NtARF11 positively regulates cadmium tolerance in tobacco by inhibiting expression of the nitrate transporter NtNRT1.1.

Heavy metal cadmium (Cd) is widespread in contaminated soil and an important factor limiting plant growth. NO3- (nitrate) affects Cd uptake and thus changes Cd tolerance in plants; however, the underlying molecular regulatory mechanisms have not yet been elucidated. Here, we analyzed a novel gene, NtARF11 (auxin response factor), which regulates Cd tolerance in tobacco via the NO3- uptake pathway, through experiments with NtARF11-knockout and NtARF11-overexpression transgenic tobacco lines. NtARF11 was highly expressed under Cd stress in tobacco plants. Under Cd stress, overexpression of NtARF11 enhanced Cd tolerance in tobacco compared to that in wild-type tobacco, as shown by the low Cd concentration, high chlorophyll concentration, and low accumulation of reactive oxygen species in NtARF11-overexpressing tobacco. Moreover, low NO3- concentrations were observed in NtARF11-overexpressing tobacco plants. Further analyses revealed direct binding of NtARF11 to the promoter of the nitrate transporter NtNRT1.1, thereby negatively regulating its expression in tobacco. Notably, NtNRT1.1 knockout reduced NO3- uptake, which resulted in low Cd concentrations in tobacco. Altogether, these results demonstrate that the NtARF11-NtNRT1.1 module functions as a positive regulator of Cd tolerance by reducing the Cd uptake in tobacco, providing new insights for improving Cd tolerance of plants through genetic engineering.

Dynamic changes in fungal communities and functions in different air-curing stages of cigar tobacco leaves.

Introduction: Air curing (AC) plays a crucial role in cigar tobacco leaf production. The AC environment is relatively mild, contributing to a diverse microbiome. Fungi are important components of the tobacco and environmental microbiota. However, our understanding of the composition and function of fungal communities in AC remains limited. Methods: In this study, changes in the chemical constituents and fungal community composition of cigar tobacco leaves during AC were evaluated using flow analysis and high-throughput sequencing. Results: The moisture, water-soluble sugar, starch, total nitrogen, and protein contents of tobacco leaves exhibited decreasing trends, whereas nicotine showed an initial increase, followed by a decline. As determined by high-throughput sequencing, fungal taxa differed among all stages of AC. Functional prediction showed that saprophytic fungi were the most prevalent type during the AC process and that the chemical composition of tobacco leaves is significantly correlated with saprophytic fungi. Conclusion: This study provides a deeper understanding of the dynamic changes in fungal communities during the AC process in cigar tobacco leaves and offers theoretical guidance for the application of microorganisms during the AC process.

Nitrogen addition enhances tree radial growth but weakens its recovery from drought impact in a temperate forest in northern China.

Forest growth in the majority of northern China is currently limited by drought and low nitrogen (N) availability. Drought events with increasing intensity have threatened multiple ecosystem services provided by forests. Whether N addition will have a detrimental or beneficial moderation effect on forest resistance and recovery to drought events was unclear. Here, our study focuses on Pinus tabulaeformis, which is the main plantation forest species in northern China. We investigated the role of climate change and N addition in driving multi-year tree growth with an 8-year soil nitrogen fertilization experiment and analyzing 184 tree ring series. A moderate drought event occurred during the experiment, providing an opportunity for us to explore the effects of drought and N addition on tree resistance and recovery. We found that N addition was beneficial for increasing the resistance of middle-aged trees, but had no effect on mature trees. The recovery of trees weakened significantly with increasing N addition, and the reduction in fine root biomass caused by multiyear N addition was a key influencing factor limiting recovery after moderate drought. Our study implies that the combined effect of increasing drought and N deposition might increase the risk of pine forest mortality in northern China.

Shifting of nutrient limitation dominates the recovery of aboveground net primary productivity of mixed forests in northeastern China after selective logging.

The primary productivity of temperate forests is commonly limited by nitrogen (N) supply, which may be aggravated by the removal of trees. After selective logging, whether and the mechanism by which the N limitation can be alleviated by the rapidly increasing nutrient turnover during the recovery processes, which is important for improving carbon sequestration in temperate forests, remain unclear. We investigated the effect of nutrient limitation (leaf N:Pcom: the leaf N:P ratio at the community level) on plant community productivity by selecting 28 forest plots including seven forest recovery periods (at the sites logged 6, 14, 25, 36, 45, 55, and 100 years ago) following low-intensity selective logging (13-14 m3/ha) and one unlogged treatment by measuring the soil N concentration, soil phosphorus (P) concentration, leaf N concentration, leaf P concentration, and the aboveground net primary productivity (ANPP) of 234 plant species. The plant growth in temperate forests was limited by N, but the P limitation was observed at the sites logged 36 years ago, which showed a transition pattern of plant growth from N limitation to P limitation during the forest recovery process. Meanwhile, a robust linear trend in the community ANPP was observed with the increase in the community leaf N:P ratio, which suggests the enhancement in community ANPP with the release of N limitation after selective logging. Nutrient limitation (leaf N:Pcom) had a significant direct effect (56.0 %) on the community ANPP and showed a higher independent contribution (25.6 %) to the variation in the community ANPP than the soil nutrient supply and even the changes in species richness. Our results suggested that selective logging alleviated the N limitation, but a shift toward P limitation should also be highly regarded in learning the changes in carbon sequestration during the recovery processes.

Melatonin alleviates cadmium toxicity and abiotic stress by promoting glandular trichome development and antioxidant capacity in Nicotiana tabacum.

Melatonin is a well-known signaling molecule that mediates a range of physiological activities and various stress reactions in plants. We comprehensively tested the effect of melatonin on the development of root hairs and glandular trichomes and found that melatonin pretreatment of tobacco seeds significantly increased the length of root hairs. Furthermore, melatonin-treated tobacco exhibited significantly higher density of trichomes and larger glandular heads on long-stalk glandular trichomes than untreated plants, which resulted in enhanced secretion in glandular trichomes. Exogenous melatonin enhanced the aphid resistance of plants by facilitating the accumulation of cembranoids in the glandular trichomes and alleviated cadmium toxicity by increasing the Cd-exudation capacity of long glandular trichomes. Metabolic analysis indicated that the contents of 108 metabolites significantly changed upon melatonin treatment, with the contents of those that are directly/indirectly involved in melatonin metabolism changing the most. Further, KEGG pathway analysis suggested that the metabolic pathways of amino acids, reducing sugar, secondary metabolites, indole alkaloid biosynthesis, purine, pyrimidine, and ABC transporters were greatly influenced by exogenous melatonin application. Moreover, metabolisms of melatonin-related antioxidants and pyrimidine nucleoside antibiotics were enhanced after melatonin treatment. Melatonin improved tobacco resistance to high salinity, drought, and extreme temperature stresses, as indicated by improved photosynthetic and antioxidant capacities in treated vs. untreated plants. This study lays a foundation for the comprehensive application of melatonin to increase the stress tolerance of plants.

Allocation Strategies of Carbon, Nitrogen, and Phosphorus at Species and Community Levels With Recovery After Wildfire.

Plant stoichiometry and nutrient allocation can reflect a plant's adaptation to environmental nutrient changes. However, the allocation strategies of carbon (C), nitrogen (N), and phosphorus (P) between leaf and fine root in response to wildfire have been poorly studied. Our primary objective was to elucidate the trade-off of elemental allocation between above- and belowground parts in response to the soil nutrient changes after a wildfire. We explored the allocation sloping exponents of C, N, and P between leaf and fine root at the species and community levels at four recovery periods (year 2, 10, 20, and 30) after moderately severe wildfire and one unburned treatment in boreal forests in Great Xing'an Mountains, northeast China. Compared with the unburned treatment, leaf C concentration decreased and fine root C increased at year 2 after recovery. The leaf N concentration at year 10 after recovery was higher than that of unburned treatment. Plant growth tended to be limited by P concentration at year 10 after recovery. Nutrient allocation between leaf and fine root differed between species and community levels, especially in the early recovery periods (i.e., 2 and 10 years). At the community level, the nutrient concentrations of the leaf changed more as compared to that of the fine root at year 2 after recovery when the fine root nutrients changed more than those of the leaf. The different C, N, and P allocation strategies advanced the understanding of plant adaptation to soil nutrient changes during the postfire ecosystem restoration.

Mutation of NtNRAMP3 improves cadmium tolerance and its accumulation in tobacco leaves by regulating the subcellular distribution of cadmium.

Cadmium (Cd) is a harmful element that affects plant growth and development. Genetic improvements could be applied for enhancing Cd tolerance and accumulation in plants. Here, a novel Cd stress-induced gene, NtNRAMP3, was identified in tobacco. We constructed two NtNRAMP3-knockout (KO) tobacco lines using the CRISPR/Cas9 system, which enhanced Cd tolerance and Cd accumulation in tobacco leaves compared with those in the wildtype (WT). Subcellular localization analysis suggested that NtNRAMP3 is a tonoplast protein and GUS (ß-glucuronidase) histochemical analysis showed that NtNRAMP3 is highly expressed in the conductive tissue of leaves. NtNRAMP3-KO tobacco showed reduced Cd translation from vacuole to cytosol in leaves compared with the WT, and its vacuolar Cd concentration was significantly higher (20.78-22.81%) than that in the WT; in contrast, Cd concentration in the cytosol was reduced by 13.72-20.15%, preventing chlorophyll degradation and reducing reactive oxygen species accumulation in the leaves. Our findings demonstrate that NtNRAMP3 is involved in regulating Cd subcellular distribution (controlling Cd transport from vacuoles to the cytosol) and affects Cd tolerance and its accumulation in tobacco. This provides a key candidate gene to improve the phytoremediation efficiency of plants via genetic engineering.

Variations and controlling factors of soil denitrification rate.

The denitrification process profoundly affects soil nitrogen (N) availability and generates its byproduct, nitrous oxide, as a potent greenhouse gas. There are large uncertainties in predicting global denitrification because its controlling factors remain elusive. In this study, we compiled 4301 observations of denitrification rates across a variety of terrestrial ecosystems from 214 papers published in the literature. The averaged denitrification rate was 3516.3 ± 91.1 µg N kg-1  soil day-1 . The highest denitrification rate was 4242.3 ± 152.3 µg N kg-1  soil day-1 under humid subtropical climates, and the lowest was 965.8 ± 150.4 µg N kg-1 under dry climates. The denitrification rate increased with temperature, precipitation, soil carbon and N contents, as well as microbial biomass carbon and N, but decreased with soil clay contents. The variables related to soil N contents (e.g., nitrate, ammonium, and total N) explained the variation of denitrification more than climatic and edaphic variables (e.g., mean annual temperature (MAT), soil moisture, soil pH, and clay content) according to structural equation models. Soil microbial biomass carbon, which was influenced by soil nitrate, ammonium, and total N, also strongly influenced denitrification at a global scale. Collectively, soil N contents, microbial biomass, pH, texture, moisture, and MAT accounted for 60% of the variation in global denitrification rates. The findings suggest that soil N contents and microbial biomass are strong predictors of denitrification at the global scale.

Changes in plant nutrient utilization during ecosystem recovery after wildfire.

Wildfire is the primary natural disturbance in boreal forest ecosystems. It substantially changes soil nutrient conditions and plant nutrient dynamics. After a wildfire, various plant strategies of nutrient utilization are fundamental to ecosystem recovery processes. Stability of plant nutrients reflects the ability of plants possessing relatively constant elemental concentrations in the face of nutrient changes, which can be calculated by the value of "nutrient homeostasis". However, the mechanism of how nutrient homeostasis mediates plant community recovery in post-fire ecosystems remains unknown. The dominant tree species that survived after fire and the new emergence of regenerated tree species are the important components of a plant community during the recovery process. Our primary objective was to elucidate the nutrient homeostasis trade-off between dominant and regenerated species over years after recovery. Five treatments, namely, 2 year, 10 year, 20 year, 30 years after moderate burning severity, and unburned forests, were designed in the boreal forests of Great Xing'an Mountains, Northeast China. Compared with unburned forests, wildfire lowered the average value of homeostasis of plant nutrients (N and P). Moreover, the mean homeostasis value of the dominant species (i.e., Larix gmelinii) was higher than that of the regenerated species (i.e., Betula platyphylla). The slope of relationship between nutrient homeostasis and recovery years of the regenerated species was higher than that of the dominant species, suggesting that the nutrient homeostasis in the regenerated species recovered more quickly than dominant species after recovery. Compared with the dominant species, changes in the regenerated species' homeostasis can explained more to the changes of species diversity during the years after recovery. This study revealed plant nutrient adaptation in different species and different plant organs with years after wildfire and highlighted the importance of nutrient homeostasis in plant adaptation strategies and the recovery of plant community.

Changes of lignin biosynthesis in tobacco leaves during maturation.

Lignin is one of the most valuable renewable industrial materials. To elucidate the mechanism via which lignin is synthesised, we compared the lignin content, leaf hardness, cell wall thickness of palisade tissue, and gene expression patterns of lignin biosynthetic enzymes in three tobacco (Nicotiana tabacum L.) varieties during maturation. The results consistently showed that during maturation, the accumulation of lignin gradually increased in tobacco leaves, reaching a peak at full maturity (45 days after topping), and then gradually decreased. Similarly, the transcript level analysis revealed that the gene expression pattern of NtPAL, NtC4H, NtCCoAOMT and NtCOMT were relatively high, and consistent with the lignin content changes. Thus, the four genes may play regulatory roles in the synthesis of tobacco lignin. Analysis of tissue expression patterns of the lignin synthesis-related gene showed that the NtPAL, NtC4H, Nt4CL, NtHCT, NtCCoAOMT, NtCOMT, NtCCR, NtCAD, and NtPAO were all expressed in stems, roots, and leaves. NtC3H and NtF5H were specifically expressed in stems and roots, and not in leaves. Consistently, the NtC3H promoter induced high GUS expression in stems and petioles, marginal in roots, and no GUS activity in leaves. These results provide insights into molecular regulation of lignin biosynthesis in tobacco.

Vital roles of soil microbes in driving terrestrial nitrogen immobilization.

Nitrogen immobilization usually leads to nitrogen retention in soil and, thus, influences soil nitrogen supply for plant growth. Understanding soil nitrogen immobilization is important for predicting soil nitrogen cycling under anthropogenic activities and climate changes. However, the global patterns and drivers of soil nitrogen immobilization remain unclear. We synthesized 1350 observations of gross soil nitrogen immobilization rate (NIR) from 97 articles to identify patterns and drivers of NIR. The global mean NIR was 8.77 ± 1.01 mg N kg-1  soil day-1 . It was 5.55 ± 0.41 mg N kg-1  soil day-1 in croplands, 15.74 ± 3.02 mg N kg-1  soil day-1 in wetlands, and 15.26 ± 2.98 mg N kg-1  soil day-1 in forests. The NIR increased with mean annual temperature, precipitation, soil moisture, soil organic carbon, total nitrogen, dissolved organic nitrogen, ammonium, nitrate, phosphorus, and microbial biomass carbon. But it decreased with soil pH. The results of structural equation models showed that soil microbial biomass carbon was a pivotal driver of NIR, because temperature, total soil nitrogen, and soil pH mostly indirectly influenced NIR via changing soil microbial biomass. Moreover, microbial biomass carbon accounted for most of the variations in NIR among all direct relationships. Furthermore, the efficiency of transforming the immobilized nitrogen to microbial biomass nitrogen was lower in croplands than in natural ecosystems (i.e., forests, grasslands, and wetlands). These findings suggested that soil nitrogen retention may decrease under the land use change from forests or wetlands to croplands, but NIR was expected to increase due to increased microbial biomass under global warming. The identified patterns and drivers of soil nitrogen immobilization in this study are crucial to project the changes in soil nitrogen retention.

Label-free fluorescent sensor for one-step lysozyme detection via positively charged gold nanorods.

In the article, a simple and label-free strategy was proposed for the sensitive detection of lysozyme based on the fluorescence quenching of positively charged gold nanorods ((+)AuNRs) to DNA-templated silver nanoclusters (DNA/AgNCs). To construct the sensor, a DNA template was designed with a C-rich sequence at the 5'-terminal for the synthesis of AgNCs, while a lysozyme binding aptamer (LBA) at the 3'-terminal for the recognition of lysozyme, and such DNA/AgNCs was used as the fluorescence probe. Meantime, the fluorescence signal of such DNA/AgNCs can be quenched based on the electrostatic adsorption of them with (+)AuNRs, due to the surface energy transfer. In the presence of lysozyme, the specific binding happened between the LBA section of DNA/AgNCs and lysozyme, inducing the reduction of the total charge of DNA/AgNCs and weakening the adsorption of them with (+)AuNRs, which directly resulting in the recovery of the fluorescence signal. Besides, the fluorescence signal recovery of DNA/AgNCs has a linear positive proportional relationship with lysozyme concentration in the range of 10 pM-2.0 nM under the optimal conditions. Moreover, a satisfactory recovery (99.6-107.2%) was obtained while detecting lysozyme in human serum samples. Graphical abstract A simple and label-free strategy was proposed for the sensitive detection of lysozyme based on the fluorescence quenching of positively charged gold nanorods ((+)AuNRs) to DNA-templated silver nanoclusters (DNA/AgNCs).

N-P utilization of Acer mono leaves at different life history stages across altitudinal gradients.

The relationship between plants and the environment is a core area of research in ecology. Owing to differences in plant sensitivity to the environment at different life history stages, the adaptive strategies of plants are a cumulative result of both their life history and environment. Previous research on plant adaptation strategies has focused on adult plants, neglecting saplings or seedlings, which are more sensitive to the environment and largely affect the growth strategy of subsequent life stages. We compared leaf N and P stoichiometric traits of the seedlings, saplings, and adult trees of Acer mono Maxim and different altitudes and found significant linear trends for both life history stages and altitude. Leaf N and P content by unit mass were greatly affected by environmental change, and the leaf N and P content by unit area varied greatly by life history stage. Acer mono leaf N-P utilization showed a significant allometric growth trend in all life history stages and at low altitudes. The adult stage had higher N-use efficiency than the seedling stage and exhibited an isometric growth trend at high altitudes. The N-P utilization strategies of A. mono leaves are affected by changing environmental conditions, but their response is further dependent upon the life history stage of the plant. Thus, this study provides novel insights into the nutrient use strategies of A. mono and how they respond to the environmental temperature, soil moisture content along altitude and how these changes differ among different life history stages, which further provide the scientific basis for the study of plant nutrient utilization strategy on regional scale.

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Fire is an important ecological factor in boreal coniferous forest, which directly affects plant taxonomic and functional diversity and consequently forest succession. We analyzed the changes of soil nutrient contents, leaf functional traits, taxonomic and functional diversity of a Larix gmelinii community under different fire intensity levels (low, medium, heavy). The forest stand had been naturally recovered for 12 years in Yakeshi area. The results showed that fire significantly reduced soil total nitrogen content, but did not affect soil total phosphorus content. Low and medium fire intensities could maintain higher species diversity and functional diversity of the community. Medium fire disturbance significantly increased species diversity of the community, with maximum species richness index, Shannon index, Simpson index and Pielou evenness index. Fire disturbance reduced functional richness and functional divergence of the community, which were the largest in medium and low fire intensity respectively (except unburned). Functional evenness and Rao qua-dratic entropy index increased after forest fire disturbance, and the largest was in the low intensity burned area. With the increases of fire intensity, leaf dry matter content, leaf tissue density and leaf total phosphorus content showed a significantly increased trend, while specific leaf area, leaf water content, leaf total nitrogen content and leaf N:P showed the trend unburned ＞ medium fire ＞ low fire ＞ heavy fire, leaf thickness increased first and then decreased. Fire intensity had significant effects on leaf functional traits and functional diversity of forest community. Moderate fire distur-bance could promote forest restoration.

Overexpression of OsPT8 Increases Auxin Content and Enhances Tolerance to High-Temperature Stress in Nicotiana tabacum.

Temperature is a primary factor affecting the rate of plant development; as the climate warms, extreme temperature events are likely to increasingly affect agriculture. Understanding how to improve crop tolerance to heat stress is a key concern. Wild plants have evolved numerous strategies to tolerate environmental conditions, notably the regulation of root architecture by phytohormones, but the molecular mechanisms of stress resistance are unclear. In this study, we showed that high temperatures could significantly reduce tobacco biomass and change its root architecture, probably through changes in auxin content and distribution. Overexpression of the OsPT8 phosphate transporter enhanced tobacco tolerance to high-temperature stress by changing the root architecture and increased the antioxidant ability. Molecular assays suggested that overexpression of OsPT8 in tobacco significantly increased the expression of auxin synthesis genes NtYUCCA 6, 8 and auxin efflux carriers genes NtPIN 1,2 under high-temperature stress. We also found that the expression levels of auxin response factors NtARF1 and NtARF2 were increased in OsPT8 transgenic tobacco under high-temperature stress, suggesting that OsPT8 regulates auxin signaling in response to high-temperature conditions. Our findings provided new insights into the molecular mechanisms of plant stress signaling and showed that OsPT8 plays a key role in regulating plant tolerance to stress conditions.

Genome-wide identification and expression analysis of HSP90 gene family in Nicotiana tabacum.

BACKGROUND: Heat shock proteins 90 (HSP90s) are a highly conserved protein family of cellular chaperones widely found in plants; they play a fundamental role in response to biotic and abiotic stresses. The genome-wide analysis of HSP90 gene family has been completed for some species; however, it has been rarely reported for the tobacco HSP90 genes. RESULTS: In this study, we systematically conducted genome-wide identification and expression analysis of the tobacco HSP90 gene family, including gene structures, evolutionary relationships, chromosomal locations, conserved domains, and expression patterns. Twenty-one NtHSP90s were identified and classified into eleven categories (NtHSP90-1 to NtHSP90-11) based on phylogenetic analysis. The conserved structures and motifs of NtHSP90 proteins in the same subfamily were highly consistent. Most NtHSP90 proteins contained the ATPase domain, which was closely related to conserved motif 2. Motif 5 was a low complexity sequence and had the function of signal peptide. At least 6 pairs of NtHSP90 genes underwent gene duplication, which arose from segment duplication and tandem duplication events. Phylogenetic analysis showed that most species expanded according to their own species-specific approach during the evolution of HSP90s. Dynamic expression analysis indicated that some NtHSP90 genes may play fundamental roles in regulation of abiotic stress response. The expression of NtHSP90-4, NtHSP90-5, and NtHSP90-9 were up-regulated, while NtHSP90-6, and NtHSP90-7 were not induced by ABA, drought, salt, cold and heat stresses. Among the five treatments, NtHSP90s were most strongly induced by heat stress, and weakly activated by ABA treatment. There was a similar response pattern of NtHSP90s under osmotic stress, or extreme temperature stress. CONCLUSIONS: This is the first genome-wide analysis of Hsp90 in N. tabacum. These results indicate that each NtHSP90 member fulfilled distinct functions in response to various abiotic stresses.

Genome-wide identification and characterization of Hsp70 gene family in Nicotiana tabacum.

Heat shock proteins 70 (Hsp70) constitute a highly conserved protein family of cellular chaperones widely distributed in plants, where they play a fundamental role in response to biotic and abiotic stress. Until now, genome-wide analyses of the Hsp70 gene family have been conducted for some species. However, reports about Hsp70 genes in Nicotiana tabacum are scarce. In this study, we systematically conducted genome-wide identification and expression analysis of the Hsp70 gene family in tobacco, including gene structure, classification, evolutionary relationships, promoters, and transcript levels in response to abiotic stress treatments. In all, 61 Hsp70 members were identified and classified into six groups that were mapped onto 18 chromosomes, where most were distributed on both ends of the chromosome. The conserved structures and motifs of NtHsp70 proteins in the same subfamily were highly consistent. At least 15 pairs of NtHsp70 genes underwent gene duplication by segment and tandem duplications. Most NtHsp70 proteins contained N-terminal hexokinase conserved motifs. Phylogenetic analysis showed that most species expanded according to their own species-specific approach during the evolution of Hsp70s. Tissue-specific expression analysis indicated that all NtHsp70 genes were involved in at least one or more abiotic stress responses, highlighting the wide participation of NtHsp70 genes in environmental adaptation. This is the first genome-wide analysis of Hsp70 in N. tabacum. These results indicate that each NtHsp70 member fulfilled distinct functions in response to various abiotic stresses.

NtMYB12 Positively Regulates Flavonol Biosynthesis and Enhances Tolerance to Low Pi Stress in Nicotiana tabacum.

Phosphorus (P) is an essential macronutrient for plant growth and development. The concentration of flavonol, a natural plant antioxidant, is closely related to phosphorus nutritional status. However, the regulatory networks of flavonol biosynthesis under low Pi stress are still unclear. In this study, we identified a PFG-type MYB gene, NtMYB12, whose expression was significantly up-regulated under low Pi conditions. Overexpression of NtMYB12 dramatically increased flavonol concentration and the expression of certain flavonol biosynthetic genes (NtCHS, NtCHI, and NtFLS) in transgenic tobacco. Moreover, overexpression of NtMYB12 also increased the total P concentration and enhanced tobacco tolerance of low Pi stress by increasing the expression of Pht1-family genes (NtPT1 and NtPT2). We further demonstrated that NtCHS-overexpressing plants and NtPT2-overexpressing plants also had increased flavonol and P accumulation and higher tolerance to low Pi stress, showing a similar phenotype to NtMYB12-overexpressing transgenic tobacco under low Pi stress. These results suggested that tobacco NtMYB12 acts as a phosphorus starvation response enhancement factor and regulates NtCHS and NtPT2 expression, which results in increased flavonol and P accumulation and enhances tolerance to low Pi stress.

Characterization of wheat TaSnRK2.7 promoter in Arabidopsis.

MAIN CONCLUSION: Expression of TaSnRK2.7 promoter is strongly induced under abiotic stress and could be used as a valuable tool for improving plant stress resistance via transgenic techniques. The sucrose non-fermenting 1-related protein kinase 2 (SnRK2) gene family plays pivotal roles in response to abiotic stresses (drought, salinity and cold). Here, we studied the expression of five wheat TaSnRK2.7 promoter-5'-deletion constructs (- 2547, - 1621, - 806, - 599, and - 254) fused to beta-glucuronidase (GUS) in Arabidopsis. Tissue-expression analysis revealed that the - 254 to ATG fragment was sufficient for inducing GUS expression in hypocotyls. Additionally, the - 806 to - 599 and - 2547 to - 1621 fragments contained leaf- and root-specific elements, respectively. Deletion analysis showed that these fragments were unresponsive to ABA treatment, suggesting that TaSnRK2.7 participates in an ABA-independent signaling pathway. Assays examining stress responses of constructs demonstrated that the - 599 to - 254 and - 806 to - 599 fragments contained elements responsive to abiotic and osmotic stress, respectively. The TaSnRK2.7 promoter contained enhancers from - 806 to - 254 and - 2547 to - 1621, while the - 1621 to - 806 fragment contained negative regulatory elements that restrict root and leaf gene expression in response to abiotic stress. Furthermore, under drought and salt stress, the TaSnRK2.7 promoter conferred greater gene expression in leaves than the rd29A promoter, even though both were induced by abiotic stress. These findings enhance our understanding of the molecular mechanisms behind TaSnRK2.7 action, which should prove useful in transgenic studies investigating stress-induced gene expression.