TMCO1 Is an ER Ca(2+) Load-Activated Ca(2+) Channel.

Maintaining homeostasis of Ca(2+) stores in the endoplasmic reticulum (ER) is crucial for proper Ca(2+) signaling and key cellular functions. The Ca(2+)-release-activated Ca(2+) (CRAC) channel is responsible for Ca(2+) influx and refilling after store depletion, but how cells cope with excess Ca(2+) when ER stores are overloaded is unclear. We show that TMCO1 is an ER transmembrane protein that actively prevents Ca(2+) stores from overfilling, acting as what we term a "Ca(2+) load-activated Ca(2+) channel" or "CLAC" channel. TMCO1 undergoes reversible homotetramerization in response to ER Ca(2+) overloading and disassembly upon Ca(2+) depletion and forms a Ca(2+)-selective ion channel on giant liposomes. TMCO1 knockout mice reproduce the main clinical features of human cerebrofaciothoracic (CFT) dysplasia spectrum, a developmental disorder linked to TMCO1 dysfunction, and exhibit severe mishandling of ER Ca(2+) in cells. Our findings indicate that TMCO1 provides a protective mechanism to prevent overfilling of ER stores with Ca(2+) ions.

A novel role of RNase L in the development of nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and affects about 25% of the population globally. NAFLD has the potential to cause significant liver damage in many patients because it can progress to nonalcoholic steatohepatitis (NASH) and cirrhosis, which substantially increases disease morbidity and mortality. Despite the key role of innate immunity in the disease progression, the underlying molecular and pathogenic mechanisms remain to be elucidated. RNase L is a key enzyme in interferon action against viral infection and displays pleiotropic biological functions such as control of cell proliferation, apoptosis, and autophagy. Recent studies have demonstrated that RNase L is involved in innate immunity. In this study, we revealed that RNase L contributed to the development of NAFLD, which further progressed to NASH in a time-dependent fashion after RNase L wild-type (WT) and knockout mice were fed with a high-fat and high-cholesterol diet. RNase L WT mice showed significantly more severe NASH, evidenced by widespread macro-vesicular steatosis, hepatocyte ballooning degeneration, inflammation, and fibrosis, although physiological and biochemical data indicated that both types of mice developed obesity, hyperglycemia, hypercholesterolemia, dysfunction of the liver, and systemic inflammation at different extents. Further investigation demonstrated that RNase L was responsible for the expression of some key genes in lipid metabolism, inflammation, and fibrosis signaling. Taken together, our results suggest that a novel therapeutic intervention for NAFLD may be developed based on regulating the expression and activity of RNase L.

Whole-genome resequencing reveals new mutations in candidate genes for Beichuan-white goat prolificacya.

In this study, we evaluated the copy number variation in the genomes of two groups of Beichuan-white goat populations with large differences in litter size by FST method, and identified 1739 genes and 485 missense mutations in the genes subject to positive selection. Through functional enrichment, ITGAV, LRP4, CDH23, TPRN, RYR2 and CELSR1 genes, involved in embryonic morphogenesis, were essential for litter size trait, which received intensive attention. In addition, some mutation sites of these genes have been proposed (ITGAV: c.38C > T; TPRN: c.133A > T, c.1192A > G, c.1250A > C; CELSR1: c.7640T > C), whose allele frequencies were significantly changed in the high fecundity goat group. Besides, we found that new mutations at these sites altered the hydrophilicity and 3D structure of the protein. Candidate genes related to litter size in this study and their missense mutation sites were identified. These candidate genes are helpful to understand the genetic mechanism of fecundity in Beichuan white goat, and have important significance for future goat breeding.

Populus euphratica plant cadmium resistance 2 mediates Cd tolerance by root efflux of Cd ions in poplar.

KEY MESSAGE: Populus euphratica PePCR2 increases Cd resistance by functioning as a Cd extrusion pump and by mediating the expression of genes encoding other transporters. Cadmium (Cd) is a non-essential, toxic metal that negatively affects plant growth. Plant cadmium resistance (PCR) proteins play key roles in the response to heavy metal stress. In this study, we isolated the gene PePCR2 encoding a plant PCR from Populus euphratica. PePCR2 gene transcription was induced by Cd, and its transcript level peaked at 24 h after exposure, at a level approximately 18-fold higher than that at 0 h. The PePCR2 protein was localized to the plasma membrane. Compared with yeast cells harboring the empty vector, yeast cells expressing PePCR2 showed enhanced Cd tolerance and a lower Cd content. Compared with wild-type (WT) plants, poplar overexpressing PePCR2 showed higher Cd resistance. Net Cd2+ efflux measurements showed that Cd2+ efflux from the roots was 1.5 times higher in the PePCR2-overexpressing plants than in WT plants. Furthermore, compared with WT plants, the PePCR2-overexpressing plants showed increased transcript levels of ABCG29, HMA5, PDR2, YSL7, and ZIP1 and decreased transcript levels of NRAMP6, YSL3, and ZIP11 upon exposure to Cd. These data show that PePCR2 increased Cd resistance by acting as a Cd extrusion pump and/or by regulating other Cd2+ transporters to decrease Cd toxicity in the cytosol. The results of this study identify a novel plant gene with potential applications in Cd removal, and provide a theoretical basis for reducing Cd toxicity and protecting food safety.

Potential candidate genes and pathways related to cytoplasmic male sterility in Dianthus spiculifolius as revealed by transcriptome analysis.

KEY MESSAGE: We introduced the candidate gene DsHSP70 into Arabidopsis thaliana, resulting in male gametophyte sterility and abnormal degeneration of sepals and petals. Cytoplasmic male sterility (CMS) is a useful tool for hybrid production. However, the regulatory mechanism of CMS in Dianthus spiculifolius remains unclear. In this study, we investigated whether male-sterile line of D. spiculifolius has a malformed tapetum and fails to produce normal fertile pollen. RNA sequencing technology was used to compare the gene expression patterns of the D. spiculifolius male-sterile line and its male fertility maintainer line during anther development. A total of 12,365 differentially expressed genes (DEGs) were identified, among which 1765 were commonly expressed in the S1, S2 and S3 stages. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these DEGs were mainly involved in oxidation-reduction processes, signal transduction and programmed cell death. Additionally, weighted correlation network analysis (WGCNA) showed that three modules may be related to male sterility. A putative regulatory pathway for the male sterility traits was constructed based on the reproductive development network. After introducing the candidate DsHSP70 gene into Arabidopsis thaliana, we found that overexpressing plants showed anther abortion and shorter filaments, and accompanied by abnormal degeneration of sepals and petals. In summary, our results identified potential candidate genes and pathways related to CMS in D. spiculifolius, providing new insights for further research on the mechanism of male sterility.

Characteristic of the components and the metabolism mechanism of goat colostrum: a review.

Colostrum contains large number of nutrients that promote the growth, differentiation, and biological functions for goat kids early somatic cells, which is crucial to meet the nutritional demands, immune function, and the health of goat kids later growth. Great attention has been given not only to nutritional ingredient differences between colostrum and normal milk, but also to function differences, and their effect on the physical and sensory properties of goat kid's growth performance and health status. This paper reviews the research progress of goat colostrum in recent years, mainly including the colostrum yield, components, i.e., proteins, lactose, and immunoglobulin, as well as the influence factor, i.e., number of lactation and littler size, nutrition during the gestation, and breeding environment. In addition, this review aims to summarize the synthesis and secretion mechanisms, and the digestion and absorption mechanism of goat colostrum. We conclude that even though the composition and physicochemical properties of goat colostrum are highly dynamic and variable, and the digestion and absorption mechanism has not been made fully clear until now, direct feed microbial (DFM) may be a promising alternative for improving the quality of colostrum that should be further explored for their practical usage.

Effect of direct-fed microbials on growth performance, blood biochemical indices, and immune status of female goats.

The effect of direct-feed microbial (DFM) treatment on body weight, serum biochemical indexes, serum immunoglobulins, and serum cytokines was studied. The study was a completely randomized design with 20 growing females Beichuan white goats, weighing 25.11 ± 1.96 kg, divided into two groups of 10 goats per treatment. Goats were offered (1) 10 mL saline solution (Control group) (2) or 10 mL microbials solution (DFM group) on days 0 and 7 for two times. No effect on final body weight and body size was observed between DFM and control group (p > 0.05). DFM treatment had greater serum total protein, globulin, and albumin/globulin ratio than the control treatment (p < 0.05). The concentrations of IgA, IgG, IgM, INF-γ, and IL-2 in DFM group were significantly higher than those in the control group on days 7, 14, and 21 (p < 0.05), and the highest content was detected on day 14 of the experiment. The concentrations of IgA, IgG, IgM, IL-2, INF-γ, INF-α, IL-4, and IL-5 in DFM group on day 14 were higher than those on day 0 (p < 0.05). In conclusion, DFM enhanced serum immunoglobulins and cytokines without affecting body weight, body size, and normal serum metabolism.

Effects of lactation number and litter size on the chemical composition and immune components of goat colostrum.

This study aimed to evaluate the effect of the number of lactations and litter size on the chemical composition, immunoglobulins, and cytokines of goat colostrum. The experiment was conducted at the Animal Research Base, Mianyang Academy of Agricultural Sciences, from February to March 2021. After delivery, 48 colostrum samples were obtained every 24 h by manual milking from both udders. The contents of colostrum proteins, IgA, and IgM increased markedly up to 48 h postpartum, reaching 250 and 1250 µg/mL, respectively (p < 0.01 compared with 0 h). However, the total Ig and IgG contents dropped quickly at 48 h postpartum to around 4.5 and 6 mg/mL, respectively, and continued to do so until 96 h postpartum (p < 0.01). As for litter size, the colostrum DM, fat, total Ig, IgG, INF-γ, and IL-2 of twin-birth goats were higher than those of single-birth goats at 0 h postpartum. Moreover, the colostrum of multiparous goats contained higher total Ig, IgA, IgG, and INF-γ concentrations than that of primiparous goats at 0 h postpartum (p < 0.01). However, the colostrum INF-α and IL-5 contents of multiparous goats were lower than those of primiparous goats at 0 h postpartum (p < 0.05). Available information indicates that colostrum secretion takes place until 48 h postpartum and that the effect of litter size and lactation number on colostrum quality is observed at 0 h postpartum.

Changes in colostrum ingredients of Hu sheep, as well as the missense mutation genes associated with colostrum yield.

This study focused on the changes in the composition and immune evolution in milk from birth to 144 h postpartum and the genes associated with the colostrum yield of Hu sheep. Twelve Hu sheep, which were bred carefully under animal health standards and have a litter size of two kids and similar gestation length (149 ± 1 days), were used. Lambs were transferred into their own cots to avoid interference. The compositional content (i.e., fat, protein, and lactose) and some other properties, including daily colostrum yield, DM, and SNF, were determined. In addition, immunity molecules (IgG, IgA, and IgM concentrations) received remarkable attention. The DM, SNF, fat, and protein contents were higher in the first days postpartum and then dropped quickly from the time of birth to 144 h postpartum. However, the lactose content displayed an increasing pattern and reached normal milk percentage at 48 h. The highest IgG (103.17 mg/mL), IgA (352.82 µg/mL), and IgM (2.79 mg/mL) colostrum concentrations were observed at partum, decreased quickly, and finally stabilized. The change law of concentration of IgA and IgM in colostrum are the same with IgG. Furthermore, the whole-genome resequencing was performed, and a missense variant locus in the SRC gene and two missense locus variants in the HIF1A gene were significantly associated with the colostrum yield of sheep by using the whole-genome selection signal detection analysis. In conclusions, colostrum contains abundant nutrients especially immunoglobulin, and the HIF1A gene may be used as candidate genes for colostrum yield, which has important information as a basic knowledge for the Hu sheep breeding program.

Picture-word order compound protein interaction: Predicting compound-protein interaction using structural images of compounds.

Identifying potential associations between proteins and compounds is significant and challenging in the drug discovery process. Existing deep-learning-based methods tend to treat compounds and proteins as sequences or graphs. Inspired by the rapid development of computer vision technology, we argue that more abundant characterizations can be extracted from the images of compounds than from their sequences or graphs. Therefore, we propose an interaction model named picture-word order compound protein interaction (PWO-CPI) which learns the representation from structural images of compounds and protein sequences by using convolutional neural network (CNN). The experiments show that PWO-CPI outperforms state-of-the-art CPI prediction models. We also perform drug-drug interaction (DDI) experiments to validate the strong potential of structural formula images of molecular structures as molecular features. In addition, with the aid of generative adversarial networks, the visualization of image features demonstrates PWO-CPI can learn compound structural features implicitly and automatically.

Plant cadmium resistance 6 from Salix linearistipularis (SlPCR6) affects cadmium and copper uptake in roots of transgenic Populus.

Phytoextraction in phytoremediation is one of the environmentally friendly methods used for restoring soils contaminated by heavy metals (HMs). The screening and identification of HM-resistant plants and their regulatory genes associated with HM ion transport are the key research aims in this field. In this study, a plant cadmium (Cd) resistance (PCR) gene family member, SlPCR6, was identified in roots of Salix linearistipularis, which exhibits strong HM resistance. The results revealed that SlPCR6 expression was induced in S. linearistipularis roots in response to Cd stress. Furthermore, SlPCR6 was mainly localized on the plasma membrane. Compared with the wild type, SlPCR6 overexpression reduced the Cd and copper (Cu) contents in the transgenic poplar (84 K) and increased its Cd and Cu resistance. The roots of transgenic poplar seedlings had lower net Cd and Cu uptake rates than wild type roots. Further investigation revealed that the transcript levels of multiple HM ion transporters were not significantly different between the roots of the wild type and those of the transgenic poplar. These results suggest that SlPCR6 is directly involved in Cd and Cu transport in S. linearistipularis roots. Therefore, SlPCR6 can serve as a candidate gene to improve the phytoextraction of the HMs Cd and Cu through genetic engineering.

AaZFP3, a Novel CCCH-Type Zinc Finger Protein from Adonis amurensis, Promotes Early Flowering in Arabidopsis by Regulating the Expression of Flowering-Related Genes.

CCCH-type zinc finger proteins (ZFP) are a large family of proteins that play various important roles in plant growth and development; however, the functions of most proteins in this family are uncharacterized. In this study, a CCCH-type ZFP, AaZFP3, was identified in the floral organ of Adonis amurensis. Quantitative real-time PCR (qPCR) analysis revealed that AaZFP3 was widely expressed in the flowers of A.amurensis. Subcellular localization analysis showed that the AaZFP3 protein was mainly localized to the cytoplasm in tobacco and Arabidopsis. Furthermore, the overexpression of AaZFP3 promoted early flowering in Arabidopsis under both normal and relatively low-temperature conditions. RNA-sequencing and qPCR analyses revealed that the expression of multiple key flowering-time genes was altered in transgenic Arabidopsis overexpressing AaZFP3 compared to wild-type. Of these genes, FLOWERING LOCUS T (AtFT) expression was most significantly up-regulated, whereas FLOWERING LOCUS C (AtFLC) was significantly down-regulated. These results suggest that the overexpression of AaZFP3 promotes early flowering in Arabidopsis by affecting the expression of flowering-time genes. Overall, our study indicates that AaZFP3 may be involved in flowering regulation in A.amurensis and may represent an important genetic resource for improving flowering-time control in other ornamental plants or crops.

RNA-Binding Protein MAC5A Is Required for Gibberellin-Regulated Stamen Development.

The development of floral organs is coordinated by an elaborate network of homeotic genes, and gibberellin (GA) signaling is involved in floral organ development; however, the underlying molecular mechanisms remain elusive. In the present study, we found that MOS4-ASSOCIATED COMPLEX 5A (MAC5A), which is a protein containing an RNA-binding motif, was involved in the development of sepals, petals, and stamens; either the loss or gain of MAC5A function resulted in stamen malformation and a reduced seed set. The exogenous application of GA considerably exacerbated the defects in mac5a null mutants, including fewer stamens and male sterility. MAC5A was predominantly expressed in pollen grains and stamens, and overexpression of MAC5A affected the expression of homeotic genes such as APETALA1 (AP1), AP2, and AGAMOUS (AG). MAC5A may interact with RABBIT EARS (RBE), a repressor of AG expression in Arabidopsis flowers. The petal defect in rbe null mutants was at least partly rescued in mac5a rbe double mutants. These findings suggest that MAC5A is a novel factor that is required for the normal development of stamens and depends on the GA signaling pathway.

LGCCT: A Light Gated and Crossed Complementation Transformer for Multimodal Speech Emotion Recognition.

Semantic-rich speech emotion recognition has a high degree of popularity in a range of areas. Speech emotion recognition aims to recognize human emotional states from utterances containing both acoustic and linguistic information. Since both textual and audio patterns play essential roles in speech emotion recognition (SER) tasks, various works have proposed novel modality fusing methods to exploit text and audio signals effectively. However, most of the high performance of existing models is dependent on a great number of learnable parameters, and they can only work well on data with fixed length. Therefore, minimizing computational overhead and improving generalization to unseen data with various lengths while maintaining a certain level of recognition accuracy is an urgent application problem. In this paper, we propose LGCCT, a light gated and crossed complementation transformer for multimodal speech emotion recognition. First, our model is capable of fusing modality information efficiently. Specifically, the acoustic features are extracted by CNN-BiLSTM while the textual features are extracted by BiLSTM. The modality-fused representation is then generated by the cross-attention module. We apply the gate-control mechanism to achieve the balanced integration of the original modality representation and the modality-fused representation. Second, the degree of attention focus can be considered, as the uncertainty and the entropy of the same token should converge to the same value independent of the length. To improve the generalization of the model to various testing-sequence lengths, we adopt the length-scaled dot product to calculate the attention score, which can be interpreted from a theoretical view of entropy. The operation of the length-scaled dot product is cheap but effective. Experiments are conducted on the benchmark dataset CMU-MOSEI. Compared to the baseline models, our model achieves an 81.0% F1 score with only 0.432 M parameters, showing an improvement in the balance between performance and the number of parameters. Moreover, the ablation study signifies the effectiveness of our model and its scalability to various input-sequence lengths, wherein the relative improvement is almost 20% of the baseline without a length-scaled dot product.

Arabidopsis cold-regulated plasma membrane protein Cor413pm1 is a regulator of ABA response.

Cold-regulated (COR) genes are considered downstream functional genes in the cold-response pathway. However, we identified a plasma membrane-type, AtCor413pm1, as a regulatory gene for the abscisic acid (ABA) response, and found that ABA induced it predominantly in Arabidopsis roots, vasculature, stipules, and guard cells. Differentially expressed genes combined with qPCR analysis revealed the expressions of three ABA-responsive genes (AtDTX50, AtABR1, and AtCIPK20) were significantly altered in the ABA-treated atcor413pm1 mutant, compared to the wild-type. Furthermore, the ABA-induced transient Ca2+ oscillation in the plasma membrane of atcor413pm1 roots was different from that observed in the wild-type. Our results revealed that AtCor413pm1 might play a role in the cross-talk between the ABA and stress response pathways.

Identification and Characterization of MIKCc-Type MADS-Box Genes in the Flower Organs of Adonis amurensis.

Adonis amurensis is a perennial herbaceous flower that blooms in early spring in northeast China, where the night temperature can drop to -15 °C. To understand flowering time regulation and floral organogenesis of A. amurensis, the MIKCc-type MADS (Mcm1/Agamous/ Deficiens/Srf)-box genes were identified and characterized from the transcriptomes of the flower organs. In this study, 43 non-redundant MADS-box genes (38 MIKCc, 3 MIKC*, and 2 Mα) were identified. Phylogenetic and conserved motif analysis divided the 38 MIKCc-type genes into three major classes: ABCDE model (including AP1/FUL, AP3/PI, AG, STK, and SEPs/AGL6), suppressor of overexpression of constans1 (SOC1), and short vegetative phase (SVP). qPCR analysis showed that the ABCDE model genes were highly expressed mainly in flowers and differentially expressed in the different tissues of flower organs, suggesting that they may be involved in the flower organ identity of A. amurensis. Subcellular localization revealed that 17 full-length MADSs were mainly localized in the nucleus: in Arabidopsis, the heterologous expression of three full-length SOC1-type genes caused early flowering and altered the expression of endogenous flowering time genes. Our analyses provide an overall insight into MIKCc genes in A. amurensis and their potential roles in floral organogenesis and flowering time regulation.

Pingyangmycin inhibits glycosaminoglycan sulphation in both cancer cells and tumour tissues.

Pingyangmycin is a clinically used anticancer drug and induces lung fibrosis in certain cancer patients. We previously reported that the negatively charged cell surface glycosaminoglycans are involved in the cellular uptake of the positively charged pingyangmycin. However, it is unknown if pingyangmycin affects glycosaminoglycan structures. Seven cell lines and a Lewis lung carcinoma-injected C57BL/6 mouse model were used to understand the cytotoxicity of pingyangmycin and its effect on glycosaminoglycan biosynthesis. Stable isotope labelling coupled with LC/MS method was used to quantify glycosaminoglycan disaccharide compositions from pingyangmycin-treated and untreated cell and tumour samples. Pingyangmycin reduced both chondroitin sulphate and heparan sulphate sulphation in cancer cells and in tumours. The effect was persistent at different pingyangmycin concentrations and at different exposure times. Moreover, the cytotoxicity of pingyangmycin was decreased in the presence of soluble glycosaminoglycans, in the glycosaminoglycan-deficient cell line CHO745, and in the presence of chlorate. A flow cytometry-based cell surface FGF/FGFR/glycosaminoglycan binding assay also showed that pingyangmycin changed cell surface glycosaminoglycan structures. Changes in the structures of glycosaminoglycans may be related to fibrosis induced by pingyangmycin in certain cancer patients.

The N-terminal and third transmembrane domain of PsCor413im1 are essential for targeting to chloroplast envelope membrane.

Cold-regulated (COR) genes, located downstream of the C-repeat binding factors (CBFs) in cold signaling pathways, play a central role in plant response to cold stress. In our previous studies, a Cor413 chloroplast envelope membrane protein, PsCor413im1, was identified from the cold-tolerant plant Phlox subulata. Its overexpression enhanced cold tolerance and altered AtCor15 expression in Arabidopsis. In the present study, the function of PsCor413im1 was further investigated. Transmission electron microscope observation showed that the chloroplast envelope membrane of cold-treated transgenic Arabidopsis seedlings was more stable than that of cold-treated wild-type seedlings. Subcellular localization of green fluorescent protein as a marker revealed that the N-terminal and putative third transmembrane domain (TMD) of PsCor413im1 were essential for its targeting of the chloroplast envelope membrane. Furthermore, overexpression of PsCor413im1 fragments containing N-terminal and third TMD also altered the expression of AtCor15 genes in Arabidopsis. Overall, our results suggest that PsCor413im1 may stabilize the chloroplast envelope membrane under cold stress, and its N-terminal and third TMD are important for its targeting capability and function.

Comparison of cadmium uptake and transcriptional responses in roots reveal key transcripts from high and low-cadmium tolerance ryegrass cultivars.

Cadmium (Cd), which seriously affects plant growth and crop production, is harmful to humans. Previous studies revealed ryegrass (Lolium multiflorum Lam.) exhibits Cd tolerance, and may be useful as a potential hyperaccumulator because of its wide distribution. In this study, the physiological and transcriptional responses of two ryegrass cultivars [i.e., high (LmHC) and low (LmLC) Cd tolerance] to Cd stress were investigated and compared. The Cd tolerance of LmHC was greater than that of LmLC at various Cd concentrations. The uptake of Evans blue dye revealed that Cd-induced root cell mortality was higher in LmLC than in LmHC after a 12-h Cd treatment. Furthermore, the content and influx rate of Cd in LmLC roots were greater than in LmHC roots under Cd stress conditions. The RNA sequencing and quantitative real-time PCR data indicated that the Cd transport regulatory genes (ABCG37, ABCB4, NRAMP4, and HMA5) were differentially expressed between the LmLC and LmHC roots. This expression-level diversity may contribute to the differences in the Cd accumulation and translocation between LmLC and LmHC. These findings may help clarify the physiological and molecular mechanisms underlying ryegrass responses to Cd toxicity. Additionally, ryegrass may be able to hyperaccumulate toxic heavy metals during the phytoremediation of contaminated soil.

Heterologous Expression of Nitrate Assimilation Related-Protein DsNAR2.1/NRT3.1 Affects Uptake of Nitrate and Ammonium in Nitrogen-Starved Arabidopsis.

Nitrogen (N) is an essential macronutrient for plant growth. Plants absorb and utilize N mainly in the form of nitrate (NO3-) or ammonium (NH4+). In this study, the nitrate transporter DsNRT3.1 (also known as the nitrate assimilation-related protein DsNAR2.1) was characterized from Dianthus spiculifolius. A quantitative PCR (qPCR) analysis showed that the DsNRT3.1 expression was induced by NO3-. Under N-starvation conditions, the transformed Arabidopsis seedlings expressing DsNRT3.1 had longer roots and a greater fresh weight than the wild type. Subcellular localization showed that DsNRT3.1 was mainly localized to the plasma membrane in Arabidopsis root hair cells. Non-invasive micro-test (NMT) monitoring showed that the root hairs of N-starved transformed Arabidopsis seedlings had a stronger NO3- and NH4+ influx than the wild-type seedlings, using with NO3- or NH4+ as the sole N source; contrastingly, transformed seedlings only had a stronger NO3- influx when NO3- and NH4+ were present simultaneously. In addition, the qPCR analysis showed that the expression of AtNRT2 genes (AtNRT2.1-2.6), and particularly of AtNRT2.5, in the transformed Arabidopsis differed from that in the wild type. Overall, our results suggest that the heterologous expression of DsNRT3.1 affects seedlings' growth by enhancing the NO3- and NH4+ uptake in N-starved Arabidopsis. This may be related to the differential expression of AtNRT2 genes.