This thesis evaluated the role of Serine/Arginine-rich proteins, also known as SR proteins, in addition to LAMMER kinases in the cold acclimation response using Brassica napus and Arabidopsis thaliana. Transcription profile analyses of SR and LAMMER kinase genes in Brassica napus and BnCBF overexpressor lines showed that exposure to low temperatures led to increased transcript levels for nine SR genes and two kinases. BnCBF overexpression was found to exacerbate this response. This was associated with increases in SR protein abundance and phosphorylation status, suggesting that SR proteins play an essential role in cold acclimation. These findings prompted further studies to assess the role of SR proteins and kinases in the cold acclimation induced adjustment of photosynthesis, the acquisition of freezing tolerance and the transcriptional profile of CBF, SPS and COR genes, which play an important role in the transcriptional cascade allowing plants to undergo cold acclimation. Using Arabidopsis loss-of-function mutants of SR proteins and AME3 LAMMER kinase, it was shown that At-RSZ22 and At-SR45 are indispensable in the regulation of photosynthesis under non-acclimated and cold acclimation conditions. At-RSZ22a, At-SCL30 and At-RS41 were then proposed to play a crucial role in the cold acclimation induced adjustment of photosynthetic performance. Moreover, the deletion of At-AME3 kinase not only jeopardized the cold acclimation induced adjustment of photosynthetic performance, but also the acquisition of freezing tolerance. This was associated with attenuation of the transcription profile of key cold responsive genes and protein abundance of COR15 A/B and dehydrins. These findings prompted further physiological characterization of ame3 mutants, and the elucidation of Serine/Arginine-rich proteins capable of interacting with this LAMMER kinase of interest. Under cold stress and acclimation conditions, the deletion of At-AME3 LAMMER kinase impeded Photosystem I physiology and state-1 state-2 transitions. These findings were associated with decreases in Photosystem II and Photosystem I protein abundance. Yeast 2-hybrid assays showed that six SR proteins are capable of physically interacting with AME3. Taken together, the results of this study demonstrate that At-RSZ22 and At-SR45 are essential in the photosynthetic performance of Arabidopsis, that At-RSZ22a, At-SCL30 and At-RS41 play an essential role in the cold acclimation induced recovery of photosynthetic performance, and that At-AME3 plays an essential role in the cold acclimation response.

Wheat (Triticum aestivum L.) is occasionally exposed to low temperature during flowering and huge economic losses can occur especially in some key production countries such as Australia. Although it is generally predicted that there will be a rise in global temperature there are still predicted to be risks associated with low temperature for temperate crops. Post head emergence frost damage remains a major constraint to increasing wheat production. Five Iraqi varieties (Abu-Ghariab, Fatah, Sham6, IPA95 and IPA99) were screened for their frost hardiness (LT50). Abu-Ghariab was chosen for further investigations along with the European cv. Claire because the acclimated frost hardiness level in these two varieties (LT50 -8.07 and -8.01°C for Claire and Abu-Ghariab respectively) was found to be significantly lower than the other varieties in this study. Several techniques were employed including REC%, IR thermography and molecular analysis of cold acclimation in order to characterise the frost resistance of those two varieties. REC% revealed that both Claire and Abu-Ghariab could tolerate some freezing when the spikes were just visible (ZCK 51-60) with an acclimation shift of the LT50 of -1.6 and -2.11°C respectively but this was not apparent at later growth stages. Based on molecular analysis, cold acclimation was shown to be activated at ZCK 51-60 in both varieties. Cbf14 was expressed after 8 hours exposure to acclimatising temperatures (4°C) and then declined to a low, but still up-regulated level in both varieties and this led to expression of the COR15a protein. These molecular changes correlated with the frost tolerance recorded at ZCK 51-60. It was concluded that the possibility existed to up-regulate cold acclimation after spike emergence if there was enough environmental stimulus. Molybdenum (Mo) was demonstrated to work synergistically with low temperature in increasing the expression of Cbf14 and COR15a. The European wheat cv. Claire showed a higher capacity (-8.14°C) to be acclimated than Iraqi wheat (-7.40°C) under the effect of both Mo and acclimation temperatures. Mo alone increased the expression of Cbf14 in both varieties but did not increase the frost tolerance. Observations of ice nucleation using an infrared thermography (IR) revealed that supercooling is highly likely in spikes and some spikes avoided frost damage even when the temperature fell to -12°C. It was observed that the proportion of frozen spikes was 22.8% while the remainder supercooled. Spraying plants with distilled water was not effective in facilitating ice nucleation in wheat spikes. Observations also revealed that spikes that did freeze started freezing at temperatures of -4 to -5 °C close to temperature of the putative constitutive frost hardiness of un-acclimated wheat and it is suggested that this may reflect that many laboratory freezing experiments may not actually freeze until -5°C. The anthesis stage of wheat was found to be the most vulnerable stage and it needs to be given more attention in terms of research to up-regulate cold acclimation. Frost damage to wheat during flowering continues to be a serious problem in certain production areas and therefore continued effort in characterising and finding suitable solutions to this are imperative.

<p>Although low temperature plays an important role in determining agricultural yield, little is known about the effect on the underlying biochemical and physiological processes that influence plant growth. Photosynthesis and respiration are central to plant growth and both processes are heavily affected by temperature. However, many plants have the ability to cope with low temperature and resume growth by cold acclimating.</p><p>We have shown that enhancement of carbon fixation, an increased flux of carbon into sucrose and the recovery of diurnal export is crucial for the recovery of functional carbon metabolism at low temperature in Arabidopsis thaliana. The recovery of efflux is governed by increased expression of sucrose transporters along with changes in vascularisation. We also demonstrate the importance of controlling the flux of metabolites between the chloroplast and the cytosol by regulating the expression of AtTPT.</p><p>We further investigated the difference in response between leaves developed at low temperature but originating from warm grown Arabidopsis and leaves from plants grown from seed at low temperature. We were able to distinguish factors that respond specifically to low temperature from those that are connected to the actual stress. Substantial difference could be seen in the different metabolomes. One conclusion drawn is that the increase in sucrose reported at low temperature is an essential feature for life in the cold. </p><p>In an extended study we were able to transfer some of the key factor of cold acclimation in Arabidopsis to other species. The study included forbs, grasses and evergreen trees/shrubs showed that there are striking similarities in the extent and biochemical changes that underpin acclimation among the different functional groups.</p><p>Low temperature does not only influence growth of the leaves, perennial organs such as the corm of the ornamental plant Crocus vernus is also affected. However in these plants low temperature has a positive effect on the final size of the corm. We were able to show that this enhanced growth was an affect of increased cell size and thus increased sink capacity, which ultimately delays leaf senescence</p>

Although low temperature plays an important role in determining agricultural yield, little is known about the effect on the underlying biochemical and physiological processes that influence plant growth. Photosynthesis and respiration are central to plant growth and both processes are heavily affected by temperature. However, many plants have the ability to cope with low temperature and resume growth by cold acclimating. We have shown that enhancement of carbon fixation, an increased flux of carbon into sucrose and the recovery of diurnal export is crucial for the recovery of functional carbon metabolism at low temperature in Arabidopsis thaliana. The recovery of efflux is governed by increased expression of sucrose transporters along with changes in vascularisation. We also demonstrate the importance of controlling the flux of metabolites between the chloroplast and the cytosol by regulating the expression of AtTPT. We further investigated the difference in response between leaves developed at low temperature but originating from warm grown Arabidopsis and leaves from plants grown from seed at low temperature. We were able to distinguish factors that respond specifically to low temperature from those that are connected to the actual stress. Substantial difference could be seen in the different metabolomes. One conclusion drawn is that the increase in sucrose reported at low temperature is an essential feature for life in the cold. In an extended study we were able to transfer some of the key factor of cold acclimation in Arabidopsis to other species. The study included forbs, grasses and evergreen trees/shrubs showed that there are striking similarities in the extent and biochemical changes that underpin acclimation among the different functional groups. Low temperature does not only influence growth of the leaves, perennial organs such as the corm of the ornamental plant Crocus vernus is also affected. However in these plants low temperature has a positive effect on the final size of the corm. We were able to show that this enhanced growth was an affect of increased cell size and thus increased sink capacity, which ultimately delays leaf senescence

It is well known that photosynthetic health impacts the overall fitness of the mature plant. This study aims to determine photosynthetic vigour of spring wheat cultivars during field development as well as their biomass composition at maturity to determine which cultivars/varieties would be optimum for cellulosic ethanol production. Additionally, specimens were grown at non-acclimating (20˚C), cold acclimating (5˚C), non-acclimating high CO2 (20˚C/750 µmol mol-1 CO2) and cold-acclimating high CO2 (5˚C/750 µmol mol-1 CO2) to resolve photosynthetic responses to different environments. Plants were photoinhibited under high irradiance (5 fold growth irradiance) and low temperature (5˚C) while photochemical efficiency of PSII was monitored throughout using chlorophyll fluorescence imaging. Vegetative production was monitored using normalised difference vegetation index. De-epoxidation of xanthophyll photoprotective pigments were also recorded using HPLC and photochemical reflectance index. Additionally, carbon assimilation rate was recorded with infra-red gas analysis methods. It was discovered that no one wheat cultivar demonstrated any photosynthetic advantage in the field or under photoinhibitory conditions. However, photosynthetic differences were observed between wheat grown in different environments. Plants that were cold-acclimated or grown under high CO2 were more resilient to photoinhibitory stress. This was also reflected by most cold-acclimated cultivars having increased triose phosphate utilization, electron transport and zeaxanthin induction. Plants acclimated to high CO2 at room temperature also displayed increased electron transport and triose phosphate utilization but had decreased zeaxanthin induction. It is hypothesized increased excitation pressure in cold acclimated and high CO2 cultivars allowed for their increase in the development of photoinhibitory tolerance.

It is well known that photosynthetic health impacts the overall fitness of the mature plant. This study aims to determine photosynthetic vigour of spring wheat cultivars during field development as well as their biomass composition at maturity to determine which cultivars/varieties would be optimum for cellulosic ethanol production. Additionally, specimens were grown at non-acclimating (20˚C), cold acclimating (5˚C), non-acclimating high CO2 (20˚C/750 µmol mol-1 CO2) and cold-acclimating high CO2 (5˚C/750 µmol mol-1 CO2) to resolve photosynthetic responses to different environments. Plants were photoinhibited under high irradiance (5 fold growth irradiance) and low temperature (5˚C) while photochemical efficiency of PSII was monitored throughout using chlorophyll fluorescence imaging. Vegetative production was monitored using normalised difference vegetation index. De-epoxidation of xanthophyll photoprotective pigments were also recorded using HPLC and photochemical reflectance index. Additionally, carbon assimilation rate was recorded with infra-red gas analysis methods. It was discovered that no one wheat cultivar demonstrated any photosynthetic advantage in the field or under photoinhibitory conditions. However, photosynthetic differences were observed between wheat grown in different environments. Plants that were cold-acclimated or grown under high CO2 were more resilient to photoinhibitory stress. This was also reflected by most cold-acclimated cultivars having increased triose phosphate utilization, electron transport and zeaxanthin induction. Plants acclimated to high CO2 at room temperature also displayed increased electron transport and triose phosphate utilization but had decreased zeaxanthin induction. It is hypothesized increased excitation pressure in cold acclimated and high CO2 cultivars allowed for their increase in the development of photoinhibitory tolerance.

Atsparumas šalčiui ir ištvermingumas žiemą – pagrindiniai veiksniai, lemiantys augalų geografinį pasiskirstymą. Be to, šie veiksniai turi didžiulę įtaką kultūrinių augalų derlingumui. Braškės, kurios yra tarp svarbiausių uoginių augalų pasaulyje ir Lietuvoje, stipriai pašąla esant nepastoviai sniego dangai, vis dažniau pasitaikančiomis permainingomis žiemojimo sąlygomis. Nepakankamas braškių ištvermingumas žiemą, aiškių kriterijų, kurie leistų per trumpesnį nei kelių metų laikotarpį atrinkti ištvermingas veisles nebuvimas, apsunkina naujų, adaptyvių, derlingų, kokybiškomis uogomis, braškių veislių sukūrimą ir introdukciją. Atsparumo šalčiui mechanizmai intensyviai tirti naudojant modelinius augalus, tačiau žinių apie žiemojančių kultūrinių augalų, kurių įvairovė labai didelė, ištvermingumo žiemą mechanizmus ir jo genetinį reguliavimą labai trūksta. Atsparių šalčiui augalų atrankai vykdyti reikalingos žinios apie augalų užsigrūdinimą, apie atsparumą šalčiui lemiančius veiksnius ir juos kontroliuojančius genus. Tiriant augalus kontroliuojamomis sąlygomis in vitro, galima ištirti atskirų veiksnių įtaką užsigrūdinimui ir atsparumui šalčiui, atsiribojant nuo, neišvengiamo natūraliomis sąlygomis, daugelio įvairių veiksnių poveikio. Braškių užsigrūdinimas ir atsparumas šalčiui svarbūs ne vien tik atrankai. Genetinių išteklių išsaugojimas ateities kartoms - svarbus šiuolaikinės biologijos uždavinys. Didelių kolekcijų išsaugojimą palengvina ir sąnaudas sumažina, augalų laikymas... [toliau žr. visą tekstą] / Cold hardiness and resistance in winter are key factors determining the geographical distribution of plants. In addition, these factors have a huge impact on harvest of cultural plants. Strawberries, which are among the most important berry plants in the world and in Lithuania, are vulnerable to cold because of unstable snow cover, which is more and more often occurring during variable wintering conditions. Insufficient resistance of strawberries in winter, absence of clear criteria, which enable to select the most resistant varieties during a shorter time than the few year period, impedes the development and introduction of new, adaptive, fertile, and of high-quality berries strawberry cultivars. The mechanisms of cold hardiness were intensively investigated using model plants. However, there is a lack of knowledge about the genetic regulation mechanisms of winterhardiness of cultural plants. For the selection of resistant to cold plants, the knowledge about plants cold acclimation and factors determining cold resistance and controlling genes is necessary. Investigating the plants under controlled conditions in vitro, it is possible to investigate the influence of individual factors on acclimation and cold hardiness by disassociating from the influence of many different factors, which is unavoidable under natural conditions. The cold acclimation and cold hardiness of strawberries are important not only for breeding. Preservation of plant genetic resources for future... [to full text]

Human beings have been exposed to different cold conditions throughout time, and have through cold acclimation developed mechanisms to survive in these conditions. Cold acclimation can be elicited through exposure to natural cold climates, or artificially induced in a laboratory to study the body’s response to repeated cold exposures. Several studies looking at the effects of cold acclimation in humans have been conducted during the last 50 years, and have reported that cold acclimation can lead to a change in skin and core temperature, heat production and shivering. An accurate quantification of shivering thermogenesis (ST) during cold acclimation has not been done before, and most previous measurements of shivering during cold acclimation have been inaccurate and inadequate. In this study a Liquid Condition Suits (LCS) was used to elicit cold acclimation (10°C, 2hr daily, for 4 weeks) while an accurate measurement of the effect of cold acclimation on changes in muscle activity was conducted. In CHAPTER 2, results showed that four weeks of cold acclimation at 10°C did not change skin and core temperature, heat production or ST. The effects on shivering pattern and fuel selection were also analysed, but no effects of cold acclimation could be observed. These measurements were a part of a larger study, in which the effects of cold acclimation on changes in BAT were the main outcome measures. These data showed that an increase in BAT volume (45%) and activity (120%) were the only observed effects of cold acclimation. In CHAPTER 3, we set out to assess if changes in shivering from pre to post cold acclimation are associated with changes in BAT volume, and if the amount of BAT a participant possesses prior to cold acclimation can be used to predict changes in shivering intensity during cold acclimation. The interindividual variability in changes in thermal responses, heat production, shivering and BAT volume occurring between subjects during four weeks of cold acclimation was also addressed in this section.

The ability of the heart and skeletal muscles to remodel to environmental demands, their plasticity, is of interest when studying animals’ adaptation to environment changes. Temperature variation due to seasonal change seems to lead to the development of a cold acclimated phenotype in small birds. To endure cold conditions a higher metabolism is required for shivering thermogenesis in aerobic skeletal muscles. This in turn leads to several physiological changes, including a heart and muscle hypertrophy as well as an increased oxygen carrying capacity of the blood. In this study were Red Junglefowls (Gallus gallus) bred indoors and outdoors and physiological aspects such as body size, growth rate, relative size of heart and skeletal muscles (pectoralis major and gastrocnemius maximus) as well as hematocrit and hemoglobin concentrations of the blood were compared between the groups. Observed significant differences included a slower growth rate in fowls bred outdoors, 2.5 (0.7) g/day than indoors, 3.8 (0.4) g/day, as well as a larger relative size of the heart and gastrocnemius muscle. The average relative size of the heart was more than twice as big in fowls bred outdoors, 0.97 (0.08) %, than indoors, 0.40-42 (0.05) %. The average relative size of the gastrocnemius muscle for the fowl bred outdoor was significantly larger than for fowl bred indoors (0.95 (0.11) %, vs. 0.58-0,63 (0.09) %). In addition, fowl bred outdoors showed an increased capacity for oxygen transportation, with blood hematocrit values of 43 (3) % higher than 35-37 (3) % for the indoor animals. Fowls bred outdoors also showed higher hemoglobin concentrations in the blood, 127 (7) g/l, than fowls bred indoors, 113 (7) g/l. Findings indicate a cold acclimated phenotype among the outdoor bred fowls.

Human beings have been exposed to different cold conditions throughout time, and have through cold acclimation developed mechanisms to survive in these conditions. Cold acclimation can be elicited through exposure to natural cold climates, or artificially induced in a laboratory to study the body’s response to repeated cold exposures. Several studies looking at the effects of cold acclimation in humans have been conducted during the last 50 years, and have reported that cold acclimation can lead to a change in skin and core temperature, heat production and shivering. An accurate quantification of shivering thermogenesis (ST) during cold acclimation has not been done before, and most previous measurements of shivering during cold acclimation have been inaccurate and inadequate. In this study a Liquid Condition Suits (LCS) was used to elicit cold acclimation (10°C, 2hr daily, for 4 weeks) while an accurate measurement of the effect of cold acclimation on changes in muscle activity was conducted. In CHAPTER 2, results showed that four weeks of cold acclimation at 10°C did not change skin and core temperature, heat production or ST. The effects on shivering pattern and fuel selection were also analysed, but no effects of cold acclimation could be observed. These measurements were a part of a larger study, in which the effects of cold acclimation on changes in BAT were the main outcome measures. These data showed that an increase in BAT volume (45%) and activity (120%) were the only observed effects of cold acclimation. In CHAPTER 3, we set out to assess if changes in shivering from pre to post cold acclimation are associated with changes in BAT volume, and if the amount of BAT a participant possesses prior to cold acclimation can be used to predict changes in shivering intensity during cold acclimation. The interindividual variability in changes in thermal responses, heat production, shivering and BAT volume occurring between subjects during four weeks of cold acclimation was also addressed in this section.