Temperature Stress In Plants Research Articles

A high temperature responsive UDP-glucosyltransferase gene OsUGT72F1 enhances heat tolerance in rice and Arabidopsis.

OsUGT72F1 enhances heat tolerance in plants by improving ROS scavenging and modifying multiple metabolic pathways, under the regulation of transcription factors OsHSFA3 and OsHSFA4a. High temperature is one of the most critical environmental constraints affecting plant growth and development, ultimately leading to yield losses in crops such as rice (Oryza sativa L.). UDP (uridine diphosphate)-dependent glycosyltransferases (UGTs) are believed to play crucial roles in coping with environmental stresses. However, the functions for the vast majority of UGTs under high temperature stress remain largely unknown. In this study, we isolated and characterized a high temperature responsive UDP-glycosyltransferase gene OsUGT72F1 in rice. Our findings demonstrated that overexpression of OsUGT72F1 enhanced heat-stress tolerance, while the mutant plants displayed a sensitive phenotype under the same stress conditions. Ectopic expression of OsUGT72F1 in Arabidopsis thaliana also conferred improved heat tolerance to the plants. Further investigation revealed that OsUGT72F1 reduced the generation of reactive oxygen species (ROS) and boosted the activity of antioxidant enzymes, thereby alleviatingoxidativedamage under heat-stress conditions. Moreover, transcriptomic analysis indicated that the action of OsUGT72F1 leads to the upregulation of multiple metabolic pathways including phenylpropanoid biosynthesis, zeatin biosynthesis, and flavonoid biosynthesis. In addition, the upstream regulatory mechanism of the OsUGT72F1 gene has been identified. We found that the transcription factors OsHSFA3 and OsHSFA4a can bind to the OsUGT72F1 promoter and enhance its transcription level. Together, this study revealed that the glycosyltransferase gene OsUGT72F1 plays a vital role in the adaptive adjustment of high temperature stress in plants, revealing a new heat tolerance pathway and providing a promising gene candidate for the breeding of heat-resistant crop varieties.

Intragenic cytosine methylation modification regulates the response of SUCLα1 to lower temperature in Solanaceae

The tricarboxylic acid cycle (TCAC) is a fundamental metabolic process governing matter and energy in plant cells, playing an indispensable role. However, its involvement in responding to low temperature stress in potato remains poorly understood. Previous studies have identified succinyl-CoA ligase (SUCL), which catalyzes the phosphorylation of TCAC substrates, as a gene associated with lower temperatures. Nevertheless, its function in potato's response to lower temperatures remains unclear. Phylogenetic analysis has revealed that Solanum tuberosum possesses α and β subunits of SUCL, which cluster with those of Solanum lycopersicum, Nicotiana tabacum and Nicotiana benthamiana. Further investigation has shown that StSUCLα1 is predominantly located within mitochondria. Low temperatures induce methylation modification alterations at 11 intragenic cytosine sites and lead to changes in StSUCLα1 expression levels. Correlation analysis suggests that alterations in intragenic cytosine methylation sites of SUCLα1 may be associated with MET1. Knocking down NbSUCLα1, the homologous gene of StSUCLα1 in N. benthamiana, results in increased susceptibility to low temperature stress in plants. In summary, we have confirmed that SUCLα1 is a key gene modulated by intragenic cytosine methylation in response to lower temperatures, providing a novel target for genetic breeding aimed at enhancing potato tolerance to low temperature stress.

Calibrating Agro-Hydrological Model under Grazing Activities and Its Challenges and Implications

Recently, the Agricultural Policy Extender (APEX) model was enhanced with a grazing module, and the modified grazing database, APEXgraze, recommends sustainable livestock farming practices. This study developed a combinatorial deterministic approach to calibrate runoff-related parameters, assuming a normal probability distribution for each parameter. Using the calibrated APEXgraze model, the impact of grazing operations on native prairie and cropland planted with winter wheat and oats in central Oklahoma was assessed. The existing performance criteria produced four solutions with very close values for calibrating runoff at the farm outlet, exhibiting equifinality. The calibrated results showed that runoff representations had coefficients of determination and Nash–Sutcliffe efficiencies >0.6 in both watersheds, irrespective of grazing operations. Because of non-unique solutions, the key parameter settings revealed different metrics yielding different response variables. Based on the least objective function value, the behavior of watersheds under different management and grazing intensities was compared. Model simulations indicated significantly reduced water yield, deep percolation, sediment yield, phosphorus and nitrogen loadings, and plant temperature stress after imposing grazing, particularly in native prairies, as compared to croplands. Differences in response variables were attributed to the intensity of tillage and grazing activities. As expected, grazing reduced forage yields in native prairies and increased crop grain yields in cropland. The use of a combinatorial deterministic approach to calibrating parameters offers several new research benefits when developing farm management models and quantifying sensitive parameters and uncertainties that recommend optimal farm management strategies under different climate and management conditions.

The Roles of Circadian Clock Genes in Plant Temperature Stress Responses.

Plants monitor day length and memorize changes in temperature signals throughout the day, creating circadian rhythms that support the timely control of physiological and metabolic processes. The DEHYDRATION-RESPONSE ELEMENT-BINDING PROTEIN 1/C-REPEAT BINDING FACTOR (DREB1/CBF) transcription factors are known as master regulators for the acquisition of cold stress tolerance, whereas PHYTOCHROME INTERACTING FACTOR 4 (PIF4) is involved in plant adaptation to heat stress through thermomorphogenesis. Recent studies have shown that circadian clock genes control plant responses to temperature. Temperature-responsive transcriptomes show a diurnal cycle and peak expression levels at specific times of throughout the day. Circadian clock genes play essential roles in allowing plants to maintain homeostasis by accommodating temperature changes within the normal temperature range or by altering protein properties and morphogenesis at the cellular level for plant survival and growth under temperature stress conditions. Recent studies revealed that the central oscillator genes CIRCADIAN CLOCK ASSOCIATED 1/LATE ELONGATED HYPOCOTYL (CCA1/LHY) and PSEUDO-RESPONSE REGULATOR5/7/9 (PRR5/7/9), as well as the EVENING COMPLEX (EC) genes REVEILLE4/REVEILLE8 (REV4/REV8), were involved in the DREB1 pathway of the cold signaling transcription factor and regulated the thermomorphogenesis gene PIF4. Further studies showed that another central oscillator, TIMING OF CAB EXPRESSION 1 (TOC1), and the regulatory protein ZEITLUPE (ZTL) are also involved. These studies led to attempts to utilize circadian clock genes for the acquisition of temperature-stress resistance in crops. In this review, we highlight circadian rhythm regulation and the clock genes involved in plant responses to temperature changes, as well as strategies for plant survival in a rapidly changing global climate.

LNKs-RVEs complex ticks in the circadian gating of plant temperature stress responses

Recently, Kidokoro et al. found that protein complex LNK3,4-RVE4,8 and LNK1,2-RVE4,8 of the circadian clock modulates plant cold- and high-temperature tolerance, respectively. Here, we reviewed the discovery of LNKs, the dynamically formed morning-phased clock complexes, and their critical role on endogenous circadian rhythms. In addition, we summarized the research work on LNKs with the interacting proteins RVEs, CCA1 in temperature responses and discussed how the circadian clock confer increased fitness via gating the rhythmic expression of their target genes.

The role of ethylene in plant temperature stress response.

Temperature influences the seasonal growth and geographical distribution of plants. Heat or cold stress occur when temperatures exceed or fall below the physiological optimum ranges, resulting in detrimental and irreversible damage to plant growth, development, and yield. Ethylene is a gaseous phytohormone with an important role in plant development and multiple stress responses. Recent studies have shown that, in many plant species, both heat and cold stress affect ethylene biosynthesis and signaling pathways. In this review, we summarize recent advances in understanding the role of ethylene in plant temperature stress responses and its crosstalk with other phytohormones. We also discuss potential strategies and knowledge gaps that need to be adopted and filled to develop temperature stress-tolerant crops by optimizing ethylene response.

A very long-chain fatty acid enzyme gene, PxHacd2 affects the temperature adaptability of a cosmopolitan insect by altering epidermal permeability

Temperature fluctuations pose challenges to poikilotherms, such as insects, especially under climate change conditions. Very long-chain fatty acids (VLCFAs) form important structural components of membranes and epidermal surfaces, so play important roles in adaptation to temperature stress in plants. It has been unclear whether VLCFAs are involved in epidermis formation and thermal resistance in insects. In this study, we focused on the 3-hydroxy acyl-CoA dehydratase 2 (Hacd2), an important enzyme in the synthesis pathway of VLCFAs, in a cosmopolitan pest, the diamondback moth, Plutella xylostella. Hacd2 was cloned from P. xylostella and the relative expression pattern was identified. Epidermal permeability increased with the decreased VLCFAs in the Hacd2-deficient P. xylostella strain, which was constructed by using the CRISPR/Cas9 system. Survival and fecundity of the Hacd2-deficient strain was significantly lower than that of the wildtype strain when subject to desiccating environmental stress. Hacd2 mediates thermal adaptability in P. xylostella by changing epidermal permeability so is likely to be key to its remaining a major pest species under predicted climate change conditions.

Identification of Thermo-Sensitive Chloroplast Development Gene TSCD5 Required for Rice Chloroplast Development under High Temperature

Chloroplasts are critical organelles for photosynthesis and play significant roles in plant growth and metabolism. High temperature is one of abiotic stresses affecting the growth and development of plants, involving chlorophyll biosynthesis and chloroplast development. It is well known that the methyl erythritol 4-phosphate (MEP) pathway is vital to photosynthesis and plant growth, and 1-deoxy-D-xylulose-5-phosphate synthase (DXS) is the enzyme that catalyze the first step of the MEP pathway. Although DXS has been widely studied in microbes and plants, no DXS gene has been identified in rice. Here, a novel thermo-sensitive chlorophyll-deficient 5 (tscd5) mutant was isolated in rice with decreased chlorophyll contents, impaired chloroplasts, and albino leaves at high temperature (35 °C). Fine mapping and DNA sequencing of TSCD5 found a missense mutation (G to A) in the sixth exon of LOC_Os05g33840 in tscd5. The TSCD5 gene encodes a 1-deoxy-D-xylulose-5-phosphate synthase 1 (OsDXS1) localized in chloroplast. Complementation tests and overexpression assay demonstrated that the mutation in LOC_Os05g33840 caused the tscd5 phenotype. qRT-PCR of TSCD5 showed it was constitutively expressed in all tissues, and its transcript amounts were reduced in tscd5 under high temperature. Here, TSCD5 is verified to be crucial to chloroplast development under high temperature in rice, which may facilitate the elucidation of the molecular mechanisms which underlie acclimation to high temperature stress in plants.

Cell Cycle Regulation in Response to Temperature Stress in Plants

식물들은 온도변화에 민감하기에 지속적인 온도 스트레스는 식물의 생산력을 감소시킨다. 그러므로 연구자들은 식물 전 생육단계에서의 온도 변화 반응 메커니즘을 밝히는 것이 매우 중요하다. 식물들은 최초로 받았던 스트레스를 몸으로 기억하며, 이는 더 강한 스트레스가 왔을 때에도 더 효율적으로 방어체계를 구축하는데 도움을 준다. 이러한 방어 기작 중 하나로 단백질과 전사인자를 이용한 세포주기조절 유전자들을 변화시키는 것이 있다. 식물체 성장을 유도하는 유전자 및 메커니즘과 연관이 있는 크로마틴 재구성 요소는 온도 변화하에서 선택적 유전자 발현을 통하여 크로마틴의 구조를 변화시키고 수정한다. 세포주기조절에 중요한 역할을 하는 싸이클린과 싸이클린 의존 효소는 온도 변화에서 세포주기를 조절하고 재배열 시킨다. 메틸레이션과 히스톤 공유 변형 역시 여러 스트레스에 반응하여 잘못 만들어진 DNA를 재구성 한다. 온도 변화에 따른 비정상적 세포주기들도 DNA 수리 공정을 통하여 재배열 된다. 하지만 아직 여전히 세포주기와 식물 방어 기작과의 연관성이 많이 밝혀진 것은 없다. 그렇기에 연구자들은 여전히 식물체 내에서의 스트레스 방어기작과 세포주기조절의 관계를 연구하고 있으며 이러한 연구는 식물체 방어기작의 효율성 및 최종 산물의 증가를 이끌 수 있을 것이다.

Heat Stress After Pollination Reduces Kernel Number in Maize by Insufficient Assimilates.

Global warming has increased the occurrence of high temperature stress in plants, including maize, resulting in decreased the grain number and yield. Previous studies indicate that heat stress mainly damages the pollen grains and thus lowered maize grain number. Other field studies have shown that heat stress after pollination results in kernel abortion. However, the mechanism by which high temperature affect grain abortion following pollination remains unclear. Hence, this study investigated the field grown heat-resistant maize variety “Zhengdan 958” (ZD958) and heat-sensitive variety “Xianyu 335” (XY335) under a seven-day heat stress treatment (HT) after pollination. Under HT, the grain numbers of XY335 and ZD958 were reduced by 10.9% (p = 0.006) and 5.3% (p = 0.129), respectively. The RNA sequencing analysis showed a higher number of differentially expressed genes (DEGs) between HT and the control in XY335 compared to ZD958. Ribulose diphosphate carboxylase (RuBPCase) genes were downregulated by heat stress, and RuBPCase activity was significantly lowered by 14.1% (p = 0.020) in XY335 and 5.3% (p = 0.436) in ZD958 in comparison to CK. The soluble sugar and starch contents in the grains of XY335 were obviously reduced by 26.1 and 58.5%, respectively, with no distinct change observed in ZD958. Heat stress also inhibited the synthesis of grain starch, as shown by the low activities of metabolism-related enzymes. Under HT, the expression of trehalose metabolism genes in XY335 were upregulated, and these genes may be involved in kernel abortion at high temperature. In conclusion, this study revealed that post-pollination heat stress in maize mainly resulted in reduced carbohydrate availability for grain development, though the heat-resistant ZD958 was nevertheless able to maintain growth.

Dynamics and thermal sensitivity of ribosomal RNA maturation paths in plants.

Ribosome biogenesis is a constitutive fundamental process for cellular function. Its rate of production depends on the rate of maturation of precursor ribosomal RNA (pre-rRNA). The rRNA maturation paths are marked by four dominant rate-limiting intermediates with cell-type variation of the processivity rate. We have identified that high temperature stress in plants, while halting the existing pre-rRNA maturation schemes, also transiently triggers an atypical pathway for 35S pre-rRNA processing. This pathway leads to production of an aberrant precursor rRNA, reminiscent of yeast 24S, encompassing 18S and 5.8S rRNA that do not normally co-occur together at sub-unit levels; this response is elicited specifically by high and not low temperatures. We show this response to be conserved in two other model crop plant species (Rice and Tomato). This pathway persists even after returning to normal growth conditions for 1 hour and is reset between 1-6 hours after stress treatment, likely, due to resumption of normal 35S pre-rRNA synthesis and processing. The heat-induced ITS2 cleavage-derived precursors and stalled P-A2-like precursors were heterogeneous in nature with a fraction containing polymeric (A) tails. Furthermore, high temperature treatment and subsequent fractionation resulted in polysome and precursor rRNA depletion.

Physiological and Molecular Biology of High Temperature Stress in Plants

During the past few years, climate change induced by global warming had caused the appearance of extreme high temperatures worldwide, which had resulted in devastating damage to crop production. High Temperature Stress (HTS) is becoming an increasingly significant problem for agricultural production. Recent studies have elucidated the complex regulatory networks and versatile metabolites involved in HTS tolerance. Here, we provided an overview of current knowledge regarding the adverse effect of HTS on plant growth and development, the impairment of HTS on photosynthesis and membrane system, the role of carbohydrate metabolism, accumulation of osmo-protectants and secondary metabolites, the induced production of Reactive Oxygen Species (ROSs) and ROS detoxification system, and the synthesis of protective proteins like Heat Shock Proteins (HSPs) in HTS tolerance. Furthermore, the role of different phytohormones in plant response to HTS were discussed and epigenetic modifications are reported to be one of the three major signaling pathways associated with HTS response in plants, through the development of a ‘stress memory’ that is generated by hypomethylation to improve the plant’s survival under recurring HTS conditions. These physiological and molecular knowledge underlying plant response to cope with HTS will be helpful for the future directions of breeding crop tolerance to HTS using these factors or other strategies for agricultural applications.

Expression dynamics of glyoxalase genes under high temperature stress in plants

Heat stress, owing to the recent climate change events, has emerged as one of the major environmental factors posing a grave threat to plant survival and crop productivity. Oxidative damage due to increased generation of reactive oxygen and dicarbonyl species in response to heat, is one of the serious consequences of high temperature stress. Methylglyoxal (MG), is one such cytotoxic dicarbonyl metabolite whose levels are known to increase in response to high temperature and various other stress conditions in plants. In fact, it is considered as a general consequence of stress in plants. The synergistic co-activation of reactive oxygen species scavenging and the MG detoxification pathways thus, plays a key role in regulation of stress responses for plant acclimation. Glyoxalase enzymes are the major MG detoxifying proteins in the living organisms. In plants, glyoxalases are encoded as multigenic families, known to be involved in plant growth and development, although, chiefly associated with multiple stress inducible behaviour and tolerance. While their role in salinity, drought and heavy metal stresses has been well-studied, their significance under high temperature stress has received limited coverage. In this review, we emphasize on the dynamic role of glyoxalases under high temperature stress for conferring thermotolerance. Through analysis of heat responsive transcriptomes and proteomes of various plants, we provide evidence for a hitherto, unexplored role of glyoxalases as a mechanism of mounting thermotolerance responses. We also, show that ‘priming’ induced stress/cross stress tolerance in plants involves the activation of glyoxalase pathway.

In silico cloning and bioinformatics analysis of HSP21 in Manihot esculenta

HSP21 gene is a key gene to respond high temperature stress in plant and plays an important role in preventing protein denaturation, protecting cell structure and maintaining normal growth and development. Therefore, cloning HSP21 gene is the basis for revealing the molecular mechanism of resistance to high temperature stress in cassava. To obtain cassava HSP21 homologous gene and analyze the properties of predicted protein, electronic cloning technology was used to assemble and derivate new gene in this study, and bioinformatics analysis method was used to analyze the primary to highest structure, hydrophilicity/hydrophobicity, signal peptide, protein homology and phylogenetic evolution of expressed protein. HSP21 gene was 969 bp, its open reading frame was 705 bp, and the predicted protein contains 234 amino acids. The predicted protein is a non-transmembrane protein that is alkaline and hydrophilic, and is mainly localized in the chloroplast. Through multiple sequence alignment and phylogenetic analysis, it was found that the cassava HSP21 protein has high homology with other plants such as Hevea brasiliensis, Ricinus communis, and Jatropha curcas. The results could provide reference for the study of cloning and transformation of this gene.

Responses of young wheat plants to moderate heat stress

Moderate heat stress may provide protection against a subsequent severe high temperature stress in plants. However, the exact mechanisms of heat acclimation of wheat are still poorly understood. In the present work, two wheat varieties Ellvis and Soissons were exposed to a moderate elevated temperature at 30 °C, and the changes of certain protective mechanisms were investigated. Although the differences in the proline level between the genotypes were not substantial, it was approx. 2–3 times higher in the heat-treated plants than in the controls. After exposure to moderate elevated temperature, the activities of ascorbate peroxidase and catalase were also induced. Similarly, the amount of the free salicylic acid also increased after moderate heat stress, independently on the genotypes. The amount of the main polyamines, namely, putrescine, spermidine, and spermine either did not change or decreased after the same period. However, heat acclimation increased the level of 1,3-diaminopropane, in parallel with a polyamine oxidase gene, TaPAO. While the expression level of the peroxisomal polyamine oxidase gene TaperPAO hardly changed, TaPAO showed a substantial increase after 1 day, especially in Soissons, and at the end of the heat treatment was still significantly higher than in the controls. These suggest that signalling processes related to polyamine metabolisms or salicylic acid-related processes might also contribute to the higher heat tolerance induced by moderate heat stress. The variations in recorded measurements were mainly temperature dependent, and the effect of genotype was less pronounced than the effect of moderate heat treatment itself.

Does Application of Brassinosteroids mitigate the Temperature Stress in Plants?

Does Application of Brassinosteroids mitigate the Temperature Stress in Plants?

Deep sequencing of small RNAs reveals ribosomal origin of microRNAs in Oryza sativa and their regulatory role in high temperature

MicroRNAs are small noncoding regulatory RNAs which control gene expression by mRNA degradation or translational repression. They are significant molecular players regulating important biological processes such as developmental timing and stress response. We report here the discovery of miRNAs derived from ribosomal DNA using the small RNA datasets of 16 deep sequencing libraries of rice. Twelve putative miRNAs were identified based on highly stringent criteria of novel miRNA prediction. Surprisingly, 10 putative miRNAs (mi_7403, mi_8435, mi_12675, mi_4266, mi_4758, mi_4218, mi_8200, mi_4644, mi_14291, mi_16235) originated from rDNA of rice chromosome 9. Expression analysis of putative miRNAs and their target genes in heat tolerant and susceptible rice cultivars in control and high temperature treated seedlings revealed differential regulation of rDNA derived miRNAs. This is the first report of rDNA derived miRNAs in rice which indicates their role in gene regulation during high temperature stress in plants. Further studies in this area will open new research challenges and opportunities to broaden our knowledge on gene regulation mechanisms.

H2O2 mediates ALA-induced glutathione and ascorbate accumulation in the perception and resistance to oxidative stress in Solanum lycopersicum at low temperatures

BackgroundLow temperature is a crucial factor influencing plant growth and development. The chlorophyll precursor, 5-aminolevulinic acid (ALA) is widely used to improve plant cold tolerance. However, the interaction between H2O2 and cellular redox signaling involved in ALA-induced resistance to low temperature stress in plants remains largely unknown. Here, the roles of ALA in perceiving and regulating low temperature-induced oxidative stress in tomato plants, together with the roles of H2O2 and cellular redox states, were characterized.ResultsLow concentrations (10–25 mg·L− 1) of ALA enhanced low temperature-induced oxidative stress tolerance of tomato seedlings. The most effective concentration was 25 mg·L− 1, which markedly increased the ratio of reduced glutathione and ascorbate (GSH and AsA), and enhanced the activities of superoxide dismutase, catalase, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase. Furthermore, gene expression of respiratory burst oxidase homolog1 and H2O2 content were upregulated with ALA treatment under normal conditions. Treatment with exogenous H2O2, GSH, and AsA also induced plant tolerance to oxidative stress at low temperatures, while inhibition of GSH and AsA syntheses significantly decreased H2O2-induced oxidative stress tolerance. Meanwhile, scavenging or inhibition of H2O2 production weakened, but did not eliminate, GSH- or AsA- induced tomato plant tolerance to oxidative stress at low temperatures.ConclusionsAppropriate concentrations of ALA alleviated the low temperature-induced oxidative stress in tomato plants via an antioxidant system. The most effective concentration was 25 mg·L− 1. The results showed that H2O2 induced by exogenous ALA under normal conditions is crucial and may be the initial step for perception and signaling transmission, which then improves the ratio of GSH and AsA. GSH and AsA may then interact with H2O2 signaling, resulting in enhanced antioxidant capacity in tomato plants at low temperatures.

Roles of C-Repeat Binding Factors-Dependent Signaling Pathway in Jasmonates-Mediated Improvement of Chilling Tolerance of Postharvest Horticultural Commodities

C-repeat binding factor- (CBF-) dependent signaling pathway is proposed to be a key responder to low temperature stress in plant. Jasmonates (JAs), the endogenous signal molecules in plant, participate in plant defense against (a)biotic stresses; however, the mechanism has not been fully clarified so far. With the progress made in JAs biopathway, signal transduction, and their relationship with CBF-dependent signaling pathway, our knowledge of the roles of the CBF-dependent signaling pathway in JAs-mediated improvement of chilling tolerance accumulates. In this review, we firstly briefly review the chilling injury (CI) characteristics of postharvest horticultural commodities, then introduce the biopathway and signal transduction of JAs, subsequently summarize the roles of the CBF-dependent signaling pathway under low temperature stress, and finally describe the linkage between JAs signal transduction and the CBF-dependent signaling pathway.

Distinct transcriptional and processing regulations control miR167a level in tomato during stress

ABSTRACTBesides their definite role in plant developmental processes miR167 also serve as mediator of stress response. Although differential expression of miR167 occurs during stresses, the regulatory-mechanism of biogenesis remained elusive. Therefore, using tomato as the model plant we have explored the mechanism of regulation of miR167a expression during stresses. Fungus or virus infections and exposure to cold stress raised the level of miR167a expression. Whereas, salt, drought and heat treatments resulted in the downregulation, indicating different stresses activated alternative mechanisms for miR167a regulation. Interestingly, the relative expression level of precursors in control versus temperature stressed plants differed from the pattern observed in the mature miR167a expression, suggesting that both transcriptional and processing regulation were important for biogenesis. The promoter-regulatory sequence of the major isoform MIR167a harbours several development and stress-related regulatory sites. Accordingly, promoter assays using transient transformation and transgenic tobacco plants proved stress-dependent regulation of the promoter. Further analyses corroborated the role of tomato DREB2A protein in the transcriptional regulation during temperature stress. Finally, in vitro assays established the importance of processing factors in cold-stress dependent efficient processing of MIR167a precursors. These data confirm distinct role of transcriptional and processing machinery in stress-influenced regulation of tomato miR167a biogenesis.