Abiotic Conditions Research Articles

ZmABF4-ZmVIL2/ZmFIP37 module enhances drought tolerance in maize seedlings.

Drought, as a primary environmental factor, imposes significant constraints on developmental processes and productivity of plants. PHDs were identified as stress-responsive genes in a wide range of eukaryotes. However, the regulatory mechanisms governing PHD genes in maize under abiotic stress conditions are still largely unknown and require further investigation. Here, we identified a mutant, zmvil2, in the EMS mutant library with a C to T mutation in the exon of the Zm00001d053875 (VIN3-like protein 2, ZmVIL2), resulting in premature termination of protein coding. ZmVIL2 belongs to PHD protein family. Compared to WT, zmvil2 mutant exhibited increased sensitivity to drought stress. Consistently, overexpression of ZmVIL2 enhances drought resistance in maize. Y2H, BiFC, and Co-IP experiments revealed that ZmVIL2 directly interacts with ZmFIP37 (FKBP12-interacting protein of 37). zmfip37 knockout mutants also exhibit decreased drought tolerance. Interestingly, we demonstrated that ZmABF4 directly binds to the ZmVIL2 promoter to enhance its activity in yeast one hybrid (Y1H), electrophoretic mobility shift assay (EMSA) and dual luciferase reporter assays. Therefore, we uncovered a novel model ZmABF4-ZmVIL2/ZmFIP37 that promotes drought tolerance in maize. Overall, these findings have enriched the knowledge of the functions of PHD genes in maize and provides genetic resources for breeding stress-tolerant maize varieties.

Genome-wide identification and expression analysis of the Eriobotrya japonica TIFY gene family reveals its functional diversity under abiotic stress conditions

BackgroundPlant-specific TIFY proteins are widely found in terrestrial plants and play important roles in plant adversity responses. Although the genome of loquat at the chromosome level has been published, studies on the TIFY family in loquat are lacking. Therefore, the EjTIFY gene family was bioinformatically analyzed by constructing a phylogenetic tree, chromosomal localization, gene structure, and adversity expression profiling in this study.ResultsTwenty-six EjTIFY genes were identified and categorized into four subfamilies (ZML, JAZ, PPD, and TIFY) based on their structural domains. Twenty-four EjTIFY genes were irregularly distributed on 11 of the 17 chromosomes, and the remaining two genes were distributed in fragments. We identified 15 covariate TIFY gene pairs in the loquat genome, 13 of which were involved in large-scale interchromosomal segmental duplication events, and two of which were involved in tandem duplication events. Many abiotic stress cis-elements were widely present in the promoter region. Analysis of the Ka/Ks ratio showed that the paralogous homologs of the EjTIFY family were mainly subjected to purifying selection. Analysis of the RNA-seq data revealed that a total of five differentially expressed genes (DEGs) were expressed in the shoots under gibberellin treatment, whereas only one gene was significantly differentially expressed in the leaves; under both low-temperature and high-temperature stresses, there were significantly differentially expressed genes, and the EjJAZ15 gene was significantly upregulated under both low- and high-temperature stress. RNA-seq and qRT-PCR expression analysis under salt stress conditions revealed that EjJAZ2, EjJAZ4, and EjJAZ9 responded to salt stress in loquat plants, which promoted resistance to salt stress through the JA pathway. The response model of the TIFY genes in the jasmonic acid pathway under salt stress in loquat was systematically summarized.ConclusionsThese results provide a theoretical basis for exploring the characteristics and functions of additional EjTIFY genes in the future. This study also provides a theoretical basis for further research on breeding for salt stress resistance in loquat. RT-qPCR analysis revealed that the expression of one of the three EjTIFY genes increased and the expression of two decreased under salt stress conditions, suggesting that EjTIFY exhibited different expression patterns under salt stress conditions.

Introgression of Δ1-pyrroline-5-carboxylate synthetase (PgP5CS) confers enhanced resistance to abiotic stresses in transgenic tobacco.

Δ1-pyrroline-5-carboxylate synthetase (P5CS) is one of the key regulatory enzymes involved in the proline biosynthetic pathway. Proline acts as an osmoprotectant, molecular chaperone, antioxidant, and regulator of redox homeostasis. The accumulation of proline during stress is believed to confer tolerance in plants. In this study, we cloned the complete CDS of the P5CS from pearl millet (Pennisetum glaucum (L.) R.Br. and transformed into tobacco. Three transgenic tobacco plants with single-copy insertion were analyzed for drought and heat stress tolerance. No difference was observed between transgenic and wild-type (WT) plants when both were grown in normal conditions. However, under heat and drought, transgenic plants have been found to have higher chlorophyll, relative water, and proline content, and lower malondialdehyde (MDA) levels than WT plants. The photosynthetic parameters (stomatal conductance, intracellular CO2 concentration, and transpiration rate) were also observed to be high in transgenic plants under abiotic stress conditions. qRT-PCR analysis revealed that the expression of the transgene in drought and heat conditions was 2-10 and 2-7.5 fold higher than in normal conditions, respectively. Surprisingly, only P5CS was increased under heat stress conditions, indicating the possibility of feedback inhibition. Our results demonstrate the positive role of PgP5CS in enhancing abiotic stress tolerance in tobacco, suggesting its possible use to increase abiotic stress-tolerance in crops for sustained yield under adverse climatic conditions.

Heterologous expression of the maize transcription factor ZmbHLH36 enhances abiotic stress tolerance in Arabidopsis

AbstractBasic helix-loop-helix (bHLH) transcription factors are widely distributed in eukaryotes, and in plants, they regulate many biological processes, such as cell differentiation, development, metabolism, and stress responses. Few studies have focused on the roles of bHLH transcription factors in regulating growth, development, and stress responses in maize (Zea mays), even though such information would greatly benefit maize breeding programs. In this study, we cloned the maize transcription factor gene ZmbHLH36 (Gene ID: 100193615, GRMZM2G008691). ZmbHLH36 possesses conserved domains characteristic of the bHLH family. RT-qPCR analysis revealed that ZmbHLH36 was expressed at the highest level in maize roots and exhibited different expression patterns under various abiotic stress conditions. Transgenic Arabidopsis (Arabidopsis thaliana) plants heterologously expressing ZmbHLH36 had significantly longer roots than the corresponding non-transgenic plants under 0.1 and 0.15 mol L−1 NaCl treatment as well as 0.2 mol L−1 mannitol treatment. Phenotypic analysis of soil-grown plants under stress showed that transgenic Arabidopsis plants harboring ZmbHLH36 exhibited significantly enhanced drought tolerance and salt tolerance compared to the corresponding non-transgenic plants. Malondialdehyde contents were lower and peroxidase activity was higher in ZmbHLH36-expressing Arabidopsis plants than in the corresponding non-transgenic plants. ZmbHLH36 localized to the nucleus when expressed in maize protoplasts. This study provides a systematic analysis of the effects of ZmbHLH36 on root growth, development, and stress responses in transgenic Arabidopsis, laying a foundation for further analysis of its roles and molecular mechanisms in maize.

Long‐term environmental flows restore benthic invertebrate communities in a highly regulated river

The construction of dams and other in‐stream structures for human use has altered river ecosystems worldwide. Dams degrade rivers, and environmental flow releases aim to mitigate flow alterations and restore rivers downstream of dams by returning elements of the pre‐regulation flow regime, resulting in an ecosystem that resembles a more natural state. We examined the responses of benthic invertebrate communities in riffles and pool‐edge habitats to an environmental flow regime (EFR) in the highly regulated Snowy River over 17 years (2000–2016). Benthic invertebrate communities significantly changed in response to prolonged increases in the magnitude of all components of the flow regime. The composition of the benthos in the regulated river became more dissimilar to pre‐EFR communities and more closely resembled those of nearby unregulated rivers. These changes were apparent in both riffle and pool‐edge habitats and were most pronounced when flow magnitudes were highest. Scrapers increased as flows increased, and in contrast, the relative abundances of gatherers declined with prolonged high flows and could be attributed to increased daily flows and high flow events mobilizing fine sediment and scouring benthic substrates. These results indicated that the EFR altered abiotic conditions or resources that previously limited invertebrate abundances, facilitating changes to the benthic community. Community changes we observed were in response to a flow regime that remained drastically lower than natural, demonstrating that restoration targets can be achieved with small flow increases that mimic natural flow patterns.

Reproductive traits of the deep-sea shrimp Plesionika williamsi (decapoda: Pandalidae) from the eastern-central atlantic

The reproductive aspects of all Plesionika species are relatively well known worldwide, except for the deepest species of the genus, Plesionika williamsi, for which little information is available throughout its range. The ovarian maturity, sex ratio, brood size, and size-depth distribution of the deep-sea shrimp Plesionika williamsi (Pandalidae) in the Canary Islands (eastern-central Atlantic) were analysed. Ovigerous females were observed all year round, with the highest number of ovigerous females recorded between July and October. The presence of a greater number of ovigerous females during the summer may reflect a high local availability of food or the optimal abiotic conditions, which are factors with a strong influence on reproduction. The presence of non-ovigerous mature females throughout the year indicates that their resting period in the reproductive cycle occurs asynchronously. The physiological size at first sexual maturity was 19.24 mm in carapace length (CL) and the length at first sexual maturity was estimated at 23.15 mm CL. Estimates of size at first sexual maturity based on ovigerous females describe the result of the reproduction process, whereas size at first sexual maturity based on ovarian maturity deals with physiological preparation for reproduction. The modal size class of egg production was 24–30 mm CL, which yielded 83.22% of the population egg production. Plesionika williamsi is an iteroparous species that can produce small eggs during egg extrusion. The mean number of external embryos carried by females was 3048 and can be considered a true approximation of the number of larvae released in each batch, which depend on the conditions existing in each system. The shallower individuals are associated with a depth stratum that represents the boundary between two water masses present in the Canary Islands. The increase in size with depth is related to the presence of submarine volcanic canyons, which constitutes a flow channel of surface organic matter to depth.

Multiple‐stressor effects on leaf litter decomposition in freshwater ecosystems: A meta‐analysis

Abstract Recent years have witnessed a surge in research on the effects of multiple stressors in freshwater ecosystems. While studies have increased, the synthesis of their findings into a broader understanding of ecosystem‐level effects remains an ongoing endeavour. Leaf litter decomposition, a frequently investigated and pivotal ecosystem function in freshwaters, is sensitive to changes in abiotic conditions and biotic communities, and therefore susceptible to multiple‐stressor effects. Here, we synthesize findings from 27 manipulative experimental studies encompassing 61 responses of litter decomposition to paired stressors such as warming, nutrient enrichment and emerging pollutants in freshwater ecosystems. We calculated the individual and overall interaction effect sizes resulting from two stressors occurring simultaneously. Furthermore, we analysed the effect of moderator variables in the size and direction of interaction effect sizes using a meta‐analytical approach. Although the vote‐counting method showed additive interactions to dominate individual observations (91.8%), weighted random‐effects meta‐analysis revealed an overall antagonistic interaction between stressors (i.e. the cumulative effect of paired stressors on litter decomposition was less than the sum of their single effects). Our results emphasized the influence of experimental characteristics such as macroinvertebrate involvement, habitat type (lentic vs. lotic) and litter quality (assumed from plant mycorrhizal association) in shaping the responses of litter decomposition to multiple stressors. Our meta‐analysis highlights the need to incorporate local ecological complexities in manipulative experiments to improve predictions of multiple‐stressor effects on biodiversity and ecosystem functions. The present study underscores the importance of considering biotic interactions and adopting the metacommunity framework in conservation and restoration actions to support the management of freshwater ecosystems in an era of rapid global change. Read the free Plain Language Summary for this article on the Journal blog.

Modulating ‘Xinomavro’ (Vitis vinifera L.) Vine Growth and Berry Composition: A Comparative Analysis of Rootstock Effects

Rootstocks serve as a strategic tool for grapevine adaptation to specific biotic and abiotic conditions and for managing vine growth, grape yield, and berry composition in commercial vineyards. This study investigates the influences of four different rootstock varieties (101-14 MGt, 3309 C, 110 R, and 140 Ru) on the viticultural performance of ‘Xinomavro’ vines, a prominent Greek red winegrape varietal. By conducting a two-year field experiment using various rootstocks, we assessed parameters related to water status, vegetative growth, yield, and berry composition. Our results revealed that rootstock selection has a significant impact on vine development, especially in terms of berry size and the concentrations of secondary metabolites. Principal component analysis confirmed the complex interaction between rootstock vigor and vine productivity. This study underscores the importance of rootstock variety in manipulating grapevine characteristics, particularly for the ‘Xinomavro’ variety, in response to regional climatic conditions.

Diel activity patterns of a canopy-inhibiting beetle community (Coleoptera) in a Neotropical rainforest

Diel activity is one main feature of animal‘s behavior and is often an intrinsic trait characterizing distinct taxonomic groups. Abiotic conditions such as temperature may influence the diel activity patterns of arthropod communities associated with a particular ecosystem or habitat. Similarly, biotic factors, such as resource availability, affect arthropod activity. In addition, diel activity is thought to be an important factor in niche partitioning of arthropod communities. As part of a larger beetle survey in a lowland tropical rainforest in southern Venezuela, I analyzed the diel activity of an arboreal beetle community collected from 23 canopy-tree species over a cumulative year. Diel activity was observed in 535 beetle species, comprising 5,948 individuals, using a canopy crane installed in the study area. Of the 535 beetle species, 198 (37%) showed diurnal activity, and 281 (52.5%) showed nocturnal activity. In contrast, the proportions of nocturnal (n = 2,024, 34%) and diurnal (n = 1,983, 33.3%) individuals were balanced. Most of the observed beetles occurred only during the activity phase in their host trees. This particularly applies to extrafloral nectary- and flower-visiting beetle species. Flowering trees attracted different proportions of diurnal and nocturnal species according to flowering syndrome, whereas extrafloral nectaries were mainly visited at night. Thus, the beetle communities associated with single tree species showed distinct compositions of nocturnal and diurnal species.

Bioprospecting of endophytic fungi from semidesert candelilla (Euphorbia antisyphilitica Zucc): Potential for extracellular enzyme production.

Endophytic microbial communities colonize plants growing under various abiotic stress conditions. Candelilla (Euphorbia antisyphilitica Zucc.) is a shrub that develops functionally in arid and semi-arid zones of Mexico; these conditions generate an association between the plant and the microorganisms, contributing to the production of enzymes as a defense mechanism for resistance to abiotic stress. The objective of this research was to isolate and identify endophyte fungi of candelilla and bioprospection of these endophytic fungi for enzyme production using candelilla by-products. Fungi were isolated and identified using ITS1/ITS4 sequencing. Their potency index (PI) was evaluated in producing endoglucanase, xylanase, amylase, and laccase. Fermentation was carried out at 30°C for 8 days at 200 rpm, with measurements every 2 days, using candelilla by-products as substrate. All fungi exhibited higher cellulase, amylase, and laccase activities on the 2nd, 6th, and 8th day of fermentation, respectively, of fermentation. The fungus Aspergillus niger ITD-IN4.1 showed the highest amylase activity (246.84 U/mg), the genus Neurospora showed the highest cellulase activity, reaching up to 13.45 FPU/mg, and the strain Neurospora sp. ITD-IN5.2 showed the highest laccase activity (3.46 U/mg). This work provides the first report on the endophytic diversity of E. antisyphilitica and its potential role in enzyme production.

Deep population structure linked to host vernalization requirement in the barley net blotch fungal pathogen.

Invasive fungal pathogens pose a substantial threat to widely cultivated crop species, owing to their capacity to adapt to new hosts and new environmental conditions. Gaining insights into the demographic history of these pathogens and unravelling the mechanisms driving coevolutionary processes are crucial for developing durably effective disease management programmes. Pyrenophora teres is a significant fungal pathogen of barley, consisting of two lineages, Ptt and Ptm, with global distributions and demographic histories reflecting barley domestication and spread. However, the factors influencing the population structure of P. teres remain poorly understood, despite the varietal and environmental heterogeneity of barley agrosystems. Here, we report on the population genomic structure of P. teres in France and globally. We used genotyping-by-sequencing to show that Ptt and Ptm can coexist in the same area in France, with Ptt predominating. Furthermore, we showed that differences in the vernalization requirement of barley varieties were associated with population differentiation within Ptt in France and at a global scale, with one population cluster found on spring barley and another population cluster found on winter barley. Our results demonstrate how cultivation conditions, possibly associated with genetic differences between host populations, can be associated with the maintenance of divergent invasive pathogen populations coexisting over large geographic areas. This study not only advances our understanding of the coevolutionary dynamics of the Pt-barley pathosystem but also prompts further research on the relative contributions of adaptation to the host versus adaptation to abiotic conditions in shaping Ptt populations.

Molecular Mechanisms of CBL-CIPK Signaling Pathway in Plant Abiotic Stress Tolerance and Hormone Crosstalk.

Abiotic stressors, including drought, salt, cold, and heat, profoundly impact plant growth and development, forcing elaborate cellular responses for adaptation and resilience. Among the crucial orchestrators of these responses is the CBL-CIPK pathway, comprising calcineurin B-like proteins (CBLs) and CBL-interacting protein kinases (CIPKs). While CIPKs act as serine/threonine protein kinases, transmitting calcium signals, CBLs function as calcium sensors, influencing the plant's response to abiotic stress. This review explores the intricate interactions between the CBL-CIPK pathway and plant hormones such as ABA, auxin, ethylene, and jasmonic acid (JA). It highlights their role in fine-tuning stress responses for optimal survival and acclimatization. Building on previous studies that demonstrated the enhanced stress tolerance achieved by upregulating CBL and CIPK genes, we explore the regulatory mechanisms involving post-translational modifications and protein-protein interactions. Despite significant contributions from prior research, gaps persist in understanding the nuanced interplay between the CBL-CIPK system and plant hormone signaling under diverse abiotic stress conditions. In contrast to broader perspectives, our review focuses on the interaction of the pathway with crucial plant hormones and its implications for genetic engineering interventions to enhance crop stress resilience. This specialized perspective aims to contribute novel insights to advance our understanding of the potential of the CBL-CIPK pathway to mitigate crops' abiotic stress.

The spatiotemporal context of individual specialization in resource use and environmental associations.

1. Individual niche specialization is widespread in natural populations and has key implications for higher levels of biological organization. This phenomenon, however, has been primarily quantified in resource niche axes, overlooking individual variation in environmental associations (i.e. abiotic conditions organisms experience). 2. Here, we explore what we can learn from a multidimensional perspective of individual niche specialization that integrates resource use and environmental associations into a common framework. 3. By combining predictions from theory and simple simulations, we illustrate how (i) multidimensional intraspecific niche variation and (ii) the spatiotemporal context of interactions between conspecifics scale up to shape emergent patterns of the population niche. 4. Contemplating individual specialization as a multidimensional, unifying concept across biotic and abiotic niche axes is a fundamental step towards bringing this concept closer to the n-dimensional niche envisioned by Hutchinson.

A carbon dot toolbox for managing biotic and abiotic stresses in crop production systems

AbstractThe productivity of global crop production is under threat caused by various biotic and abiotic adverse conditions, such as plant diseases and pests, which are responsible for 20%–40% of global crop losses estimated at a value of USD 220 billion, and can be further exacerbated by climate change. Agricultural industries are calling for game‐changer technologies to enable productive and sustainable farming. Carbon dots (C‐dots) are carbon‐based nanoparticles, smaller than 50 nm, exhibiting unique opto‐electro‐properties. They have been shown to have positive impact on managing diverse biotic and abiotic stresses faced by the crops. Owing to their versatile carbon chemistry, the surface functionalities of C‐dots can be readily tuned to regulate plant physiological processes. This review is focussed on establishing the correlations between the physiochemical properties of C‐dots and their impacts on plants growth and health. The summary of the literature demonstrates that C‐dots hold great promise in improving plant tolerance to heat, drought, toxic chemicals, and invading pathogens.image

Experimental evidence that root-associated fungi improve plant growth at high altitude.

Unravelling how species communities change along environmental gradients requires a dual understanding: the direct responses of the species to their abiotic surroundings and the indirect variation of these responses through biotic interactions. Here, we focus on the interactive relationships between plants and their symbiotic root-associated fungi (RAF) along stressful abiotic gradients. We investigate whether variations in RAF community composition along altitudinal gradients influence plant growth at high altitudes, where both plants and fungi face harsher abiotic conditions. We established a translocation experiment between pairs of Bistorta vivipara populations across altitudinal gradients. To separate the impact of shifting fungal communities from the overall influence of changing abiotic conditions, we used a root barrier to prevent new colonization by RAF following translocation. To characterize the RAF communities, we applied DNA barcoding to the root samples. Through the utilization of joint species distribution modelling, we assessed the relationship between changes in plant functional traits resulting from experimental treatments and the corresponding changes in the RAF communities. Our findings indicate that RAF communities influence plant responses to stressful abiotic conditions. Plants translocated from low to high altitudes grew more when they were able to associate with the resident high-altitude RAF compared to those plants that were not allowed to associate with the resident RAF. We conclude that interactions with RAF impact how plants respond to stressful abiotic conditions. Our results provide experimental support that interactions with RAF improve plant stress tolerance to altitudinal stressors such as colder temperatures and less nutrient availability.

Genome-wide identification and expression analysis of the SET domain-containing gene family in potato (Solanum tuberosum L.)

Genes containing the SET domain can catalyse histone lysine methylation, which in turn has the potential to cause changes to chromatin structure and regulation of the transcription of genes involved in diverse physiological and developmental processes. However, the functions of SET domain-containing (StSET) genes in potato still need to be studied. The objectives of our study can be summarized as in silico analysis to (i) identify StSET genes in the potato genome, (ii) systematically analyse gene structure, chromosomal distribution, gene duplication events, promoter sequences, and protein domains, (iii) perform phylogenetic analyses, (iv) compare the SET domain-containing genes of potato with other plant species with respect to protein domains and orthologous relationships, (v) analyse tissue-specific expression, and (vi) study the expression of StSET genes in response to drought and heat stresses. In this study, we identified 57 StSET genes in the potato genome, and the genes were physically mapped onto eleven chromosomes. The phylogenetic analysis grouped these StSET genes into six clades. We found that tandem duplication through sub-functionalisation has contributed only marginally to the expansion of the StSET gene family. The protein domain TDBD (PFAM ID: PF16135) was detected in StSET genes of potato while it was absent in all other previously studied species. This study described three pollen-specific StSET genes in the potato genome. Expression analysis of four StSET genes under heat and drought in three potato clones revealed that these genes might have non-overlapping roles under different abiotic stress conditions and durations. The present study provides a comprehensive analysis of StSET genes in potatoes, and it serves as a basis for further functional characterisation of StSET genes towards understanding their underpinning biological mechanisms in conferring stress tolerance.

Exogenous melatonin enhances heat stress tolerance in sweetpotato by modulating antioxidant defense system, osmotic homeostasis and stomatal traits

Heat stress hinders the growth and productivity of sweetpotato plants, predominantly through oxidative damage to cellular membranes. Therefore, the development of efficient approaches for mitigating heat-related impairments is essential for the long-term production of sweetpotatoes. Melatonin has been recognised for its capacity to assist plants in dealing with abiotic stress conditions. This research aimed to investigate how different doses of exogenous melatonin influence heat damage in sweetpotato plants. Heat stress drastically affected shoot and root fresh weight by 31.8 and 44.5%, respectively. This reduction resulted in oxidative stress characterised by increased formation of hydrogen peroxide (H2O2) by 804.4%, superoxide ion (O2•−) by 211.5% and malondialdehyde (MDA) by 234.2%. Heat stress also reduced chlorophyll concentration, photosystem II efficiency (Fv/Fm) by 15.3% and gaseous exchange. However, pre-treatment with 100 μmol · L-1 melatonin increased growth and reduced oxidative damage to sweetpotato plants under heat stress. In particular, melatonin decreased H2O2, O2•− and MDA by 64.8%, 42.7% and 38.2%, respectively. Melatonin also mitigated the decline in chlorophyll levels and improved stomatal traits, gaseous exchange and Fv/Fm (13%). Results suggested that the favorable outcomes of melatonin treatment can be associated with elevated antioxidant enzyme activity and an increase in non-enzymatic antioxidants and osmo-protectants. Overall, these findings indicate that exogenous melatonin can improve heat stress tolerance in sweetpotatoes. This study will assist researchers in further investigating how melatonin makes sweetpotatoes more resistant to heat stress.

Selection of a suitable reference gene for gene-expression studies in Trichomonas gallinae under various biotic and abiotic stress conditions

Trichomonas gallinae, a globally distributed protozoan parasite, significantly affects the pigeon-breeding industry. T. gallinae infection mainly causes yellow ulcerative nodules on the upper respiratory tract and crop mucosa of pigeons, impeding normal breathing and feeding and ultimately causing death. Real-time quantitative PCR (qPCR) is a crucial technique for gene-expression analysis in molecular biology. Reference-gene selection for normalization is critical for ensuring this technique’s accuracy. However, no systematic screening or validation of T. gallinae reference genes has been reported. This study quantified the transcript levels of ten candidate reference genes in T. gallinae isolates with different genotypes and culture conditions using qPCR. Using the geNorm, NormFinder, and BestKeeper algorithms, we assessed these reference genes’ stabilities and ranked them using RankAggreg analysis. The most stable reference gene was tubulin beta chain (TUBB), while the widely used reference genes TUBG and GAPDH demonstrated poor stability. Additionally, we evaluated these candidate reference genes’ stabilities using the T. gallinae TgaAtg8 gene. On using TUBB as a reference gene, TgaAtg8′s expression profiles in T. gallinae isolates with different genotypes remained relatively consistent under various culture conditions. Conversely, using ACTB as a reference gene distorted the data. These findings provide valuable reference-gene-selection guidance for functional gene research and gene-expression analysis in T. gallinae.

Effects of nitrogen deficiency and drought stresses on miRNA expressions in Arabidopsis thaliana

MicroRNAs are now known to have an important role in regulating gene expression of eukaryotic organisms. miRNA research in plants gained importance after the discovery that several stress factors alter certain miRNA expressions, which subsequently regulate their target gene expressions and affect development and growth of plants. In this study, two of the widely studied abiotic stress conditions for plants, nitrogen deficiency and drought were used individually and as a combined stress treatment on Arabidopsis thaliana callus tissues to observe the change of expressions in certain miRNAs, when multiple stressors are encountered. Combined stress strongly inhibited callus growth compared to other conditions, while strongly altering certain miRNA expressions. Compared to control, in 7-day stress treated callus, miR165a-3p,b, miR319a,b, miR396b-5p, miR399d and miR827 showed significant downregulation for all stress treatments, while 7-day N deficiency caused miR167 upregulation. Stress treatments for 7 days mostly downregulated miR167c-5p, miR319c, miR399a, miR399e expressions except for the N deficient samples. After 14 days of stress, miR165a-3p,b, miR396a-5p, miR399b, miR399d were downregulated. During 14-day drought and combined stress, miR399a and miR396b-5p expressions were upregulated, respectively. The differences observed in this study between stress responses of 7 and 14-day-long treatments are believed to be valuable to further elucidate the associated molecular mechanisms of miRNAs, determination and validation of miRNA targets, and how plants respond to stress conditions via various genetic regulations.

Non-indigenous species and their realized niche in tidepools along the South-East Pacific coast

Non-indigenous species (NIS) have the potential to colonize and become established in a wide range of coastal habitats. Species with broad environmental tolerances can quickly adapt to local conditions and expand their niches along environmental gradients, and even colonize habitats with extreme abiotic conditions. Here we report and document the distribution of eight marine NIS (four seaweed and four invertebrate species) found in tidepools along a 3,000 km latitudinal gradient along the Pacific coast of Chile (18.4°S to 41.9°S). The seaweed NIS Codium fragile, Capreolia implexa, Schottera nicaeensis and Mastocarpus latissimus were mostly distributed towards high latitudes (i.e., more southerly locations), where temperatures in tidepools were low. The invertebrate NIS Anemonia alicemartinae, Ciona robusta, Bugula neritina and Bugulina flabellata were more common towards low latitudes, where high temperatures were registered in the tidepools. Across the intertidal gradient, seaweed NIS were mostly found in pools in the mid and low intertidal zone, while invertebrate NIS occurred mostly in pools from the mid and upper intertidal zones. The realized niche spaces of NIS (based on the Outlying Mean Index, OMI) in the study area were mainly influenced by environmental conditions of temperature and salinity (along the latitudinal and intertidal gradients), while other tidepool characteristics (depth, surface area, exposition, and complexity) only had minor effects. Five of the eight NIS exhibited a realized niche space coinciding with the average tidepool environmental conditions, while marginal niches were occupied by species with affinities for specific temperatures and salinities along the latitudinal and intertidal gradients. Our results indicate that physiological tolerances to environmental factors play a fundamental role in the distribution of seaweed and invertebrate NIS in tidepools along the Chilean coast. This study confirms that tidepools offer suitable conditions for some seaweed and invertebrate NIS, potentially facilitating their invasion into new natural habitats.