Transcript Levels Of Stress-responsive Genes Research Articles

Are tomato plants co-exposed to heat and salinity able to ensure a proper carbon metabolism? – An insight into the photosynthetic hub

Abiotic stress combinations, such as high temperatures and soil/water salinization, severely threaten crop productivity worldwide. In this work, an integrative insight into the photosynthetic metabolism of tomato plants subjected to salt (100 mM NaCl) and/or heat (42 °C; 4 h/day) was performed. After three weeks, the stress combination led to more severe consequences on growth and photosynthetic pigments than the individual stresses. Regarding the photochemical efficiency, transcript accumulation and protein content of major actors (CP47 and D1) were depleted in all stressed plants, although the overall photochemical yield was not negatively affected under the co-exposure. Gas-exchange studies revealed to be mostly affected by salt (single or combined), which harshly compromised carbon assimilation. Additionally, transcript levels of stress-responsive genes (e.g., HsfA1 and NHX2) were differentially modulated by the single and combined treatments, suggesting the activation of stress-signature responses. Overall, by gathering an insightful overview of the main regulatory hub of photosynthesis, we show that the impacts on the carbon metabolism coming from the combination of heat and salinity, two major conditioners of crop yields, were not harsher than those of single stresses, indicating that the growth impairment might be attributed to a proficient distribution of resources towards defense mechanisms.

Loss-of-Function Mutation of ACTIN-RELATED PROTEIN 6 (ARP6) Impairs Root Growth in Response to Salinity Stress.

H2A.Z-containing nucleosomes have been found to function in various developmental programs in Arabidopsis (e.g., floral transition, warm ambient temperature, and drought stress responses). The SWI2/SNF2-Related 1 Chromatin Remodeling (SWR1) complex is known to control the deposition of H2A.Z, and it has been unraveled that ACTIN-RELATED PROTEIN 6 (ARP6) is one component of this SWR1 complex. Previous studies showed that the arp6 mutant exhibited some distinguished phenotypes such as early flowering, leaf serration, elongated hypocotyl, and reduced seed germination rate in response to osmotic stress. In this study, we aimed to investigate the changes of arp6 mutant when the plants were grown in salt stress condition. The phenotypic observation showed that the arp6 mutant was more sensitive to salt stress than the wild type. Upon salt stress condition, this mutant exhibited attenuated root phenotypes such as shorter primary root length and fewer lateral root numbers. The transcript levels of stress-responsive genes, ABA INSENSITIVE 1 (ABI1) and ABI2, were found to be impaired in the arp6 mutant in comparison with wild-type plants in response to salt stress. In addition, a meta-analysis of published data indicated a number of genes involved in auxin response were induced in arp6 mutant grown in non-stress condition. These imply that the loss of H2A.Z balance (in arp6 mutant) may lead to change stress and auxin responses resulting in alternative root morphogenesis upon both normal and salinity stress conditions.

Heterologous expression of ZmNF-YA12 confers tolerance to drought and salt stress in Arabidopsis

Drought and salinity are serious environmental factors limiting the growth and productivity of plants worldwide. Therefore, it is necessary to develop ways to improve drought and salinity stress tolerance in plants. In this study, a drought-responsive nuclear factor Y subunit A gene, ZmNF-YA12, was cloned from maize. qPCR revealed ZmNF-YA12 transcript in all vegetative and reproductive tissues, with higher levels in young roots. Expression analyses of maize revealed that ZmNF-YA12 was induced by abscisic acid (ABA), jasmonic acid (JA), and abiotic stresses, including dehydration, high salinity, cold, and polyethylene glycol (PEG) treatment. The heterologous expression of ZmNF-YA12 in Arabidopsis plants resulted in increased root length and better plant growth than in wild-type (WT) plants under conditions of mannitol, salt, and JA stress on 1/2 MS medium. Transgenic Arabidopsis showed improved tolerance to drought and salt stresses in soil, and higher proline content and lower malondialdehyde (MDA) content than WT controls. The transgenic plants also maintained higher peroxidase (POD) activities than WT plants under conditions of NaCl stress. A yeast two-hybrid experiment demonstrated that ZmNF-YA12 interacted with ZmNF-YC1 and ZmNF-YC15. Moreover, the transcript levels of stress-responsive genes (RD29A, RD29B, RAB18, and RD22) were markedly increased in transgenic lines under conditions of drought and salt stress. These observations suggested that the ZmNF-YA12 gene may confers drought and salt stress tolerance by regulating stress-related genes or interacting with ZmNF-YC1 and ZmNF-YC15, and has potential applications in molecular breeding with maintenance of production under conditions of stress.

Red-seaweed biostimulants differentially alleviate the impact of fungicidal stress in rice (Oryza sativa L.)

Red seaweed-derived biostimulants facilitate plant health and impart protection against abiotic stress conditions by their bioactive compounds and plant nutrients. The potency of red seaweed biostimulants (LBS6 and LBD1) on rice cv. IR-64 in response to fungicides induced stress was investigated in this study. Foliar application of LBS6 maintained the stomatal opening and leaf temperature under the fungicidal stress condition. Reactive Oxygen Species (ROS) such as hydrogen peroxide and superoxide radicals were significantly reduced in LBS6-treated stressed plants. After applying seaweed biostimulants, ROS production was stabilized by antioxidants viz., CAT, APX, SOD, POD, and GR. LBS-6 application increased the Ca+ and K+ levels in the stressed plants, which perhaps interacted with ROS and stomatal opening signalling systems, respectively. In the rice plants, fungicidal stress elevated the expression of stress-responsive transcriptional factors (E2F, HSFA2A, HSFB2B, HSFB4C, HSFC1A, and ZIP12). A decline in the transcript levels of stress-responsive genes was recorded in seaweed treated plants. For the first time, we present an integrative investigation of physicochemical and molecular components to describe the mechanism by which seaweed biostimulants in rice improve plant health under fungicidal stress conditions.

Alpha-ketoglutarate-dependent dioxygenase homolog 10B, an N6 -methyladenosine mRNA demethylase, plays a role in salt stress and abscisic acid responses in Arabidopsis thaliana.

N6 -methyladenosine (m6 A) is an abundant methylation mark in eukaryotic mRNAs. It is installed and removed by methyltransferases ("writers") and demethylases ("erasers"), respectively. A recent study has demonstrated that alpha-ketoglutarate-dependent dioxygenase homolog 10B (ALKBH10B) is an mRNA m6 A eraser affecting floral transition in Arabidopsis thaliana. However, the roles of m6 A eraser proteins, including ALKHB10B, in plant adaptation to abiotic stresses are largely unknown. In this study, we aimed to determine the role of ALKBH10B in the response of A. thaliana to abiotic stresses and abscisic acid (ABA). The m6 A level increased in response to salt stress, and m6 A levels in alkbh10b mutants were higher than those in the wild-type under both normal and salt stress conditions. Germination of alkbh10b mutant seeds was markedly delayed under salt stress but not under dehydration, cold, or ABA conditions. Seedling growth and survival rate of alkbh10b mutants were enhanced under salt stress. Notably, salt-tolerant phenotypes of alkbh10b mutants were correlated with decreased levels of several m6 A-modified genes, including ATAF1, BGLU22, and MYB73, which are negative effectors of salt stress tolerance. In response to ABA, both seedling and root growth of alkbh10b mutants were inhibited via upregulating ABA signaling-related genes, including ABI3 and ABI4. Collectively, these findings indicate that ALKBH10B-mediated m6 A demethylation affects the transcript levels of stress-responsive genes, which are important for seed germination, seedling growth, and survival of Arabidopsis thaliana in response to salt stress or ABA.

The bZIP Transcription Factor GmbZIP15 Negatively Regulates Salt- and Drought-Stress Responses in Soybean.

Soybean (Glycine max), as an important oilseed crop, is constantly threatened by abiotic stress, including that caused by salinity and drought. bZIP transcription factors (TFs) are one of the largest TF families and have been shown to be associated with various environmental-stress tolerances among species; however, their function in abiotic-stress response in soybean remains poorly understood. Here, we characterized the roles of soybean transcription factor GmbZIP15 in response to abiotic stresses. The transcript level of GmbZIP15 was suppressed under salt- and drought-stress conditions. Overexpression of GmbZIP15 in soybean resulted in hypersensitivity to abiotic stress compared with wild-type (WT) plants, which was associated with lower transcript levels of stress-responsive genes involved in both abscisic acid (ABA)-dependent and ABA-independent pathways, defective stomatal aperture regulation, and reduced antioxidant enzyme activities. Furthermore, plants expressing a functional repressor form of GmbZIP15 exhibited drought-stress resistance similar to WT. RNA-seq and qRT-PCR analyses revealed that GmbZIP15 positively regulates GmSAHH1 expression and negatively regulates GmWRKY12 and GmABF1 expression in response to abiotic stress. Overall, these data indicate that GmbZIP15 functions as a negative regulator in response to salt and drought stresses.

Overexpression of HbMBF1a, encoding multiprotein bridging factor 1 from the halophyte Hordeum brevisubulatum, confers salinity tolerance and ABA insensitivity to transgenic Arabidopsis thaliana

Key messageHbMBF1a was isolated and characterized in H. brevisubulatum, and overexpressed HbMBF1a could enhance the salt tolerance and ABA insensitivity in Arabidopsis thaliana. The transcript levels of stress-responsive genes were significantly increased in the transgenic lines under salt and ABA conditions.Salinity is an abiotic stress that considerably affects plant growth, yield, and distribution. Hordeum brevisubulatum is a halophyte that evolved to become highly tolerant to salinity. Multiprotein bridging factor 1 (MBF1) is a transcriptional coactivator and an important regulator of stress tolerance. In this study, we isolated and characterized HbMBF1a based on the transcriptome data of H. brevisubulatum grown under saline conditions. We overexpressed HbMBF1a in Arabidopsis thaliana and compared the phenotypes of the transgenic lines and the wild-type in response to stresses. The results indicated that HbMBF1a expression was induced by salt and ABA treatments during the middle and late stages. The overexpression of HbMBF1a in A. thaliana resulted in enhanced salt tolerance and ABA insensitivity. More specifically, the enhanced salt tolerance manifested as the increased seed germination and seedling growth and development. Similarly, under ABA treatments, the cotyledon greening rate and seedling root length were higher in the HbMBF1a-overexpressing lines, suggesting the transgenic plants were better adapted to high exogenous ABA levels. Furthermore, the transcript levels of stress-responsive genes were significantly increased in the transgenic lines under salt and ABA conditions. Thus, HbMBF1a is a positive regulator of salt and ABA responses, and the corresponding gene may be useful for producing transgenic plants that are salt tolerant and/or ABA insensitive, with few adverse effects. This study involved a comprehensive analysis of HbMBF1a. The results may provide the basis and insight for the application of MBF1 family genes for developing stress-tolerant crops.

Over-expression of the Brachypodium ASR gene, BdASR4, enhances drought tolerance in Brachypodium distachyon.

BdASR4 expression was up-regulated during abiotic stress and hormone treatments. Plants over-expressing BdASR4 improved drought tolerant. BdASR4 may regulate antioxidant activities and transcript levels of stress-related and abscisic acid-responsive genes. Abiotic stress conditions negatively affect plant growth and developmental processes, causing a reduction in crop productivity. The abscisic acid-, stress-, ripening-induced (ASR) proteins play important roles in the protection of plants from abiotic stress. Brachypodium distachyon L. is a well-studied monocot model plant. However, ASR proteins of Brachypodium have not been widely studied. In this study, five ASR genes of Brachypodium plant were cloned and characterized. The BdASR genes were expressed in response to various abiotic stresses and hormones. In particular, BdASR4 was shown to encode a protein containing a nuclear localization signal in its C-terminal region, which enabled protein localization in the nucleus. To further examine functions of BdASR4, transgenic Brachypodium plants harboring BdASR4 were generated. Over-expression of BdASR4 was associated with strong drought tolerance, and plants over-expressing BdASR4 preserved more water and displayed higher antioxidant enzyme activities than did the wild-type plants. The transcript levels of stress-responsive genes, reactive oxygen species scavenger-associated genes, and abscisic acid-responsive genes tended to be higher in transgenic plants than in WT plants. Moreover, plants over-expressing BdASR4 were hypersensitive to exogenous abscisic acid at the germination stage. Taken together, these findings suggest multiple roles for BdASR4 in the plant response to drought stress by regulating antioxidant enzymes and the transcription of stress- and abscisic acid-responsive genes.

Abiotic stress tolerance and ABA responses of transgenic Glycine max plants with modulated RACK1 expression

The receptor for activated C kinase 1 (RACK1) belongs to a protein subfamily that contains a tryptophan-aspartic acid-domain (WD) repeat structure. In this study, a soybean RACK1 gene (GmRACK1) was cloned. The amino acid sequence of GmRACK1 had seven WD repeats, which contained typical glycine-histidine (GH) and WD dipeptides. Tissue-specific expression showed that GmRACK1 is expressed at differential levels in all examined tissues and strongly down-regulatd by abscisic acid (ABA) and abiotic stress. Subcellular localization suggested that GmRACK1 is located in the plasma membrane, cytoplasm, and nucleus. In response to drought and salt stress, the GmRACK1-RNAi lines showed significantly higher dry weight of whole plants, chlorophyll content, and survival rate of soybean seedlings than wild-type and homozygous (OE) lines. GmRACK1-RNAi plants were observed to be more sensitive to ABA-mediated seed germination and root growth. Furthermore, we found that the level of ABA and transcript levels of stress-responsive genes were clearly up-regulated under drought and salt stress conditions in GmRACK1-RNAi plants. Consistent with the accumulation of reactive oxygen species (ROS), elevated electrolyte leakage and malondialdehyde, and lower expression of ROS-scavenging genes were found in the GmRACK1-OE lines. The GmRACK RNAi lines exhibited increased tolerance to drought and salt stress compared with the wild-type and OE lines. In addition, yeast two-hybrid assays showed that GmRACK1 could interact with WNK8, eIF6, and RbohC-N. These results therefore indicate that GmRACK1 responds to drought and salt stress through ABA signaling and the regulation of cellular ROS production in plants.

Arabidopsis AtNAP functions as a negative regulator via repression of AREB1 in salt stress response.

AtNAP , an Arabidopsis NAC transcription factor family gene, functions as a negative regulator via transcriptional repression of AREB1 in salt stress response. AtNAP is an NAC family transcription factor in Arabidopsis and is known to be a positive regulator of senescence. However, its exact function and underlying molecular mechanism in stress responses are not well known. Here, we investigated functional roles of AtNAP in salt stress response. AtNAP expression significantly increased at the seedling stage, with higher expression in both shoots and roots under NaCl, mannitol, and ABA treatments. T-DNA insertional loss-of-function mutants of AtNAP were more tolerant to salt stress than wild type (WT), whereas AtNAP-overexpressing transgenic plants (OXs) were more sensitive to salt stress than WT during germination, seedling development, and mature plant stage. Transcript levels of stress-responsive genes in the ABA-dependent pathway, such as AREB1, RD20, and RD29B, were significantly higher and lower in atnap mutants and AtNAP OXs, respectively, than in WT under salt stress conditions, suggesting that AtNAP might negatively regulate the expression of those genes under salt stress conditions. Indeed, AtNAP repressed the promoter activity of AREB1 under normal and salt stress conditions. These results indicate that AtNAP functions as a negative regulator in the salt stress response. Our results, together with previous studies, suggest that AtNAP functions as a negative regulator in osmotic stress responses, whereas it functions as a positive regulator in senescence.

Proteomic profiling of γ-ECS overexpressed transgenic Nicotiana in response to drought stress

The contribution of Glutathione (GSH) in drought stress tolerance is an established fact. However, the proteins which are directly or indirectly related to the increased level of GSH in response to drought stress are yet to be known. To explore this, here, transgenic tobacco plants (NtGp11) overexpressing gamma-glutamylcysteine synthetase (γ-ECS) was tested for tolerance against drought stress. NtGp11 conferred tolerance to drought stress by increased germination rate, water retention, water recovery, chlorophyll, and proline content compared with wild-type plants. Semi-quantitative RT-PCR analysis revealed that the transcript levels of stress-responsive genes were higher in NtGp11 compared with wild-type in response to drought stress. Two-dimensional gel electrophoresis (2-DE) coupled with MALDI TOF-TOF MS/MS analysis has been used to identify 43 differentially expressed proteins in response to drought in wild-type and NtGp11 plants. The results demonstrated the up-accumulation of 58.1% of proteins among which 36%, 24%, and 20% of them were related to stress and defense, carbon metabolism and energy metabolism categories, respectively. Taken together, our results demonstrated that GSH plays an important role in combating drought stress in plants by inducing stress related genes and proteins like HSP70, chalcone synthase, glutathione peroxidase, thioredoxin peroxidase, ACC oxidase, and heme oxygenase I.

Time course analysis of RNA stability in human placenta.

BackgroundEvaluation of RNA quality is essential for gene expression analysis, as the presence of degraded samples may influence the interpretation of expression levels. Particularly, qRT-PCR data can be affected by RNA integrity and stability. To explore systematically how RNA quality affects qRT-PCR assay performance, a set of human placenta RNA samples was generated by two protocols handlings of fresh tissue over a progressive time course of 4 days. Protocol A consists of a direct transfer of tissue into RNA-stabilizing solution (RNAlater™) solution. Protocol B uses a dissection of placenta villosities before bio banking. We tested and compared RNA yields, total RNA integrity, mRNA integrity and stability in these two protocols according to the duration of storage.ResultsA long time tissue storage had little effect on the total RNA and mRNA integrity but induced changes in the transcript levels of stress-responsive genes as TNF-alpha or COX2 after 48 h. The loss of the RNA integrity was higher in the placental tissues that underwent a dissection before RNA processing by comparison with those transferred directly into RNA later™ solution. That loss is moderate, with average RIN (RNA Integration Numbers) range values of 4.5–6.05, in comparison with values of 6.44–7.22 in samples directly transferred to RNAlater™ (protocol A). Among the house keeping genes tested, the B2M is the most stable.ConclusionThis study shows that placental samples can be stored at + 4°C up to 48 h before RNA extraction without altering RNA quality. Rapid tissue handling without dissection and using RNA-stabilizing solution (RNAlater™) is a prerequisite to obtain suitable RNA integrity and stability.