhealthy cats in a clinical situation should be performed

An effect of eyestalk ablation on antennular function in the spiny lobster, Panulirus argus

1. Unilateral removal of the eyestalk (optic ganglia and medulla terminalis) in the Bermuda spiny lobster, Panulirus argus , disrupts normal initiation of feeding activity via chemo-tactile stimulation of the antennule on the side of the ablation. This deficit may be permanent for it has lasted without apparent remission for over five months and two molts. Unilateral eyestalk ablation also produces a temporary increase in antennular cleaning activity directed toward the antennule on the side of the ablation. This effect seems to last for less than 16 days. Unilateral eyestalk ablation does not appreciably disturb spontaneous antennular movements or responses to mechanical stimulation of the antennule on either side. Nor does it destroy the ability of the lobster to give differential responses to mechanical and chemo-tactile stimuli. Most lobsters recover normal sensory and motor functions in antennules that regenerate after amputation of the distal segment and sensory flagellae. In about 1/3 of the animals, however, some form of sensory or motor abnormality is evident in the regenerated antennule. These deficits are ascribed to occasional failure of regenerating neurons to reform appropriate central connections. They do not resemble the deficits following eyestalk ablation. The medulla terminalis is tentatively proposed as the portion of the nervous system critical for normal antennular function that is removed by eyestalk ablation. There seem to be similarities between the effects of eyestalk ablation in the Crustacea as described here and the effects following destruction of portions of the corpora pedunculata in insects. 1. Einseitige Entfernung des Augenstieles (optische Ganglien und Medulla terminalis) bei der Bermuda-Stachellanguste, Panulirus argus , unterbricht auf der operierten Seite den Beginn der normalen Freß-tätigkeit, die durch chemische und taktile Reizung der Antennula eingeleitet wird. Diese Störung kann irreversibel sein, denn sie überdauerte in über 5 Monaten zwei Häutungen.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47096/1/359_2004_Article_BF00340474.pd

A developmental perspective on the evolution of the nervous system.

The evolution of nervous systems in animals has always fascinated biologists, and thus multiple evolutionary scenarios have been proposed to explain the appearance of neurons and complex neuronal centers. However, the absence of a robust phylogenetic framework for animal interrelationships, the lack of a mechanistic understanding of development, and a recapitulative view of animal ontogeny have traditionally limited these scenarios. Only recently, the integration of advanced molecular and morphological studies in a broad range of animals has allowed to trace the evolution of developmental and neuronal characters on a better-resolved animal phylogeny. This has falsified most traditional scenarios for nervous system evolution, paving the way for the emergence of new testable hypotheses. Here we summarize recent progress in studies of nervous system development in major animal lineages and formulate some of the arising questions. In particular, we focus on how lineage analyses of nervous system development and a comparative study of the expression of neural-related genes has influenced our understanding of the evolution of an elaborated central nervous system in Bilateria. We argue that a phylogeny-guided study of neural development combining thorough descriptive and functional analyses is key to establish more robust scenarios for the origin and evolution of animal nervous systems

Brain architecture in the terrestrial hermit crab Coenobita clypeatus (Anomura, Coenobitidae), a crustacean with a good aerial sense of smell

<p>Abstract</p> <p>Background</p> <p>During the evolutionary radiation of Crustacea, several lineages in this taxon convergently succeeded in meeting the physiological challenges connected to establishing a fully terrestrial life style. These physiological adaptations include the need for sensory organs of terrestrial species to function in air rather than in water. Previous behavioral and neuroethological studies have provided solid evidence that the land hermit crabs (Coenobitidae, Anomura) are a group of crustaceans that have evolved a good sense of aerial olfaction during the conquest of land. We wanted to study the central olfactory processing areas in the brains of these organisms and to that end analyzed the brain of <it>Coenobita clypeatus </it>(Herbst, 1791; Anomura, Coenobitidae), a fully terrestrial tropical hermit crab, by immunohistochemistry against synaptic proteins, serotonin, FMRFamide-related peptides, and glutamine synthetase.</p> <p>Results</p> <p>The primary olfactory centers in this species dominate the brain and are composed of many elongate olfactory glomeruli. The secondary olfactory centers that receive an input from olfactory projection neurons are almost equally large as the olfactory lobes and are organized into parallel neuropil lamellae. The architecture of the optic neuropils and those areas associated with antenna two suggest that <it>C. clypeatus </it>has visual and mechanosensory skills that are comparable to those of marine Crustacea.</p> <p>Conclusion</p> <p>In parallel to previous behavioral findings of a good sense of aerial olfaction in C. clypeatus, our results indicate that in fact their central olfactory pathway is most prominent, indicating that olfaction is a major sensory modality that these brains process. Interestingly, the secondary olfactory neuropils of insects, the mushroom bodies, also display a layered structure (vertical and medial lobes), superficially similar to the lamellae in the secondary olfactory centers of <it>C. clypeatus</it>. More detailed analyses with additional markers will be necessary to explore the question if these similarities have evolved convergently with the establishment of superb aerial olfactory abilities or if this design goes back to a shared principle in the common ancestor of Crustacea and Hexapoda.</p