Aquaporin Genes Research Articles

Genome‐wide characterization of aquaporins in Spirodela polyrhiza and expression of aquaporin gene family under high‐temperature stress

As one of the most conserved classes of gene families, aquaporins (AQP) genes play an essential role in water transport and stress resistance in organisms. However, little information was available for their evolutionary and functional roles in aquatic plant species. In the present study, gene sequences, gene structure, phylogeny, conserved motif and promoter of the AQP genes in Spirodela polyrhiza were investigated to explore the mechanisms of osmoregulation underwater. A total of 24 putative AQP genes divided into four clades representing four distinct AQPs subfamilies were identified. The gene structure and intron numbers of the AQP genes were conserved among subfamilies. The Froger's residues, conserved Ar/R selectivity filters and conserved NPA domain showed high differences among different subfamilies. Moreover, the promoter profiling of SpAQP genes for conserved cis‐acting regulatory elements revealed enrichment of cis‐elements involved in light response and stress responsiveness, reflecting their varying gene expression pattern. Our results demonstrated that the AQP genes in the S. polyrhiza were similar to those in the Zostera marina and might have experienced adaptive evolution to water. Transcriptome results showed that the SpTIP2‐3, SpPIP2‐2, SpPIP1‐3, SpTIP4‐3 and SpNIP5‐2 were expressed significantly among different treatments indicating they might play important roles under temperature stress. These data offer valuable sight into understanding the adaptation to the water of the S. polyrhiza. They may further inform our understanding of the evolution of water tolerance in S. polyrhiza.

Cadmium promotes the absorption of ammonium in hyperaccumulator Solanum nigrum L. mediated by ammonium transporters and aquaporins

Cadmium (Cd) is a toxic heavy metal affecting the normal growth of plants. Nitrate (NO3−) and ammonium (NH4+) are the primary forms of inorganic nitrogen (N) absorbed by plants. However, the mechanism of N absorption and regulation under Cd stress remains unclear. This study found that: (1) Cd treatment affected the biomass, root length, and Cd2+ flux in Solanum nigrum seedling roots. Specifically, 50 μM Cd significantly inhibited NO3− influx while increased NH4+ influx compared with 0 and 5 μM Cd treatments measured by non-invasive micro-test technology. (2) qRT-PCR analysis showed that 50 μM Cd inhibited the expressions of nitrate transporter genes, SnNRT2;4 and SnNRT2;4-like, increased the expressions of ammonium transporter genes, SnAMT1;2 and SnAMT1;3, in the roots. (3) Under NH4+ supply, 50 μM Cd significantly induced the expressions of the aquaporin genes, SnPIP1;5, SnPIP2;7, and SnTIP2;1. Our results showed that 50 μM Cd stress promoted NH4+ absorption by up-regulating the gene expressions of NH4+ transporter and aquaporins, suggesting that high Cd stress can affect the preference of N nutrition in S. nigrum.

Mitigation of the salinity stress in rapeseed (Brassica napus L.) productivity by exogenous applications of bio-selenium nanoparticles during the early seedling stage

In recent years, much attention has been directed toward using nanoparticles (NPs) as one of the most effective strategies to improve plant growth, especially under salt stress conditions. Further research has been conducted to develop NPs using various chemical ways; accordingly, knowledge about the beneficial effect of bioSeNPs in rapeseed is obscure. Selenium (Se) is a vital micronutrient with a series of physiological and antioxidative properties. Seed priming is emerging as a low-cost, efficient, and environment-friendly seed treatment in nanotechnology. The current study was carried out to examine the promising effects of nanopriming via bioSeNPs on the expression level of aquaporin genes, seed microstructure, seed germination, growth traits, physiochemical attributes, and minerals uptake of two rapeseed cultivars under salinity stress conditions. Our investigation monitored the positive effects of bioSeNPs on the expression level of aquaporin genes (BnPIP1-1 and BnPIP2-1) and water uptake during the seed imbibition (4 and 8 h of priming), which indicated higher imbibition potential and germination promotion with bioSeNPs application (most effective at 150 μmol/L). The total performance index was significantly enhanced with nano-treatments in rapeseed seedlings. Collectively, nano-application improved seed microstructure, seed germination, and photosynthetic efficiency directly correlated with higher seedlings biomass, especially with a higher concentration of bioSeNPs. The enhancement in α-amylase and free amino acid contents in nanoprimed seeds resulted in rapid seed germination. Moreover, bioSeNPs increased the osmotic adjustment and enhanced the efficiency of the plant's defense system by improving the activity of enzymatic and non-enzymatic antioxidants, thus enhancing ROS scavenging under salt stress. The obtained results may indicate the strengthening of seed vigor, improving seedling growth and physiochemical attributes via bioSeNPs. Our findings displayed that bioSeNPs modulated the Na+ and K+ uptake, which improved the rapeseed growth and showed a close relationship with the low contents of toxic Na+ ion; thus, it prevented oxidative damage due to salt stress. This comprehensive data can add more knowledge to understand the mechanisms behind plant-bioSeNPs interaction and provide physiological evidence for the beneficial roles of nanopriming using bioSeNPs on rapeseed germination and seedling development under salinity stress conditions. Such studies can be used to develop simple prepackaged nano primer products, which can be used before sowing to boost seed germination and crop productivity under stress conditions.

Genome-Wide Characterization of the Aquaporin Gene Family in Radish and Functional Analysis of RsPIP2-6 Involved in Salt Stress.

Aquaporins (AQPs) constitute a highly diverse family of channel proteins that transport water and neutral solutes. AQPs play crucial roles in plant development and stress responses. However, the characterization and biological functions of RsAQPs in radish (Raphanus sativus L.) remain elusive. In this study, 61 non-redundant members of AQP-encoding genes were identified from the radish genome database and located on nine chromosomes. Radish AQPs (RsAQPs) were divided into four subfamilies, including 21 plasma membrane intrinsic proteins (PIPs), 19 tonoplast intrinsic proteins (TIPs), 16 NOD-like intrinsic proteins (NIPs), and 5 small basic intrinsic proteins (SIPs), through phylogenetic analysis. All RsAQPs contained highly conserved motifs (motifs 1 and 4) and transmembrane regions, indicating the potential transmembrane transport function of RsAQPs. Tissue- and stage-specific expression patterns of AQP gene analysis based on RNA-seq data revealed that the expression levels of PIPs were generally higher than TIPs, NIPs, and SIPs in radish. In addition, quantitative real-time polymerase chain reaction (qRT-PCR) revealed that seven selected RsPIPs, according to our previous transcriptome data (e.g., RsPIP1-3, 1-6, 2-1, 2-6, 2-10, 2-13, and 2-14), exhibited significant upregulation in roots of salt-tolerant radish genotype. In particular, the transcriptional levels of RsPIP2-6 dramatically increased after 6 h of 150 mM NaCl treatment during the taproot thickening stage. Additionally, overexpression of RsPIP2-6 could enhance salt tolerance by Agrobacterium rhizogenes-mediated transgenic radish hairy roots, which exhibited the mitigatory effects of plant growth reduction, leaf relative water content (RWC) reduction and alleviation of O2– in cells, as shown by nitro blue tetrazolium (NBT) staining, under salt stress. These findings are helpful for deeply dissecting the biological function of RsAQPs on the salt stress response, facilitating practical application and genetic improvement of abiotic stress resistance in radish.

Analysis of photosynthetic ability and related physiological traits in nodal leaves of grape

Fruiting shoots are comprised of leaves, fruits, and branches, which determine the grape yield and the quality of produce. Fruiting shoots are also the important structural unit for studying the relationship between the source and sink. The number of leaves, leaf area, photosynthetic activities, and density are directly or indirectly affecting the ability of the fruiting shoot to fix CO2 and provide a carbon source for the fruit growth and development. ‘Summer Black’ is used as the main test material and observed certain differences in appearance, microstructure, photosynthetic pigments, relative water content (RWC), organic acids, carbohydrates, mineral elements, and photosynthetic capacity at different nodal leaves. The net photosynthetic rate (A) was significantly positively correlated with leaf shape parameters (the ratio between branch point to lobe tip and distal sinus to petiolar junction), leaf structure thickness, RWC, carotenoids, soluble sugars, and organic acids, but it was significantly negatively correlated with intercellular CO2 concentration (Ci), Chlorophyll b and phosphorus content. The variations in magnesium, water, glucose, and fructose content in leaves matched the expression patterns of aquaporin, magnesium transporter, and sugar metabolism-related genes in various nodal leaves. In addition, the physiological traits and photosynthetic capacity at different nodal leaves of ‘Shine Muscat’ and ‘Kyoho’ were similar to those of ‘Summer Black’. The results of this study will provide a reference for the differences in leaf sink–source effects at different nodes and provide a basis for reasonably removing leaves.

Genome-Wide Identification and Expression Analyses of the Aquaporin Gene Family in Passion Fruit (Passiflora edulis), Revealing PeTIP3-2 to Be Involved in Drought Stress.

Aquaporins (AQPs) in plants can transport water and small molecules, and they play an important role in plant development and abiotic stress response. However, to date, a comprehensive study on AQP family members is lacking. In this study, 27 AQP genes were identified from the passion fruit genome and classified into four groups (NIP, PIP, TIP, SIP) on the basis of their phylogenetic relationships. The prediction of protein interactions indicated that the AQPs of passion fruit were mainly associated with AQP family members and boron protein family genes. Promoter cis-acting elements showed that most PeAQPs contain light response elements, hormone response elements, and abiotic stress response elements. According to collinear analysis, passion fruit is more closely related to Arabidopsis than rice. Furthermore, three different fruit ripening stages and different tissues were analyzed on the basis of the transcriptome sequencing results for passion fruit AQPs under drought, high-salt, cold and high-temperature stress, and the results were confirmed by qRT-PCR. The results showed that the PeAQPs were able to respond to different abiotic stresses, and some members could be induced by and expressed in response to multiple abiotic stresses at the same time. Among the three different fruit ripening stages, 15 AQPs had the highest expression levels in the first stage. AQPs are expressed in all tissues of the passion fruit. One of the passion fruit aquaporin genes, PeTIP3-2, which was induced by drought stress, was selected and transformed into Arabidopsis. The survival rate of transgenic plants under drought stress treatment is higher than that of wild-type plants. The results indicated that PeTIP3-2 was able to improve the drought resistance of plants. Our discovery lays the foundation for the functional study of AQPs in passion fruit.

An aquaporin gene MdPIP1;2 from Malus domestica confers salt tolerance in transgenic Arabidopsis

Aquaporins are known as water channel proteins. In this study, an aquaporin gene MdPIP1;2 was cloned from Malus domestica cv. Qinguan encoding a protein of 289 amino acids that formed the typical structure of aquaporin by six transmembrane domains, two asparagine–proline–alanine motifs, aromatic/arginine filter, and Forger's position. MdPIP1;2 was highly expressed in the water-sensitive or water-requiring tissues, and upregulated by salt and PEG stresses. MdPIP1;2 transgenic Arabidopsis exhibited enhanced salt stress tolerance with less Na + accumulation, lower malondialdehyde (MDA) content, lower electrolyte leakage (EL) level, and higher superoxide dismutase (SOD) and peroxidase (POD) activities compared with WT plants. Additionally, transcriptome analysis indicated MdPIP1;2 transgenic Arabidopsis could present healthier growth and development condition probably through regulating morphological structures and accumulating specific secondary metabolites under salt stress. Our results are a useful reference for better understanding the biological function of aquaporin in apple tree, especially in plant response to abiotic stress.

Comparative study on the expression profile of Aquaporin5 (AQP5) gene in skinfibroblast cells of native goats enabling their water utilization efficiency

Lack of availability of good quality and quantity of water affects the physiology of goats. Goats may adapt to thrive well in water deficit areas by making alterations in the expressions of aquaporin (AQP) genes for their efficient water use. The present study was carried out in skin fibroblast cells of Barbari and Sirohi goats. Under in vitro conditions, during different temperatures and osmotic treatments, variations in relative expression of AQP3 genes were observed. At higher temperatures the relative expression of AQP3 genes in Sirohi breed was found to be significantly higher (p<0.01) than Barbari breed. However, during hyperosmotic concentrations at normal temperatures the relative expression of AQP3 mRNA was significantly ((p<0.05) increased in both the breeds. During a high temperature combination with hyperosmotic concentration in cell culture of skin fibroblast cell the relative expression of AQP3 mRNA was increased in Sirohi breed (p<0.01) whereas it decreased in Barbari goat’s breed. The variations in AQP3 gene expression during water stress condition and during hyperosmotic concentrations of skin fibroblasts cell culture suggests the positive involvement of AQP3 gene in maintaining the water balance in the body.

Identification and Characterization of Salt- and Drought-Responsive AQP Family Genes in Medicagosativa L.

Aquaporins (AQP) are distributed ubiquitously in plants, and they play important roles in multiple aspects of plant growth and development, as well as in plant resistance to various environmental stresses. In this study, 43 MsAQP genes were identified in the forage crop Medicago sativa. All the MsAQP proteins were clustered into four subfamilies based on sequence similarity and phylogenetic relationship, including 17 TIPs, 14 NIPs, 9 PIPs and 3 SIPs. Analyses of gene structure and conserved domains indicated that the majority of the deduced MsAQP proteins contained the signature transmembrane domains and the NPA motifs. Analyses on cis-acting elements in the promoter region of MsAQP genes revealed the presence of multiple and diverse stress-responsive and hormone-responsive cis-acting elements. In addition, by analyzing the available and comprehensive gene expression data of M. truncatula, we screened ten representative MtAQP genes that were responsive to NaCl or drought stress. By analyzing the sequence similarity and phylogenetic relationship, we finally identified the corresponding ten salt- or drought-responsive AQP genes in M. sativa, including three MsTIPs, three MsPIPs and four MsNIPs. The qPCRs showed that the relative expression levels of these ten selected MsAQP genes responded differently to NaCl or drought treatment in M. sativa. Gene expression patterns showed that most MsAQP genes were preferentially expressed in roots or in leaves, which may reflect their tissue-specific functions associated with development. Our results lay an important foundation for the future characterization of the functions of MsAQP genes, and provide candidate genes for stress resistance improvement through genetic breeding in M. sativa.

Molecular Characterization of Aquaporins Genes from the Razor Clam Sinonovacula constricta and Their Potential Role in Salinity Tolerance

Aquaporins (AQPs) play crucial roles in osmoregulation, but the knowledge about the functions of AQPs in Sinonovacula constricta is unclear. In this study, Sc-AQP1, Sc-AQP8, and Sc-AQP11 were identified from S. constricta, and the three Sc-AQPs are highly conserved compared to the known AQPs. The qRT-PCR analysis revealed that the highest mRNA expressions of Sc-AQP1, Sc-AQP8, and Sc-AQP11 were detected in the gill, digestive gland, and adductor muscle, respectively. In addition, the highest mRNA expression of Sc-AQP1 and Sc-AQP11 was detected in the D-shaped larvae stage, whereas that of SC-AQP8 was observed in the umbo larvae stage. The mRNA expression of Sc-AQP1, Sc-AQP8, and Sc-AQP11 significantly increased to 12.45-, 12.36-, and 27.44-folds post-exposure of low salinity (3.5 psu), while only Sc-AQP1 and Sc-AQP11 significantly increased post-exposure of high salinity (35 psu) (p < 0.01). The fluorescence in situ hybridization also showed that the salinity shift led to the boost of Sc-AQP1, Sc-AQP8, and Sc-AQP11 mRNA expression in gill filament, digestive gland, and adductor muscle, respectively. Knockdown of the Sc-AQP1 and Sc-AQP8 led to the decreased osmotic pressure in the hemolymph. Overall, these findings would contribute to the comprehension of the osmoregulation pattern of AQPs in S. constricta.

Genome assembly of the numbat (Myrmecobius fasciatus), the only termitivorous marsupial.

The numbat (Myrmecobius fasciatus) is an endangered Australian marsupial, and the last surviving member of the Myrmecobiidae family. The numbat regularly undergoes torpor and is unique amongst marsupials as it is the only diurnal and termitivorous species. Here we sequenced the first draft genome of the numbat using 10× Genomics Chromium linked-read technology, resulting in a 3.42 Gbp genome with a scaffold N50 of 223 kbp. A global transcriptome from liver, lung and tongue was also generated to aid genome annotation, identifying 21,465 protein-coding genes. To investigate adaptation to the numbat's termitivorous diet and arid/semi-arid range, we interrogated the most highly expressed transcripts within the tongue and manually annotated taste, vomeronasal and aquaporin gene families. Antimicrobial proteins and proteins involved in digestion were highly expressed in the tongue, alongside umami taste receptors. However, sweet taste receptors were not expressed in this tissue, which combined with the putative contraction of the bitter taste receptor gene repertoire in the numbat genome, may indicate a potential evolutionary adaptation to their specialised termitivorous diet. Vomeronasal and aquaporin gene repertoires were similar to other marsupials. The draft numbat genome is a valuable tool for conservation and can be applied to population genetics/genomics studies and to investigate the unique biology of this interesting species.

Silicon Nanoparticles Enhance Ginger Rhizomes Tolerance to Postharvest Deterioration and Resistance to Fusarium solani.

Postharvest deterioration of ginger rhizome caused by microorganisms or wound infections causes significant economic losses. Fusarium solani is one of the important causal agents of prevalent ginger disease soft rot across the world. The massive and continuous use of chemical fungicides in postharvest preservation pose risks to human health and produce environmental contamination. Hence, new alternative tools are required to reduce postharvest deterioration and extend the postharvest life of ginger. In this study, the use of silicon nanoparticles (SiNPs) on the storability of ginger rhizomes during postharvest storage and their resistance to Fusarium solani was investigated. The results showed that 50, 100, and 150 mg L−1 of SiNPs increased the firmness of the ginger rhizome during storage but decreased the decay severity, water loss, total color difference, and the reactive oxygen species (ROS; H2O2 and superoxide anion) accumulation. Specifically, 100 mg L−1 (SiNP100) demonstrated the best effect in the extension of postharvest life and improved the quality of the ginger rhizomes. SiNP100 application increased the activities of antioxidant enzymes (SOD and CAT) and the total phenolics and flavonoid contents, thereby reducing the ROS accumulation and malondialdehyde (MDA) content. Meanwhile, SiNP100 treatment negatively impacts the peroxidase (POD) and polyphenol oxidase (PPO) activities, which may have contributed to the lower level of lignin and decreased total color difference. SiNP100 likely decreased water loss and the transfer of water by altering the expression of aquaporin genes. Moreover, SiNP100 modulated the expression of lignin synthesis and phytopathogenic responses genes including MYB and LysM genes. Furthermore, SiNP100 inhibited Fusarium solani by preventing the penetration of hyphae into cells, thus decreasing the severity of postharvest pathogenic decay. In summary, this study revealed the physiology and molecular mechanisms of SiNPs-induced tolerance to postharvest deterioration and resistance to disease, which provides a foundation for using SiNPs resources as a promising alternative tool to maintain ginger quality and control postharvest diseases.

Pearl Millet Aquaporin Gene PgPIP2;6 Improves Abiotic Stress Tolerance in Transgenic Tobacco.

Pearl millet [Pennisetum glaucum (L) R. Br.] is an important cereal crop of the semiarid tropics, which can withstand prolonged drought and heat stress. Considering an active involvement of the aquaporin (AQP) genes in water transport and desiccation tolerance besides several basic functions, their potential role in abiotic stress tolerance was systematically characterized and functionally validated. A total of 34 AQP genes from P. glaucum were identified and categorized into four subfamilies, viz., plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), and small basic intrinsic proteins (SIPs). Sequence analysis revealed that PgAQPs have conserved characters of AQP genes with a closer relationship to sorghum. The PgAQPs were expressed differentially under high vapor pressure deficit (VPD) and progressive drought stresses where the PgPIP2;6 gene showed significant expression under high VPD and drought stress. Transgenic tobacco plants were developed by heterologous expression of the PgPIP2;6 gene and functionally characterized under different abiotic stresses to further unravel their role. Transgenic tobacco plants in the T2 generations displayed restricted transpiration and low root exudation rates in low- and high-VPD conditions. Under progressive drought stress, wild-type (WT) plants showed a quick or faster decline of soil moisture than transgenics. While under heat stress, PgPIP2;6 transgenics showed better adaptation to heat (40°C) with high canopy temperature depression (CTD) and low transpiration; under low-temperature stress, they displayed lower transpiration than their non-transgenic counterparts. Cumulatively, lower transpiration rate (Tr), low root exudation rate, declined transpiration, elevated CTD, and lower transpiration indicate that PgPIP2;6 plays a role under abiotic stress tolerance. Since the PgPIP2;6 transgenic plants exhibited better adaptation against major abiotic stresses such as drought, high VPD, heat, and cold stresses by virtue of enhanced transpiration efficiency, it has the potential to engineer abiotic stress tolerance for sustained growth and productivity of crops.

Relationships between aquaporins gene expression and nutrient concentrations in melon plants (Cucumis melo L.) during typical abiotic stresses

Melon ( Cucumis melo L.), a member of the Cucurbitaceae, in Mediterranean regions is usually affected by abiotic stresses like salinity, nutrients deficiency or high temperature. These abiotic stresses have been shown to produce the modulation of gene expression as a response to the altered conditions. Among these genes, aquaporins (transmembrane proteins) stand out due to their vital function as transporters of water and different solutes. For this reason, the aim of this work was to study the expression levels of all (31) aquaporins of melon plants (CmAQPs) after exposure to salinity (50 mM NaCl), nutrient deficiency (10% Hoagland solution) or high temperature (40 ºC for 1 h/day) and relate them with nutrient content, water relations and hydraulic conductance. There were general decreases in plant nutrient concentrations, especially in the root (Fe, K, Mn or Zn), while the concentrations of some elements for each stress (B, Ca, Mg, Mo or Si) increased. Physiological parameters were regulated depending on the treatment, showing the important role of hydric physiology regulation in the whole melon plant response to the different stresses. For most of the aquaporins, their expression decreased in the root (PIP2;1, PIP2;5 and PIP2;6 within the PIPs; most of the TIPs; NIP6;1, NIP7;1; SIP1;1) in all three treatments, while other aquaporins were over-expressed, such as the PIP1s (high temperature treatment), PIP2;2 (nutrient deficiency and high temperature treatments) and TIP1;1 (salinity and high temperature treatments) and NIP5;1 (nutrients deficiency treatment). The leaf aquaporin expression levels were less affected. This study shows that CmAQPs expression is modified differently in response to distinct abiotic stresses and that this is related to plant water relations and nutrients levels. • The expression of the water-transporting aquaporins, PIP2;1 , PIP2;5 and PIP2;6 decreased as response to abiotic stress. • The differences of TIPs and NIPs between stresses revealed the importance of the nutrient balance in relation to resistance. • Salinity stress decreased the expression of all root aquaporins for preserving water and nutrients inside the root cells. • Nutrient deficit, decreased in the expression of aquaporins genes in the roots and leaves, affecting directly cell turgor. • High temperature stress modify the expression level of aquaporins in relation to some specific nutrients.

EuPIP1;2, a Plasma Membrane Aquaporin Gene from Eucommia ulmoides, Enhances Drought and Salt Tolerance in Transgenic Tobacco

Aquaporins are a specific type of membrane channel proteins that efficiently transport water molecules and other small molecular substrates in plants. In this study, we isolated the plasma membrane aquaporin gene EuPIP1;2 from Eucommia ulmoides, an important medicinal plant in China. The EuPIP1;2 protein was localized on the plasma membrane. Quantitative RT-PCR analysis revealed that EuPIP1;2 was constitutively expressed in all analyzed tissues, with the highest expression levels detected in the fruit and root. Overexpression of EuPIP1;2 in transgenic tobacco enhanced plant tolerance of drought and salinity. Under drought and salt stress, the transgenic lines exhibited higher percentage germination, longer roots, and enhanced percentage survival compared with wild-type plants. The contents of malonaldehyde and proline suggested that EuPIP1;2 improved drought and salt tolerance in transgenic lines by reducing damage to membranes and improving osmotic adjustment. We predict that EuPIP1;2 could be applied to improve drought and salt tolerance in transgenic plants.

Integrative Transcriptome and Proteome Analysis Reveals the Absorption and Metabolism of Selenium in Tea Plants [Camellia sinensis (L.) O. Kuntze

Certain tea plants (Camellia sinensis) have the ability to accumulate selenium. In plants, the predominant forms of bioavailable Se are selenite (SeO32–) and selenate (SeO42–). We applied transcriptomics and proteomics to hydroponically grown plants treated with selenite or selenate for 48 h in the attempt to elucidate the selenium absorption and assimilation mechanisms in tea. A total of 1,844 differentially expressed genes (DEGs) and 691 differentially expressed proteins (DEPs) were obtained by comparing the Na2SeO3 and Na2SeO4 treatments against the control. A GO analysis showed that the genes related to amino acid and protein metabolism and redox reaction were strongly upregulated in the plants under the Na2SeO3 treatment. A KEGG pathway analysis revealed that numerous genes involved in amino acid and glutathione metabolism were upregulated, genes and proteins associated with glutathione metabolism and ubiquinone and terpenoid-quinone biosynthesis were highly expressed. Genes participating in DNA and RNA metabolism were identified and proteins related to glutathione metabolism were detected in tea plants supplemented with Na2SeO4. ABC, nitrate and sugar transporter genes were differentially expressed in response to selenite and selenate. Phosphate transporter (PHT3;1a, PHT1;3b, and PHT1;8) and aquaporin (NIP2;1) genes were upregulated in the presence of selenite. Sulfate transporter (SULTR1;1 and SULTR2;1) expression increased in response to selenate exposure. The results of the present study have clarified Se absorption and metabolism in tea plants, and play an important theoretical reference significance for the breeding and cultivation of selenium-enriched tea varieties.

Molecular Characterization and Gene Expression Analysis of Aquaporin in Haemaphysalis qinghaiensis.

Aquaporins (AQPs) are important functional proteins and are widely present in the cell membrane of almost all organisms, mediating transmembrane transport of liquid and other solutes. Much is known about the molecular characterization of AQPs in other tick species; however, nothing is known about them in Haemaphysalis qinghaiensis. In this study, we first sequenced the transcript variants of AQPs in H. qinghaiensis (HqAQPs), analyzed the biological structure features of AQPs, and investigated the pattern of gene expression of the AQP gene of H. qinghaiensis in different tick tissues and stages to predict their biological functions. In conclusion, four AQP transcript variants (i.e., HqAQP1-1, HqAQP1-2, HqAQP1-3, and HqAQP1-4) of H. qinghaiensis were found, and the sequences were comparable with its orthologs from the reported tick species. Gene expression of AQPs in different tick tissues and stages showed the higher expression level in salivary glands and gut of adult female, as well as in the female and nymph than in Malpighian tubules, ovary, male, larvae, and egg. Further studies will be performed to evaluate the function of HqAQPs against H. qinghaiensis infestation on animals.

Early and abrupt salinity reduction impacts European eel larval culture.

Reducing water salinity towards iso-osmotic conditions is a common practice applied in euryhaline fish farming to limit osmoregulation costs and enhance growth. In this respect, the present study investigated the timing of salinity reduction in an abrupt manner during European eel (Anguilla anguilla) larval culture by examining associated impacts on morphological and molecular levels. Larvae from 3 different parental combinations (families) were reared at constant 36 psu for 6days (control) or subjected to a direct reduction to 18 psu on 1, 2, or 3days post-hatch. Overall, salinity reduction enhanced growth and survival, resulting from more efficient energy resource utilization. In the control group, expression of growth-related igf2 remained constant, demonstrating a steady growth progression, while igf1 expression increased over time only for the salinity reduced treatments, potentially qualifying as a useful biomarker for growth performance. Even though each parental combination seems to have a different capacity to cope with salinity alterations, as observed by family-driven water-transport-related aquaporin (aqp1, aqp3) gene expression, it could be inferred that the abrupt salinity change is generally not stressful, based on non-upregulated heat shock proteins (hsp70, hsp90). However, the applied salinity reduction (irrespective of timing) induced the development of pericardial edema. As such, we conclude that despite the positive effect of salinity reduction on early growth and survival, the long-term benefit for eel larval culture lies in establishing a protocol for salinity reduction, at a precise developmental time point, without causing pericardial malformations.

Impact of Single and Combined Salinity and High-Temperature Stresses on Agro-Physiological, Biochemical, and Transcriptional Responses in Rice and Stress-Release.

Here, for the first time, we aimed to identify in rice the key mechanisms and processes underlying tolerance to high-temperature (HT) or salt stress (SS) alone, the co-occurrence of both stresses, and recovery using physiological and biochemical measurements and gene expression analysis. We also investigated whether recovery from the two stressors depended on the relative intensities/relief of each stressor. Wild type (‘Yukinkomai’) rice plants were found to be more susceptible to salinity or heat applied individually. SS leads to a depletion of cellular water content, higher accumulation of Na+, and alterations in photosynthetic pigments. The stress-tolerant cultivar ‘YNU31-2-4’ (YNU) displayed a lower Na+/K+ ratio, higher water content in cells and improved photosynthetic traits, antioxidant system, and expression of defence genes. Strikingly, the SS + HT combination provided a significant level of protection to rice plants from the effects of SS alone. The expression pattern of a selected set of genes showed a specific response and dedicated pathways in plants subjected to each of the different stresses, while other genes were explicitly activated when the stresses were combined. Aquaporin genes were activated by SS, while stress-related (P5CS, MSD1, HSPs, and ions transporters) genes were shaped by HT. Hierarchical clustering and principal component analyses showed that several traits exhibited a gradually aggravating effect as plants were exposed to the combined stresses and identified heat as a mitigating factor, clearly separating heat + salt-stressed from salt-non-heat-stressed plants. Furthermore, seedling recovery was far more dependent on the relative intensities of stressors and cultivars, demonstrating the influence of one stressor over another upon stress-release. Taken together, our data show the uniqueness and complexity of the physiological and molecular network modules used by rice plants to respond to single and combined stresses and recovery.

Two Aquaporin Genes, GhPIP2;7 and GhTIP2;1, Positively Regulate the Tolerance of Upland Cotton to Salt and Osmotic Stresses.

Aquaporins (AQPs) facilitate the transport of water and small molecules across intrinsic membranes and play a critical role in abiotic stresses. In this study, 111, 54, and 56 candidate AQP genes were identified in Gossypium hirsutum (AD1), Gossypium arboreum (A2), and Gossypium raimondii (D5), respectively, and were further classified into five subfamilies, namely, plasma intrinsic protein (PIP), tonoplast intrinsic protein (TIP), nodulin 26-like intrinsic protein (NIP), small basic intrinsic protein (SIP), and uncategorized X intrinsic protein (XIP). Transcriptome analysis and quantitative real-time PCR (qRT-PCR) revealed some high-expression GhPIPs and GhTIPs (PIP and TIP genes in G. hirsutum, respectively) in drought and salt stresses. GhPIP2;7-silenced plants decreased in the chlorophyll content, superoxide dismutase (SOD) activity, and peroxidase (POD) activity comparing the mock control (empty-vector) under 400 mM NaCl treatment, which indicated a positive regulatory role of GhPIP2;7 in salt tolerance of cotton. The GhTIP2;1-silenced cotton plants were more sensitive to osmotic stress. GhTIP2;1-overexpressed plants exhibited less accumulation of H2O2 and malondialdehyde but higher proline content under osmotic stress. In summary, our study elucidates the positive regulatory roles of two GhAQPs (GhPIP2;7 and GhTIP2;1) in salt and osmotic stress responses, respectively, and provides a new gene resource for future research.