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Plants can not escape from biotic and abiotic stress factors such as, extreme temperatures, high light intensity, drought, UV radiation, heavy metals, and pathogen attack. Plants have versatile defens systems against such stress conditions. In this study, the role of glutathione S-transferases (GSTs) in cold stress conditions were examined. Glutathione S-transferases are the enzymes that detoxify natural and exogenous toxic compounds by conjugation with glutathione. Glutathione, an endogenous tripeptide, is important as reducing agent, nucleophilic scavenger, and alleviate the chemical toxicity in the plants by the reaction of GSTs. Glutathione conjugates can be transported to the vacuoles or apoplast and are generally much less toxic than the parent compounds. In plants there are four distinct families of the soluble GSTs, namely Phi (F), Type I / Zeta (Z), Type II / Tau (U), Type III / Theta (T), Type IV. By contrast with the mammalian families of GST, relatively little is known about the plant GST families. Up to date, there is not any study on GST isolation and characterization from Turkish red pine, in this respect, this study well play a frontier role the future research dealing with this topic. In this study, some properties of Turkish red pine GST activity towards CDNB (1-chloro-2,4 dinitrobenzene) were examined. The average specific activity of Turkish red pine GST towards CDNB was found as 200&plusmn / 50 (Mean&plusmn / SE, n= 18) nmole/min/mg cytosolic protein. GSTs in cytosol prepared from Turkish red pine needles retained its activity without loss for four weeks at -80&amp / #61616 / C. The rate of conjugation reactions were linear up to 0.8mg of Turkish red pine cytosolic protein and 0.4 mg cytosolic protein was routinely used. The Turkish red pine GST showed its maximum activity at pH 8.0 in 25 mM phosphate buffer and 42 &amp / #730 / C. The measurements were carried out at room temperature (RT) of 25 &amp / #61616 / C. Turkish red pine GST seemed to be saturated at 1 mM CDNB and 1 mM GSH concentrations. The Vmax and Km values of Turkish red pine GST for CDNB was 416nmole/min/mg protein and 0,8 mM, respectively, and for GSH 106.4 nmole/min/mg protein and 0.10 mM, respectively. Turkish red pine cytosol was applied on DEAE-Sepharose fast flow column but almost no purification was achieved with respect GST activity. In order to examine the effects of cold stress on Turkish red pine GST activity, the GST activity was determined in 240 seedlings at &ndash / 3&amp / #61616 / , 0&amp / #61616 / and 13 &amp / #61616 / C environmental temperatures. It was observed that GST activity was the highest at -3&amp / #730 / C and the lowest at 13&amp / #730 / C in both cold resistant and sensitive families with the exception of Yaylaalan and &Ccedil / ameli.

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Indiana University-Purdue University Indianapolis (IUPUI) / Abiotic stresses, such as cold, are serious agricultural problems resulting in substantial crop and revenue losses. Soybean (Glycine max) is an important worldwide crop for food, feed, fuel, and other products. Soybean has long been considered to be cold-intolerant and incapable of cold acclimation. In contrast to these reports, this study demonstrates that cold acclimation improved freezing tolerance in the domestic soybean cultivar ‘Williams 82’ with 50% enhancement of freezing tolerance after 5.2 +\- 0.6 days of cold exposure. Decreases in light dependent photosynthetic function and efficiency accompanied cold treatment. These decreases were due to an increase in photon dissipation likely driven by a decrease in plastoquinone (PQ) pool size limiting electron flow from photosystem II (PSII) to photosystem I (PSI). Cold-induced damage to operational photosynthesis began at 25 minutes of cold exposure and maximal photosynthesis was disrupted after 6 to 7 hours of cold exposure. Cold exposure caused severe photodamage leading to the loss of PSII reaction centers and photosynthetic efficiency. Comparisons of eight cultivars of G. max demonstrated a weak correlation between cold acclimation and northern cultivars versus southern cultivars. In the non-domesticated soybean species Glycine soja, the germination rate after cold imbibition was positively correlated with seedling cold acclimation potential. However, the overall cold acclimation potential in G. soja was equal to that of domestic soybean G. max reducing the enthusiasm for the “wild” soybean as an additional source of genetic diversity for cold tolerance. Despite being relatively cold intolerant, the soybean genome possesses homologs of the major cold responsive CBF/DREB1 transcription factors. These genes are cold-induced in soybean in a similar pattern to that of the cold tolerant model plant species Arabidopsis thaliana. In Arabidopsis, EIN3, a major component of the ethylene signaling pathway, is a negative transcriptional regulator of CBF/DREB1. In contrast to AtEIN3 transcript levels which do not change during cold treatment in Arabidopsis, we observed a cold-dependent 3.6 fold increase in GmEIN3 transcript levels in soybean. We hypothesized that this increase could prevent effective CBF/DREB1 cold regulation in soybean. Analysis of our newly developed cold responsive reporter (AtRD29Aprom::GFP/GUS) soybean transgenic lines demonstrated that inhibition of the ethylene pathway via foliar sprays (AVG, 1-MCP, and silver nitrate) resulted in significant cold-induced GUS activity. Transcripts of GmEIN3A;1 increased in response to ethylene pathway stimulation (ACC and ethephon) and decreased in response to ethylene pathway inhibition in the cold. Additionally, in the cold, inhibition of the ethylene pathway resulted in a significant increase in transcripts of GmDREB1A;1 and GmDREB1A;2 and stimulation of the ethylene pathway led to a decrease in GmDREB1A;1 and GmDREB1B;1 transcripts. To assess the physiological effects of these transcriptional changes; electrolyte leakage, lipid oxidation, free proline content, and photosynthesis were examined. Improvement in electrolyte leakage, a measure of freezing tolerance, was seen only under silver nitrate treatment. Only 1-MCP treatment resulted in significantly decreased lipid oxidation. Transcripts for CBF/DREB1 downstream targets (containing the consensus CRT/DRE motifs) significantly decreased in plants treated with ethylene pathway stimulators in the cold; however, ethylene pathway inhibition generally produced no increase over basal cold levels. To identify if GmEIN3A;1 was capable of binding to GmDREB1 promoters, the negative regulator GmEIN3A;1 and the positive regulator GmICE1A were cloned and expressed in Escherichia coli (E. coli). Preliminary binding results indicated that GmEIN3A;1 can bind to a double stranded section of the GmDREB1A;1 promoter containing putative EIN3 and ICE1 binding sites. GmICE1A is capable of binding to the same section of the GmDREB1A;1 promoter, though only when single stranded. Additional experiments will be required to demonstrate that GmEIN3A;1 and GmICE1A are capable of binding to the GmDREB1A;1 promoter and this work provides the tools to answer these questions. Overall, this work provides evidence that the ethylene pathway transcriptionally inhibits the CBF/DREB1 pathway in soybean through the action of GmEIN3A;1. Yet when GmCBF/DREB1 transcripts are upregulated by ethylene pathway inhibition, no consistent change in downstream targets was observed. These data indicate that the limitation in cold tolerance in soybean is due to a yet unidentified target downstream of CBF/DREB1 transcription.

Living at the northern limits of its geographic range, the Florida manatee is particularly susceptible to cold stress-related mortality during the winter months, with most deaths occurring in the lower two-thirds of the state. Contributing to this cold stress susceptibility is the manatee's limited physiological and behavioral responses available when thermally stressed. While capable of migrating south in response to falling water temperatures, manatees must still find warm water when ambient river temperature drops below 20°C for more than a few days. This is in part due to the species low metabolic rate, limited capacity for thermogenesis, and limited ability to raise its metabolic rate. Prolonged exposure to cold temperatures may result in cold stress syndrome, which involves a number of potentially life-threatening, if not fatal physiological changes. Survival during the winter months is therefore, dependent upon the manatee's ability to balance basic physiological needs, primarily the need to forage and to obtain fresh water with the need to stay warm. When identifying which animals are most susceptible and where, analyses of statewide manatee mortality records from 1996 through 2011 (n = 823) indicated that, size and location matter. Medium to large-sized calves accounted for the majority of documented death from cold stress (46.6%), while subadults and small calves were the least represented size classes (14.3 % and 9.5%, respectively). Adults slightly outnumbered subadults (15.8%). Males outnumbered females in all size classes but gender differences were not statistically significant. With regards to location, two areas of the state, the southwest and central east coasts showed the highest incidents of cold stress-related mortality. Both are regions with no primary, natural warm-water springs and whose principal warm-water refugia are power plant effluents. Brevard County on the central east coast is the area most at risk during cold weather events accounting for more than 25% of all cold stress deaths statewide. Warm-water sites within this region are few and relatively underrepresented in the literature in an area well-studied in terms of manatee abundance and distribution relative to the operational power plant. Results from cold stress data analyses emphasize the importance of identifying and characterizing the physical attributes of both known and suspected secondary warm-sites used by manatees in this region for both long and short term protection of the species, and its critical habitat. Three locations within Brevard County identified as passive thermal basins (PTBs), and classified as secondary warm-water sites, have been documented supporting in excess of 100 manatees on numerous occasions, and during winters of varying severity. Unique in physical appearance, distance to forage, hydrology including thermal profiles, and when it was used by manatees, each site challenged the accepted definitions and criteria of what constitutes an acceptable and appropriate warm-water site. Through analyses of photo-identification records, site fidelity at two of these warm-water sites, the Berkeley Canal and the Desoto Canal, was established for a minimum of 20 highly identifiable animals, 15 of which used adjacent sites within the same year, and 6 that used both sites but during different years. Observations of daily use patterns within the sites supported optimization of thermoregulation through adjustments in both vertical and horizontal movement, the latter of which seemed to follow the path of the sun. Manatees using the sites also made use of bottom sediment presumably to stay warm at all three locations. Temperature data indicated that water temperatures monitored in the sediment at secondary sites were some of the highest in the county. The predictable movements during all but the coldest weather fronts indicated that manatees utilized these sites during the early morning and afternoon hours when ambient river temperatures were coldest, gradually returning to the river to feed as ambient temperatures began to rise later in the afternoon. The availability of PTBs in proximity to primary warm-water sites within the region may provide an important component needed for manatees to successfully balance the need to forage with the need to stay warm by providing a network that allows for more efficient foraging while reducing exposure to sub-critical ambient river temperatures. The challenge of balancing the need to forage and to maintain homeostasis in the face of thermal stress is complex. This complexity was best approached and better understood through use of a manatee energetics model. The model was designed to facilitate simulation of an unlimited number of different case scenarios involving the exposure of virtually created manatees to a variety of winter conditions as might be experienced by real manatees in a natural system. Sixty-four different simulations were run using six virtual manatees of differing ages, gender, physical parameters, and knowledge of warm-water sites. Simulations were conducted using actual winter water temperature data from Brevard secondary sites and the ambient river from both a mild and a severe winter season. Outcomes, measured as changes in physical parameters indicative of body condition (i.e. mass, percent body fat, blubber depth, girths, etc.), showed that all else being equal, calves in the 2 year-old range fared poorly in all scenarios when compared to individuals of larger size. Subadults fared better than larger adults. This outcome illustrates the complex relationship between size, energy requirements and the synergistic effects of body mass, body fat and blubber thickness on SA:V ratio. Model outcomes agree closely with manatee cold stress mortality analyses predicting that medium to large-sized calves are most susceptible to CS, followed by adults, then subadults. Because all models are simplifications of complex systems, the manatee energetics model is not without its flaws and limitations. The current version of the model could not predict the point at which cold stress mortality would occur. However, a cold stress warning system incorporated into the design alerts the user if potential CSS is likely based on changing physical parameters. Another limitation was the inability of the model to account for the behavioral plasticity of individual subjects since virtual manatees respond to water temperatures based on the user defined rules. A number of additional limitations related to gaps in existing manatee data the gaps were identified and defined. Despite these gaps, the model is designed to allow for incorporation of additional interactions, feedback loops and relevant data as it becomes available and as additional physiological interactions and energy requirements are more clearly defined. Sensitivity analyses, a feature of the model that allowed for modifications in a number of physical as well as environmental parameters, provided an otherwise unlikely opportunity to see how incremental changes in input values, specifically the starting values for mass, percent body fat and blubber depth affected the model's outcome. Ultimately the goal of the model was to facilitate a better understanding of complex relationships by challenging our preconceived understanding of the manatee and its environment.

<p> As the effects of climate change become more evident, the development of effective strategies for improving livestock climatic adaptation and the long-term sustainability of animal food production have become key priorities around the world, including in the US. Together with nutrition, infrastructure, and management practices, genetically improving animals is an effective and lasting alternative to simultaneously improve productive efficiency and climatic adaptation of animals. Genetic improvement requires basic understanding of the genomic architecture of the indicator traits of interest and the availability of large-scale datasets. Understanding the role of evolution and selection (both natural and artificial) on shaping animal genomes is of paramount importance for the optimization of breeding programs and conservation of genetic resources. In addition, properly quantifying environmental stress and individual animal responses to thermal stress are still important challenges in breeding programs. Thus, the identification of optimal statistical methods and traits that better capture key biological mechanisms involved in the heat stress response has the potential to enable more accurate selection for thermal tolerant individuals. Therefore, this thesis aimed to investigate complementary topics related to thermal tolerance in livestock species based on genomic information. A total of 946 genotypes from 34 cattle breeds, as well as Datong yak (<em>Bos grunniens</em>) and Bali (<em>Bos javanicus</em>) populations, adapted to divergent climatic conditions, were used to investigate the genetic diversity and unravel genomic regions potentially under selection for thermal tolerance, with a focus on Chinese local cattle breeds and yak. Different signature of selection analyses and a comprehensive description of genetic diversity in 32 worldwide cattle and Datong yak populations was presented. Moderate genetic diversity was observed within each Chinese cattle population. However, these results highlighted the need to adopt strategies to avoid further reduction in the genetic diversity of these populations. Several candidate genes were identified as potentially under selection for thermal tolerance, and important biological pathways, molecular functions, and cellular components were identified, which contribute to our understanding of the genetic background of thermal tolerance in <em>Bos</em> species. Secondly, 8,992 genotyped individuals were used to provide a comprehensive description of genotype-by-environment interaction effects, defining optimal environmental variables based on public weather station data, and critical periods to evaluate heat tolerance for various reproduction, growth, and body composition traits in US Large White pigs. The period of 30 days before the measurement date was suggested to analyze genotype-by-environment interaction for off-test weight, muscle depth, and backfat thickness. While for number of piglets weaned and weaning weight, the suggested period ranged from the last trimester of gestation until weaning. This same population was used to access the genomic predictive ability of heat tolerance based on routinely-measured traits and explore candidate regions involved in the biological mechanisms that underlie heat stress response in pigs. Genotype-by-environment interaction was identified for most of the traits evaluated, and moderate (>0.36 ± 0.05) breeding values prediction accuracy were achieved using genomic information. Lastly, various behavioral, anatomical, and physiological indicators of heat stress were measured in a population of 1,645 multiparous Large White x Landrace lactating sows. This dataset was used to identify the best statistical models and estimate genomic-based genetic parameters for 23 indicators of heat stress, including automatically-measured vaginal temperature, skin surface temperatures, respiration efficiency, respiration rate, panting score, body condition scores, hair density, body size, and ear measurements. All the traits evaluated are heritable, with heritability estimates ranging from 0.04 ± 0.01 to 0.40 ± 0.09. The genetic correlations among these traits ranged from -0.49 (between repeated records of vaginal temperature measured at 0800 hours and caliper body condition score) to 1.0 (between repeated records of vaginal temperature measured at 0800 hours and single record of vaginal temperature measured at 0800 hours; and between repeated records of vaginal temperature measured at 1200 hours and single record of vaginal temperature measured at 1200 hours). These findings indicate that genetic progress for thermotolerance in pigs can be achieved through direct indicators of heat stress in selection schemes. However, special attention is needed due to complex relationship between these traits as evidenced by their genetic correlations. In conclusion, this thesis provides important information to be used when designing breeding strategies for improving thermal tolerance in cattle and pigs, important genomic regions and metabolic pathways that are important for understanding the biological mechanism regulating thermal tolerance, as well as future directions for investigations in the area of livestock climatic adaptation.</p>

Background Patients’ cold exposure is a neglected problem in prehospital emergency care. Cold stress increases pain and anxiety and contributes to fear and an overall sense of dissatisfaction. When left untreated, cold stress disturbs vital body functions until ultimately reaches hypothermia. Aim The overall aim was to investigate patients’ experiences of thermal comfort and reactions to cold exposure in prehospital emergency care and to evaluate the effects of an intervention using active warming from underneath. Method Study I: Persons (n=20) injured in a cold environment in the north of Sweden were interviewed. Active heat was given to 13 of them. Study II: In wintertime, 62 patients were observed during prehospital emergency care. The field study was based on observations, questions about thermal discomfort, vital signs, and temperature measurements. Study III: Healthy young persons (n=23) participated in two trials each. Data were collected inside and outside a cold chamber. In one trial, the participants were lying on a regular ambulance stretcher and in a second trial on a stretcher supplied with a heated mattress. Outcomes were the Cold Discomfort Scale (CDS), back, finger, and core body temperature, four statements from the State-TraitAnxiety-Inventory (STAI), vital signs, and short notes about their experiences of the two stretchers. Study IV: A quantitative intervention study was conducted in prehospital emergency care in the north of Sweden. The patients (n=30) in the intervention group were transported in an ambulance supplemented with a heated mattress on the stretcher, whereas only a regular stretcher was used in the ambulance for the patients (n=30) in the control group. Outcomes were the CDS, finger, core body, and air temperature, and questions about cold experiences. Results Study I: Patients suffered more because of the cold than from the pain of their injuries. The patients were in a desperate need of heat. Study II: Patients are exposed to cold stress due to cold environments. There was a significant decrease from the first measurement in finger temperature of patients who were indoors when the ambulance arrived, compared to the measurement taken in the ambulance. In the patient compartment of the ambulance, 85% of the patients had a finger temperature below the comfort zone and almost half of them experienced the patient compartment in the ambulance to be cold. The regular mattress surface temperature at the ambulance ranged from -22.3 to 8.4 ºC. Study III: A statistical increase of the participants’ back temperature was found between those lying on the heated mattress compared to those lying on the regular mattress. The heated mattress was experienced as warm, comfortable, providing security, and easy to relax on. Study IV: Thermal comfort increased for the patients in the intervention group and decreased in the control group. A significant higher proportion of the participants rated the stretcher as cold to lie on in the control group compared to the intervention group. Conclusion The ambulance milieu is too cold to provide thermal comfort. Heat supply from underneath increased comfort and might prevent cold stress and hypothermia

As RNA helicases da família DEAD-box são enzimas que alteram as estruturas secundárias do RNA e auxiliam a formação de complexos ribonucleoproteicos, e são muito importantes em processos basais como a degradação dos RNAs e a biogênese dos ribossomos. A &#945;-proteobactéria criotolerante Caulobacter crescentus é um modelo experimental interessante para compreender o papel destas enzimas em baixa temperatura. A caracterização fenotípica de linhagens mutantes simples de quatro RNA helicases DEAD-box permitiu demonstrar que a RNA helicase RhlE é necessária para o crescimento em baixa temperatura. Os mutantes duplos mostraram redução do crescimento à temperatura normal e em baixa temperatura, com perda de viabilidade e alterações morfológicas. Através de ensaios de complementação cruzada mostrou-se que seus papéis fisiológicos são até certo ponto redundantes. O mutante rhlE também apresentou redução na formação de biofilme. A medida da expressão relativa dos genes que codificam as RNA helicases mostrou um aumento na expressão de três destes genes em estresse frio, e a análise dos perfis ribossomais mostrou a possível participação destas três RNA helicases na biogênese do ribossomo. / The RNA helicases of the DEAD-box family are enzymes that modify RNA secondary structures and help the formation of ribonucleoprotein complexes, and are very important in basal processes such as RNA degradation and ribosome biogenesis. The cryotolerant &#945;-proteobacterium Caulobacter crescentus is an interesting experimental model to understand the role of these enzymes at low temperature. The phenotypic characterization of strains with single mutations of four DEAD-box RNA helicases showed that RNA helicase RhlE is required for growth at low temperature. The double mutants showed reduction in growth at both normal and low temperatures, with loss of viability and morphological changes. Through cross-complementation assays it has been shown that their physiological roles are to some extent redundant. The rhlE mutant also showed reduction in biofilm formation. The relative expression of the genes encoding the RNA helicases showed an increase in the expression of three of these genes under cold stress, and the analysis of the ribosomal profiles showed the possible participation of these three RNA helicases in ribosome biogenesis.

Os cofatores de ferro-enxofre [Fe-S] estão entre os mais versáteis e antigos cofatores enzimáticos encontrados na natureza. As células têm explorado as propriedades eletrônicas e estruturais destes cofatores inorgânicos para uma ampla variedade de atividades incluindo a transferência de elétrons, a catálise e a ativação de substratos. Um grande número de proteínas está envolvido na biogênese dos cofatores [Fe-S], e este processo pode ser dividido em três etapas principais: (i) formação do enxofre elementar, (ii) montagem do cofator [Fe-S], e (iii) inserção do cofator em apoproteínas. As plantas realizam fotossíntese e respiração, dois processos que requerem proteínas Fe-S, sendo os únicos organismos em que a biossíntese destas proteínas é compartimentalizada. Diversos fatores afetam o desenvolvimento das plantas, entre eles, a temperatura baixa, fator limitante à produtividade e à distribuição geográfica das plantas, incluindo Eucalyptus grandis, uma espécie com grande importância econômica. Devido a esse fato, foi realizada uma análise transcricional dos genes codificados pelo sistema ISC de biossíntese de cofatores [Fe-S] NFS1, ISA1 e ISU1 de E. grandis por meio de PCR quantitativa (RT-qPCR), após plântulas desta espécie serem submetidas ao tratamento de frio. O gene NFS1 teve sua expressão reprimida nas primeiras 48 horas de tratamento, porém, após esse período observa-se um aumento da expressão gênica em relação à condição controle. O genes ISU1 e ISA1 apresentaram maior expressão gênica nas primeiras duas horas de tratamento, diminuindo drasticamente logo após este período. Foi verificado um aumento na quantidade relativa de Fe e S nos nas plântulas submetidas ao tratamento de frio, indicando um possível aumento na quantidade de cofatores [Fe-S] requeridos para o reestabelecimento da homeostase celular. As bactérias, por sua vez, desenvolveram pelo menos três sistemas de biossíntese, altamente conservados, que estão envolvidos na formação dos cofatores [Fe-S], sendo estes NIF, ISC e SUF. Em muitas proteobactérias, a regulação da produção de cofatores [Fe-S] pelos sistemas ISC e SUF é controlada por uma única proteína, IscR, pertencente à família de reguladores Rrf2. A proteína IscR possui um domínio de ligação ao DNA na região N-terminal e um segundo domínio de ligação de cofatores com três resíduos de cisteínas (Cys) altamente conservados. A ligação de um cofator do tipo [2Fe-2S] reprime a transcrição do seu próprio promotor in vitro. O genoma de Azotobacter vinelandii não inclui um sistema SUF completo e, portanto, permite o estudo dos efeitos da regulação de IscR não relacionada a SUF. No presente trabalho, objetivamos analisar a expressão do operon isc em linhagens selvagens e mutantes para IscR de A. vinelandii por meio das técnicas de sequenciamento do transcritoma e RT-qPCR. As substituições das Cys92, Cys104, His107 e a deleção de 120 pb da região codificadora do segundo domínio de IscR levaram à indução de um aumento da expressão de todo o operon isc. Notou-se também uma diferença fenotípica clara no tamanho das colônias portadoras das substituições de Cys e His, sendo estas menores em relação à linhagem selvagem. As substituições das Cys98 e Cys111, ou ainda a dupla substituição Cys98/111 não levaram a alteração da expressão do operon. A ligação ou não do cofator [Fe-S] é, portanto, responsável pela regulação do operon isc em A. vinelandii, bem como, de outros operons codificadores de proteínas envolvidas em cadeias tranportadoras de elétrons. / The iron-sulfur clusters [Fe-S] are among the oldest and most versatile enzyme cofactors found in nature. The cells have explored the structural and electronic properties of these inorganic clusters for a wide variety of activities including electron transfer, catalysis and activation of substrates. A large number of proteins is involved in the biogenesis of the [Fe-S] clusters, and this process can be divided into three main steps: (i) formation of elemental sulfur, (ii) assembly of the [Fe-S] cluster and (iii) insertion into apoproteins. Plants perform photosynthesis and respiration, two processes that require Fe-S protein, and in these organisms the synthesis of these proteins is compartmentalized. Several factors affect the development of plants, among them, the low temperature is a limiting factor to productivity and geographical distribution of plants, including Eucalyptus grandis, a specie with great economic importance. Due to this fact, we performed a transcriptional analysis by quantitative PCR (RT-qPCR) of the genes encoded by the E. grandis [Fe-S] cluster ISC system NFS1, ISA1 and ISU1 after seedlings were submitted to the chilling treatment. The NFS1 gene expression is repressed in the first 48 hours of treatment, but after this period there was an increase in gene expression relating to the control condition. The genes ISU1 and ISA1 showed higher gene expression in the first two hours of treatment, followed by a sharp decrease. There was an increase in the relative amount of Fe and S in the seedlings subjected to cold treatment, indicating a possible increase in the amount of [Fe-S] clusters, required for the reestablishment of cellular homeostasis. Bacteria have developed at least three synthesis systems, highly conserved, which are involved in the formation of Fe-S proteins, NIF, ISC and SUF. In many proteobacteria, the regulation of clusters production by ISC and SUF is controlled by a single protein, IscR, belonging to the Rrf2 regulators family. The protein IscR has a DNA binding site at the N-terminal domain and second cofactors binding domain with three cysteine residues (Cys) highly conserved. The binding of a [2Fe-2S] cluster represses the transcription of its own promoter in vitro. The genome of Azotobacter vinelandii does not include a full SUF system and thus permits the study of the effects of IscR regulation unrelated to SUF. In this study, the aim was to analyze the expression of isc operon in wild type and mutant strains of A. vinelandii IscR by the techniques of the transcriptome sequencing and qRT-PCR. The replacement of Cys92, Cys104, His107 and a deletion of 120 bp region encoding the second IscR domain led to an increased expression of the whole isc operon. It also showed a clear phenotypic difference in colonies size in the strains carrying the substitutions of His and Cys, it was smaller compared to the wild type strain. The replacement of Cys98 and Cys111, or the double substitution Cys98/111 not led to an altered operon expression. The [Fe-S] cluster binding or not, is therefore responsible for the regulation of the isc operon in A. vinelandii as well as of other operons encoding proteins involved in electron tranport chains.